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DEPARTMENTS Department of Oncology .. Department of Environmental Health Sciences ... Department of Neuroscience .. Department of Cardiovascular Research .. Department of Molecular Biochemistry and Pharmacology ... Department of Epidemiology...... Department of Public Health............................................................................. LABORATORIES AND CENTERS Laboratory of Regulatory Policies ..... Centre of Computer Science Engineering. Italian Cochrane Center .. The Catullo and Daniela Borgomainerio Center Library ....

125 159 185 235 261 269 273 281 283

NEGRI BERGAMO LABORATORIES

DEPARTMENTS Department of Molecular Medicine .. 289 Department of Biomedical Engineering. 309 Laboratory of Biology and Therapy of Metastasis.. 327

ALDO and CELE DACCO CENTER

DEPARTMENT Department of Renal Medicine. 331 LABORATORIES AND CENTERS Rare Diseases Documentation and Research.................................................... 359 International Relations Office of rare Diseases........................................................ 371 T he Transplant Research Center. 377

EDUCATIONAL ACTIVITIES STAFF

All the staff of the Institute is listed on its website www.marionegri.it

379 383

PUBLICATIONS
A comprehensive list of the Institutes publications is available on the www.marionegri.it website Section Publications

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Edited by Isabella Bordogna printed May 2011

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PREFACE
This booklet provides a brief description of the research and training work done at the Mario Negri Institutes in Milan and Bergamo. It is divided by departments, and in some cases by single laboratories. Details of the results are provided in the text itself, so here we shall just draw a few general remarks. This is our first year in the new Milan Headquarters: during this time we devoted most of our efforts improving the new available technologies. Particularly, we have carried out translational research following the mouse clinic patterns. Nuclear magnetic resonance, micro cat scanner, ultrasound, Doppler, photon microscope are some of the methodologies used to do research on human diseases in mice, following the techniques utilized in medical clinic. This will allow to transfer data in more effective ways, to study more in depth and to use a lower number of animals in experimental research. Imaging will play an important role, thanks to the installation of electronic microscopy, of contrast, time- lapse microscopy and atomic force microscopy. New important technologies will also allow us to develop our proteomics research. The tendency nowadays is towards widespread use of molecular biology techniques, especially for studying the mechanism of action of drugs. In vitro studies are an essential basis for thorough investigations although a significant number of in vivo experiments are still needed as this is the only mean we have of validating in vitro findings and establishing models that resemble human diseases as closely as possible. This has led to a substantial increase in the use of transgenic animals. The Institute is still concentrating on its traditional research lines: oncology, neurosciences, cardiovascular and renal diseases, organ transplants, rare diseases with strong cell biology and molecular biochemistry connotations. Significant work has been carried out on the environment and health as a whole. Experimental, clinical and epidemiological research on rare diseases and orphan drugs is growing all the time. The Mario Negri Institute strives to develop a multifaceted approach to all these research themes, ranging from basic research, to pharmacokinetics, pharmacology, controlled clinical trials, epidemiological analysis and whenever possible the epidemiology of services. So far we have published more than 11.000 articles on peer reviewed scientific journals. If research is to continue young scientists must be continually trained. Working in the laboratory not only gives them an outlet for their ideas, but enables them to obtain worthwhile qualifications by taking part in the Institutes training schemes, which are recognised by the Lombardy Region in Italy, or by working for a Ph.D. awarded by the Open University, UK . Training courses are also available on biomedical statistics, for general practitioners, family pediatricians, and clinical trial nurses. A vital part of the Institutes work involves providing information, at all levels. This is done, in particular, through the Rare Diseases Information Center, the Center for Information on Medicinal Drugs and the Website (www.marionegri.it). We also provide information to medical doctors, nurses, patients associations and lay people, through the media and through a recently developed website: www.partecipasalute.it. Because research is going through a very difficult time it is vital to be supported by the Government, by private and public bodies as well as by foundations and private citizens. Silvio Garattini Director

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Mario Negri

INSTITUTE FOR

PHARMACOLOGICAL RESEARCH

Milan

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departments and laboratories

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DEPARTMENT OF ONCOLOGY
STAFF

Chief

Maurizio DINCALCI, M.D.

Oncological Studies Office and Documentation

Scientific Documentalist Stefania FILIPPESCHI, Chemist

Laboratory of Cancer Pharmacology

Head Maurizio DINCALCI, M.D.

Biophysics Unit
Head Paolo UBEZIO, Phys.D.

Flow Citometry Unit

Head Eugenio ERBA, Biochem.D

Cancer Clinical Pharmacology Unit

Head Massimo ZUCCHETTI, Chem.Pharm.D.

Laboratory of Molecular Pharmacology

Head Massimo BROGGINI, Ph.D.

DNA Repair Unit

Head Giovanna DAMIA, M.D.

Laboratory of Biology and Treatment of Metastases

Head Raffaella GIAVAZZI, Biol.Sci.D., Ph.D.

Tumor Angiogenesis Unit

Head Unit located in Bergamo Giulia TARABOLETTI, Biol.Sci.D.

Molecular Cancer Therapeutics Unit

Head Maria Rosa BANI, Biol.Sci.D., Ph.D.

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Laboratory for the development of new pharmacological strategies

Head Valter TORRI, M.D.

Laboratory of Clinical Trials

Head Irene FLORIANI, Dr.Sci.Biol., Dr.Stat., Ph.D.

Clinical Trials Informatics and Management Unit

Head Davide POLI, Phys.D.

Laboratory of Translational and Outcome Research in Oncology

Head Giovanni APOLONE, M.D.

Gynecology Oncology Unit

Head Roldano FOSSATI, M.D.

CERP: Center for the Evaluation and Research on Pain

Head Giovanni APOLONE, M.D. Oscar CORLI, M.D.

Laboratory of Medical Research and Consumer Involvement

Head Paola MOSCONI, Biol.Sci.D.

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CURRICULA
Maurizio D'Incalci obtained his Medical Degree cum Laude from the University of Milan in 1977. After specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of Genoa, he worked in the Laboratory of Molecular Pharmacology of the National Cancer Institute in Bethesda, MD, USA. Since 1986 he has been chief of the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since 1996 he has become chief of the Department of Oncology at the Mario Negri Institute. He has been President of the Pharmacology and Molecular Mechanisms Group of the European Organization for Research and Treatment of Cancer (EORTC). From 1994 to 1997 he was Chairman of the New Drug Development Coordinating Committee and from 1997 to 2000 he was chairman of the Research Division of the EORTC. He has been member of the Board of the EORTC from April 2000 to 2003. Since 1995 he is member of the Board of Directors of the Nerina and Mario Mattioli Onlus Foundation. From 1997 he is the Preclinical Coordinator of the Southern Europe New Drug Organization (SENDO) and since 2005 the Chairman of the New Agents Committee (NAC). From 2006 he is president of the Scientific Committee of the Mario Negri Gynecologic Oncology group (MaNGO). From 2007 he is member of the Scientific Committee of the Italian Association for Cancer Research (AIRC). From 2009 he is member of the Board of Directors of the Italian Cancer Society (SIC). From 2010 he is member of the Scientific Committees of the ABO (Application of Biotechnologies in Oncology) Foundation, a national foundation for cancer research, and of the Buzzi Unicem Onlus Foundation for the research, diagnosis and cure of malignant mesothelioma. FHe is on the editorial board of many international cancer-related scientific journals and from September 2000 to December 2010 he is Editor for Experimental Oncology of the European Journal of Cancer. Dr D'Incalci is author of more than 440 papers on cancer chemotherapy published in peer reviewed international journals, and of several chapters in books on cancer chemotherapy.
Selected publications Frapolli R., Tamborini E., Virdis E., Bello E., Tarantino E., Marchini S., Grosso F., Sanfilippo R., Gronchi A., Tercero J.C., Peloso G., Casali P., Pilotti S., DIncalci M. Novel models of myxoid liposarcoma xenografts mimicking the biological and pharmacological features of human tumors. Clinical Cancer Res., 16(20): 4958-4967 (2010). Germano G., Frapolli R., Simone M., Tavecchio M., Erba E., Pesce S., Pasqualini F., Grosso F., Sanfilippo R., Casali P., Gronchi A., Virdis E., Tarantino E., Pilotti S., Greco A., Nebuloni M., Galmarini C.M., Tercero J.C., Mantovani A., DIncalci M., Allavena P. Anti-tumor and anti-inflammatory effects of trabectedin on human myxoid liposarcoma cells. Cancer Res., 70(6): 2235-2244 (2010). Frapolli R, Zucchetti M, Sessa C, Marsoni S, Vigano' L, Locatelli A, Rulli E, Compagnoni A, Bello E, Pisano C, Carminati P, D'Incalci M. Clinical pharmacokinetics of the new oral camptothecin gimatecan: The inter-patient variability is related to (1)-acid glycoprotein plasma levels. Eur. J. Cancer, 46: 505-516 (2010) Forni C, Minuzzo M, Virdis E, Tamborini E, Simone M, Tavecchio M, Erba E, Grosso F, Gronchi A, Aman P, Casali P, D'Incalci M, Pilotti S, Mantovani R. Trabectedin (ET-743) promotes differentiation in myxoid liposarcoma tumors. Mol Cancer Ther, 8 : 449-457 (2009). Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M, Dell'Anna T, Apolone G, Broggini M, D'Incalci M. Analysis of gene expression in early-stage ovarian cancer Clin Cancer Res, 14 : 7850-7860 (2008). Grosso F., Jones R.L. Demetri G.D., Judson I.R., Blay J.Y., Le Cesne A., Sanfilippo R., Casieri P., Collini P., Dileo P., Spreafico C., Stacchiotti S., Tamborini E., Tercero J.C., Jimeno J., DIncalci M., Gronchi A., Fletcher J.A., Pilotti S., Casali P.G. Efficacy of Trabectedin (ET-743) in advanced pre-treated myxoid liposarcomas. Lancet Oncology, 2007; 8:595-602.

Giovanni Apolone, got his Medical degree in 1982 (Pavia, Italy) and his post-doctoral specializations in Internal Medicine in 1987 (Pavia, Italy) and Pharmacological Research (1992). He is Head of the Laboratory of Translational and Outcome Research. He is also Vice-President of the Ethics Committee of the European Institute of Oncology in Milan (Italy). His main fields of interest are: Methodological, ethical and regulatory aspects of clinical research, with special emphasis on oncology and the cancer pain. Health care evaluation with special emphasis on oncology; Development and validation of case-mix and patient-reported outcome measures;

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Education and health promotion research and programs. He is author or co-author of more than 270 publications, most in International peer-reviewed Journals. Mean Impact Factor, computed on peer-reviewed papers: 3.2. H-index (from ISI-Web of knowledge, March 2010): 30. Number of citations: 4111. Average citation per item: 32.37. Number of papers with >50 citations: 19.
Selected publications Greco M T, Corli O, Montanari M, Deandrea S, Zagonel V, Apolone G, CPOR SG Investigators. Epidemiology and pattern of care of Breakthrough cancer Pain (BTcP) in a longitudinal sample of cancer patients. Results from the CPORSG. Clin J Pain 2010, e-pub. Mannucci E, Petroni M L, Villanova N, Rotella C M, Apolone G, Marchesini G, QUOVADIS Study Group. Clinical and psychological correlates of health-related quality of life in obese patients. Health Qual Life Outcomes 2010 8 : 90. Knudsen K A, Brunelli C, Kaasa S, Apolone G, Corli O, Montanari M, Fainsinger R, Aass N, Fayers P, Caraceni A, Klepstad P, European Palliative Care Research Collaborative (EPCRC), European Pharmacogenetic Study (EPOS).Which variables are associated with pain intensity and treatment response in advanced cancer patients? Implications for a future classification system for cancer pain. Eur J Pain 2010, e-pub. Gacci M, Corona G, Apolone G, Lanciotti M, Tosi N, Giancane S, Masieri L, Serni S, Maggi M, Carini M. Influence of serum testosterone on urinary continence and sexual activity in patients undergoing radical prostatectomy for clinically localized prostate cancer. Prostate Cancer Prostatic Dis 2010 13 : 168-172. Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone M V, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta M G, Riva E. Prevalence, incidence and types of mild anemia in the elderly: the " Health and Anemia" population-based study. Haematologica 2010 95 : 18491856.

Massimo Broggini followed the faculty of Science of the University of Milan, got the specialization in Biochemistry at Mario Negri Institute, and the PhD degree at the Open University, London,UK. He worked in the laboratory of Molecular Pharmacology of the National Cancer Institute of Bethesda, Md, in 1986. From 1991 he is the head of the Molecular Pharmacology Unit of the Mario Negri Institute and from 1999 he his the head of the Laboratory of Molecular Pharmacology of the same Institute. His main fields of interest are the study of the mechanism of action of new anticancer agents, the search of altered proteins and genes in human cancer and the study of oncosuppressor genes. He is member of the "Pharmacology and Molecular Mechanisms Group" of the European Organisation for the Research and Treatment of Cancer (EORTC) and of the American Association for Cancer Research. He is in the Editorial board of the European Journal of Cancer. He is author of more than 100 articles published in international journals.
Selected publications Floriani I, Garassino MC, Broggini M, Veronese S, Marsoni S, Marabese M, Farina G, Scanni A. Role of cetuximab in the treatment of patients with NSCLC: are we throwing out the baby with the bath water? J Clin Oncol. 2010 ;28:467. Mazzoletti M, Broggini M. PI3K/AKT/mTOR inhibitors in ovarian cancer. Curr Med Chem. 2010;17(36):4433-47. Sala G, Dituri F, Raimondi C, Previdi S, Maffucci T, Mazzoletti M, Rossi C, Iezzi M, Lattanzio R, Piantelli M, Iacobelli S, Broggini M, Falasca M. Phospholipase Cgamma1 is required for metastasis development and progression. Cancer Res. 2008 Dec 15;68(24):10187-96. Marrazzo E, Marchini S, Tavecchio M, Alberio T, Previdi S, Erba E, Rotter V, Broggini M. The expression of the DeltaNp73beta isoform of p73 leads to tetraploidy. Eur J Cancer. 2009 Feb;45(3):443-53. Epub 2008 Nov 12. Falasca M, Chiozzotto D, Godage HY, Mazzoletti M, Riley AM, Previdi S, Potter BV, Broggini M, Maffucci T. A novel inhibitor of the PI3K/Akt pathway based on the structure of inositol 1,3,4,5,6-pentakisphosphate. Br J Cancer. 2010 Jan 5;102(1):104-14. PubMed PMID: 20051961; Ganzinelli M, Carrassa L, Crippa F, Tavecchio M, Broggini M, Damia G. Checkpoint kinase 1 down-regulation by an inducible small interfering RNA expression system sensitized in vivo tumors to treatment with 5-fluorouracil. Clin Cancer Res. 2008 Aug 15;14(16):5131-41.

Irene Floriani got her degree in Biological Sciences at the University of Milan in 1988, her degree in Biostatistics and Experimental Statistics at the University of Milan in 2003 and her phD in Life Sciences at Open University of London (UK) in 2005. After ten-year experience in pharmaceutical industry, in 2002 she became Head of the Biometry and Data Management Unit of the Laboratory of Clinical Research in Oncology and since 2006 she is Head of Laboratory of Clinical Trials. She is President of the Ethics Committee of the Ospedale SantAnna of Como, Vice-President of that of the Fondazione IRCCS Istituto Neurologico Carlo Besta of Milan, and member of further two Ethics Committees. Her main fields of interest are: statistical aspects of methodology of clinical research with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature and Methodological aspects of

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diagnostic test evaluation.

Selected publications Floriani I, Garattini S, Torri V. Looking for efficiency rather than efficacy in randomized controlled trials in oncology. Ann Oncol. 2010 Jul; 21(7):13911393. Luciani A, Ascione G, Bertuzzi C, Marussi D, Codec C, Di Maria G, Caldiera SE, Floriani I, Zonato S, Ferrari D, Foa P. Detecting disabilities in older patients with cancer: comparison between comprehensive geriatric assessment and vulnerable elders survey-13. J Clin Oncol. 2010 Apr 20;28(12):2046-50. Floriani I, Torri V, Rulli E, Garavaglia D, Compagnoni A, Salvolini L, Giovagnoni A. Performance of imaging modalities in diagnosis of liver metastases from colorectal cancer: a systematic cancer: a systematic review and metaanalysis. J Magn Reson Imaging. 2010 Jan;31(1):19-31. Floriani I, Garassino MC, Broggini M, Veronese S, Marsoni S, Marabese M, Farina G, Scanni A. Role of Cetuximab in the Treatment of Patients With NSCLC: Are We Throwing out the Baby With the Bath Water? J Clin Oncol. 2010 Jun 28. Loupakis F, Ruzzo A, Cremolini C, Vincenzi B, Salvatore L, Santini D, Masi G, Stasi I, Canestrari E, Rulli E, Floriani I, Bencardino K, Galluccio N, Catalano V, Tonini G, Magnani M, Fontanini G, Basolo F, Falcone A, Graziano F. KRAS codon 61, 146 and BRAF mutations predict resistance to cetuximab plus irinotecan in KRAS codon 12 and 13 wild-type metastatic colorectal cancer. Br J Cancer 2009 101 : 715-721 Floriani I, Santini D, Torri V, Cremolini C, Falcone A, Loupakis F. Do we need biopsies of metastases for colorectal cancer patients? Br J Cancer 2009 101 : 374-375

Raffaella Giavazzi got her Biological Sciences degree (1979) at the University of Milan, and her PhD in Pharmacology at the Mario Negri Institute of Milan (1984), followed by a specialization in pharmacology (1994) at the University of Milan. From 1981 to 1983 she was a Fellow in the Cancer Metastasis and Treatment Laboratory, NCI-FCRDC, Frederick, MD, and from 1983 to 1985 Assistant Professor at the Department of Cell Biology of M.D. Anderson Hospital and Tumour Institute, University of Texas System Cancer Centre in Houston, TX. Raffaella Giavazzis research interests are in the field of tumour biology and pharmacology. Specifically, she is studying aspects related to the metastatic process and angiogenesis. She is involved in the preclinical evaluation of new therapeutic strategies against cancer focusing on the angiogenesis inhibitors and combination therapies. From 1986 to 1993 she was Head of the Cancer Metastasis Treatment Unit and since 1993 she has been the Head of the Laboratory of Biology and Treatment of Metastasis at Mario Negri Institute for Pharmacological Research. She is also adjutant Professor in Oncology, Medical School-University of Brescia, member of the Teaching Committee for the PhD course in Physiology-Pharmacology-Molecular and Cellular Toxicology-University of Siena, member of the Executive Committee at SENDO (Southern Europe New Drug Development Organization) and member of the Executive Committee of the European Association for Cancer Research (EACR). She was consulting scientist for the NCI-Drug Therapeutics Program, USA (1996-2006); She is a member of the American Association for Cancer Research (AACR), International Metastases Research Society, EORTC-Screening and Pharmacology Group, European Association for Cancer Research (EACR), Italian Cancer Society (SIC) of which she was President (2006-2007). She is on the Editorial Board of international scientific journals such as the European Journal of Cancer, Journal of Clinical & Experimental Metastasis, and The International Journal of Biological Markers. She has published approximately 200 articles on peer reviewed scientific journals.
Selected publications Borgia B., Rsli C., Fugmann T., Schliemann C., Cesca M., Neri D., Giavazzi R. A proteomic approach for the identification of vascular markers of liver metastasis. Cancer Research, 70(1):309-18, 2010. Rsli C., Borgia B., Schliemann C., Gunther M., Wunderli-Allenspach H., Giavazzi R., Neri D. Comparative analysis of the membrane protome of closely related metastatic and non-metastatic tumor cells. Cancer Research, 69(13):5406-14, 2009. Cesca M., Frapolli R., Berndt A., Scarlato V., Richter P., Kosmehl H., DInclaci M., Ryan A.J., Giavazzi R. The effects of vandetanib on paclitaxel tumor distribution and antitumor activity in a xenograft model of human ovarian carcinoma. Neoplasia, 11(11):1155-64, 2009. Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, 2008. Giavazzi R., Bani M.R.,Taraboletti G.: Tumorhost interaction in the optimization of paclitaxel-based combination therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:48188, 2007 Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R., Giavazzi R. The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor, causing apoptosis of endothelial cells and inhibition of angiogenesis. Clin. Cancer Research 12(6):1839-49, 2006.

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Rybak J.N., Ettore A., Kaissling B., Giavazzi R., Neri D., Elia G. In vivo protein biotinylation for identification of organ-specific antigens accessible from the vasculature. Nature Methods 2(4):291-98, 2005.

Paola Mosconi got her Biological Science degree (Milan 1982) and the specialisation in Pharmacological Research (Milan 1984). Paola Mosconi is involved in several national projects on issues pertaining the patient involvement in care aspects and outcome research. She published more than 300 articles in leading national and international journals (111), as well as books on issues related to her main areas of interest.

Significant experiences has been coordinated: development of research projects and strategies to involve patients or consumer associations in health debate, and clinical research, consensus conferences training for consumers on quality of information, and methodological aspects of clinical research; studies for estimate the type of information on diseases and treatments received by patients, mainly in cancer patients; set-up of websites targeted on consumers/patients www.partecipasalute.it, www.paincare.it, www.fondazionemattioli.it; projects on the assessment of Quality of Life in randomised clinical trials or in epidemiological survey; translation and cultural adaptation of questionnaires for Quality of Life; evaluation of the consumers satisfaction with the health services and the care received. Paola Mosconi has participated as teacher, or coordinator, to the realization of training course on Methodological aspects of clinical research or Evaluation of quality of life for health care professionals and representatives of voluntary associations.
Major present functions - President of the Ethical Committees of the AUSL Bologna - Elected member of Associazione per la Ricerca sulla Efficacia della Assistenza Sanitaria - Italian Cochrane Centre - Founding member of Europa Donna Italia, the European breast cancer coalition
Selected publications Mosconi P, Colombo C Fostering a strategic alliance between patients associations and health care professionals J Ambul Care Manage 2010, 33 (3): 223-30. Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta MG, Riva E Prevalence, incidence and types of mild anemia in the elderly: the "Health and Anemia" population-based study Haematologica 2010; 95(11): 1849-1856 Mosconi P, Donati S, Colombo C, Mele A, Liberati A, Satolli R. The Consensus Conference WorkingGroup. Informing women about hormone replacement therapy: the Consensus conference statement. BMC Woman Health Journal 2009; 9:14 doi:10.11886/1472-6874-9-14 Mosconi P, Colombo C, Satolli R, Liberati A. PartecipaSalute, an Italian project to involve lay people, patients associations and scientific-medical representatives on the health debate. Health Expectations 10: 194-204, 2007. Mosconi P, Poli P, Giolo A, Apolone G. How health consumers feel about clinical research: a questionnaire survey. European Journal of Public Health 15: 372-379, 2005. Mosconi P, Buchanan M, Kyriakides S, Fernandez-Marcos A, Horvatin J, O'Connell D, Zernik N, on behalf of EUROPA DONNA. EUROPA DONNA: has strength in its heterogeneity. European J Cancer 40: 1145-1149, 2004.

Valter Torri got his Medical degree in 1985 and the specialization in medical Oncology in 1989 at the University of Milano. Education: 1985: MD Degree with full honors cum Laude, University of Milano; 1988 Post-Doctoral Degree in Pharmacological Research, Mario Negri Institute, Milano; 1989 Post-Doctoral Degree in Medical Oncology, University of Milano; 1989-1991 Research Fellow at the Biometric Research Branch of Cancer Treatment Evaluation Program, NCI, Bethesda, MD (USA) Areas of Interest: Statistical aspects of clinical research methodology with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature; Methodological aspects of diagnostic test evaluation. Present Position: Head of Laboratory of Clinical Research In Oncology, Oncology Department, Mario

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Negri Institute, Milano. Chronology of Professional Appointments: 1983-1985: Clinical research Fellow in Internal Medicine at the University Hospital, University of Milan; 1985-1989: Research assistant at the Clinical Trial Unit of the Laboratory of Clinical Epidemiology, Mario Negri Institute for Pharmacological Research, Milano; 1989-1991: Research fellow at the Biometric Research Branch of Cancer Treatment Evaluation Program, NCI, Bethesda, MD (USA); 1994: Head of Biometric Unit of the Laboratory of Cancer Clinical Epidemiology, Oncology Department, Mario Negri Institute for Pharmacological Research, Milano, Italy; 1995 Vice Director of the Italian Cochrane Center; 2001: Head of Laboratory of Clinical Research In Oncology, Oncology Department, Mario Negri Institute, Milano. 2006: Head of Laboratory for the development of new pharmacological strategies , Oncology Department, Mario Negri Institute, Milano. Member of Consiglio Direttivo Nazionale dellAssociazione Italiana di Oncologia Medica Member of Independent data monitoring committee of International Randomised Clinical trials in NSCLC and ovarian carcinoma Co-author of more than 160paper published on peer reviewed journals and of 5 chapters of scientific books relative to clinical research methodology for therapeutic and diagnostic studies.
Selected publications Rossi A, Garassino MC, Cinquini M, Sburlati P, Di Maio M, Farina G, Gridelli C, Torri V. Maintenance or consolidation therapy in small-cell lung cancer: a systematic review and meta-analysis. Lung Cancer. 201;70(2):119-28. Sargent DJ, Marsoni S, Monges G, Thibodeau SN, Labianca R, Hamilton SR, French AJ, Kabat B, Foster NR, Torri V, Ribic C, Grothey A, Moore M, Zaniboni A, Seitz JF, Sinicrope F, Gallinger S. Defective mismatch repair as a predictive marker for lack of efficacy of fluorouracil-based adjuvant therapy in colon cancer. J Clin Oncol. 2010 Jul 10;28(20):3219-26. Epub 2010 May 24. Erratum in: J Clin Oncol. 2010 ; 28(30):4664. 3. Floriani I, Garattini S, Torri V. Looking for efficiency rather than efficacy in randomized controlled trials in oncology. Ann Oncol. 2010 21(7):1391-3. Epub 2010 May 25. PubMed PMID: 20501504. Laghi L, Bianchi P, Miranda E, Balladore E, Pacetti V, Grizzi F, Allavena P, Torri V, Repici A, Santoro A, Mantovani A, Roncalli M, Malesci A. CD3+ cells at the invasive margin of deeply invading (pT3-T4) colorectal cancer and risk of postsurgical metastasis: a longitudinal study. Lancet Oncol. 2009; 10 (9): 877-84. Garassino MC, Borgonovo K, Rossi A, Mancuso A, Martelli O, Tinazzi A, Di Cosimo S, La Verde N, Sburlati P, Bianchi C, Farina G, Torri V. Biological and clinical features in predicting efficacy of epidermal growth factor receptor tyrosine kinase inhibitors: a systematic review and meta-analysis. Anticancer Res. 2009; 29 (7): 2691-701. Torri V: Clinical trials and data management In: Oxford textbook of oncology, 2nd. ed. Vol. 1. Oxford Univ. Press, Oxford; 2002 : 1123-1134

Maria Rosa Bani got her Biological Sciences degree at the University of Milan in 1998 attaining the Italian Government Qualification to practice as Biologist in 1990. She obtained the specialization in Pharmacological Research from the Department of Education of the Regional Government of Lombardia in 1991 and the specialization in Biomedical Research from the Department of Education of the Regional Government of Abruzzo in 1993. In 2005 she was awarded the degree of Doctor of Philosophy (PhD), Discipline of Life Sciences of the Open University Research School (UK). From 1991 to 1995 she was a Post Doctoral Fellow at the Cancer Research Division, Sunnybrook Health Science Centre, University of Toronto (Canada); from 2000 to 2001 she was Guest Scientist at the Advance Technology Centre, National Cancer Institute, National Institute of Health (USA). From 1996, she was a Fellow Research Scientist at the Mario Negri Institute for Pharmacological Research, Laboratory of Biology and Treatment of Metastasis and she became a staff research scientist in 2003. Since 2004 she was appointed Head of the Molecular Cancer Therapeutics Unit in the same laboratory. She has been the Scientific Manager of STROMA (since 2004) and ADAMANT (since2008), two Integrated Projects funded in the 6th and 7th Framework Programs of the European Commission. She is a member of the American Association for Cancer Research (AACR), the European Association for Cancer Research (EACR) and the Italian Cancer Society (SIC). Maria Rosa Bani research interests are in the field of cancer biology and therapeutics, with a focus on molecular studies of the malignant progression and on the preclinical efficacy of therapeutics modalities. She is co-author of 36 peer reviewed publications, 2 book chapters and 67 abstracts of which 15 selected for oral presentations at international meetings.
Selected publications Silini A., Ghilardi C., Ardinghi C., Bernasconi S., Carraro F., Naldini A., Bani M.R., Giavazzi R. Protease-activated receptor-1 (PAR-1) promotes the motility of human melanomas and is associated to their metastatic phenotype. Clinical Experimental Metastasis, 27 (1) : 43-53, 2010

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Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, 2008. Giavazzi R., Bani M.R.,Taraboletti G.: Tumorhost interaction in the optimization of paclitaxel-based combination therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:48188, 2007. Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R. & Giavazzi R. The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor, causing apoptosis of endothelial cells and inhibition of angiogenesis. Clinical Cancer Research 12: 1839-1849, 2006. Bani M.R., Nicoletti M.I., Alkharouf N.W., Ghilardi C., Petersen D., Erba E., Sausville E.A., Liu E.T. and Giavazzi R. Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts. Molecular Cancer Therapeutics 3: 111-121, 2004. Taraboletti G., Sonzogni L., Vergani V., Hosseini G., Ceruti R., Ghilardi C., Bastone A., Toschi E., Borsotti P., Scanziani E., Giavazzi R., Pepper M.S., Stetler-Stevenson W.G., Bani M.R. Post-transcriptional stimulation of endothelial cell matrix metalloproteinases 2 and 1 by endothelioma cells. Experimental Cell Research 258 : 384-394, 2000.

Giovanna Damia obtained her Medical Degree cum Laude from the University of Milan in 1985. After specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of Milan, she worked as a post-doctoral fellow in the Laboratory of Experimental Immunology of the National Cancer Institute, Frederick, USA. She worked as a research fellow in the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since April 2003 she has become chief of the DNA Repair Unit at the Mario Negri Institute. From 1992 to1995 she has been consultant of the General Secretariat of the Progetto Finalizzato CNR "Applicazioni Cliniche della Ricerca Oncologica". Since September 2005 she is Deputy Editor for Experimental Oncology of the European Journal of Cancer. Her main fields of interest are: mechanism of action of anticancer drugs, cell cycle checkpoints and natural compounds.
Selected publications Damia G, D'Incalci M. Genetic instability influences drug response in cancer cells. Curr Drug Targets. 2010 Oct;11(10):1317-24. Carrassa L, Montelatici E, Lazzari L, Zangrossi S, Simone M, Broggini M, Damia. G. Role of Chk1 in the differentiation program of hematopoietic stem cells. Cell Mol Life Sci. 2010 May;67(10):1713-22. Carrassa L, Sanchez Y, Erba E, Damia G. U2OS cells lacking Chk1 undergo aberrant mitosis and fail to activate the spindle checkpoint. J Cell Mol Med. 2009 Aug;13(8A):1565-76. Ganzinelli M, Carrassa L, Crippa F, Tavecchio M, Broggini M, Damia G. Checkpoint kinase 1 down-regulation by an inducible small interfering RNA expression system sensitized in vivo tumors to treatment with 5-fluorouracil. Clin Cancer Res. 2008 Aug 15;14(16):5131-41. Tavecchio M, Simone M, Erba E, Chiolo I, Liberi G, Foiani M, D'Incalci M, Damia G. Role of homologous recombination in trabectedin-induced DNA damage. Eur J Cancer. 2008 Mar;44(4):609-18. Epub 2008 Feb 19. Damia G, D'Incalci M. Targeting DNA repair as a promising approach in cancer therapy. Eur J Cancer. 2007 Aug;43(12):1791-801.

Eugenio Erba has obtained his Biological and Biochemistry Analysis Degree at the University of Urbino. He worked as a research fellow in the Laboratory of Cancer Chemotherapy at the Mario Negri Institute and since 1984 he is head of the Flow Cytometry Unit in the Department of Oncology at the Mario Negri Institute of Milan. He has worked as a visiting fellow in the Department of Istochemistry and Cytochemistry of the University of Leiden, The Netherlands in 1983. Since 1997 he is Teacher of Post-Graduate Studies in Cytometry at the University of Milan and Co-ordinator and Teacher of PostGraduate Studies in Cytometry for the Italian Cytometry Group. He has been President of the Italian Cytometry Group from 1999 to 2001. Since 2001 he is member of the Executive Board of the Italian Cytometry Group. Scientific areas of interest: studies on the mechanism of action of different compounds with provided antitumoral activity evaluating the mechanism of cell death and cell cycle phase perturbations induced on different human cancer cell lines by using flow cytometry. Co-ordinator of working-group in a quality control study on flow cytometric DNA content analysis in human tumors.
Selected publications Urru S.A.M., Veglianese P., De Luigi A., Fumagalli E., Erba E., Gonella Diaza R., Carr A., Davoli E., Borsello T., Forloni G., Pengo N., Monzani E., Cascio P., Cenci S., Sitia R., Salmona M. A new fluorogenic peptide determines proteasome activity in single cells. J.Med.Chem., 53: 7452-7460 (2010). Germano G., Frapolli R., Simone M., Tavecchio M., Erba E., Pesce S., Pasqualini F., Grosso F., Sanfilippo R., Casali P., Gronchi A., Virdis E., Tarantino E., Pilotti S., Greco A., Nebuloni M., Galmarini C.M., Tercero J.C., Mantovani A., DIncalci M., Allavena P. Anti-tumor and anti-inflammatory effects of trabectedin on human myxoid liposarcoma cells. Cancer Res., 70(6): 2235-2244 (2010).

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C. Forni, M Minuzzo, E. Virdis, E. Tamburini, M. Simone, M. Tavecchio, E. Erba, F. Grosso, A. Gronchi, P.Aman, P. Casali, M. DIncalci, S. Pilotti , R. Mantovani. Trabectedin (ET-743) promotes differentiation in myxoid liposarcoma tumors. Mol. Ca. Ther. 8(2), 449-57, 2009 E. Marrazzo, S. Marchini, M. Tavecchio, T. Alberio, S. Previdi, E. Erba, V. Rotter, M. Broggini The expression of the Np73 isoform of p73 leads to tetraploidy. Eur J Ca 45, 443-53, 2009 M.Tavecchio, M. Simone, E.Erba, I. Chiolo, G. Liberi, M. Foiani, M. DIncalci, G. Damia. Role of homologous recombination in trabectedin-induced DNA damage. Eur. J. Ca 44:609-618 (2008) Paulis M., Bensi M., Orioli D., Mondello C., Mazzini G., DIncalci M., Falcioni C., Radaelli E., Erba E., Raimondi E., De Carli L. Transfer of a Human Chromosomal Vector from a Hamster Cell Line to a Mouse Embryonic Stem Cell Line. Stem Cell , 25:2543-2550 (2007)

Roldano Fossati got his Medical Degree cum Laude from the University of Milan in 1980, his PostDoctoral Degree in Endocrinolgy cum Laude from the University of Verona in 1983 and his PostDoctoral Degree in Medical Statistics from the University of Milan in 1992. He has been consultant at the Mario Negri Institute since 1983 and, at present, he is head of the Gynecology and Oncology Unit of the Laboratory of Translational and Outcome Research. Areas of Interest: Statistical and methodologic aspects of clinical research with focus on Controlled Clinical Trials in Oncology; Systematic Overview of the medical literature.
Selected publications Hogberg T, Signorelli M, de Oliveira CF, Fossati R, Lissoni AA, Sorbe B, Andersson H, Grenman S, Lundgren C, Rosenberg P, Boman K, Tholander B, Scambia G, Reed N, Cormio G, Tognon G, Clarke J, Sawicki T, Zola P, Kristensen G. Sequential adjuvant chemotherapy and radiotherapy in endometrial cancer--results from two randomised studies. Eur J Cancer. 2010 Sep;46(13):2422-31. Epub 2010 Jul 7. Signorelli M, Lissoni AA, Cormio G, Katsaros D, Pellegrino A, Selvaggi L, Ghezzi F, Scambia G, Zola P, Grassi R, Milani R, Giannice R, Caspani G, Mangioni C, Floriani I, Rulli E, Fossati R. Modified Radical Hysterectomy Versus Extrafascial Hysterectomy in the Treatment of Stage I Endometrial Cancer: Results From the ILIADE Randomized Study. Ann Surg Oncol. 2009 Oct 16 Andrea Alberto Lissoni, Nicoletta Colombo, Antonio Pellegrino, Gabriella Parma, Paolo Zola, Dionyssios Katsaros, Stefania Chiari, Alessandro Buda, Fabio Landoni, Michele Peiretti, Tiziana DellAnna, Robert Fruscio, Mauro Signorelli, Roberto Grassi, Irene Floriani, Roldano Fossati , Valter Torri, Eliana Rulli. A phase II, randomized trial of neoadjuvant chemotherapy comparing a three-drug combination of paclitaxel, ifosfamide and cisplatin (TIP) versus paclitaxel and cisplatin (TP) followed by radical surgery in patients with locally advanced squamous cell cervical carcinoma: the Snap02 Italian Collaborative Study. Annals of Oncology, 20:660-665;2009 Fruscio R, Colombo N, Lissoni AA, Garbi A, Fossati R, Ieda' N, Torri V, Mangioni C.A phase II randomised clinical trial comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and epirubicin in newly diagnosed advanced epithelial ovarian cancer: long-term survival analysis. Br J Cancer. 2008 Feb 5; Maggi R, Lissoni A, Spina F, Melpignano M, Zola P, Favalli G, Colombo A, Fossati R. Adjuvant chemotherapy vs radiotherapy in high-risk endometrial carcinoma: results of a randomised trial. Br J Cancer. 2006 Aug 7;95(3):266-71 Maggioni A, Benedetti Panici P, Dell'anna T, Landoni F, Lissoni A, Pellegrino A, Rossi RS, Chiari S, Campagnutta E, Greggi S, Angioli R, Manci N, Calcagno M, Scambia G, Fossati R, Floriani I, Torri V, Grassi R, Mangioni C.Randomised study of systematic lymphadenectomy in patients with epithelial ovarian cancer macroscopically confined to the pelvis. Br J Cancer. 2006 Sep 18;95(6):699-704.

Davide Poli got his degree in Physics at the University of Milan in 2007 and his specialization in Biochemical Research Technician" at the Mario Negri Institute for Pharmacological Research in 2004. Since 2010 he is Head of Clinical Trials Informatics and Management Unit of the Clinical Trials Laboratory. His areas of interest are: design of eCRF in Clinical Trials, new electronic aspects of Clinical Research especially towards technologies of Web-based Electronic Data Capture, methodology and data management aspects in Clinical Research.
Selected Publications Ocular Hypertension Treatment Study Group, European Glaucoma Prevention Study Group (EGPS), Poli D The accuracy and clinical application of predictive models for primary open-angle glaucoma in ocular hypertensive individuals Ophthalmology, Volume 115, Number 11 pp. 2030-2036, November 2008 Porcu L, Poli D, Torri V, Rulli E, Cropalato di Tullio M, Cinquini M, Bajetta E, Labianca R, Di Costanzo F, Nitti D, Floriani I Impact of recent legislative bills regarding clinical research on Italian ethics committee activity Journal of Medical Ethics 2008, Volume 34, pp. 747-750 Gordon MO, Torri V. Miglior S, Beiser JA, Floriani I, Miller JP, Gao F, Adamsons I, Poli D, D'Agostino RB, Kass MA. A validated prediction model for the development of primary open-angle glaucoma in individuals with ocular hypertension.

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Ophthalmology, Volume 114, Number 1, pp. 10-19, January 2007 The European Glaucoma Prevention Study (EGPS) Group, Poli D Results of the European Glaucoma Prevention Study. Ophthalmology, Volume 112, Number 3, pp. 366-375, March 2005 Icon, Torri V, Floriani I, Poli D, Santillo M, Buda A Paclitaxel plus platinum-based chemotherapy versus conventional platinum-based chemotherapy in women with relapsed ovarian cancer: The ICON4/AGO-OVAR-2.2 Trial. Lancet 2003; 361: 2099-2106

Giulia Taraboletti got her degree cum laude in Biological Sciences at the University of Pavia (Pavia, Italy) in 1983, and the specialization in Pharmacological Research at the Mario Negri Institute, Milano, Italy in 1986. From 1986 to 1988 she was a post-doctoral fellow at the Laboratory of Pathology, NCI, NIH, Bethesda, MD, and from 1988-1995 research scientist at Mario Negri Institute in Bergamo, Italy. Since 1995 she is Head of the Unit of Tumor Angiogenesis, at Mario Negri Institute, in Bergamo. Research interests include tumor angiogenesis, endogenous inhibitors of angiogenesis (thrombospondin-1) and preclinical studies of antiangiogenic and vascular disrupting compounds, including tubulin-targeting agents. She is member of Metatasis Research Society (MRS), American Association for Cancer Research (AACR), European Association for Cancer Research (EACR), and the Italian Society of Oncology (SIC). She is on the editorial board of European Journal of Cancer and Current Cancer Therapy Reviews.
Selected publications Colombo G, Margosio B, Ragona L, Neves M, Bonifacio S, Annis DS, Stravalaci M, Tomaselli S, Giavazzi R, Rusnati M, Presta M, Zetta L, Mosher DF, Ribatti D, Gobbi M, Taraboletti G. Non-peptidic thrombospondin-1-mimics as fibroblast growth factor-2 inhibitors: an integrated strategy for the development of new antiangiogenic compounds. J Biol Chem, 285: 8733-8742, 2010. Bonezzi K., Taraboletti G., Borsotti P., Bellina F., Rossi R., Giavazzi R. Vascular disrupting activity of tubulin-binding 1,5-diaryl-1H-imidazoles. J Med Chem 52, 79067910, 2009. Margosio B, Rusnati M, Bonezzi K, Cordes B-lA, Annis DS, Urbinati C, Giavazzi R, Presta M, Ribatti D, Mosher DF, and Taraboletti G. Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain. Int J Biochem Cell Biol 40: 700-709, 2008. Giavazzi R., Bani M.R.,Taraboletti G. Tumorhost interaction in the optimization of paclitaxel-based

combination therapies with vascular targeting compounds. Cancer Metastasis Rev, 26:48188, 2007. Martinelli M., Bonezzi K., Riccardi E., Kuhn E., Frapolli R., Zucchetti M., Ryan A.J., Taraboletti G., Giavazzi R.
Sequence dependent antitumour efficacy of the vascular disrupting agent ZD6126 in combination with paclitaxel. Br J Cancer 97:888-94, 2007. Margosio B., Marchetti D., Vergani V., Giavazzi R., Rusnati M., Presta M., and Taraboletti G. Thrombospondin-1 as a scavenger for matrix-associated fibroblast growth factor-2. Blood 102: 4399-4406, 2003.

Paolo Ubezio got his B.Sc. degree in Physics at the University of Milan, in 1982, and the specialisation in Pharmacological Research Specialist" at the Mario Negri Institute for Pharmacological Research in 1986. Main activities are: i) Computer simulation of tumor proliferation during/after treatments using models based on the cell cycle; ii) Development of new methods and data analysis tools in flow cytometry and in time-lapse imaging of living cells; iii) Optimization of anticancer drug scheduling. Since 1991 is Head of the Unit of Biophysics at the Mario Negri Institute
Selected publications Ubezio, P.; Lupi, M., Branduardi, D.; Cappella, P., Cavallini, E., Colombo, V., Matera, G., Natoli, C., Tomasoni, D., DIncalci, M. (2009) Quantitative assessment of the complex dynamics of G1, S and G2M checkpoint activities. Cancer Res. 69: 5234-5240 Valentini, G., DAndrea, C., Ferrari, R., Pifferi, A., Cubeddu, R., Martinelli, M., Natoli, C., Ubezio, P. and Giavazzi R. (2008) In-vivo measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. Photochem. Photobiol. 84:1249-1256. Ubezio, P. and Cameron, D. (2008) Cell killing and resistance in pre-operative breast cancer chemotherapy. BMC Cancer 8:201. Basse, B., Ubezio, P. (2007) A generalised age and phase structured model of human tumour cell populations both umperturbed and exposed to a range of cancer therapies. Bull. Math. Biol. 69:1673-90. Lupi, M., Matera, G., Natoli, C., Colombo, V., Ubezio, P. (2007) The Contribution of p53 in the Dynamics of Cell Cycle Response to DNA Damage Interpreted by a Mathematical Model. Cell Cycle 6:943-950. Lupi, M., Matera, G., Branduardi, D., D'Incalci M. and Ubezio, P. (2004) Cytostatic and cytotoxic effects of topotecan decoded by a novel mathematical simulation approach. Cancer Res. 64: 2825-2832.

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Massimo Zucchetti obtained his Chem. Pharm. Degree from the University of Milan in 1982. After specializing in Pharmacology at the Mario Negri Institute of Milan (1988), he worked in the Laboratory of Clinical Pharmacology of Department of Oncology at San Giovanni Hospital, Bellinzona, Switzerland (1988-1990). Since 1996 he has been chief of the Cancer Clinical Pharmacology Unit at the Mario Negri Institute. He is member of the Pharmacology and Molecular Mechanisms Group of the European Organization for Research and Treatment of Cancer (EORTC) from 1988 up to date. His main field of interest are: - Clinical pharmacology, phase I and Phase II studies - Analysis of drugs, development of new analytical method, pharmacokinetic and pharmacodynamic studies in humans in GCP and GLP conditions - Pharmacokinetic, toxicokinetic and metabolic studies in animals - Pharmacokinetic drug interaction Dr Zucchetti is author of more than 90 papers on pre-clinical and clinical cancer chemotherapy published in peer reviewed international journals.
Selected publications Sala F., Marangon E., Bagnati R., Livi V., Cereda R., DIncalci M., Zucchetti M. Development and validation of a HPLC-MS/MS method for the determination of the novel proteasome inhibitor CEP-18770 in human plasma and its application in a clinical pharmacokinetic study. J Mass Spectrom. 2010; 45: 1309-15. Frapolli R., Zucchetti M., Sessa C., Marsoni SA., Vigan L., Locatelli A., Rulli E., Compagnoni A., Bello E., Pisano C., Carminati P., DIncalci M. Clinical pharmacokinetics of the new oral camptothecin gimatecan: the intra-patients variability is related to 1-acid glycoprotein plasma levels. Eur J Cancer 2010, 66:635-41 Sala F., Zucchetti M., Bagnati R., DIncalci M., Pace S., Capocasa F., Marangon E. Development and validation of a HPLC-MS/MS m,ethod for the determination of ST1926, a novel oral antitumor agent, adamantyl retinoid derivative, in human plasma of patients partecipatig in a phase I study. J Chromatogr B: Anal. Tecnol. Biomed. Life Sci. 2009; 31: 1826. Gambacorti-Passerini CB, Tornaghi L, Marangon E, Franceschino A, Pogliani EM, D'Incalci M, Zucchetti M.. Imatinib concentrations in human milk Blood. 2007 Feb 15;109(4):1790. Marangon E, Sala F, Caffo O, Galligioni E, D'Incalci M, Zucchetti M. Simultaneous determination of gemcitabine and its main metabolite, dFdU, in plasma of patients with advanced non-small-cell lung cancer by high-performance liquid chromatography-tandem mass spectrometry. J Mass Spectrom. 2008 Feb;43(2):216-23. Frapolli R., Marangon E., Zaffaroni M., Colombo T., Falcioni C., Bagnati R., Simone M., DIncalci M., Manzotti C., Fontana G., Morazzoni P., Zucchetti M. Pharmacokinetics and metabolism in mice of IDN 5390 (13-(N-Boc-3-ibutylisoserinoyl)-C-7,8-seco-10-deacetylbaccatin III), a new oral C-seco-taxane derivative with antiangiogenic property effective on paclitaxel-resistant tumors. Drug Metabolism and Disposition, 34(12):2028-2035 (2006).

ACTIVITIES
The Oncology Department comprises three preclinical experimental laboratories (Laboratory of Cancer Pharmacology, Laboratory of Molecular Pharmacology and Laboratory of Biology and Treatment of Metastases) and four laboratories dealing with clinical research and clinical trials (Laboratory for the Development of New Pharmacological Strategies, Laboratory of Clinical Trials, Laboratory of Translational and Outcome Research in Oncology and Laboratory for Medical Research and Consumer Involvement). The Oncology department hosts the coordination center of two networks of hospitals that carry on clinical research in gynecologic cancer (MaNGO: Mario Negri Gynecologic Oncology) and in cancer pain (CPOR-SG: Cancer Pain Outcome Research Study Group) and a center for cancer pain assessment and research (CERP:Center for the Evaluation and Research on Pain). In some cases research projects are carried out by single laboratories or research units, in other cases by collaborations between different laboratories of the Oncology Department or other departments, or other groups outside the Institute (see National and International Collaborations). Preclinical laboratories focus on the discovery and development of new antitumor and antimetastatic drugs and their new combinations; on tumor biology, not only to acquire new scientific knowledge, but particularly as a base for more selective therapeutic approaches and to identify and evaluate experimental models for discovering and studying new drugs or treatments. Clinical new drug development involves close participation in the activity of SENDO (Southern

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Europe New Drug Organization) and studies driven by the Laboratory of Cancer Pharmacology, the Laboratory of Molecular Pharmacology and the Laboratory of Biology and Treatment of Metastases. The Laboratory for the Development of New Pharmacological Strategies, the Laboratory of Clinical Trials, the Laboratory of Translational and Outcome Research in Oncology and the Laboratory for Medical Research and Consumer Involvement are involved in the evaluation of the effects of new therapeutic modalities in phase I/II and in phase III comparative and effectiveness outcome studies. Outcome Research implies organizing trials to clarify the results of certain health care practices and interventions in clinical practice. Observational (surveys) and outcome research (effectiveness) studies are carried out, in collaboration with regional and national health authorities and other scientific associations. At the preclinical and clinical level there are studies of various human tumors, with particular emphasis on ovarian tumors and more recently on soft tissue sarcomas.

MAIN FINDINGS
At nanomolar concentrations, Trabectedin affects the regulatory mechanisms of the transcription. Cells that are deficient in Homologous Recombination DNA Repair -e.g. with mutations of BRCA1 or BRCA2 genes- are hypersensitive to the drug Nucleotide excision repair deficient cells that are hypersensitive to UV rays and to other DNA damaging drugs are resistant to Trabectedin. The selective activity of Trabectedin against human myxoid liposarcoma appears related to the drug ability to modulate the transcription of genes involved in adipocytic differentiation. Trabectedin modulates the transcription of genes involved in pro-inflammatory mechanisms that are potentially relevant for tumor growth and progression and inhibits the production of cytokines and chemokines by macrophages that are tumor associated. New sarcoma experimental models have been obtained. They will be useful to investigate new drugs for these diseases. Use of mathematical models of tumor growth and anticancer treatment to interpret experimental data and to manage the complexity of underlying biological phenomena. A new method enabling to perform dynamical measures of cell cycle checkpoint activities in response to anticancer treatments. Gene profiling analysis shows specific molecular signatures according to the histotype and prognosis of stage I ovarian carcinoma. The expression of a truncated form of p63 (DNp63) increases with the increased malignancy of ovarian cancer. Patients expressing high levels of DNp63 have a worst prognosis. DNp63 represents therefore a new potential target for selective therapies in this malignancy. CHK1 downregulation by specific inhibitors or siRNA, increases the antitumor activity of 5fluorouracil in vivo. This effect is particularly evident in p53 deficient tumors indicating that this combination increases the selectivity of this anticancer agent. An anthracycline derivative, Nemorubicin, has a peculiar mechanism of action and is active against tumors resistant to drugs such as cisplatin. A mechanism of resistance against this drug has been identified, which involves the abrogation of the expression of a nucleotide excision repair gene.

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The use of combinations of PI3K/akt/mTOR inhibitors acting at different sites of the same target, induces a pronounced antitumor effect. Mechanistically there is a selective inhibition of the translation of proteins involved in the cellular growth. Mutations in the K-RAS gene have a different impact on the response to treatment that is dependent from the type of aminoacid substitution present at codon 12. The growth of breast cancer cells in the bones is slowed down by selective c-met inhibitors. Human umbilical cord-derived stem cells express checkpoints proteins only in specific differentiation stages. It is likely that this is related to the different susceptibility of the cells. Identification of genes differentially expressed by tumor isolated endothelial cells, in respect of healthy tissue endothelial cells. For some of them we demonstrated their expression in human tumors, in which the protein was associated to vasculature and stromal component.. The vascular endothelial growth factor (VEGF) released by cancer modifies the gene expression of tumoral microenvironment and response to treatment: in this condition a number of differentially expressed genes has been identified. We are currently studying, their role in tumor progression and in affecting the response to therapy. Soluble VEGFC levels (the main mediator in lymphoangiogenesis) in plasma and ascites correlate with progression and invasion of ovarian cancer. Preclinical studies are in progress to assess the antitumor and antimetastatic properties of selective inhibitors of the VEGF/VEGFRs pathway. A new antiangiogenic domain of thrombospondin (a physiological inhibitor of angiogenesis) that binds the angiogenic factor FGF-2 has been identified and characterized. Non peptidic small molecules, mimetic of this domain, have been identified and are studied as potential inhibitors of angiogenesis. A series of proteins preferentially expressed in liver metastases has been identified in vivo tissue perfusions through mass spectrometric analysis. These proteins could be a potential target for selective therapies. Preclinical pharmacokinetic and pharmacodynamic studies have been used to support the biological and pharmacological rationale for the treatment regimens, which combine chemotherapeutics and inhibitors of angiogenesis. The website of the project PartecipaSalute (www.partecipasalute.it) has a very innovative character in comparison with the other health Italian sites because it introduces and develops information in a very active way with specific instruments. A randomized phase III trial has shown that pelvic systematic lymphadenectomy in early endometrial cancer does not improve overall survival. As pelvic systematic lymphadenectomy is not devoid of side effects, this trial will spare patients with early endometrial cancer important early and late surgical morbidities. A twin trial in ovarian carcinoma, published in 2005, came to similar conclusions. Results from a systematic review of literature and from a prospective epidemiologic study

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suggest that an important proportion of patients with cancer pain (up to 43%) receive an analgesic treatments that is not appropriate with the intensity of pain Results from a survey carried out on a national level on a sample of 1801 patients with cancer pain confirm that in Italy a relevant part of cancer patients does not receive an appropriate information about their prognosis: physicians reported that according to their knowledge only 31% received information about their prognosis. An independent survey carried out in a Northern Italian Region confirmed this finding: among 550 patients treated at home for cancer pain with palliative care , only 58% were classified to be fully aware of their prognosis. An observational longitudinal study carried out in 110 Italian centers and involving about 1800 patients with metastatic cancer and pain have documented that that in terms of analgesics effectiveness, that each drugs prescribed by investigators (morphine, fentanyl, buprenorphine and oxycodone) were able to reduce the intensity of pain of about 2 points on a 11-eleven point numerical rating scale (p<0.001). The application of specific pe-planned algorithm identified about 30% cases who were classified as non-responders. Preliminary analyses documented some differences between drugs in terms of size of the analgesic effect, dosages required and side effects reported. A randomized phase III trial has shown that a modified radical (Piver-Rutledge class II) hysterectomy does not improve survival and locoregional control compared to the standard extrafascial (Piver-Rutledge class I) hysterectomy in patients with stage I endometrial cancer . This trial has enrolled 520 patients who have been followed-up for over 5 years The training and information activity organized with the associations of citizens & patients in the framework of the PartecipaSalute project has been finalized to the organization of the Parita task Participate to the research project with the associations. Parita is organised to discuss with the scientific community the grey areas of the medical assistance and clinical research identified from the patients and their associations, and to develop specific protocols for future research programs.

NATIONAL COLLABORATIONS
Agenzia Sanitaria Regionale (ASR), Bologna Agenzia Italiana del Farmaco (AIFA), Roma Alleanza Contro il Tumore Ovarico (ACTO) Azienda Sanitaria Locale, Rimini Azienda Sanitaria Locale, Vercelli Assessorato Sanit, Regione Emilia Romagna Associazione Italiana di Oncologia Medica (AIOM) Azienda Sanitaria Unica Regionale, Regione Marche

Azienda Ospedaliera di Reggio Emilia Arcispedale S. Maria Nuova Azienda Ospedaliera San Gerardo, Universit Milano-Bicocca, Monza
Casa Sollievo della Sofferenza, San Giovanni Rotondo (IRCCS) CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano CNR IGBE, Pavia CNR, Istituto di Chimica del Riconoscimento Molecolare, Milano Cochrane Collaboration

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ENEA Centro Ricerche, Unit di Tossicologia e Scienze Biomediche, Roma Fondazione IRCCS Istituto Nazionale dei Tumori (INT), Milano Fondazione Istituto FIRC di Oncologia Molecolare (IFOM), Milano Fondazione LUVI, Milano Fondazione Nerina e Mario Mattioli Onlus, Milano Fondazione Salvatore Maugeri, Pavia Fondazione SmithKline (FSK), Milano Fondo Edo Tempia, Laboratorio di Farmacogenomica, Biella I.A.S.I., Roma Istituti Ospitalieri di Cremona Istituto Clinico Humanitas, Rozzano MI Istituto Dermopatico dell'Immacolata, Roma Istituto Ortopedico Galeazzi, Milano Istituti Ortopedici Rizzoli, Bologna

Istituto di Endocrinologia ed Oncologia Sperimentale (IEOS), CNR, Napoli Istituto

Europeo di Oncologia (IEO), Milano Istituto di Fisica, Politecnico di Milano Istituto di Genetica Molecolare CNR, Sezione di Istochimica e Citometria, Pavia Istituto Nazionale per la Ricerca sul Cancro (IST), Genova Istituto Nazionale Tumori Fondazione G. Pascale, Napoli Istituto Neurologico Carlo Besta, Milano Istituto Regina Elena, Roma Istituto Superiore di Sanit, Roma Laboratorio Cell factory, Policlinico di Milano Ospedale Fatebenefratelli e Oftalmico, Milano Ospedale San Matteo, Pavia Ospedale Santa Chiara, Trento Regione Toscana Rete Oncologica Lombarda (ROL), Milano Spedali Civili di Brescia Universit Cattolica del Sacro Cuore, Roma Universit di Bari Universit di Brescia Universit di Catania Universit di Chieti Universit di Milano Universit di Modena e Reggio Emilia Universit di Monza Universit di Padova Universit di Siena Universit di Torino Universit La Sapienza, Roma Zadig, Agenzia di Giornalismo scientifico

INTERNATIONAL COLLABORATIONS
ADAMANT Consortium, IP 7th FP, EC ARCAGY (Association de Recherche sur les Cancers Gyncologiques), France Breakthrough Breast Cancer Center, Instutite of Cancer Reasearch, London, U.K.

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Cancer Biomarkers and Prevention Group, University of Leicester, U.K. Cancer Research UK, London, U.K. EORTC, Brussels, Belgium EUROPA DONNA European Agency for the Evaluation of Medicinal Products (EMEA), London, U.K. European Association for Palliative Care (EAPC) European Network of Gynaecological Oncology Trials groups (ENGOT)Eusoma (European Society of Breast Cancer Specialist) Florence, Italy Executive Board of GCIG (Gynecologic Cancer Intergroup) Frontier science & technology Research Foundation Southern Europe (FSE), Switzerland Genome Institute of Singapore (GIS), Singapore German Cancer Research Center, Division of Toxicology and Cancer Risk Factors, Heidelberg, Germany Goteborg University, Lundberg Laboratory for Cancer Research, Goteborg, Sweden Gynecologic Cancer Intergroup (GCIG) Helios Klinikum Erfurt GmbH, Institute of Pathology, Germany Institute of Pathology, Friedrich Schiller University, Jena, Germany Istituto Oncologico della Svizzera Italiana Johns Hopkins University, USA Ludwig Institute for Cancer Research, London, U.K. National Cancer Center, Singapore Stony Brook University, NY, USA Massachusetts General Hospital and Harvard Medical School, USA MD Anderson Cancer Center, Houston, Texas, USA MRC, London, U.K. National Cancer Institute (NCI), Bethesda and Frederick, MD, USA Ospedale San Giovanni, Bellinzona, Switzerland Paterson Institute for Cancer Research, Manchester, U.K. Southern Europe New Drug Organization (SENDO), Milan, Italy Swiss Federal Institute of Technology, Zurich, Switzerland The Sackler Institute, University College London, U.K. Tumor Biology and Metastasis Institute of Cancer Research, Sutton, U.K. University College, London Medical School, London, U.K. University of Birmingham, U.K. University of Cincinnati, USA University of Crete Medical School, Greece University of Newcastle, U.K. University of Pau, France University of Wisconsin, Madison, WI, USA Kyoto University, Japan Weizmann Institute of Science, Israel

EDITORIAL BOARD MEMBERSHIP

American Journal of Cancer Research (Maurizio DIncalci) Attualit in Senologia (Paola Mosconi) British Journal of Cancer (Maurizio DIncalci) Chemotherapy (Maurizio DIncalci) Clinical Experimental Metastasis (Raffaella Giavazzi) Current Opinion in Oncologic, Endocrine and Metabolic Drugs (Maurizio DIncalci)

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Current Cancer Therapy Reviews (Raffaella Giavazzi, Giulia Taraboletti) European Journal of Cancer (Maurizio DIncalci, Giovanna Damia, Raffaella Giavazzi, Massimo Broggini e Giulia Taraboletti) Frontiers in Pharmacology (Maurizio DIncalci) Health and Quality of Life Outcomes (Giovanni Apolone, Paola Mosconi) International Journal of Biological Markers (Raffaella Giavazzi) International Journal for Quality in Health Care (Giovanni Apolone) Journal of Ambulatory Care and Management (Giovanni Apolone) Journal of B.U.ON. (Maurizio DIncalci) Journal of Cancer Microenvironment (Raffaella Giavazzi) Journal of Chemotherapy (Raffaella Giavazzi) Journal of Medicine and the Person (Giovanni Apolone) Journal of Preventive Medicine anf Hygiene (Giovanni Apolone) Molecular Cancer Therapeutics (Maurizio DIncalci) Oncology Research (Maurizio DIncalci) Tumori (Maurizio DIncalci, Raffaella Giavazzi) www.PartecipaSalute.it (Paola Mosconi)

PEER REVIEW ACTIVITIES

Acta Orthopaedica, American Journal of Pathology, Annals of Hematology, Annals of Oncology, Anti-cancer Drugs, Biochemical Pharmacology, BioMed Central Editorial, British Journal of Cancer, British Journal of Pharmacology, British Medical Journal, Cancer Chemotherapy and Pharmacology, Cancer Detection and Prevention, Cancer Letters, Cancer Research, Carcinogenesis, Chemico-Biological Interactions, Clinical & Experimental Metastasis, Clinical Cancer Research, Cytometry, European Journal of Cancer, European Journal of Immunology, Faseb Journal, Gynecologic Oncology, Health and Quality of Life Outcomes, Health Expectations, European Journal of Neurology, Intensive Care Medicine, International Journal of Biological Markers, International Journal of Cancer, International Journal of Gynecological Cancer, International Journal for Quality in Health Care, Journal of Ambulatory Care and Management, Journal of Biological Chemistry, Journal of Biological Markers, Journal of Cell Biochemistry, Journal of Cellular and Molecular Medicine, Journal of Chemotherapy, Journal of Clinical Oncology, Journal of Experimental Therapeutics and Oncology, Journal of Medicinal Chemistry, Journal of Medicine and the Person, Journal of the National Cancer Institute, Journal of Neurology, Journal of Nucleic Acids, Journal of Preventive Medicine and Hygiene, Journal of the National Cancer Institute, Leukemia, Molecular Cancer Therapeutics, Molecular Medicine, Nature Biotechnology, Nature Reviews, Oncology Research, PharmacoEconomics, PLoS ONE, Psycho-Oncology, Quality of Life Research, Science, The Patient: patient-centered outcomes research, Tumori, ZEG Centre for Epidemiology & Health Research.

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Ethical Committee, Centro di Riferimento Oncologico, Aviano PN, Italy Ethical Committee, Ente Ospedaliero San Paolo, Milan, Italy Comitato Etico Fondazione CNAO - Centro Nazionale di Adroterapia Oncologica, Pavia Comitato Etico Fondazione IRCCS Istituto Nazionale dei Tumori (INT), Milano

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Comitato Etico Istituto Clinico Humanitas, Rozzano, MI Ethical Committee, Istituto Europeo di Oncologia, Milan, Italy Ethical Committee, Istituto Neurologico Carlo Besta, Milan, Italy Ethical Committee, Istituto Scientifico Eugenio Medea, Bosisio Parini, Lecco, Italy Ethical Committee, Ospedale San Gerardo, Monza, Milan, Italy Ethical Committee, Ospedale SantAnna, Como, Italy Ethical Committee, Ospedale della Valtellina e Valchiavenna, Sondrio, Italy Ethical Committee, IRCCS MultiMedica, Sesto San Giovanni, Milan, Italy Ethical Committee, Azienda USL di Bologna, Italy Executive Board of GCIG (Gynecologic Cancer Intergroup) Comitato Scientifico, Fondazione Buzzi Unicem Onlus Comitato Strategico e di Studio per la Leucemia Linfoblastica Acuta (CSS - LLA) Comitato Tecnico-Scientifico, Alleanza Contro il Tumore Ovarico (ACTO), Milano Scientific Committee, Associazione Italiana Ematologia e Oncologia Pediatrica, Monza, Milan, Italy Scientific Committee, Pezcoller Foundation, Trento, Italy Technical-Scientific Commitee, Associazione Italiana per la Ricerca sul Cancro, Milan, Italy Board of Directors, Fondazione Nerina e Mario Mattioli Onlus, Milan, Italy Board of Directors, Societ Italiana di Cancerologia (SIC) Board of Directors, Societ Italiana di Citometria (GIC) Directional Council Areas- Centro Cochrane Italiano (CCI), Milan National Advisory Board 8th World Congress of Psycho-Oncology Developmental Therapeutics Program, National Cancer Institute (NCI) Decision Network and Executive Committee, South Europe New Drug Organization (SENDO) Executive Board, Europa Donna Executive Committee, European Association for Cancer Research (EACR) Fondazione Attilia Pofferi, Pistoia, Italy NHS R&D National Coordinating Centre for Health Technology Assessment, UK Pezcoller Foundation-ECCO Award University Medical School of Siena, Italy

EVENT ORGANIZATION
Investigators meeting: Studio clinico randomizzato e controllato, in aperto, per comparare lefficacia analgesica di percorsi terapeutici effettuati con ossicodone, fentanyl e buprenorfina verso morfina, in pazienti con dolore associato a cancro di intensit moderata-severa, a partire dal momento in cui iniziano il trattamento con 3 scalino della scala analgesica del WHO. STUDIO CERP, Milan, April 13, 2010. Conference: International Clinical Trials Day 2010. Quale ricerca per quale salute? La centralit di cittadini & pazienti nel dibattito sulla dimensione sociale della salute. Milan, May 19, 2010. Meeting: Nuovi farmaci in ginecologia oncologica e 7 Assemblea Gruppo MaNGO, Milan, June 18-19, 2010. Meeting: V edizione percorso di formazione PartecipaSalute - Orientarsi in salute e sanit. Rimini, October-September 2010.

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Conference: XXVIII Conferenza Nazionale di Citometria - Scuola Nazionale di Citometria, Corsi residenziali di formazione e aggiornamento. Urbino, September 29 October 2 2010. Conference: IV edizione percorso di formazione PartecipaSalute - LAccademia del Cittadino. Milan - Montecatini Terme, October 2009-March 2010. Conference: Informarsi, conoscere e partecipare per migliorare la qualit della vita - Il caso di asma, diabete di tipo 2 e cancro al seno. Milan, November 24, 2010. Conference: Tumore Ovarico Primo incontro pazienti, ricercatori e clinici. Alleanza Contro il Tumore Ovarico (ACTO). Milan, December 3, 2010.

CONFERENCE AND WORKSHOP CONTRIBUTIONS

Conference: International Conference in memory of Judah Folkman - Update on angiogenesis: translational research. Rome (Italy), January 15-16, 2010. Inhibitors of angiogenesis in preclinical models. Conference: La chemioterapia metronomica: quale razionale e quali evidenze. Brescia (Italy), February 26, 2010. Inibitori dellangiogenesi in combinazione con chemioterapici: nuove strategie terapeutiche. Conference: CONTACI. Biella (Italy), March 20, 2010. Raccontaci: i percorsi di diagnosi e cura, lesperienza di Ascoltaci: esperienze di pazienti, noi contiamo su di voi. Meeting: XIV Riunione Scientifica Annuale Italian Sarcoma Group. Pordenone (Italy), March 18-19, 2010. Ricerca traslazionale e sviluppo di farmaci per i sarcomi. Conference: Stress ossidativo e infiammazione nel malato oncologico. Milan (Italy), March 27, 2010. Chemioterapici antineoplastici di origine naturale vegetale nellera delle terapie mirate. Meeting: 4 Meeting Internazionale: Imaging Metabolico PET per una moderna Radioterapia. Reggio Emilia (Italy), April 16-17, 2010. Health Technology Assessment: modelli, strutture ed esperienze. Ipotesi, opportunit e limiti di: Ricerca di Base. Course: 9 Corso di formazione avanzata Cellule staminali tumorali: il vero bersaglio nella cura dei tumori. Pavia (Italy), April 19-23, 2010. Come il paradigma di cellula staminale tumorale (e sua nicchia) indirizza la strategia terapeutica nella cura dei tumori. Conference: 5th International Conference on Thrombosis and Hemostasis Issues in Cancer. Stresa (Italy), April 23-25, 2010. Biology of blood coagulation. Simposyum: First EFIC Symposium Societal Impact Pain (SIP). Brussels (Belgium), May 4-5, 2010. Pain undertreatment in oncological patients.

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Workshop: SIICA - Angiogenesi: basi molecolari ed implicazioni terapeutiche III. Pontignano, Siena (Italy), May 10-12, 2010. F8-IL2 immunocytokine in combination with chemotherapy on melanoma xenograft. Identifying rational combination therapies with inhibitors of angiogenesis and chemotherapy in cancer treatment. PRSS3/Trypsinogen IV, a potential marker of tumor endothelium and its role in angiogenesis. Tubulin acetylation as a possible mechanism of the antiangiogenic activity of paclitaxel: implications for combination therapies with HDAC inhibitors. Design of antiangiogenic agents based on the endogenous inhibitor thrombospondin-1 Congress: V Giornata Novarese di Studio: Malattie dellapparato uro-genitale maschile e femminile. Santa Margherita Ligure (Italy), May 15, 2010. Dalla diagnosi alla terapia: ruolo e valore dell'HTA (Health Tecnology Assessment). Course: 3rd Course of in vivo Preclinical Assays in Cancer Therapy. Paris (France), May 17-19, 2010. Testing anti-angiogenesis drugs in vivo. Congress: 10 Congresso Nazionale Associazione Italiana di Miologia. Milan (Italy), June 3-5, 2010. La scala SF-36: Perch altri questionari?. Meeting: Annual Meeting ASCO. Chicago (USA), June 4-8, 2010. Effect of tumor-specific KRAS mutational status on impact of anti-EGFR therapy in non-small cell lung cancer (NSCLC) Conference: Gordon Research Conference: Molecular Mechanisms in Lymphatic Function and Disease. Lucca (Italy), June 13-18, 2010. VEGFC and VEGFR3 in ovarian tumor progression. Meeting: MaNGO - Nuovi farmaci in ginecologia oncologica. Milan (Italy), June 18-19, 2010. Angiogenesi e anti-angiogenesi. Meeting: 21st Meeting of the European Association for Cancer Research. Oslo (Norway), June 26-29, 2010. Inflammation and cancer. The FGF-2 binding domain of thrombospondin-1: functional characterization and exploitation to design antiangiogenic compounds. Differences in the stroma of human ovarian carcinoma xenografts endowed with different angiogenic phenotypes. Conference: Thrombospondins and matricellular proteins in tissue organization and homeostasis. FASEB summer research conferences. Snowmass Village (USA), July 18-23, 2010. Rational design of antiangiogenic agents based on thrombospondin-1. Conference: 9th Global Conference - Making Sense of: Health, Illness and Disease, Oxford (UK), September 11, 2010. Self-rated health: the impact of symptoms in a gender perspective. Conference: MRS-AACR Joint Conference on Metastasis and the Tumor Microenvironment. Philadelphia (USA), September 12-15, 2010. Molecular profile of the stroma from human ovarian carcinoma xenografts equipped with different angiogenic phenotypes.

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Conference: XXVIII Conferenza Nazionale di Citometria - Scuola Nazionale di Citometria, Corsi residenziali di formazione e aggiornamento. Urbino (Italy), September 29 October 2, 2010. Proliferazione cellulare e apoptosi. Meeting: 52nd Meeting della Societ Italiana di Cancerologia (SIC). Rome (Italy), October 4-7, 2010. Cancer modeling. Combinations of chemotherapy and Bevacizumab in a xenograft model of human ovarian carcinoma. Congress: ESMO. Milan (Italy), October 8, 2010. Metabolic approach to the enhancement of antitumor effect of chemotherapy: a key role for carnitine system (Simposio Sigma-Tau Cancer: the bioenergetic challenge) Sorafenib activity in NSCLC cell lines seems related to a different type of K-RAS mutations KRAS mutations differing for aminoacid substitution confer different drug sensitivity and resistance in NSCLC. Congress: 2010 Joint Colloquium of the Cochrane and Campell Collaborations. Keystone (Colorado, USA), October 18-22, 2010. Integrating and deriving evidence, experiences and preferences (IN-DEEP): developing research-based health information applicable to decision making and self-management by people with multiple sclerosis. Training for patients and citizens representatives: evaluating the PartecipaSalute courses. Meeting: I Cantieri ravennati Eticit e laicit nella ricerca biomedica. Ravenna (Italy), October 23, 2010. La ricerca biomedica indipendente. Congress: The International Event & Conference on Biotechnologies - Biotechs promises for the future: turning challenges into opportunities. Milan (Italy), October 26-28, 2010. Angiogenesis inhibitors: from preclinical studies to clinical development. Meeting: EORTC-NCI-ASCO Annual Meeting on Molecular Markers in Cancer. Brussels (Belgium), October 27-29, 2010. Role of different types of K-Ras mutations in determining drug sensitivity and tumor behavior in non-small cell lung cancer. Congress: Le biblioteche per pazienti in Italia, esperienze a confronto. Reggio Emilia (Italy), October 29-30, 2010. Fare informazione attraverso la partecipazione: lesempio del progetto PartecipaSalute. Course: Introductory PhD course in translational medicine. Milan (Italy), November 8-12, 2010. How to develop an anti-tumor drug, a few examples of success. Congress: 3rd European Public Health Conference Integrated Public Health. Amsterdam (The Netherlands), November 1013, 2010. A community health program to promote womens knowledge of menopause and hormone therapy. Simposyum: 22nd EORTC-NCI-AACR Symposium. Berlin (Germany), November 16-19, 2010. Myxoid liposarcoma tumors with different chimera subtypes xenografted in nude mice are characterized by different response to trabectedin and gene expression profile. Paclitaxel enhances therapeutic efficacy of F8-antibody mediated delivery of Interleukin-2 to xenografted melanoma cancer.

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E-3810, an inhibitor of the VEGF and FGF family receptors, inhibits the FGF-dependent growth of tumor cells Changes in the transcription regulation mediated by FUS-CHOP gene in a myxoid liposarcoma cell line resistant to trabectedin Pharmacokinetics (Pk) and pharmacodynamics (Pd) of the novel proteasome inhibitor Cep18770 during a phase I trial in patients with solid tumor, non-hodgkin lymphoma or multiple myeloma. Congress: 27 Congresso Nazionale SIMG. Florence (Italy), November 25-27, 2010. Il Progetto SIMG sulla valutazione e gestione del dolore cronico nella Medicina Generale. Studio di un intervento randomizzato. Course: Highlight in oncologia ginecologica. Santa Margherita Ligure (Italy), November 25-27, 2010. Molecular Box I Molecular Box II Congress: Tumore Ovarico Primo incontro pazienti, ricercatori e clinici. Alleanza Contro il Tumore Ovarico (ACTO). Milan, December 3, 2010. Cellule tumorali staminali dellovaio. Un nuovo progetto di ricerca in collaborazione Ieo/Ifom e Istituto Mario Negri per lidentificazione dei fattori alla base della sensibilit o resistenza ai farmaci.

GRANTS AND CONTRACTS

ABO Project SpA Arcispedale Santa Maria Nuova di Reggio-Emilia Associazione Italiana Otorinolaringologi Ospedalieri (AOOI) Associazione Volontarimini - Associazione per lo Sviluppo del Volontariato della Provincia di Rimini Azienda Sanitaria Locale, Reggio Emilia Agenzia Italiana del Farmaco (AIFA) Amgem SpA, Milano AIRC Associazione Italiana per la Ricerca sul Cancro ArQule USA Astra Zeneca UK AVAPO (Associazione Volontari Assistenza Pazienti Oncologici) Azienda Ospedaliera Fatebenefratelli e Oftalmico- Milano Boehringer CIPOMO (Collegio Italiano dei Primari Oncologi Medici Ospedalieri) CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano Compagnia di San Paolo Elsevier Science Ltd EOS SpA European Commission - 7th Framework Programme (ADAMANT) European Union (EPOC) FIRB-MIUR Fondo per gli Investimenti della Ricerca di Base-Ministero Istruzione Universit e Ricerca

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FIRC Fondazione Italiana per la Ricerca sul Cancro Fondazione Cassa di Risparmio delle Province Lombarde Fondazione IRCCS Istituto Nazionale dei Tumori, Milano Fondazione Lilly Fondazione Nerina e Mario Mattioli Onlus Fondazione SmithKline, Roma FSE Frontier Southern Europe GISCAD (Gruppo Italiano Studi di Carcinomi Apparato Digerente) GlaxoSmithKline, Verona Grunenthal Italia, Milano Indena SpA Istituto Europeo di Oncologia (IOSI) Istituto Oncologico Svizzera Italiana Istituto Superiore di Sanit Italfarmaco SpA Merck Sharp & Dome Ministero della Salute Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA Oncoethix OTD - Oncology Therapeutic Development s.a.r.l. Pfizer Italia Pharma Mar, SA Regione Emilia Romagna Regione Lombardia Regione Toscana Formas Sanofi-Aventis Pharma SENDO-Tech Srl Sia-ssb SpA Sigma-Tau SpA SIMG Firenze Universit Federico II Napoli (Dipartimento di Endocrinologia ed Oncologia molecolare e clinica)

SCIENTIFIC PUBLICATIONS (2010)

Frapolli R, Zucchetti M, Sessa C, Marsoni S, Vigano' L, Locatelli A, Rulli E, Compagnoni A, Bello E, Pisano C, Carminati P, D'Incalci M Clinical pharmacokinetics of the new oral camptothecin gimatecan: The inter-patient variability is related to (1)-acid glycoprotein plasma levels Eur J Cancer 2010 46 : 505-516 Mosconi P, Taricco M, Bergamini M, Bosisio Fazzi L, Colombo Cinzia, Patrucco V, Corti M, Giobbe D, Guerreschi M, Magnarella M R, Sallemi G Family burden of severe brain injury: The Italian experience with families and volunteer associations Patient 2010 E-pub Silini Antonietta, Ghilardi A, Ardinghi C, Bernasconi S, Naldini A, Bani M R, Giavazzi R Protease-activated receptor-1 (PAR-1) promotes the motility of human melanomas and is associated to their metastatic phenotype Clin Exp Metastasis 2010 27 : 43-53 Borgia B, Roesli C, Fugmann T, Schliemann C, Cesca M, Neri D, Giavazzi R A proteomic approach for the identification of vascular markers of liver metastasis Cancer Res 2010 70 : 309-318

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Andreis F, Rizzi A, Mosconi P, Braun C, Rota L, Meriggi F, Mazzocchi M, Zaniboni A Quality of life in colon cancer patients with skin side effects: preliminary results from a monocentric cross sectional study Health Qual Life Outcomes 2010 8 : 40 Azzariti A, Porcelli L, Simone G M, Quatrale A E, Colabufo N A, Berardi F, Perrone R, Zucchetti M, D'Incalci M, Xu J M, Paradiso A Tyrosine kinase inhibitors and multidrug resistance proteins: interactions and biological consequences Cancer Chemother Pharmacol 2010 65 : 335-346 Greco M T, Corli O, Montanari M, Deandrea S, Zagonel V, Apolone G, CPOR SG Investigators Epidemiology and pattern of care of Breakthrough cancer Pain (BTcP) in a longitudinal sample of cancer patients. Results from the CPOR-SG Clin J Pain 2010 E-pub De Pangher V, Recchia L, Cafferata M, Porta C, Siena S, Giannetta L, Morelli F, Oniga F, Bearz A, Torri V, Cinquini M Malignant peritoneal mesothelioma: a multicenter study on 81 cases Ann Oncol 2010 21 : 348-353 Previdi S, Maroni P, Matteucci E, Broggini M, Bendinelli P, Desiderio M A Interaction between human-breast cancer metastasis and bone microenvironment through activated hepatocyte growth factor/Met and beta-catenin/Wnt pathways Eur J Cancer 2010 46 : 1679-1691 Zucchetti M, Meco D, Di Francesco A M, Servirei T, Patriarca V, Cusano G, D'Incalci M, Forestieri D, Pisano C, Riccardi R Antitumor activity and pharmacokinetics of oral gimatecan on pediatric cancer xenografts Cancer Chemother Pharmacol 2010 66 : 635-641 Sabatino M A, Marabese M, Ganzinelli M, Caiola E, Geroni C, Broggini M Down-regulation of the nucleotide excision repair gene XPG as a new mechanism of drug resistance in human and murine cancer cells Mol Cancer 2010 9 : 259 Falasca M, Chiozzotto D, Godage H Y, Mazzoletti M, Riley A M, Previdi S, Potter B V L, Broggini M, Maffucci T A novel inhibitor of the PI3K/Akt pathway based on the structure of inositol 1,3,4,5,6-pentakisphosphate Br J Cancer 2010 102 : 104-114 Berndt A, Kollner R, Richter P, Franz M, Voigt A, Berndt Angela, Borsi L, Giavazzi R, Neri D, Kosmehl H A comparative analysis of oncofetal fibronectin and tenascin-C incorporation in tumour vessels using human recombinant SIP format antibodies Histochem Cell Biol 2010 133 : 467-475 Floriani I, Torri V, Rulli E, Garavaglia D, Compagnoni A, Salvolini L, Giovagnoni A Performance of imaging modalities in diagnosis of liver metastases from colorectal cancer: a systematic review and meta-analysis J Magn Reson Imaging 2010 31 : 19-31 Urru S A M, Veglianese P, De Luigi A, Fumagalli E, Erba E, Gonella Diaza R, Carr A, Davoli E, Borsello T, Forloni G, Pengo N, Monzani E, Cascio P, Cenci S, Sitia R, Salmona M A new fluorogenic peptide determines proteasome activity in single cells J Med Chem 2010 53 : 7452-7460 Cazzola M, Floriani I, Page C P The therapeutic efficacy of erdosteine in the treatment of chronic obstructive bronchitis: a meta-analysis of individual patient data Pulm Pharmacol Ther 2010 23 : 135-144 Pezzola S, Antonini G, Geroni C, Beria I, Colombo M, Broggini M, Mongelli N, Leboffe L, MacArthur R, Mozzi A F, Federici G, Caccuri A M Role of glutathione transferases in the mechanism of brostallicin activation Biochemistry 2010 49 : 226-235

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Carrassa L, Montelatici E, Lazzari L, Zangrossi S, Simone M, Broggini M, Damia G Role of Chk1 in the differentiation program of hematopoietic stem cells Cell Mol Life Sci 2010 67 : 1713-1722 Howells L M, Britton R G, Mazzoletti M, Greaves P, Broggini M, Brown K, Steward W P, Gescher A J, Sale S Preclinical colorectal cancer chemopreventive efficacy and p53-modulating activity of 3',4',5'-trimethoxyflavonol, a quercetin analogue Cancer Prev Res (Phila Pa) 2010 E-pub Previdi S, Malek A, Albertini V, Riva C, Capella C, Broggini M, Carbone G M, Rohr J, Catapano C V Inhibition of Sp1-dependent transcription and antitumor activity of the new aureolic acid analogues mithramycin SDK and SK in human ovarian cancer xenografts Gynecol Oncol 2010 118 : 182-188 Colombo G, Margosio B, Ragona L, Neves M, Bonifacio S, Annis D S, Stravalaci M, Tomaselli S, Giavazzi R, Rusnati M, Presta M, Zetta L, Mosher D F, Ribatti D, Gobbi M, Taraboletti G Non-peptidic thrombospondin-1 mimics as fibroblast growth factor-2 inhibitors: an integrated strategy for the development of new antiangiogenic compounds J Biol Chem 2010 285 : 8733-8742 Sansone V A, Panzeri M, Montanari M, Apolone G, Gandossini S, Rose M R, Politano L, Solimene C, Siciliano G, Volpi L, Angelini C, Palmieri A, Toscano A, Musumeci O, Mongini T, Vercelli L, Massa R, Panico M B, Grandi M, Meola G Italian validation of INQoL, a quality of life questionnaire for adults with muscle diseases Eur J Neurol 2010 E-pub Mannucci E, Petroni M L, Villanova N, Rotella C M, Apolone G, Marchesini G, QUOVADIS Study Group Clinical and psychological correlates of health-related quality of life in obese patients Health Qual Life Outcomes 2010 8 : 90 Brunelli D, Tavecchio M, Falcioni C, Frapolli R, Erba E, Iori R, Rollin P, Barillari J, Manzotti C, Morazzoni P, D'Incalci M The isothiocyanate produced from glucomoringin inhibits NF-kB and reduces myeloma growth in nude mice in vivo Biochem Pharmacol 2010 79 : 1141-1148 Naldini A, Filippi I, Moschetta M, Giavazzi R, Carraro F Interleukin-1 regulates the migratory potential of MDAMB231 breast cancer cells through the hypoxia-inducible factor-1 Eur J Cancer 2010 46 : 3400-3408 Germano G, Frapolli R, Simone M, Tavecchio M, Erba E, Pesce S, Pasqualini F, Grosso F, Sanfilippo R, Casali P, Gronchi A, Virdis E, Tarantino E, Pilotti S, Greco A, Nebuloni M, Galmarini C M, Tercero J C, Mantovani A, D'Incalci M, Allavena P Antitumor and anti-inflammatory effects of trabectedin on human mixoid liposarcoma cells Cancer Res 2010 70 : 2235-2244 Miserocchi E, Modorati G, Mosconi P, Colucci A, Bandello F Quality of life in patients with uveitis on chronic systemic immunosuppressive treatment Ocul Immunol Inflamm 2010 18 : 247-254 Frey K, Schliemann C, Schwager K, Giavazzi R, Johannsen M, Neri D The immunocytokine F8-IL2 improves the therapeutic performance of sunitinib in a mouse model of renal cell carcinoma J Urol 2010 184 : 2540-2548 D'Incalci M, Galmarini C M A review of trabectedin (ET-743): A unique mechanism of action Mol Cancer Ther 2010 E-pub Floriani I, Garassino M C, Broggini M, Veronese S, Marsoni S, Marabese M, Farina G, Scanni A Role of cetuximab in the treatment of patients with NSCLC: are we throwing out the baby with the bath water? J Clin Oncol 2010 28 : e467

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Cremolini C, Loupakis F, Ruzzo A, Perrone G, Rulli E, Vincenzi B, Tonini G, Graziano F, Onetti Muda A, Falcone A Predictors of benefit in colorectal cancer treated with cetuximab: are we getting &quot;Lost in TranslationAL&quot;? J Clin Oncol 2010 28 : e173-e174 Luciani A, Ascione G, Bertuzzi C, Marussi D, Codec C, Di Maria G, Caldiera S, Floriani I, Zonato S, Ferrari D, Foa P Detecting disabilities in older patients with cancer: comparison between comprehensive geriatric assessment and Vulnerable Elders Survey-13 J Clin Invest 2010 28 : 2046-2050 Rossi A, Garassino M C, Cinquini M, Sburlati P, Di Maio M, Farina G, Gridelli C, Torri V Maintenance or consolidation therapy in small-cell lung cancer: A systematic review and meta-analysis Lung Cancer 2010 70 : 119-128 Caffo O, Fallani S, Marangon E, Nobili S, Cassetta M I, Murgia V, Sala F, Novelli A, Mini E, Zucchetti M, Galligioni E Pharmacokinetic study of gemcitabine, given as prolonged infusion at fixed dose rate, in combination with cisplatin in patients with advanced non-small-cell lung cancer Cancer Chemother Pharmacol 2010 65 : 1197-1202 Mosconi P, Colombo Cinzia Fostering a strategic alliance between patients' associations and health care professionals J Ambul Care Manage 2010 33 : 223-230 Wale J, Colombo Cinzia, Belizan M, Nadel J International health consumers in the cochrane collaboration: fifteen years on J Ambul Care Manage 2010 33 : 182-189 Knudsen K A, Brunelli C, Kaasa S, Apolone G, Corli O, Montanari M, Fainsinger R, Aass N, Fayers P, Caraceni A, Klepstad P, European Palliative Care Research Collaborative (EPCRC), European Pharmacogenetic Study (EPOS) Which variables are associated with pain intensity and treatment response in advanced cancer patients? Implications for a future classification system for cancer pain Eur J Pain 2010 E-pub Bertuzzi F, Suzani M, Tagliabue E, Cavaletti G, Angeli R, Balgera R, Rulli E, Ferrarese C, Miglior S Diagnostic validity of optic disc and retinal nerve fiber layer evaluations in detecting structural changes after optic neuritis Ophthalmology 2010 117 : 1256-1264 Sargent D J, Marsoni S, Monges G, Thibodeau S N, Labianca R, Hamilton S R, French A J, Kabat B, Foster N R, Torri V, Ribic C, Grothey A, Moore M, Zaniboni A, Seitz J - F, Sinicrope F, Gallinger S Defective mismatch repair as a predictive marker for lack of efficacy of fluorouracil-based adjuvant therapy in colon cancer J Clin Oncol 2010 28 : 3219-3226 Pisano C, Vesci L, Milazzo F M, Guglielmi M B, Foder R, Barbarino M, D'Incalci M, Zucchetti M, Petrangolini G, Tortoreto M, Perego P, Zuco V, Orlandi A, Passeri D, Carminati P, Cavazza C, Zunino F Metabolic approach to the enhancement of antitumor effect of chemotherapy: a key role of acetyl-L-carnitine Clin Cancer Res 2010 16 : 3944-3953 Gacci M, Corona G, Apolone G, Lanciotti M, Tosi N, Giancane S, Masieri L, Serni S, Maggi M, Carini M Influence of serum testosterone on urinary continence and sexual activity in patients undergoing radical prostatectomy for clinically localized prostate cancer Prostate Cancer Prostatic Dis 2010 13 : 168-172 Corli O, Maltoni M Pain and bone metastases In :Osteo-oncology textbook Poletto Editore, Vermezzo (MI), 2010; 277-294 Damia G, D'Incalci M Genetic instability influences drug response in cancer cells Curr Drug Targets 2010 11 : 1317-1324

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NSCLC Meta-Analyses Collaborative Group, Torri V Adjuvant chemotherapy, with or without postoperative radiotherapy, in operable non-small-cell lung cancer: two meta-analyses of individual patient data Lancet 2010 375 : 1267-1277 Sala F, Marangon E, Bagnati R, Livi V, Cereda R, D'Incalci M, Zucchetti M Development and validation of a high-performance liquid chromatographytandem mass spectrometry method for the determination of the novel proteasome inhibitor CEP-18770 in human plasma and its application in a clinical pharmacokinetic study J Mass Spectrom 2010 45 : 1299-1305 Mosconi P, Donati S, Colombo Cinzia, Senatore S Role of hormone therapy in the management of menopause Obstet Gynecol 2010 116 : 442 Mazzoletti M, Broggini M PI3K/AKT/mTOR inhibitors in ovarian cancer Current Medicinal Chemistry - Anti-Cancer Agents 2010 E-pub Colombo C, Satolli R, Liberati A, Mosconi P, Giurie dei cittadini Working Group Citizens' juries in health care BMJ 2010 341 : c5141 Casciani E, Masselli G, Di Nardo G, Polettini E, Bertini L, Oliva S, Floriani I, Cucchiara S, Gualdi G MR enterography versus capsule endoscopy in paediatric patients with suspected Crohn's disease Eur Radiol 2010 E-pub Catalano M, Scandale G, Minola M, Carzaniga G, Carotta M, Perilli A, Dimitrov G, Cortellazzo A, Cinquini M Elastic properties and structure of the radial artery in patients with type 2 diabetes Diab Vasc Dis Res 2010 6 : 244-248 Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone M V, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta M G, Riva E Prevalence, incidence and types of mild anemia in the elderly: the &quot;Health and Anemia&quot; population-based study Haematologica 2010 95 : 1849-1856 Hogberg T, Signorelli M, Freire de Oliveira C, Fossati R, Lissoni A A, Sorbe B, Andersson H, Grenman S, Lundgren C, Rosenberg P, Boman K, Tholander B, Scambia G, Reed N, Cormio G, Tognon G, Clarke J, Sawicki T, Zola P, Kristensen G Sequential adjuvant chemotherapy and radiotherapy in endometrial cancer - Results from two randomised studies Eur J Cancer 2010 46 : 2422-2431 Sessa C, D'Incalci M Trabectedin in ovarian cancer:could we expect more? Ann Oncol 2010 E-pub Taraboletti G, Rusnati M, Ragona L, Colombo G Targeting tumor angiogenesis with TSP-1-based compounds: rational design of antiangiogenic mimetics of endogenous inhibitors Oncotarget 2010 E-pub Rusnati M, Urbinati C, Bonifacio S, Presta M, Taraboletti G Thrombospondin-1 as a paradigm for the development of antiangiogenic agents endowed with multiple mechanisms of action Pharmaceuticals 2010 3 : 1241-1278 Floriani I, Garattini S, Torri V Looking for efficiency rather than efficacy in randomized controlled trials in oncology Ann Oncol 2010 21 : 1391-1393 Colombo C, Mosconi P, Buratti M G, Liberati A, Donati S, Mele A, Satolli R Press coverage of hormone replacement therapy and menopause

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Eur J Obstet Gynecol Reprod Biol 2010 153 : 56-61 Labianca R, Sobrero A, Isa L, Cortesi E, Barni S, Nicolella D, Aglietta M, Lonardi S, Corsi D, Turci D, Beretta G D, Fornarini G, Dapretto E, Floriani I, Zaniboni A, GISCAD Intermittent versus continuous chemotherapy in advanced colorectal cancer: a randomised 'GISCAD' trial Ann Oncol 2010 E-pub

LAY PRESS SELECTION (2010)

Villani W, Mosconi P Il nuovo MisuraAssociazioni http://www.partecipasalute.it/cms_2/node/1574 , 21/10/2010 Mosconi P Un progetto-ricerca di crescita e sviluppo centrato sul paziente: PartecipaSalute In: Quale salute per chi, a cura di Fede Ruggeri, Franco Angeli/Sanit Editore Ottobre 2010: 127-141 Carra L, Colombo C. Linformazione in medicina: come destreggiarsi In: Quale salute per chi, a cura di Fede Ruggeri, Franco Angeli/Sanit Editore Ottobre 2010: 81-105. Mosconi P, Carra L A salute nun saccatta ma sabbusca http://www.partecipasalute.it/cms_2/node/1565 , 20/10/2010 Mosconi P Lanno del girasole pallido http://www.partecipasalute.it/cms_2/node/1549 , 14/9/2010 Colombo C Prima dellipertensione, ma gi a rischio http://www.partecipasalute.it/cms_2/node/1546 , 10/9/2010 Braun C, Mosconi P Farmaci: la lunga strada per lautorizzazione allimmissione in commercio http://www.partecipasalute.it/cms_2/node/1542, 31/8/2010 Mosconi P, Colombo C, Liberati A, Satolli R Fare empowerment con le associazioni di cittadini e pazienti. Lesperienza di PartecipaSalute. I Quaderni di Monitor 2010, 25 (Suppl 6): 124-130 Mosconi P, Marsico G, Satolli R, Casali P Attivit formative e progetti collaborativi destinati ai componenti laici dei comitati etici http://www.partecipasalute.it/cms_2/node/1455 , 16/4/2010 Mosconi P Cittadini, pazienti e partecipazione Bollettino SIFO Volume 56 N 3 (marzo-aprile 2010): 86-87 Mosconi P A proposito di farmaci equivalenti Mia Farmacia Online, marzo 2010: 26-27 Greco M T, Montanari M, Deandrea S, Corli O, Zagonel V, Caraceni A, Apolone G, CPOR SG Investigators Il dolore nel paziente con cancro: sintesi dei risultati di un progetto quinquennale Ricerca & Pratica 2010 n.153 : 95-105 Gallus S, Apolone G, Padula A, Casadei G, Motterlini N, Garattini L &quot;Quaderni di farmacoeconomia&quot; risponde ai lettori Quaderni Farmacoeconomia 2010 12 : 24-31

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Mercadante S, Amadori D, Apolone G, Arcuri E, Barbato A, Caraceni A, Maltoni M, Marchetti P, Mattia C, Varrassi G, Zagonel V, Zucco F Raccomandazioni per la gestione del Breakthrough cancer Pain (BTcP) Rivista Italiana di Cure Palliative 2010 1 : 17-23 Torri V Valutazione della risposta clinica In :Oncologia medica pratica, 3a Ed. Societ Editrice Universo, Roma, 2010; 921-930

RESEARCH ACTIVITIES Laboratory of Cancer Pharmacology Mode of action of Ecteinascidins

A project ongoing since several years is about the characterization of marine natural products possessing antitumor activity. In particular we carried on the studies on the effects of ET-743 in cells defective for some DNA repair mechanisms. Cells deficient for Homologous Recombination (HR) are very sensitive to the drug, while cells deficient for Non Homologous End-Joining (NHEJ) are only slightly more sensitive, but surpraisingly cell lines defective for Nucleotide Excision Repair (NER) are less sensitive to ET-743. Flow cytometric analysis coupled to a software of computer simulation, developed in our laboratory, has demonstrated that NER defective cells showed, after ET-743 treatment, cell cycle perturbations different than those occurring in NER proficient cells, probably for the activation of different and more efficient repair mechanisms. We study also a functional evaluation of the DNA repair mechanisms by the cell capacity to recognize and repair double helix breaks with a recently introduced test that is very sensitive to detect the phosphorylation of histone H2AX. An in vitro study is ongoing with flow cytometry and immunofluorescence techniques to evaluate in different tumor cell lines the phosphorylation level of histone H2AX in relation to the distribution of the cells in the different phases of the cell cycle and the cytotoxic effect induced after treatment with ET-743. Studies are in progress on the mechanism of action of new ET-743 derivates compounds that have shown antitumoral activity on cell lines with different DNA repair mechanisms. A new project is the study of the selective action of ET-743 on mixoid lyposarcoma, a pathology representing 10% of all soft tissue sarcomas, trying to understand if the significative antitumor effect is due to a selective action of the compound on pathogenetic alterations characteristic of this pathology. In particular we are trying to evaluate how ET-743 interact with the transcriptional modifications of specific genes due to the translocation FUS-CHOP that characterizes mixoid sarcomas or those caused by the interaction host-tumor, modifying inflammatory and angiogenetic processes. Studies are in progress to obtain cell lines and xenografts of mixoid lyposarcomas exhibiting the same molecular features of the patients tumors.

Combinations of natural products of marine origin with other anticancer drugs

We have observed additive or synergistic activity of ET-743 combined with other anticancer drugs such as cisplatin, doxorubicin, campthotecin,inhibitors of telomerase, bleomicin and varinostat.

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Analysis of cell cycle data and interactions of different drugs

The Biophysics Unit is engaged in theoretical and methodological studies aimed at a critical evaluation of current techniques of investigation of drug effects on heterogeneous cell populations. Several computing tools have been produced to simulate the cell proliferation at different levels (from molecular interactions to in vivo growth of solid tumours) and the process of measure. Collaborations are ongoing with other research groups for design and data analysis of drug combination studies in vitro. In this field, a number of computer programs have been developed, allowing comparative data analysis with the most common models of drug interaction.

Evaluation of the complexity of the response of cell populations to treatment with anticancer drugs
This project of the Biophysics Unit addresses the issue of establishing a connection between the intracellular drug interactions and the resulting cell cycle perturbations. It starts from the singlecell level of investigation to reach the cell-population level where the relevant end points of treatment efficacy are evaluated by flow cytometry and growth inhibition/cytotoxicity assays. The model adopted for data analysis and interpretation is the result of the merging of two mathematical models. One model describes the cell cycle, exploiting the results of the theory of age-structured cell population dynamics. The second model describes the response to the drug's challenge, using distinct parameters ("effect descriptors") measuring either the strength of cell cycle arrest, damage repair or cell death in every phase (G1, S and G2M). In this way, it is possible to reach a quantitative interpretation of the experimental results, overcoming the current qualitative and partial approaches to this problem, which are unable to resolve the overlapping of cytostatic and cytotoxic effects, and to establish a connection with phase-related events.

Cell cycle dYsregulation in erlotinib-based treatments decoded by flow cytometry and mathematical modeling
Epidermal growth factor receptor (EGFR) inhibitors represent one of the most promising class of anticancer compounds, some of them, like erlotinib, are already used for clinical therapy. Nevertheless, so far, the research has focused on molecular interaction of these compounds, somewhat neglecting the study of the dynamics of cell cycle perturbations and underscoring the importance of this issue for the optimization of both single and multidrug therapies. In order to fill this gap, the Biophysics Unit is currently studying in detail the time- and dosedependence of the cell cycleperturbations induced by different treatment schedules of erlotinib and gemcitabine. The results are expected to disclose the origin of the interactions between the two drugs and advance towards optimization of the combined treatment.

Anticancer Drug Effects Decoded by Time-Lapse Imaging, Flow Cytometry and Modeling in silico
We use flow cytometric (cell-population based analysis) and time-lapse imaging (single cell lineage based analysis) techniques to generate data that will be used to predict drug responses in term of the major components of cytostatic/cytotoxic actions of anticancer drugs: specific cell cycle perturbations (detecting accumulation or depletion of cells in G1, S and G2M phases) and the commitment to cell death (apoptosis). Time lapse data are currently integrated with those from single and multiparametric flow cytometric experiments, and univocally interpreted with a common computer program that renders in silico the proliferation process through the cell cycle and in the cell generations that follow one another during and after treatment. This kind of dynamic rendering establishes a connection between the available macroscopic data (time-lapse and flow cytometric) and the activity of molecular pathways which are in charge to the several functions that concur in the

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pharmacological response with individual timing and dose-dependence-, and which are not otherwisemeasurable. Final aim is to achieve a quantitative level of understanding of the dynamics of response to anticancer treatment, enabling a full appreciation of the role and relative importance of the main cellular functions contributing to the overall response. Methods and computing tools with intuitive interface developed for these tasks will be shared with the scientific community.

Clinical pharmacokinetics of E-3810 (a novel inhibitor of angiogenesis)

The phase I clinical trial started in October. In the months before we developed a new method to measure the drug in human plasma by HPLC-mass-spectrometry in order to study the drug pharmacokinetic profile in the enrolled patients. The trial showed that this compound, which is administered orally for 28 consecutive days, provides high plasma exposure reaching drug concentrations at steady state potentially pharmacologically active after one week of therapy.

Clinical pharmacology of CEP-18770 (a new proteasome inhibitor)

The study in collaboration with SENDO, that was performed in 40 patients, has defined the pharmacokinetic and the pharmacodynamic profiles of the new proteasoma inhibitor, CEP18770, in patients enrolld in the phase I clinical study. This study evidencied for this compound a very long plasma half-life assuring a prolonged drug esposition. Pharmacodynamic studies showed that in patients treated at the recommended dose CEP-18770 caused a significant inhibition (50%) of the proteasome activity.

Quality assurance program

During the year 2010 we improved the quality assurance program aims to bring the pharmacokinetic unit, inside the laboratory of Cancer Pharmacology, in compliance with Good Laboratory Practice (GLP). The first phase has been completed and also the writing of the operating procedures has been finalized. We hope in this year to end the process of development and to request GLP certification for the laboratory.

Antitumoral activity and pharmacokinetic properties of new drugs and combinations

The antitumor activity, pharmacokinetic properties and toxicity of novel anticancer drugs with specific targets (e.g. different kinase inhibitors), conventional anticancer drugs (taxanes and camptothecins) and combinations is being investigated using rodent tumors and human tumor xenografts.

Development of a software framework for a rationalized process of Microarrays analysis

It is currently active in the Institute a multidisciplinary group involved in the rationalization of the various microarray analysis aspects, with many external collaborations. The different possible analysis procedures have been discussed, compared and formalized in order to obtain a common work flow to be accepted from the scientific community. Thanks to this activity it is now possible to automate some aspects of the analysis making them faster end better reproducible for people managing the analysis itself. This activity has involved the development of the necessary software for data analysis and the implementation of a Beowulf computer cluster, using the old computers dismissed by the desktop users in order to obtain a sufficient power. External collaborations: Fondo Edo Tempia (ref.: Giovanna Chiorino) Istituto Toscano Tumori (ref: Duccio Cavalieri) Aberdeen University (ref. Tony Travis)

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Microarray data analysis for the Oncology Department

The data analysis activities concern the biology of sensible versus resistant to therapy human ovarian carcinoma and the study of the trabectedin working mechanism in human sarcoma, sensible and resistant. External collaborations: Ospedale S.Gerardo, Monza

Development of the new database handling software for the ovarian tumors bio-bank, compliant with the new privacy related laws for data handling for clinical trials and with a new reporting engine for a better data extration
External collaborations; Ospedale S.Gerardo, Monza

Development of a validated distributed database for clinical trials

This activity involves the continuous development of a validated distributed database engine for clinical trials. The system architecture is a decentralized platform (push based peer-to-peer) for data sharing for virtual organizations. External collaborations: Istituto Toscano Tumori (ref: Duccio Cavalieri) Aberdeen University (ref. Tony Travis)

Laboratory of Molecular Pharmacology G2 checkpoint and cell cycle

In the last years our laboratory has clarified the role of the Chk1 protein in the G2, S and M blocks induced by different chemotherapeutic drugs. More recently the role of chk1 in unperturbed cell cycle progression has been highlighted in some tumor cell lines. Treatment of small inhibitors of the catalytic activity of chk1 or small interference RNA (siRNA) against chk1 has been shown to cause cell death. In fact, chk1 inhibitors have been though to be used in combination with DNA damaging agents as they would interfere with the activation of the chk1 induced cell cycle checkpioint and would increase the cytotoxic activity of the anticancer agents. The new data on the activity of chk1 inhibitors in some tumor cell lines would suggest a possible use of the inhibitors as single agents. In order to better understand the mechanism at the basis of the observed cytotoxic activity of the inhibitors, we set up a high-through put screening through which we will hopefully find those proteins, whose inactivation is in synthetic lethality with chk1. We will use a siRNA library against 700 human protein kinases, described to have a key role in tumor cells. The results of this screening will allow us the identification of those proteins whose inactivation render tumor cells particularly sensitive to chk1 inhibition and could help in selection the patients that would potentially benefit from a chk1 inhibitor mono-therapy.

Characterization of new potential oncosuppressor genes

DRAGO gene, identified and cloned in our laboratory is one of the most interesting projects of the group. The characterization of the response of KO mice for DRAGO to ionising radiation is similar to normal mice. Mice KO for DRAGO have been crossed with with p53 KO mice to evaluate the potential oncosuppressive function of DRAGO. The double mutants are viable and the genotypes arising

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from the crossing are at the normal Mendelian ratio, indicating that no specific genotypes (p53;DRAGO) are favoured. In a p53KO background, removal of DRAGO gene accelerates tumor development suggesting a cooperative role of the two genes in the prevention of tumnor formation. We are currently evaluating the spectra of tumor formation to determine whether the lack of DRAGO gene preferentially increases the number of some specific histologycal type of tumor. We are characterising the function of the gene both in in vitro and in in vivo systems.

Molecular characterization of ovarian carcinoma

Micro RNAs are small non coding RNAs playing an important role in the regulation of mRNA transcription. They have been found deregulated in several human cancers. We have started a study aimed at evaluating the expression of approximately 600 microRNAs in ovarian cancer from patients with a very high response to treatment (surviving more than 10 years from diagnosis) compared to patients not responding at first line therapy. The analysis of the tumors so far characterised indicates that microRNAs are differentially expressed in the two subgroups . If confirmed in a larger sample size, that could represent a potential therapeutic target. In parallel we have characterised retrospectively, polymorphisms in genes participating in the response to damage such as mdm2, ERCC1 and XPG as possible predictors of response to treatment in patients with ovarian cancer. 420 patients have been genotyped and the allelic frequency found is the expected one for a Caucasian population. The data generated will be analysed together with the clinical parameters and with the follow-up data available for all the samples analysed.

Expression of gene involved in DNA repair in human ovarian cancer

By Real Time PCR, the expression of genes involved in DNA repair has been evaluated in 77 stage I, 81 stage III and 13 borderline samples of ovarian cancer. The genes analysed include those belonging to the nucleotide excision repair, in the fanconi anemia repair, in the base excision repair. In addition, genes important for the cellular response to damage, such as chk1 and claspin have been studied. Two were the aims of the study: 1) to understand whether there are genes differentially expressed in the three categories analysed that could help us in understanding the biology of ovarian cancer; 2) to correlate mRNA levels of the different genes with the response to treatment with the idea of finding new possible response markers. Data analysis showed that genes involved in the Fanconi Anemia pathway and some of the genes involved in checkpoints are more expressed in stage I carcinoma than stage I borderline tumors. These data might suggest that the malignant phenotype is associated with an upregulation of these genes that would endow tumor ovarian cell with higher growth and metastatic potentials. In Stage III ovarian patients a number of correlation between the expression of the repair genes and the response to therapy have been performed; however no clear cut statistically significant correlation could be found. The data, even if negative, have been obtained in a quite large sample size and we think pose some doubts on role of the expression of single gene as predictive of response, as suggested by other studies.

Inhibition of the signal mediated by PI3K/akt

Pi3K/akt axis represents one of the major altered pathway in human cancers and therefore is a good target for the development of new drugs. The laboratory has been involved in the pharmacological characterisation of new molecules able to inhibit the pathway. We have characterised the molecular mechanism at the basis of the interaction between two molecules able to inhibit mTOR (the kinase downstresam PI3K/akt) at two different portion of the protein. In vitro and in vivo data indicate that the strategy to inhibit the same target acting at different level could be an intersting strategy to shut down a transduction signal. The combination of the molecules, in fact, is able to inhibit tumor growth more than the single

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drugs, even when these are used at doubled doses. The mechanism of activity of the combination is the ability to selectively inhibit one of the downstream effectors of mTOR leading to a selective inhibiton of translation. The study combines cellular, molecular and proteomic analysis.

The already performed studies on the role of phospholipase C 1 in favouring the metastatic process, have been extended in a human breast cancer model with a strong ability to metastatize to the bones. In this model we have reproduced the systes previously utilized to inhibit phospholipase C 1 and new clones with inducible expression of phospholipase C 1 have been consequently generated. These models will be extremely useful for better understanding the role of this protein in the metastatic process. In particular we are currently evaluating whether phospholipase C 1 has a role in bone metastasis formation or if its role is confined to specific tissue districts.

Role of phospholipase C 1 in the development of metastasis

Oncosuppressors p53, p63 and p73

The p63 protein, belonging together with p73 to the p53 family, can be expressed in cells in different isoforms. Among these the DN isoform , which originates from an alternative internal promoter, seems to have a role in the development and progression of cancer. In the previous year, our laboratory the expression of p63 members in human ovarian cancer, both at early and advanced stage, has been evaluated. We have shown that the advanced stage expresses much higher levels of DNp63 than early stage tumors. Following these results, we have generated cellular clones in vitro which express a differential, inducible expression of DNp63. These clones which have been characterised for their protein expression, will allow us to define the role of this protein in the growth and response to treatment of cancer cells. In parallel we are evaluating the molecular mechanism responsible for the modulation of DNp63 levels in human ovarian cancers. To date we have been able to exclude an involvement of a chromosomal rearrangement in the area where the p63 gene is present. In this chromosomal area, in fact, we did not find duplication, amplification of genetic material both in early and advanced ovarian cancer.

Mechanisms of action of new antitumor drugs

In collaboration with the laboratory of Biology and Therapy of Metastasis, we have characterised the mechanism of action and the antitumor activity of a new antiangiogenic drug, E3810. This drug is a small molecule able to inhibit receptors playing important roles in the tumor angiogenesis processes (VEGFR, FGFR). Our studies allowed us to define that the drug has a potent antiangiogenic activity, with a broad spectrum of activity in different human tumors transplanted in immunodeficient mice. We are currently investigating combinations of E3810 with other anticancer agents to better define in which context the new drug might be included in the polychemoterapy treatments of human tumors.

Generation of new cellular systems for in vivo imaging

We have generated new cell clones derived from human cancer cells growing in vitro, which stably express fluorescent or luminescent probes which can allow us to follow in vivo the growth of primary tumors and metastasis in mice. These systems generated in human ovarian, breast and prostate cancer cell lines, can be implanted in nude mice and the growth and response to therapy followed by either optical and luminescent imaging or microTAC analysis. We have in particular set up models derived from human breast cancer, which are able to metastasis to the bone which can be evidentiated by optical imaging and microTC techniques in laboratory animals. Utilising different reporter genes, we have generated fluorescent and luminiscent human cancer cell lines which can be transplanted in immunodeficient mice. These

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cells can then be visualised in organs such as peritoneum and lungs were these cells were previously be observed only after sacrifice of the animal. The cells generated to be fluorescent or bioluminiscent will also have specific gene defects which will be useful for understanding the mechanism of action of new molecules. These systems will be particularly useful to study the antimetastatic potential of new drugs.

Studies on the bone metastatic processes

Using a model of human breast cancer cells metastatizing to the bones, we have characterised some molecular pathways involved in the colonisation and metastatic growth. In particular, we have evaluated the role of cMet receptor and of its activation both in vitro and in vivo. The in vivo model utilized develops bone metastasis following intraventricular injection of cancer cells. The bone metastasis can be visualized by optical imaging already after 10 days from cancer cell inoculum. By microTC analysis, bone osteolytic lesions can be evidentiated after 3-4 weeks from tumor cells injection. With this model we have shown that c-met plays an important role in the process and we have been able to demonstrate a cross-talk in vivo between human cancer cells and host cells. In particular we have found that inside cancer cells growing in the bone, there is HGF, the ligand of c-met, both produced by the cancer cells themselves and by the murine host cells. HGF present in the bone can therefore be activated by human cancer cells there have colonized in the bones, thus activating an autoregulatory loop able to stimulate tumor growth. The data obtained with selective c-met inhibitors or with shRNA directed against c-met, indicate that the inhibition of the HGF/c-met axis is effective in inhibiting bone metastatic growth, particularly when the two treatment modalities are combined.

Identification of cancer stem cells from ovarian cancer

This project is aimed at isolating and characterizing a possible cancer stem cell from ovarian cancers. There are increasing evidences supporting the idea that few important multipotent cancer cells, termed cancer stem cells, are among the most relevant cells to be killed in a tumor. Normally present as quiescent cells inside the tumors, they are able to rapidly generate dividing and growing cancer cells. The current hypothesis is that normally dividing cancer cells can be preferentially killed by chemotherapy while the cancer stem cells would be more difficult to kill and would be responsible for the relapse following treatment. The possibility to identify and characterize the cancer stem cell would theoretically open the way to the selection of new generation molecules able to preferentially kill these cells. Several studies have been conducted in ovarian fresh tumor samples, obtained from the Gynecological Department of Ospedale San Gerardo di Monza, directed by Prof Mangioni, that lead to the identification of a cell bearing the characteristic of a tumor initiating cells. We have almost completed the molecular and pharmacologycal characterisation of ovarina cancer stem cell. The results of this characterisation will possibly lead to the identification of specific genes that could be targeted in the ovarian tumor initiating cells with the final aim to improve the therapy of this tumor.

Determination of the impact of EGFR mutations in the activity of tyrosine kinase inhibitors in patients with NSCLC
The study on the characterisation of the response of patients with NSCLC to therapy with or without EGFR inhibitors is ongoing. The data available so far show that among the patients enrolled in the study, the percentage with mutations in the EGFR gene is 8-9%, in line with what previously reported for this tumor in the Caucasian population. The study aims also to evaluate the role of mutations in another gene, K.RAS , in determining the response to treatments. This gene is mutated in a significantly higher proportion of patients than EGFR. The mutational spectrum found in this tumor is different from that reported in other

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types of tumor, such as the colrectal cancer. These differences could be responsible for a different effect on tumor growth and response to treatment and this hypothesis will be tested by generating new cellular models harbouring different mutations of the K-RAS gene in NSCLC cell lines.

Determination of the impact of K-RAS mutations on the activity of anticancer agents.

The K-RAS gene results mutated in significantly higher percentage of NSCLC patients than EGFR. The spectrum of mutation found in NSCLC is different from that observed in other tumor types such as colorectal cancer. The different mutations could explain the different impact of K_RAS on the selction of patients for therapies. In fact in colon cancer mutation in the K_RAS gene is an exclusion criteria for treatment with anti EGFR drugs such as cetuximab. In NSCLC the role fo K-RAS is more controversial. From the available clinical data we went back to the laboratory generating isogenic cellular systems differning for the type of K-RAS mutation. In particular we have generated in NSCLC cell lines clones overexpressing the wt KRAS or mutants in which the glycine at codon 12 is substituted with aspartic acid, cysteine or valine. These mutants have indeed a different impact on the response to treatment of these cells with drugs such as cisplatin, sorafenib or taxol. Our data suggest that for the stratification of patients it is necessary to consider not only the presence of K-RAS mutation, but also the kind of mutation present which could modify the selection f the best therapeutic options.

Laboratory of Biology and Treatment of Metastases Physiologic regulation of angiogenesis

Angiogenesis - the neoformation of blood vessels from existing ones - has a critical role in tumor progression. A delicate balance between pro- and antiangiogenic factors finely tunes this process. We have long studied endogenous angiogenesis-regulatory factors, as a basis to develop new inhibitors. In particular, our studies focus on thrombospondin-1 (TSP-1), an endogenous angiogenesis inhibitor. The ability to directly bind to angiogenic factors, in particular FGF-2 (Fibroblast Growth Factor-2), reducing its bioavailability and activity is one of the manifold functions of this molecule. In a structure/function relationship analysis of different active domains of TSP-1, we have identified its binding site to FGF-2. This active sequence of TSP-1 is being used as a model to design new antiangiogenic and antineoplastic compounds.

Lymphangiogenesis in ovarian carcinoma

Lymphatic spread in ovarian cancer is an important predictor of outcome both in early and advanced stages of this cancer. To clarify the molecular mechanisms involved in the lymphatic spread of ovarian cancer we have developed animal tumor models derived from human ovarian cancer transplanted under the bursa (orthotopic xenograft), expressing luciferase and disseminating in the peritoneal cavities of immunodeficient mice. Preliminary results show that the levels of soluble VEGFC the main factor stimulating the formation of lymphatic vessels as measured in plasma and ascites of mice bearing ovarian cancer, correlates with the tumor growth as measured through optical fluorescence as well as the lymphatic invasion. Studies are in progress to assess the antitumor and antimetastatic activities of selective inhibitors of the VEGF/VEGFR pathway.

How the tumor microenvironment affects endothelial cell gene expression

Blood vessels play a major role in the tumor development, malignant progress and generation of metastasis. The understanding of qualitative and functional differences between endothelial cells (EC) lining the tumor vasculature and EC of normal blood vessels should allow the identification of

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selective markers/targets and lead to novel pharmacological interventions. We have identified gene expression difference in the tumor derived EC with respect to normal EC, through "microarray" analysis followed by validation with RealTime PCR, and in situ hybridization analysis of normal and tumor tissue. Notably the expression of some of these genes is modified by conditions simulating , in vitro, the angiogenic/tumoral microenvironment. We are studying the relevance of these gene transcripts, which are abundant in the tumor EC and whose proteins are expressed in the tumor endothelium/stroma.

The vascular Endothelial Growth Factor (VEGF) modifies the tumoral microenvironment
We have observed that the production of VEGF from the tumoral cells and its release in the tumoral microenvironment is accompanied with an altered response to chemotherapy (e.g. paclitaxel). Bevacizumab (Avastatin) an antibody directed to VEGF restores the therapeutic efficacy of paclitaxel, suggesting that the tumoral environment might play a role in modulating the sensitivity to therapy. To determine the molecular mechanism/s of tumor-VEGF modified microenvironment, we are studying the transcriptional activity of the stromal component (microdissected from the tumoral tissue), using the hybridization of micromatrix of DNA. The results of the analysis with GeneChip Mouse Genome 430 2.0 Array (Affymetrix) indicate that the stroma (microenvironment) of highly producing VEGF tumors preferentially expresses 294 genic transcripts. For some of them the expression of the protein preferentially localizing in the stromal component and/or associated to the vasculature of VEFG-rich tumors has been demonstrated. Their relevance in the tumor progression and response to treatments is under investigation.

Preclinical evaluation of inhibitors of angiogenesis and combination therapies

Antineoplastic therapies directed against the tumor vascular system may be developed accordingly to two different strategies. Antiangiogenic therapy (inhibitors of angiogenesis) prevents the formation of new vessels, while vascular disrupting agents (VDA) aim to selectively destroy the already formed tumor vessels. For the last decade we have investigated the antiangiogenic/antivascular activity of novel antitumor molecules of interest for the clinical development, in particular: a) peptides and non-peptidic small molecules, which mimic endogenous inhibitors of angiogenesis, including compounds similar to thrombospondin-1; b) small molecules inhibiting tyrosin kinase receptors, in particular VEGFRs, FGFR and PDGR, which mediate the signal with the angiogeneic growth factors; c) vascular disrupting compounds, in particular tubuline binding molecules (colchicines and combrestatins analogues) which, causing a depolimerization of microtubules, selectively deteriorate the tumor blood vessels. The study of the combination of angiogenesis inhibitors or VDA with conventional chemotherapy is one of the main interests in our laboratory. In particular, studies have been conducted and are in progress to optimize the modalities of administration of the combinations (i.e. choice of the drugs accordingly to the characteristic of the tumor, dose and treatment schedules accordingly to their mechanism of action), which are connected to the pharmacokinetic and pharmacodynamic profiles associated to the treatment efficacy.

Laboratory for the development of new pharmacological strategies

The laboratory was born out of the consideration that the advent of oncological drugs endowed with mechanisms of action different from those of traditional chemiotherapics, introduces new

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treatment opportunities. At the same time, new problems arise concerning the choice of the most appropriate and effective design for research into the clinical activity profile of these new treatments. The traditional paradigm where the choice of dose is based on the maximal tolerated toxicity, and the screening of therapeutic activity focus on tumor mass reduction, may not necessarily be suitable for the evaluation of new agents whose targets may include the extracellular compartment or specific molecular targets. The clinical development of non toxic anti tumor molecules requires a critical review of the existing models as well as of all the aspects relative to the conduction of clinical trials including: dose selection criteria, methods for determination and confirmation of pharmacological activity, and the validation of new technologies and laboratory methods. This is where the need for a profound integration of the clinical screening and the preclinical research lies. It is a prerequisite for the construction of the pharmacological rationale for the identification of the most interesting molecules, the choice of dose, the hypotheses of combination with other drugs, and of the most appropriate indicators of clinical activity. The acquisition of know how and the development and application of new designs for clinical activity studies, including the use of randomization, the introduction of groups of patients treated with placebo, and new discontinuation designs, proceed in parallel to the above. Another fundamental issue in laboratory research is the recognition that the genomic characterization of any single tumor may now play a more relevant role in drug development and treatment identification. This notwithstanding, numerous uncertainties remain regarding the role of biomarkers in drug development and in the implementation of genomic technologies in clinical trials. It is therefore necessary to improve the methodology and more biomarkers evaluation already in the early stages of research, thus shifting translational research from a simple process of correlation search to one producing knowledge regarding the predictive role of the clinical activity of the investigational treatments. Therefore, the primary focus of the laboratory is the optimization of the methods for evaluating the activity of cytotoxic drugs, but mostly for those therapies aimed at specific molecular targets, as well as the identification of factors predictive of therapeutic response. In 2010, phase II studies on activity of Panitumumab in breast cancer patients, and of ET743 in pancreatic cancer and mesothelioma patients have been initiated.

Laboratory of Clinical Trials

The Laboratory of Clinical Trials is involved in the planning, coordination and analysis of randomized clinical trials in oncology, conducted in cooperation with a network of medical oncologists. Main covered research areas are gastric, colo-rectal, breast and lung cancer. Moreover the laboratory works on a second line study in patients with pediatric glaucoma in collaboration with Azienda Ospedaliera Spedali Civili Di Brescia and supported by AIFA.

Gastric cancer
ITACAS Intergruppo Nazionale Adiuvante Gastrico study is a randomised, open-label, multicenter, trial aimed at assessing the role of adjuvant chemotherapy in the treatment of gastric cancer. It compares the efficacy and safety of a sequential treatment (campto plus flurouracil/leucovorin, followed by taxotere and cisplatin) versus flurouracil/leucovorin regimen, used as standard reference in patients with radically resected adenocarcinoma of the stomach or gastroesophageal junction. The study, sponsored by Mario Negri Institute, involves 11 oncological collaborative groups and is being conducted in more than 110 Italian experimental centers. From February 2005 to August 2009, 1107 patients have been enrolled. The follow-up ends for all patients after the achievement of the target number of events. First

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results about feasibility and tollerability will be published after the mid-2011. The expertise of ITACA-S will be followed by another study ITACA-S2 conducted in patients with adenocarcinoma of the stomach. This randomized, multicentre, 2x2 factorial study, supported by a grant of AIFA, evaluates the efficacy of a peri-operative versus a post-operative chemotherapy treatment and the efficacy of a post-surgical chemo-radiotherapy treatment versus no other treatment , irrespectively of the timing of chemotherapy. The study is in progress and the first patient is expected to be enrolled on the mid-2010.

Lung cancer
On September 2007 the accrual of a multicentre, randomized, Italian study started. The aim of this project (TAILORAIFA trial) is the optimization of second line therapy in patients with advanced NSCLC. The development of target-therapy suggested evaluating the treatment efficacy according to molecular featuresof the tumoral cell. In particular the epidermal growth factor (EGFR) is a promising target for anticancer therapy. A "tailored therapy" based on individual molecular features may result in better responses and optimization of sources and costs. The predictive value of EGFR protein expression, EGFR gene amplification and KRAS mutations in determining the effect of erlotinib as compared to chemotherapy will be assessed . TAILORAIFA trial sponsored by Azienda Ospedaliera Fatebenefratelli e Oftalmico of Milan and supported by the Agenzia Italiana del Farmaco (AIFA), has already registered more than 500 patients. Another multicentre, randomized, double-blind, randomized, placebo-controlled, phase III study conducted in patients with advanced or metastatic NSCLC is starting recruitment. It is aimed at assessing whether acetilcarnitine prolongs toxicity free survival, and reduce neurotoxicity due to platinum compounds. In fact, in patients receiving chemotherapy administered with legitimate curative intent, many toxicities can be justified to accomplish this goal in patients with metastatic cancer for whom the goal is to palliate symptoms and optimise the quality of life this is less acceptable and justified. The target number of patients is 650, randomized in approximately 30 Italian experimental centres.

Colon cancer
On June 2007 started the accrual of a randomised, phase III clinical trial aimed at identifying the best therapeutic adjuvant strategy in radically resected colon cancer patients is starting. The study, sponsored by Fondazione Giscad per la Cura dei Tumori and supported by the Agenzia Italiana del Farmaco (AIFA), will assess the following two questions: 1) Optimal duration of FOLFOX-4 regimen (3 vs 6 months) 2) Efficacy of the addition of Bevacizumab to FOLFOX-4 regimen (only in high risk stage III patients) For both questions, primary efficacy endpoint will be recurrence free survival. Another phase III clinical trial has started. This study aimed at comparing the efficacy in terms of PFS of the addition of cetuximab to FOLFIRI vs. FOLFIRI alone given as first line therapy in patients with advanced CRC KRAS wild-type. In particular, the predictive value of PTEN mutation will be assessed in determining the effect of cetuximab+FOLFIRI as compared to chemotherapy alone. The patient accrual period is planned for approximately 30 months. To assess PFS, all patients will be followed for up to 24 months after the last patient is randomized. The maximum estimated study duration is approximately 54 months. This study, sponsored by Regione Lombardia, forsees the involvement of 30 centers and the enrollment of 290 KRAS negative patients.

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At last a phase III clinical trial has been activated. This study (COMETS), sponsored by Fondazione Giscad per la Cura dei Tumori and supported by AIFA, aimed at identifying the best sequence of treatment (Irinoteca/Cetuximab followed by FOLFOX-4 vs FOLFOX-4 followed by Irinotecan/Cetuximab) in KRAS negative patients with advanced colorectal cancer after a first line chemotherapy with FOLFIRI/Bevacizumab. Primary efficacy endpoint will be overall survival. The maximum estimated study duration is approximately 52 months and about 350 patients will be enroll.

Head and neck

On March 2008 started the accrual of a randomized multicenter open label phase 3 factorial trial evaluating the overall survival in patients with locally advanced squamous cell carcinoma of head and neck treated with locoregional treatment (radiotherapy plus concomitant chemotherapy or cetuximab) with or without neoadjuvant chemotherapy. Patients will be randomized to receive 3 cycles of neoadjuvant chemotherapy (TPF) followed by radiotherapy plus concomitant chemo or cetuximab, or radiotherapy plus concomitant chemo or cetuximab alone. The primary objectives of the study are to compare the overall survival between neoadjuvant and no neoadjuvant arm and to compare the toxicity between concomitant chemoradiotherapy and radiotherapy plus Cetuximab. The study will be conducted by a multidisciplinary team, composed by oncologists, radiotherapists and othorinolaryngologists and will enroll approximately 350 patients in Italian centers.

Laboratory of Translational and Outcome Research in Oncology

The Laboratory is mainly aimed at documenting, by using either Systematic Literature Review, Randomized or Outcome Research studies, the value of new diagnostic and therapeutic interventions in oncology, paying particular attention to two critical steps: the passage from early to late clinical research (from the activity to efficacy evaluation) and from phase III to clinical practice (from efficacy to effectiveness). The principal lines of research are three: cancer pain evaluation, clinical research on gynecologic cancers and evalution of the effectiveness of complex clinical programs in oncology care. In order to facilitate the research activities and optimize the outputs, the Laboratory hosts the Coordination Centers of two multidisciplinary Groups (MANGO: Mario Negri Gynecologic Oncology and the CP-OR: Cancer Pain Outcome Research Study Groups). As from 2007 on, all the activities of research and training in the field of chronic pain has been coordinated by a dedicated center (CERP:Center for the Evaluation and Research on Pain).

The Center for Evaluation and Research on Pain (CERP)

CERP is active since early 2008. It coordinates several studies and other activities regarding chronic pain, particularly of oncologic nature. CERP is aimed at advancing the scientific knowledge in this field and at improving the quality of palliative care and pain treatment. CERP activities mainly focus on clinical research, but pre-clinical research (in vivo), information and educational activities are also considered. Main activities of CERP in 2010 were primarily focused to plan and launch the new randomized clinical trial:Studio clinico randomizzato e controllato, in aperto, per comparare lefficacia analgesica di percorsi terapeutici effettuati con ossicodone, fentanyl e buprenorfina verso morfina, in pazienti con dolore associato a cancro di intensit moderata-severa, a partire dal momento in cui iniziano il trattamento con 3 scalino della scala analgesica del WHO (an open, controlled, randomized clinical trial to compare the analgesic effectiveness of therapies with oxycodone, fentanyl, buprenorphine versus morphine, in patients with moderate-severe cancer pain at the start of the 3rd-step-WHO analgesic ladder therapy). This study also includes an ancillary pharmaco-genomic project: Valutazione, in parallelo, dellassetto genico dei

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pazienti e delle possibili correlazioni con gli effetti clinici osservati (Evaluation, in parallel to the main project, of the genetic profiles of patients and their potential correlations to observed clinical effects). - In May 2010 CERP completed the writing of the study protocol and the CRF. - In July 2010 the single option was received by the Ethic Committee of IRCCS Fondazione INT, Milan. - From September 2010 until the end of the year, the entire documentation was sent to all Centers that will participate in the study (85 Centers in total, divided into oncology wards and palliative care centers). - Along this project, CERP concluded the planning and performed the launch of the study Progetto di ricerca per la validazione di un algoritmo e di una scheda elettronica per la gestione del paziente con Dolore Cronico Non Oncologico in Medicina Generale (A research project for the validation of algorithm and electronic form for the management of patients with non-oncological chronic pain in General Medicine), in collaboration with S.I.M.G. (Italian Society of General Medicine). Regarding the educational/formative activities of CERP, in March 2010, module clinical module of the 9th Master in Cure Palliative al termine della vita (Master in Palliative Care at the End of Life) was done, in collaboration with the Universit degli Studi di Milano. The website http://crc.marionegri.it/cancerpain dedicated to all activities of CERP was completely renewed and updated.

The collaborative group in clinical gynecologic oncology named MaNGO

The Mario Negri Gynecologic Oncology group (MaNGO) is a new name for a collaborative group that has been active in clinical gynecologic oncology for several years. Infact, this group consolidated its network and logistics while running the ICONs studies which were conducted in very close partnership with researchers at the Medical Research Council, Clinical Trial Unit, UK. MaNGO was formally set up in May 2006 and is mainly representative of the northern part of Italy, although there are important sites in the central and southern part of the country too. Participating centers are either general public and private hospitals or university clinics. One of MaNGOs main statutory objectives was to foster an active collaboration with the Gynecologic Cancer Intergroup (GCIG), and the European Network of Gynaecological Oncology Trials groups (ENGOT) that represent two International Forum circulating the scientific proposals from many national collaborative groups. MaNGO group is actively involved in many international phase III trials. MaNGO has been coordinating the Italian participation to the PORTEC 3 study: this is an academic randomized phase III trial in endometrial cancer promoted by the Dutch collaborative group. MaNGO received government funds from the Italian Agency for Drugs (AIFA) supporting its national coordinating role. In 2010 the MaNGO network has been the third group in terms of number of patients enrolled into the trial. In 2010 the randomized clinical trial in ovarian cancer with a new antiangiogenic drugs (pazopanib) and internationally coordinated by the German onco-gynecologic group named AGO reached the target sample size. This protocol was emended in 2010 to prolong the maintenance therapy with pazopanib from one to two years. Another international randomized clinical trial centrally managed by AGO was launched in 2010. This trial will compare a maintainance treatment with vargatef (an angiogenesis paninhibitor) with placebo. The MaNGO group is the promoter of an ancillary study for the vargatef trial aimed at clarifying the role of angiogenic markers such as VEGFC, VEGFB, VEGFA and their receptors VEGFR3, VEGFR2) This translational study will be receiving specific funds. In 2009 MaNGO launched the TAUL study, a randomized phase II trial aimed to evaluate the efficacy of trabectedina in the treatment of patients with uterine leiomyosarcoma and, during 2010, 26 Italian sites have been activated and 9 patients randomized. During 2010

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the MaNGO group developed and implemented the INOVATYON protocol. This is an academic, international, phase III, randomized clinical trial aimed at comparing the combination of pegylated liposomal doxorubicin+carboplatin with the combination of pegylated liposomal doxorubicin and trabectedin in partially platinum sensitive ovarian carcinoma relapses.This protocol was approved in October by the Ethics Committee of the European Institute of Oncology in Milan and will be run under the aupsices of the international intergroups GCIG and ENGOT. During 2010, MaNGOs Technical-Scientific Committee met quarterly while MaNGO affiliates were conveyed at the 7 General Assembly that was held in June.

As to the third lineof activities (development and evaluation of complex health carevinterventions) two activites are at the moment ongoing on colo-rectal and breast cancer patients Colo-rectal cancer
The assessment of efficacy of screening for relapses of colorectal carcinoma has been debated for a long time, with controversial results. GILDA is an open label, international, randomised study comparing two different strategies of post surgical surveillance in colorectal cancer (Dukes B2-C stage): minimalist versus intensive. Primary endpoints of this trial are disease free survival (which is used to assess diagnostic anticipation of metastases), overall survival, health related quality of life, direct and indirect costs evaluation. At present, GILDA trial is the largest randomised study evaluating the efficacy of two follow-ups in colorectal carcinoma. The trial was closed to patient entry in September 2006 when a total of 1200 patients had been enrolled. At the end of 2008 a manuscript for an Italian journal has been prepared to present the progress of the trial, in 2009 a preliminary analysis on data available has been carried out to test the data maturity and the final analyses is planned for the first half of 2011.

Breast cancer
In the context of a multi-regional project sponsored by the Italian Ministery of Health (6 Progetto Integrato Oncologia) LaTOR is coordinating a project in which 5 different regional groups are involeved: so far, 2 multi-center RCTs testing the efficacy of different follw-up regimens in breast and uterus cancers, after diagnosis and first curative therapy , are ongoing and a third prospective study to test the predictive value of bio-markers in the prognosis of brest cancer is going to be launched.

Other research activities

LaTOR has continued on 2010 the collaboration with other laboratories of the Mario Negri Institute. The most important collaboration are those epertaining the evaluation of incidence and impact of mild anemia in elderly patients and on the attitude of smokers in the utilization of pharmacological interventions to quit smoking. Resulst of these collaboration have been extensively published on international peer-reviewed journals. Another collaboration is ongoing with a regional network of General Practitioner to evaluate the frequency and effect of determinants related to social vulnerability on indicators of care accessibility and continuity

Laboratory of Medical Research and Consumer Involvement

The Laboratory promotes various research activities aimed at developing the participation of citizens & patients and their representatives to the choices and decisions regarding health. The laboratory organize training courses and information discussion that would enable consumers to deal effectively with physicians and researchers. Also carried out by the laboratory research

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projects to evaluate the type of information provided on illness and treatment, development of internet website and information on health issues (www.partecipasalute.it); projects involving groups of patients for publication of information material; projects to assess quality of life and health.

PartecipaSalute (Participate in Health Care) - fostering a strategic alliance between consumers associations and medical community
The project, started in 2003, is coordinated by Mario Negri Institute, with the collaboration of the Italian Cochrane Centre and Zadig, an editorial and publishing company. The project develops initiatives dedicated both to patients, citizens and their organizations, and to clinicians, researchers and medical societies. The main aims of the project are to boost patients participation to health care debate and decision making processes - also through their organizations - and to increase medical societies attention to patients demands related to clinical research and dissemination of medical information. Different stakeholders are involved: patients and consumers associations, medical societies, researchers, medical journalists, experts in medical communication. The main 2010 activities are: - a five modules training course for consumers and patients representatives - called Accademia del cittadino - organized together with the Centro Gestione Rischio Clinico e Sicurezza dei Pazienti ed il Settore Equit e Accesso della Direzione Generale Diritto alla Salute della Toscana; - a two modules training course organized together with the Centro del Volontariato in Rimini; - the development of PartecipaSalute website 2.0 http://www.partecipasalute.it/cms_2/drigg_home where users can post comments, articles and share documents. Every article published on the website can be commented and rated by users. Areas restricted to participants of the training course have been created, where online forms with exercises are available. A tool to evaluate patients associations credibility and quality called Misurassociazioni - has been developed together with the advisory of training courses participants; - the development of a project called Citizens juries, a method of deliberative democracy to directly involve the citizens in healthcare decisions. It is based on the idea that many issues are best decided by a group of lay people who have no vested interests and who apply their common sense and experience, having been presented with the best possible evidence. One of the matter at issue is the availability of prostate cancer screening; - a survey on the correspondence between clinical research and patients needs addressed to 123 pediatric volunteers associations. The survey has been developed with the Mother and Child care Laboratory and dealt with unanswered needs of patients, volunteers associations knowledge of ongoing studies, their involvement in research activities, their funding, their support to research projects. Results are in press; - a survey on the transparency of patients associations and drug companies websites. The websites of 17 drug companies and 157 patients and citizens associations have been evaluated trough two forms dealing with the disclosure of sponsorships respectively delivered and received. Several issues have been considered. Supported patients associations -declared by drug companies- who have no websites have been contacted by mail.

Project ConMe Conoscere la menopausa

The Partecipasalute project together with the National Guidelines System (SNLG) based at the Istituto Superiore di Sanit organized in 2008 the Consensus conference (CC) Informing women on hormone replacement therapy in order to assess the current status of the quality of information on hormone replacement therapy (HRT) and re-visit recent research findings on its

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risks/benefits. The final recommendations concern menopause, HRT and information to convey to women (http://www.partecipasalute.it/cms/files/Documento-definitivo-consenso.pdf). The Laboratory of Medical Research and Consumer Involvement, Zadig - editorial and publishing company, and the Istituto Superiore di Sanit developed the project CONoscere la MEnopausa to evaluate the impact of training and information initiatives targeted to women and healthcare operators, focused on the CC recommendations. The project is funded by the Agenzia Italiana del Farmaco. Four regions participate in the project: Lombardia, Toscana, Lazio, Sicilia. The training and information interventions are carried out in four local healthcare agencies: ASL di Bergamo, ASL 7 di Siena, ASL RMH, ASL di Enna. Education programs on this issue are organized trough residential sessions and online modules dedicated to medical practitioners, gynecologists, pharmacists, obstetricians. A booklet for women and information material for healthcare operators are also provided. Three main outcomes will be considered: HRT prescriptions, healthcare operators and women knowledge about menopause and HRT, quality of information conveyed to women through the press. In 2010 the booklets addressed to women and information material addressed to healthcare operators have been developed, printed and sent to the healthcare agencies involved. The survey on knowledge, attitude and practice of physicians and gynecologists has been concluded. The survey on the press dealing with HRT is ongoing.

SNAP project - Smoke, Nutrition, Alcohol and Physical Activity

SNAP - Smoke, Nutrition, Alcohol and Physical Activity - is a campaign for the health with particular attention to young people between 11 and 20 years. This project, commissioned by FSE - Frontier Science & Technology Research Foundation, Southern Europe, a foundation to support independent research - in collaboration with the Mario Negri Institute, has been launched with the aim of increasing knowledge and changing opinions, attitudes and behaviors of young people on the four issues examined, with an active process of information through the distribution of written material, a website built ad hoc and a public event. Between 2008 and 2009, a prototype phase was carried out in a company furnishing, to assess the effectiveness of training-information. A questionnaire on the 4 themes SNAP, on knowledge, opinions and attitudes has been distributed to more than 500 employees before a formation-information public event and then 3 months and one year after the event. Data from the first and second survey have been analyzed and data were presented to employees through a brochure. Data from the third questionnaire will be analyzed and compared with those of the first two.

Programma 1, WP5 -Alleanza contro il cancro Servizio nazionale di accoglienza e informazione sul cancro. Gruppo per l'Informazione ACCP1 WP5
The project is coordinated by the Istituto superiore di sanit. Other partners are the Centro di Riferimento Oncologico - Aviano, AIMaC Associazione italiana malati di cancro, Istituto Nazionale dei Tumori, Istituto Europeo di Oncologia. The Laboratory of Medical Research and Consumer Involvement refers to the working group focusing on information, composed by members of Istituto Superiore Sanit, ospedale S. Giovanni Rotondo, S. Raffaele, Fondazione Pascale di Napoli, Universit di Genova - MEDINFO, AIMaC, INT di Milano, CRO di Aviano, Oncologico di Bari, Az. ospedaliera S. Andrea di Roma. The Laboratory evaluated the quality of websites dealing with breast cancer, colon rectal cancer, cervical cancer. An ad hoc evaluation form was created, considering the quality of the website and some issues related to the contents. Each website was independently evaluated by two reviewers, using the defined form. Data collected were analyzed and discussed by the Laboratory.

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Each evaluation form was included in Cignoweb, a website that publishes a database collecting websites and information materials about cancer. In 2010 the results of the project have been presented in a meeting dealing with the national cancer program and publications have been planned.

Project: inform, learn and participate in improving the quality of life. The case of asthma, type 2 diabetes and breast cancer
This project on the quality of information provided to citizens and patients, sponsored by the Smith Kline Foundation, was established with the objective of evaluating the quality of the pamphlets produced and distributed by the associations of patients of the three diseases and to produce a core of information from which associations can draw evidence-based information to produce their own brochures. During the project were contacted associations in Lombardy, Veneto, Tuscany, Emilia Romagna, Puglia and Sardinia were evaluated ad hoc board with all the pamphlets, produced or distributed by the associations contacted. In the course of 2010 a document called "core information", with the information-base needed to provide an evidence-based information, has been organised. The "core information" was discussed during a public meeting.

Follow-up in oncology setting

Two studies on follow-up have been designed and carried out in collaboration with the Laboratory of Giovanni Apolone. The first in collaboration with the Network Oncologica Piemontese regards the follow-up of patients with endometrial cancer organization for which the evidence available is not sufficient to draw a path of sure effectiveness. TOTEM study that has the characteristics of an open randomized multicenter study comparing two different modulations of visits and examinations. The second study that takes place in the context of the 6th Integrated Project Oncology (Health Ministry) provides for the comparative assessment of two follow-up for women at moderatelow risk with a diagnosis of breast cancer and lead to a randomization minimalist follow-up coordinated by the oncologist or by general practitioner. The study starts the randomization in September 2010.

Quality of life projects

No specific research projects have been carried out on quality of the life evaluation. However we have been supporting and coordinating other groups using the instruments of quality of life translated and validated by our research group, SF-36, SF-12, PGWBI. During the year the website http://crc.marionegri.it/qol has been periodically updated.

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DEPARTMENT OF ENVIRONMENTAL HEALTH SCIENCES

STAFF

Head

Roberto FANELLI, Biol.Sci.D.

Laboratory of Analytical Biochemistry

Head Chiara CHIABRANDO, Biol.Sci.D.

Laboratory of Environmental Chemistry and Toxicology

Head Emilio BENFENATI, Chem.D.

Industrial and Environmental Health Unit Head Marco LODI, Chemist

Laboratory of Food Toxicology

Head Ettore ZUCCATO, M.D.

Laboratory of Mass Spectrometry

Head Enrico DAVOLI, Anim.Sci.D.

Laboratory of Molecular Toxicology

Head Protein and Gene Biomarkers Unit Head Luisa AIROLDI, Pharm.D.

Roberta PASTORELLI, Biol.Sci.D

Departments Units
Environmental Pollutants Risk Assessment Unit Head Elena FATTORE, Biol.Sci.D Analytical Instrumentation Unit Head

Renzo BAGNATI, Chem.D.

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CURRICULA
Roberto Fanelli, Head of the Environmental Health Sciences Department since 1997, Laboratory Head 1978-97, Researcher 1975-78, Research fellow 1969-74 at the Mario Negri Institute. Doctoral Degree in Biological Sciences (University of Milan, 1973), Assistant Professor in Biochemistry at Baylor College of Medicine (Houston, Texas). Member of the Commissione Consultiva Prodotti Fitosanitari (Ministero Salute), Member of the Scientific Panel on Contaminants in the Food Chain (European Food Safety Authority, 2003-2006), Certified Italian Toxicologist. Member of the Comitato Scientifico Ente Risi. Research areas: Sources, diffusion, toxicology, human exposure and risk assessment of persistent environmental pollutants. Environmental risk of plant protection products. Development of analytical methods for identification and measurement of biomarkers in toxicology. Mechanisms of toxic action by proteomic techniques.
Selected publications: 1. Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol 2009 ; 20 : 123-130. 2. Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev 2008 ; 27 : 378-394 3. Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008 ; 116 : 1027-1032 4. Hodgson S, Thomas Laura, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L. Bone mineral density changes in relation to environmental PCB exposure. Environ Health Perspect 2008 ; 116 : 1162-1166 5. Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: 882-894 6. Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new evidence-based tool to monitor community drug abuse. Environ Health 2005; 4: 14 (http://www.ehjournal.net/content/4/1/14 2005)

Luisa Airoldi, Head of the Molecular Toxicology Laboratory since 1994, Unit Head 1987-94, Researcher 1978-87, Technician 1967-75 at the Mario Negri Institute. Doctoral Degree in Pharmacy (University of Milan, 1975), Postdoctoral fellow at the Massachusetts Institute of Technology (Cambridge, MA, 1976) and at the Northwestern University Medical School (Chicago, Il, 1977), Researcher at the Yale University Medical School (New Haven, CT, 1980-81). Research areas: Proteomics in toxicology with particular interest on the study of proteome changes in tissues and biological fluids from animals and humans after exposure to toxic compounds; clinical proteomics aimed at the identification of protein biomarkers as diagnostic tools; molecular epidemiology focused on the identification and measurement of biomarkers of exposure to environmental carcinogens and disease susceptibility.
Selected publications: 1. Gallo V, Neasham D, Airoldi L, Ferrari P, Jenab M, Boffetta P, Overvad K, Tjonneland A, Clavel-Chapelon F, Boeing H, Pala V, Palli D, Panico S, Tumino R, Arriola L, Lund E, Bueno-De-Mesquita H B, Peeters P H, Melander O, Hallmans G, Riboli E, Saracci R, Vineis P. Second-hand smoke, cotinine levels, and risk of circulatory mortality in a large cohort study of neversmokers. Epidemiology 2010 ; 21 : 207-214. 2. Airoldi L, Magagnotti C, Iannuzzi AR, Marelli C, Bagnati R, Pastorelli R, Colombi A, Santaguida S, Chiabrando C, Schiarea S, Fanelli R. Effects of cigarette smoking on the human urinary proteome. Biochem Biophys Res Commun. 2009 381: 397402. 3. Carpi D, Korkalainen M, Airoldi L, Fanelli R, Hakansson H, Muhonen V, Tuukkanen J, Viluksela M, Pastorelli R. Dioxinsensitive proteins in differentiating osteoblasts: effects on bone formation in vitro. Toxicol Sci. 2009 108: 330-43. 4. Peluso M, Airoldi L, Colombi A, Munnia A, Veglia F, Autrup H, Dunning A, Garte S, Gormally E, Malaveille C, Matullo G, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita H B, Peeters P H, Kumle M, Gonzalez C A, Martinez C, Dorronsoro M, Barricarte A, Tormo M J, Quiros J R, Berglund G, Janzon L, Jarvholm B, Day N E, Key T J, Saracci R, Kaaks R, Riboli E, Bingham S, Vineis P. Bulky DNA adducts, 4-aminobiphenyl hemoglobin adducts, diet and air pollution in a healthy European population. Br J Nutr 2008 100: 489-495. 5. Veglia F, Loft S, Matullo G, Peluso M, Munnia A, Perera F, Phillips D H, Tang D, Autrup H, Raaschou-Nielsen O, Tjonneland A, Vineis P, Genair-EPIC Investigators. DNA adducts and cancer risk in prospective studies: a pooled analysis and a meta-analysis. Carcinogenesis 2008 ; 29 : 932-936. 6. Pastorelli R, Saletta F, Campagna R, Carpi D, Dell'Osta C, Schiarea S, Vineis P, Airoldi L, Matullo G Proteome characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl Proteome Sci 2007 5: 6

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Emilio Benfenati, Head of the Laboratory of Environmental Chemistry and Toxicology since 1997, Unit Head 1987-97, Researcher 1986-87, Research fellow 1981-86 at the Mario Negri Institute. Researcher at Istituto Biochimico Italiano 1979-1981. Doctoral Degree in Chemistry (University of Milan, 1979). Member of Commissione Consultiva Prodotti Fitosanitari (Ministero Salute 1997-99), Certified Italian Chemist. Research areas: Computer-based models for chemistry and toxicology; Molecular descriptors; QSAR; Toxicity prediction; Metabolism studies; Characterization and assessment of wastes, industrial effluents, emissions from landfill and incinerator; Integration of chemical analysis and eco-toxicological data; Chemical analysis of organic compounds by mass spectrometry.
Selected publications: 1. Boriani E, Mariani A, Baderna D, Moretti C, Benfenati E, ERICA: a multiparametric toxicological risk index for the assessment of environmental healthiness, Environmental International, 2010 36 : 665674 2. Toropov A A, Toropova A P, Benfenati E, Leszczynska D, Leszczynksi J, InChI-based optimal descriptors: QSAR analysis of fullerene[C60]-based HIV-1 PR inhibitors by correlation balance, Eur J Med Chem 2010 45 : 1387-1394 3. Benfenati E, Benigni R, DeMarini D M, Helma C, Kirkland D, Martin T M, Mazzatorta P, Oudraogo-Arras G, Richard A, Schilter B, Schoonen W G E J, Snyder R D, Yang C. Predictive models for carcinogenicity and mutagenicity: frameworks, state-of-the-art, and perspectives. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 2009 ; 27 : 57-90. 4. Colombo A, Benfenati E, Mariani G, Lodi M, Marras R, Rotella G, Senese V, Fattore E, Fanelli R. PCDD/Fs in ambient air in north/east Italy: The role of a MSWI inside an industrial area. Chemosphere 2009; 77: 1224-1229 5. Zhao C, Boriani E, Chana A, Roncaglioni A, Benfenati E, A new hybrid QSAR model for the prediction of bioconcentration factors (BCF), Chemosphere 2008 73 : 1701-1707 6. Roncaglioni A, Benfenati E, In silico-aided prediction of biological properties of chemicals: oestrogen receptor-mediated effects, Chem Soc Rev 2008 37: 441-450

Chiara Chiabrando, Head of the Analytical Biochemistry Laboratory since 1997, Unit Head 1987-97, Researcher 1978-87, Research fellow 1975-78 at the Mario Negri Institute. Doctoral degree in Biological Sciences (University of Milan, 1974), Postdoctoral fellow at the Baylor College of Medicine (Houston, Texas, 1974-75). Postgraduate degree in Pharmacological Research, Mario Negri Institute (1977). Research areas: Development and application of bio-analytical methods based on mass spectrometry in the fields of biochemistry, metabolism, clinical chemistry and pharmacology. Identification and characterization of proteins and peptides of biomedical interest by proteomic approaches and mass spectrometry. Structural characterization of proteins by mass spectrometry. Proteomics in oncology. Comparative characterization of cancer cell lines secretomes by a global proteomic approach and systems biology tools.
Selected publications 1. Schiarea S, Solinas G, Allavena P, Scigliuolo GM, Bagnati R, Fanelli R, Chiabrando C. Secretome analysis of multiple pancreatic cancer cell lines reveals perturbations of key functional networks. J Proteome Res. 2010;9:4376-92. 2. Solinas G, Schiarea S, Liguori M, Fabbri M, Pesce S, Zammataro L, Pasqualini F, Nebuloni M, Chiabrando C, Mantovani A, Allavena P. Tumor-conditioned macrophages secrete migration-stimulating factor: a new marker for M2-polarization, influencing tumor cell motility. J Immunol. 2010;185:642-52. 3. Airoldi L, Magagnotti C, Iannuzzi AR, Marelli C, Bagnati R, Pastorelli R, Colombi A, Santaguida S, Chiabrando C, Schiarea S, Fanelli R. Effects of cigarette smoking on the human urinary proteome. Biochem Biophys Res Commun. 2009; 381:397402. 4. Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol 2009 ; 20 : 123-130. 5. Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008; 116: 1027-1032. 6. Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev 2008; 27: 378-394.

Enrico Davoli, Head of the Mass Spectrometry Laboratory since 1997, Unit Head 1994-97, Researcher 1989-94, Research Fellow 1985-87 at the Mario Negri Institute. Fellow at USDA, Beltville, MD 1977-78. Doctoral Degree in Animal Sciences (University of Milan, 1983), Postdoctoral fellow at the University of Nebraska (Lincoln, NE, 1987) and at the University of Colorado Health Sciences Center (Denver, CO, 1988). Postgraduate degree in Pharmacological Research, Mario Negri Institute (1988). Member of the

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American Association for Mass Spectrometry (ASMS) of the Environment and Safety Commission of IGQ and of the ETS (Emission Trading System) commission. Member of the National Biomass Research Center Scientific Committee. Environmental Applications Interest Group Coordinator (ASMS). Research areas: Development of methodology, instrumentation and software for environmental research. Studies of urban air pollution and characterization of environmental odor annoyance.
Selected Publications 1. Adani F, Tambone F, Davoli E, Scaglia B. Surfactant properties and tetrachloroethene (PCE) solubilisation ability of humic acid-like substances extracted from maize plant and from organic wastes: A comparative study. Chemosphere 2010 78 : 10171022. 2. Orzi V, Cadena E, D'Imporzano G, Artola A, Davoli E, Crivelli, M, Adani F. Potential odour emission measurement in organic fraction of municipal solid waste during anaerobic digestion: relationship with process and biological stability parameters. Bioresour Technol 2010 101 : 7330-7337. 3. Davoli E, Fattore E, Paiano V, Colombo A, Palmiotto M, Fanelli R, Rossi A N, Il Grande M. Waste management health risk assessment: A case study of a solid waste landfill in South Italy. Waste Manag 2009, DOI 10.1016/j.wasman.2009.10.013. 4. Davoli E, Bianchi G. Odour emission rates from a waste treatment plant: results from a multi year follow-up study. Chemical Engineering Transactions 2008; 15 : 95-102. 5. Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from European countries. Food Chem Toxicol 2008, 46: 1062-1067. 6. Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21 : 1998-2002

Ettore Zuccato, Head of the Food Toxicology Laboratory since 2005, Unit Head 1997-2005, Researcher 1986-97, Technician 1975-86 at the Mario Negri Institute. Doctoral degree in Medicine (University of Milan, 1986), Postdoctoral degree in Human Nutrition (1999), Postdoctoral fellow at the Kings College School of Medicine (London, UK, 1988-89). Member of the ANSISA, EMEA expert, member of the Commissione Consultiva per i Prodotti Fitosanitari, and expert for the evaluation of plant protection products for registration within the EU. Research areas: Food safety, including the study of dietary chemical contaminants, safety assessment of GMO in human nutrition, food allergens and toxicants, emerging issues in food toxicology, risk perception and risk communication to the consumers, and evaluation of plant protection products for registration within the European Union. Environmental pollution by pharmaceuticals, and monitoring of illicit drugs in surface waters to estimate community drug abuse.
Selected publications 1. Zuccato E, Castiglioni S, Bagnati R, Melis M, Fanelli R. Source, occurrence and fate of antibiotics in the Italian aquatic environment. J Hazard Mater 2010 ; 179 : 1042-1048 2. Grassi P, Fattore E, Generoso C, Fanelli R, Arvati M, Zuccato E. Polychlorobiphenyls (PCBs), polychlorinated dibenzo-pdioxins (PCDDs) and dibenzofurans (PCDFs) in fruit and vegetables from an industrial area in northern Italy. Chemosphere 2010 ; 79 : 292-298 3. Illicit drugs in the environment: Emerging contaminants and indicators of drug abuse. Castiglioni S, Zuccato E. Integrated Environmental Assessment and Management 2010 ; 6 : 186-187 4. Zuccato E, Castiglioni S. Illicit drugs in the environment. Philos Transact A Math Phys Eng Sci 2009 ; 367 : 3965-3978. 5. Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis. Environ Health Perspect 2008, 116: 1027-1032. 6. Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometry analysis of illicit drugs in wastewater and surface water. Mass Spectrom Rev, 2008, 27: 378-394.

Renzo Bagnati, Head of the Analytical Instrumentation Unit since 2005, Researcher 1992-2005, Research fellow 1986-92 at the Mario Negri Institute. Doctoral degree in Chemistry (University of Turin, 1985), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1989). Research areas: Mass spectrometry applied to the analysis of biological and environmental relevant substances (proteins, peptides, hormones, pharmaceuticals, drugs of abuse, pesticides).
Selected Publications 1. Schiarea S, Solinas G, Allavena P, Scigliuolo G, Bagnati R, Fanelli R, Chiabrando C. Secretome analysis of multiple pancreatic cancer cell lines reveals perturbations of key functional networks. J Proteome Res 2010; 9: 4376-4392. 2. Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C, Garattini E. Role of the molybdoflavoenzyme aldehyde oxidase homolog 2 in the biosynthesis of retinoic acid: generation and characterization of a knockout mouse. Mol Cell Biol 2009 ; 29 : 357-377. 3. Zuccato E, Castiglioni S, Bagnati R, Chiabrando C, Grassi P, Fanelli R. Illicit drugs, a novel group of environmental contaminants. Water Res 2008 ; 42 : 961-968.

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4. 5. 6.

Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21 : 1998-2002. Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs and their metabolites in urban wastewater by liquid chromatography-tandem mass spectrometry. Anal Chem 2006; 78: 8421-8429. Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new evidence-based tool to monitor community drug abuse. http://www.ehjournal.net/content/4/1/14 2005.

Elena Fattore, Head of the Environmental Pollutants Risk Assessment Unit since 2005, Researcher 2001-2004, Research fellow 1991-1997 at the Mario Negri Institute. Doctoral Degree in Biological Sciences (University of Milan, 1991), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1994), Postdoctoral fellow at the National Institute of Environmental Medicine, Karolinska Institutet, Stockholm (1998-2000). Member of the Working Group of External Scientific Experts to externally review the quality of the scientific outputs of the European Food Safety Authority (EFSA) in the area of activity of chemical risk assessment and connected fields. Research areas: Environmental chemistry, toxicology, assessment of human exposure and risk from environmental pollutants with emphasis on dioxins and dioxin-like compounds.
Selected publications 1. Grassi P, Fattore E, Generoso C, Fanelli R, Arvati M, Zuccato E. Polychlorobiphenyls (PCBs), polychlorinated dibenzo-pdioxins (PCDDs) and dibenzofurans (PCDFs) in fruit and vegetables from an industrial area in northern Italy. Chemosphere 2010; 79: 292-298 2. Davoli E, Fattore E, Paiano V, Colombo A, Palmiotto M, Rossi A N, Il Grande M, Fanelli R. Waste management health risk assessment: A case study of a solid waste landfill in South Italy. Waste Manag 2010; 30: 1608-1613. 3. Oberg M, Westerholm E, Fattore E, Stern N, Hanberg A, Haglund P, Wiberg K, Bergendorff A, Hakansson H. Toxicity of Bromkal 70-5DE, a technical mixture of polybrominated diphenyl ethers, following 28 d of oral exposure in rats and impact of analysed impurities. Chemosphere 2010; 80, 137-143. 4. Fattore E, Fanelli R, Dellatte E, Turrini A, Di Domenico A. Assessment of the dietary exposure to non-dioxin-like PCBs of the Italian general population. Chemosphere 2008, 73: S278-S283. 5. Hodgson S, Thomas L, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L Bone mineral density changes in relation to environmental PCB exposure. Environmental Health Perspective 2008, 116: 1162-1166. 6. Carubelli G, Fanelli R, Mariani G, Nichetti S, Crosa G, Calamari D, Fattore E. PCB contamination in farmed and wild sea bass (Dicentrarchus labrax L.) from a coastal wetland area in central Italy. Chemosphere 2007 ; 68 : 1630-1635.

Marco Lodi, Head of the Industrial and Environmental Unit since 2002, Consultant 1997-2002 at the Mario Negri Institute. General Certificate of Education in Industrial Chemistry (Milan, 1974). Member of AIDII (Italian Industrial Hygiene Association), certified by ACGIH (American Conference of Governmental Industrial Hygienist). Research areas: Emission sources, environmental diffusion, toxicology, human exposure and risk assessment of persistent environmental pollutants. Environmental risk of chemical pollution products. Development of sampling methods for environmental toxic compounds.
Selected publications 1. Colombo A, Benfenati E, Mariani G, Lodi M, Marras R, Rotella G, Senese V, Fattore E, Fanelli R. PCDD/Fs in ambient air in north/east Italy: The role of a MSWI inside an industrial area. Chemosphere 2009 ; 77 : 1224-1229. 2. Benfenati E, Azimonti G, Auteri D, Lodi M Environmental and ecological toxicology: computational risk assessment. Computational toxicology. Risk assessment for pharmaceutical and environmental chemicals John Wiley, Hoboken, NJ, 2007; 625-650 3. Grosso M, Cernuschi S, Palini E, Lodi M, Mariani G. PCDD/Fs release during normal and transient operation of a full scale MSWI plant. Organohalogen Compounds 2004; 66: 1243-1249 4. Benfenati E, Mariani G, Lodi M, Reitano G, Fanelli R. Is bioexsiccation releasing dioxins? Organohalogen Compounds 2004; 66: 955-961 5. Fattore E, Mariani G, Guzzi A, Di Guardo A, Benfenati E, Lodi M, Fanelli R. Air dioxin concentrations in Seveso area. In: Halogenated Environmental Organic Pollutants, 18th. Symp., Stockholm, Sweden, august 17-21, 1998. 1998 : 237-240 6. Benfenati E, Mariani G, Schiavon G, Lodi M, Fanelli R. Diurnal, weekly and seasonal air concentrations of PCDD and PCDF in an industrial area. Fresenius Journal Analytical Chemistry 1994; 348: 141-143

Roberta Pastorelli, Head of Protein and Gene Biomarkers Unit since 2004, Researcher 1992-2003, Research fellow 1983-92 at the Mario Negri Institute.

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Doctoral Degree in Biological Sciences (University of Milan, 1982), Postgraduate degree in Pharmacological Research, Mario Negri Institute (1986), Postdoctoral fellow at the Massachusetts Institute of Technology, Cambridge, MA (1987-89 and 1991). Research areas: Toxicoproteomic investigation of global protein expression profiles and their modulation evoked by environmental compounds in different biological compartments. Critical targets and pathways in toxicology. Pharmacogenetics: effects of genetic polymorphisms in the human population on the individual susceptibility to environmental xenobiotic and carcinogen effects.
Selected publications: 1. Airoldi L, Magagnotti C, Iannuzzi AR, Marelli C, Bagnati R, Pastorelli R, Colombi A, Santaguida S, Chiabrando C, Schiarea S, Fanelli R. Effects of cigarette smoking on the human urinary proteome. Biochem Biophys Res Commun. 2009 381: 397402. 2. Carpi D, Korkalainen M, Airoldi L, Fanelli R, Hakansson H, Muhonen V, Tuukkanen J, Viluksela M, Pastorelli R. Dioxinsensitive proteins in differentiating osteoblasts: effects on bone formation in vitro. Toxicol Sci. 2009 108: 330-43. 3. Pastorelli R, Saletta F, Campagna R, Carpi D, DellOsta C, Schiarea S, Vineis P, Airoldi L, Matullo G. Proteome characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl. Proteome Science 2007. 4. Moretti M, Dell'omo M, Villarini M, Pastorelli R, Muzi G, Airoldi L, Pasquini R. Primary DNA damage and genetic polymorphisms for CYP1A1, EPHX and GSTM1 in workers at a graphite electrode manufacturing plant. BMC Public Health. 2007 7: 270 5. Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: 882-894 6. Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C.Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: 4936-4945

ACTIVITIES
The Department works to investigate environmental factors and their effects on human health. The main research lines focus on the survey of environmental contaminants, the assessment of human exposure with related health risks, and toxicity mechanisms of pollutants. The assessment of environmental contamination is carried out not only for well-known and widespread compounds, like dioxins and PCBs, but also for new classes of "unconventional" pollutants, e.g., endocrine disruptors, potentially toxic "natural" compounds, and drugs entering the environment after human or veterinary use. The identification for the first time of illicit drugs in urban waste and river waters, led to a new original tool for the evidence-based monitoring of community drug abuse. For all these survey activities sophisticated analytical methods based on advanced mass spectrometric techniques are developed. The Department is active in the assessment of human exposure to toxic compounds in the atmosphere and the diet, which is the main source of priority pollutants (PCBs, dioxins and other endocrine disruptors). Assessment of the risk associated to contamination in real-life scenarios has recently gained much importance. In order to respond to the growing demand for information, the Department is more and more involved in toxicological and ecotoxicological risk analysis, based on studies in field and predictive models of toxicity. The toxic effects of environmental contaminants on the central nervous system are also investigated, using in vivo and in vitro animal models. Molecular epidemiology studies are used to identify genetic and/or environmental factors posing risks to human health. By this approach, we search for new useful biological markers" to identify susceptible subjects, in view of finding appropriate preventive strategies. The Department has implemented an advanced technological proteomic platform, in order to identify proteins differentially expressed in biological compartments in various experimental and clinical conditions. This approach is particularly relevant in toxicology, since it can contribute to find new biomarkers of toxicity or pathology, and to identify molecular targets and toxic effect mechanisms of pollutants and drugs. To integrate our proteomic studies, we have now introduced among our activities metabolomics, i.e., the study of small molecules, such as amino acids, carbohydrates, lipids, hormones etc., the final products of protein expression and activity which contribute to define the biochemical phenotype of a biological system.

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Mass spectrometry (MS) is a central analytical technique at the Department, where a complete set of state-of-the-art instrumentation is available, from GC-MS and LC-MS to MALDI-TOFMS. These instruments are provided with modern solutions for sample introduction (chip-based nanoLC), sample ionization (ESI, DESI and MALDI), tandem MS (MSn) by triple quadrupole and TOF-TOF instruments, high mass resolution analysis (hybrid ion trap/orbitrap).

FINDINGS/MAIN RESULTS
Evidence of new molecular players in the effects of TCDD on bone development provided by proteomics coupled to networks analysis. Resistance to the bladder carcinogen 4-aminobiphenyl in human urothelial cells is mediated by deregulation of apoptosis and membrane trafficking proteins. Bone protein profile in a murine model of osteoporosis. Identification of novel protein targets responsive to the effects of estrogens in bone. TCDD's effect on the liver proteome profile of exposed rats. Determination of a subset of rat hepatic proteins indicative of differences in dioxin susceptibility. The presence of 4-aminobiphenyl-hemoglobin adducts may help identify nonsmokers at high risk of cancers related to environmental tobacco smoke exposure. Reference values of allele and genotype frequency of several metabolic genes in 15,000 control subjects. CYP1A1 polymorphism affects lung tumor risk. Identification of CYP2C9 genetic polymorphism as a determinant of severe adverse reactions to phenytoin. On-line in silico models to predict ecotoxicity of pesticides for regulatory purposes. New in silico models, freely available on-line, to predict toxicity and ecotoxicity of chemicals for the REACH European legislation. A tool to assess if a chemical is bioaccumualive, with a high rate of accuracy, avoiding the use of the experimental fish model. A new model for identification of endocrine disruptors using molecular docking. A new index integrating risk assessment for human and ecotoxicity endpoints. A method aimed at characterizing environmental odors to identify odor sources in complex environments. Proteomic/bioinformatic workflow for comparative secretome analysis in cancer cell lines. Global proteomic profiles of pancreatic carcinoma cell lines secretomes with identification of perturbed functional networks. Albumin can become a source of potentially antigenic peptides in proteinuric nephropaties. Moderate-to-high fish consumption can result in exceeding the daily intake safety levels of PCBs and dioxin-like compounds established by the European Commission. The same food type may contain significantly different concentrations of PCBs and dioxin-like compounds, depending on the geographic origin (this may help lower the risk for the consumers by understanding and controlling the causes of the differences). The environmental levels of several drugs exceed the safety limits established for them. Environmental pollution from pharmaceuticals is a general phenomenon that can be described by controllable variables (environmental load and mass balance). Illicit drug residues and their metabolites were found in urban waste and river waters. Environmental levels can be used as a new tool to estimate illicit drugs consumption in the population. The distribution of dietary intake values of dioxins, dioxin-like PCBs and non dioxin-like PCBs was characterized for the general Italian population.

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The higher intake of PCBs due to consumption of farmed fish vs. wild fish is mainly due to the higher fat content in farmed fish. Development of novel mass spectrometric methods for the selective measurement of therapeutic and illicit drugs in environmental samples. Activation of Toll-like receptor 4 (TLR4) by endotoxin/lipopolysaccharide (LPS) induces neuroinflammatory responses and motor neuron degeneration in spinal cord primary cultures; protective role of a cyanobacteria-derived TLR4 antagonist (VB3323).

NATIONAL COLLABORATIONS
ARPA Emilia Romagna ARPA Veneto ASL di Bergamo ASL di Brescia ASL di Como ASL di Cagliari ASL di Napoli Centro Reach Srl CLIR Spa Lomellina CNR IRSA Comune di Peschiera del Garda (BS) Comune di Rosignano Marittimo (LI) Comune di SantUrbano (PD) CSRA-Asti Dipartimento delle Politiche Antidroga, Presidenza del Consiglio dei Ministri Federchimica Fondazione 'S. Maugeri' Fondazione ISI, Torino Gruppo Collaborativo sulla Suscettibilit Genetica ai Cancerogeni Ambientali (GSEC), Milano, Italia INRAN-Istituto Nazionale di Ricerca sugli Alimenti e la Nutrizione ISPO, Firenze Istituto Clinico Humanitas, Milano Istituto Superiore di Sanit I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana Metropolitana Milanese Mineracqua Ministero dell'Ambiente Ministero della Salute Ministero dello Sviluppo Economico Politecnico di Milano Politecnico di Torino Provincia di Vercelli Provincia Pordenone Rotary Club Sirmione (BS) Stazione Sperimentale dei Combustibili, Milano Universit degli Studi del Piemonte Orientale

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Universit degli Studi di Cagliari Universit degli Studi di Genova Universit degli Studi di Milano Universit degli Studi di Napoli "Federico II" Universit degli Studi di Palermo Universit degli Studi di Pavia Universit degli Studi di Perugia Universit degli Studi di Roma "La Sapienza" Universit degli Studi di Siena Universit degli Studi di Torino Universit dellInsubria, Varese Universit degli Studi di Verona

INTERNATIONAL COLLABORATIONS
BASF Agricultural Centre, Limburgerhof, Germany Centre for Environmental Policy, Imperial College, London, UK Danish Institute of Agricultural Sciences, Research Centre Foulum, Tjele, Denmark Department of Analytical and Pharmaceutical Chemistry, The Royal Danish School of Pharmacy, Denmark Department of Anatomy and Cell Biology, University of Oulu, Oulu, Finland Department of Computer Science and Engineering, University of Galati, Romania Department of Electrical and Computer Engineering, University of Patras, Greece Department of Environmental Health, National Public Health Institute, Kuopio, Finland Department of Epidemiology & Public Health, Imperial College, London, UK Department of Inland Fisheries, Institute of Freshwater Ecology and Inland Fisheries, Berlin, Germany Department of Molecular Biology, University of Bergen, Bergen, Norway Department of Organic Chemistry, Universidad de Cadiz, Cadiz, Spain Division of Endocrinology, Department of Internal Medicine, Sahlgrenska University Hospital, Gothenburg, Sweden Environmental Chemistry, IIQAB-CSIC, Barcelona, Spain Environmental Hygiene and Chemistry Department, Institute of Environmental Medicine and Hospital Epidemiology, University of Freiburg, Germany Environmental Protection Agency, US EPA - National Risk Management Research Laboratory (NRMRL), Cincinnati OH, USA European Chemicals Agency, ECHA, Helsinki, Finland European Monitoring Centre for Drugs and Drug Addiction (EMCDDA), Lisbon, Portugal Facult de Mdicine et de Pharmacie, Universit de Mons-Hainaut, Mons, Belgium Faculty of Veterinary Medicine, Utrecht University, Utrecht, Netherlands Food and Environment Research Agency, York, UK Forschungzentrum Jlich Gmbh, Jlich, Germany Helmholtz-Zentrum fr Umweltforschung UFZ, Leipzig, Germany Institute of Environmental Medicine. Karolinska Institute, Stockholm, Sweden Institute of Pharmaceutical Chemistry, University of Pcs, Pecs, Hungary Institute of Phytomedicine, Biological Control, Horticulture and Nematology, Vienna, Austria Institute of Soil Science and Plant Cultivation, Pulawy, Poland Interuniversitaeres Forchunginstitut fuer Agrarbiotechnologie, Tulln, Austria Istituto di Chimica di So Carlos, Universit di So Paulo, Brazil KnowledgeMiner Software, Berlin, Germany In Vitro Testing Industrial Platform, Tres Cantos (Madrid), Spain

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Laboratory of Chemometrics & Bioinformatics, University of Orlans, Orlans, France Laboratory of Neurobiology, Centro de Investigation Principe Felipe, Valencia, Spain Lithuanian Institute of Agricultrure, Vilnius, Lithuania Liverpool John Moores University, Liverpool, UK National Institute of Chemistry, Kemijski Institut Ljubljana, Ljubliana, Slovenia Natural Resources Research Institute, University of Minnesota, Duluth, USA National Institute for Public Health and the Environment (RIVM), Bilthoven, Netherlands Pesticide Safety Directorate, York, UK Plant Protection Institute, Hungarian Academy of Sciences, Budapest, Hungary PublicSpace Ltd, Ulverstone, UK School of Biomedical Sciences, University of Ulster, Coleraine, UK SETAC Europe, Brussels, Belgium Symlog, Paris, France Syngenta Crop Protection AG, Basel, Switzerland Technische Universitaet Dresden, Dresden, Germany TNO, Delft, Netherlands Unit of Environmental Risk and Health, Flemish Institute for Technological Research, Boeretang, Belgium. Universitat Politcnica de Catalunya, Barcelona, Spain Universitat Rovira i Virgili, Tarragona, Spain University of Tartu, Tartu, Estonia

EDITORIAL BOARD MEMBERSHIP

Journal of Environmental Science and Health, Part B (Emilio Benfenati), Journal of Environmental Science and Health, Part C (Emilio Benfenati), International Journal of Computational Intelligence (Emilio Benfenati), International Journal of Information Technology (Emilio Benfenati), International Journal of Signal Processing (Emilio Benfenati), Chemistry Central Journal (Emilio Benfenati), Current Computer Aided Drug Design (Emilio Benfenati), Advances in Chemoinformatics and Computational Methods (Emilio Benfenati), The Open Biomarkers Journal (Luisa Airoldi).

PEER REVIEW ACTIVITIES

Addiction, Analytical Chemistry, Analytical and Bioanalytical Chemistry, Chemical Biology & Drug Design, Chemical Research Toxicology, Chemometrics and Intelligent Laboratory Systems, CHEMOLAB, Chemosphere, Clinical Biochemistry, Drug and Alcohol Dependence, Environment International, Environmental Pollution, Environmental Modelling & Software, Environmental Science & Technology, International Journal of Molecular Sciences, Journal of Cellular Biochemistry, Journal of Chemical Information and Modeling, International Journal of Molecular Science, Journal Computer-Aided Molecular Design, Journal of Hazardous Materials, Molecular Cellular Proteomics, Molecular Diversity, Proteome Science, Royal Society's Philosophical Transactions, Stroke, Toxicology Letters, Waste Management, Water Research, International Journal of Environmental Analytical Chemistry, Molecular Nutrition and Food Research, Journal of Chromatography A, The Open Biomarkers Journal, Toxicological Sciences, External review of the quality of the scientific outputs of the European Food Safety Authority (EFSA).

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NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

CCPF - Commissione Consultiva Prodotti Fitosanitari (Ministero della Salute, Ministero dell'Ambiente) ECCO - European Commission Coordination EFSA - European Food Safety Authority IGQ - Commissione Ambiente e Sicurezza

EVENT ORGANIZATION
58th American Society for Mass Spectrometry Conference Finding unknowns in the environment: high resolution and MS/MS approach, Salt Lake City, UT, USA, June 1-5, 2010. OSIRIS Training Course on Integrated Testing Strategies (ITS), Istituto di Ricerche Farmacologche Mario Negri, Milano, Italy, November 3-5, 2010. Participation, as leader of the Local Organizing Committee (Emilio Benfenati), to the organization of the 21st SETAC Europe Annual Congress, Milano, Italy, May 15-19 2011.

CONFERENCE AND WORKSHOP CONTRIBUTIONS

Workshop Risk Assessment: valutazione dei rischi da inquinamento ambientale ed effetti sulla salute umana, GSISR-CNR, Milano, Italia, March 2, 2010. 7th Federation of European Neurosciences (FENS) Forum, Amsterdam, Paesi Bassi, July 3-7, 2010. 1st RISKCYCLE Workshop Risk-based management of chemicals and products in a circular economy at a global scale, Hanoi, Vietnam, May 4-5, 2010. SETAC Europe 20th Annual Meeting, Seville, Spain, May 23-27, 2010.

14th International Workshop on Quantitative StructureActivity Relationships in Environmental and Health Sciences, QSAR 2010, Montreal, Canada, May 24-28, 2010. 58th American Society for Mass Spectrometry Conference, Salt Lake City, UT, USA, June 1-5, 2010. Workshop Ecopharmacovigilance wich future?, Academie Nationale de Pharmacie, Paris, France, September 21, 2010. NOSE 2010. International Conference on Environmental Odour Control and Monitoring. Florence, Italy. September 22-24, 2010

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NATO SfP 982590 project workshop Characterization of hazardous chemical contamination From environmental chemistry and toxicology to risk assessment , Dubrovnik, Croatia, September 23 - 26, 2010. OSIRIS Training Course on Integrated Testing Strategies (ITS), Istituto di Ricerche Farmacologche Mario Negri, Milano, Italy, November 3-5, 2010. 2nd RISKCYCLE Workshop Risk of chimical additives and recycled materials: State of the art, new challenges and future trends, Shenyang, China PR, November 15-19, 2010.

GRANTS AND CONTRACTS

A2A Brescia ACEGAS S.p.A, Trieste AIIPA (Associazione Italiana Industrie Prodotti Alimentari) AMA, Roma ASL Cagliari ASL Como ASL Mantova ASL Napoli 2 ASSOFOODTEC/UCIMAC (Costruttori Italiani Macchine per Caff Espresso ed Attrezzature per Bar) Bergamo Pulita S.r.l. Bluegreen Biotech Bracco Imaging Spa Cambrex, Paullo (MI) Catanzaro Costruzioni S.r.l. CLIR S.p.A. COGEIDE S.p.A. COOP Italia CSRA Comune di Lomello (PV) Comune di Mazzano e Rezzato (BS) Comune di Rosignano Marittimo (LI) Comune di SantUrbano (PD) Consorzio Quadrifoglio S.p.A. Dipartimento delle Politiche Antidroga, Presidenza del Consiglio dei Ministri ECODECO S.r.l. EnergyGreen S.r.l. European Commission (ANTARES, ORCHESTRA, OSIRIS, RISKCYCLE) Federchimica, Milano FIAT Auto S.p.A. Fondazione CARIPLO, Milano Fondazione Italo Monzino, Milano Gruppo CSA, S.p.a. Rimini (RN) HERA S.p.A. (Holding Energia Risorse Ambiente) INDENA S.p.A. I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana Lachifarma, Zollino (LE)

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Ministero dell'Ambiente, Italia Ministero della Salute, Italia Nufarm S.A.S., Francia Oxon Italia S.p.A., Pero (MI) Politecnico di Milano Provincia di Pordenone Provincia di Vercelli Regione Lombardia SO.GE.NU.S. S.p.A Tenacta Group TM.E. S.p.A. Universit degli Studi di Milano Veolia Servizi Ambientali S.p.A.

SCIENTIFIC PUBLICATIONS (2010)

Gallo V, Neasham D, Airoldi L, Ferrari P, Jenab M, Boffetta P, Overvad K, Tjonneland A, Clavel-Chapelon F, Boeing H, Pala V, Palli D, Panico S, Tumino R, Arriola L, Lund E, Bueno-De-Mesquita H B, Peeters P H, Melander O, Hallmans G, Riboli E, Saracci R, Vineis P. Second-hand smoke, cotinine levels, and risk of circulatory mortality in a large cohort study of never-smokers. Epidemiology 2010 21 : 207-214. Schiarea S, Solinas G, Allavena P, Scigliuolo G, Bagnati R, Fanelli R, Chiabrando C. Secretome analysis of multiple pancreatic cancer cell lines reveals perturbations of key functional networks. J Proteome Res 2010 9 : 4376-4392. Solinas G, Schiarea S, Liguori M, Fabbri M, Pesce S, Zammataro L, Pasqualini F, Nebuloni M, Chiabrando C, Mantovani A, Allavena P. Tumor-conditioned macrophages secrete migration-stimulating factor: a new marker for m2-polarization, influencing tumor cell motility. J Immunol 2010 185 : 642-652. Diana V, Botti F, Fumagalli E, Calcagno E, De Paola M, Cagnotto A, Invernici G, Parati E, Curti D, Mennini T. Neural precursor-derived astrocytes of wobbler mice induce apoptotic death of motor neurons through reduced glutamate uptake. Exp Neurol 2010 225 : 163-172. Toropov A A, Toropova A P, Benfenati E. QSPR modelling of normal boiling points and octanol/water partition coefficient for acyclic and cyclic hydrocarbons using SMILES-based optimal descriptors. Central European Journal Chemistry 2010 8 : 1047-1062. Benfenati E, Gini G, Hoffmann S, Luttik R. Comparing in vivo, in vitro and in silico methods and integrated strategies for chemical assessment: problems and prospects. Altern Lab Anim 2010 38 : 153-166. Toropov A A, Toropova A P, Benfenati E, Leszczynska D, Leszczynksy J. SMILES-based optimal descriptors: QSAR analysis of fullerene-based HIV-1 PR inhibitors by means of balance of correlations. J Comput Chem 2010 31 : 381-392. Senese V, Boriani E, Baderna D, Mariani Alessandro, Lodi M, Finizio A, Testa S, Benfenati E. Assessing the environmental risks associated with contaminated sites: Definition of an Ecotoxicological Classification index for landfill areas (ECRIS). Chemosphere 2010 80 : 60-66. Toropov A A, Toropova A P, Benfenati E, Leszczynska D, Leszczynksy J. QSAR analysis of 1,4-dihydro-4-oxo-1(2-thiazolyl)-1,8-naphthyridines exhibiting anticancer activity by optimal SMILES-based descriptors. J Math Chem 2010 47 : 647-666. Lombardo A, Gonella Diaza R, Roncaglioni A, Benfenati E. CAESARs approach for alternative in silico methods for REACH. ALTEX 2010 27 : 109-116. Benfenati E. The CAESAR project for in silico models for the REACH legislation. Chem Cent J 2010 4 (Suppl 1) : I1.

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Lombardo A, Roncaglioni A, Boriani E, Milan C, Benfenati E. Assessment and validation of the CAESAR predictive model for bioconcentration factor (BCF) in fish. Chem Cent J 2010 4 (Suppl 1) : S1. Chaudhry Q, Piclin N, Cotterill J, Pintore M, Price N, Chretien J R, Roncaglioni A. Global QSAR models of skin sensitisers for regulatory purposes. Chem Cent J 2010 4 (Suppl 1) : S5. Fjodorova N, Vrachko M, Novich M, Roncaglioni A, Benfenati E. New public QSAR model for carcinogenicity. Chem Cent J 2010 4 (Suppl 1) : S3. Cassano Antonio, Manganaro A, Martin T M, Young Douglas, Piclin N, Pintore M, Bigoni D, Benfenati E. CAESAR models for developmental toxicity. Chem Cent J 2010 4 (Suppl 1) : S4. Toropov A A, Toropova A P, Benfenati E, Manganaro A. QSAR modelling of the toxicity to Tetrahymena pyriformis by balance of correlations. Mol Divers 2010 14 : 821-827. Toropov A A, Toropova A P, Benfenati E. QSAR-modeling of toxicity of organometallic compounds by means of the balance of correlations for InChI-based optimal descriptors. Mol Divers 2010 14 : 183-192. Toropov A A, Toropova A P, Benfenati E, Leszczynska D, Leszczynksy J. Use of the international chemical identifier for constructing QSPR-model of normal boiling points of acyclic carbonyl substances. J Math Chem 2010 47 : 355-369. Boriani E, Mariani A., Baderna D, Moretti C, Lodi M, Benfenati E. ERICA: A multiparametric toxicological risk index for the assessment of environmental healthiness. Environ Int 2010 36 : 665-674. Toropov A A, Toropova A P, Raska I, Benfenati E. QSPR modeling of octanol/water partition coefficient of antineoplastic agents by balance of correlations. Eur J Med Chem 2010 45 : 1639-1647. Toropov A A, Toropova A P, Benfenati E, Leszczynska D, Leszczynksy J. InChI-based optimal descriptors: QSAR analysis of fullerene[C60]-based HIV-1 PR inhibitors by correlation balance. Eur J Med Chem 2010 45 : 1387-1394. Toropova A P, Toropov A A, Lombardo A, Roncaglioni A, Benfenati E, Gini G. A new bioconcentration factor model based on SMILES and indices of presence of atoms. Eur J Med Chem 2010 45 : 4399-4402. Piir G, Sild S, Roncaglioni A, Benfenati E, Maran U. QSAR model for the prediction of bio-concentration factor using aqueous solubility and descriptors considering various electronic effects. SAR QSAR Environ Res 2010 21 : 711-729. Toropov A A, Toropova A P, Benfenati E. SMILES-based optimal descriptors: QSAR modeling of carcinogenicity by balance of correlations with ideal slopes. Eur J Med Chem 2010 45 : 3581-3587. Toropova A P, Toropov A A, Benfenati E, Leszczynska D, Leszczynksy J. QSAR modeling of measured binding affinity for fullerene-based HIV-1 PR inhibitors by CORAL. J Math Chem 2010 48 : 959-987. Vigan L, Benfenati E, Bottero S, Cevasco A, Monteverde M, Mandich A. Endocrine modulation, inhibition of ovarian development and hepatic alterations in rainbow trout exposed to polluted river water. Environ Pollut 2010 158 : 3675-3683. Adani F, Tambone F, Davoli E, Scaglia B. Surfactant properties and tetrachloroethene (PCE) solubilisation ability of humic acid-like substances extracted from maize plant and from organic wastes: A comparative study. Chemosphere 2010 78 : 1017-1022. Davoli E, Fattore E, Paiano V, Colombo A, Palmiotto M, Rossi A N, Il Grande M, Fanelli R. Waste management health risk assessment: a case study of a solid waste landfill in South Italy. Waste Manag 2010 30 : 1608-1613. Urru S A M, Veglianese P, De Luigi A, Fumagalli E, Erba E, Gonella Diaza R, Carr A, Davoli E, Borsello T, Forloni G, Pengo N, Monzani E, Cascio P, Cenci S, Sitia R, Salmona M. A new fluorogenic peptide determines proteasome activity in single cells. J Med Chem 2010 53 : 7452-7460. Orzi V, Cadena E, D'Imporzano G, Artola A, Davoli E, Crivelli M, Adani F. Potential odour emission measurement in organic fraction of municipal solid waste during anaerobic digestion: relationship with process and biological stability parameters. Bioresour Technol 2010 101 : 7330-7337.

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Bianchi G, Celeste G, Palmiotto M, Davoli E. Source identification of odours and VOCs from a composting plant by multivariate analysis of trace volatile organic compounds. Chemical Engineering Transactions 2010 23 : 279-284. Zuccato E, Castiglioni S, Bagnati R, Melis M, Fanelli R. Source, occurrence and fate of antibiotics in the Italian aquatic environment. J Hazard Mater 2010 179 : 1042-1048. Castiglioni S, Zuccato E. Illicit drugs in the environment: Emerging contaminants and indicators of drug abuse. Integrated Environmental Assessment and Management 2010 6 : 186-187. Grassi P, Fattore E, Generoso C, Fanelli R, Arvati M, Zuccato E. Polychlorobiphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in fruit and vegetables from an industrial area in northern Italy. Chemosphere 2010 79 : 292-298. Heath E, Kosjek T, Farre' M, De Alencastro F, Castiglioni S, Gans O, Langford K, Loos R, Radjenovic J, Mainero Rocca L, Budzinski H, Tsipi D, Petrovic M, Barcelo D. Second interlaboratory exercise on non-steroidal antiinflammatory drug analysis in environmental aqueous samples. Talanta 2010 81 : 1189-1196. berg M, Westerholm E, Fattore E, Stern N, Hanberg A, Haglund P, Wiberg K, Bergendorff A, Hakansson H. Toxicity of Bromkal 70-5DE, a technical mixture of polybrominated diphenyl ethers, following 28 d of oral exposure in rats and impact of analysed impurities. Chemosphere 2010 E-pub : 10.1016/j.chemosphere.2010.04.006. Sala F, Marangon E, Bagnati R, Livi V, Cereda R, D'Incalci M, Zucchetti M. Development and validation of a highperformance liquid chromatographytandem mass spectrometry method for the determination of the novel proteasome inhibitor CEP-18770 in human plasma and its application in a clinical pharmacokinetic study. J Mass Spectrom 2010 45 : 1299-1305.

LAY PRESS SELECTION (2010)

Fattore E, Davoli E, Bonati M. Il Fantasma della discarica. Il Sole 24 ore Sanit. 30 nov. 6 dic. 2010 Fattore E. ADHD ed esposizione a pesticidi organofosforici. Ricerca & Pratica 2010, 155: 204-205 Fattore E. Neurotossicit dei composti chimici sullo sviluppo. Ricerca & Pratica 2010, 155: 205-207 Fattore E. Esposizione prenatale agli IPA e QI nei bambini. Ricerca & Pratica 2010, 156: 257-260 Zuccato E, Olandese R, Antinozzi R, Castiglioni S. Stima dei consumi di sostanze stupefacenti mediante analisi delle acque reflue: confronto anni 2008 e 2009. Il case-study della citt di Como.

RESEARCH ACTIVITIES Laboratory of Molecular Toxicology Proteome Analysis

Proteome analysis includes protein separation by one- and two-dimensional gel electrophoresis, protein excision from the gel, their digestion with proteolytic enzymes and their identification by mass spectrometry (MALDI-TOF-MS, LC-ESI-MS/MS) coupled to the use of existing databases. Alternatively, peptides resulting from the digestion of protein mixtures with specific proteases are separated by two-dimensional liquid chromatography.

Toxicoproteomics
Studies are ongoing on the characterization of changes in the proteome profile induced by

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environmental toxic compounds, with the aim of obtaining protein biomarkers with the ability to differentiate two or more biological states. Proteome changes in tissues and target organs of animals, and cells treated with endocrine disruptors, estrogens, or environmental carcinogens, are related to functional changes during toxicological processes.

Clinical Proteomics
Qualitative and quantitative proteome changes resulting from the exposure to environmental toxic compounds or in pathological conditions are monitored in human biological plasma and urine. Ongoing studies aim at the characterization of protein biomarkers for early diagnosis of diseases and for the identification of therapeutic targets.

Pathways analysis
An integrated data-mining platform such as MetaCore (GeneGo Inc., USA) is used in order to map the differentially expressed proteins into biological networks and for functional interpretation of the protein data.

Metabolomics
Metabolites are the biological endpoints of gene expression and enzyme activity and include small molecules such as amino acids, carbohydrates, fatty acids, hormones, etc., that provide the metabolic phenotype of individuals. Metabolomic research focuses on the quantitative analysis of metabolites in biological fluids to link human metabolic profile variations to endogenous or exogenous pathophysiological stimuli and to genetic modifications. Selected metabolites are extracted from plasma or urine by solid phase extraction and analyzed by HPLC coupled to high-resolution mass spectrometry. The integration of proteomic and metabolomic studies will provide information that can help to better understand disease development and to identify preventive interventions.

Molecular Epidemiology
The laboratory works mainly on the measurement of biological markers used to assess human exposure to environmental toxic compounds. Our studies include DNA- and blood proteinadduct formation by several environmental carcinogens. In addition, we study whether the polymorphism of genes coding for enzymes involved in the activation and detoxification of carcinogens are determinants of adduct formation. Genotypes are detected by restriction fragment length polymorphism analysis, after the amplification by polymerase chain reaction of specific nucleotide sequences of the genes under study. The laboratory participates in an international cooperation study aimed at the collection of reference values on allele and genotype frequency of the most common metabolic enzyme polymorphisms in control populations.

Laboratory of Analytical Biochemistry Identification and characterization of proteins by mass spectrometry

Our laboratory is developing different analytical and instrumental techniques based on mass spectrometry for the identification and characterization of proteins and peptides in biological samples. This activity is mainly aimed at 1) global proteomic characterization and comparison of secretomes from human cancer cell lines; 2) profiling proteins in biological fluids for discovery and identification of biomarkers of physiopathological and toxicological relevance, 3) identifying and characterizing endogenous degradation products of proteins, 4) identifying proteins produced by cells in vitro in response to given stimuli, 5) identifying and characterizing

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biologically relevant proteins isolated from biological samples by immunoaffinity-based techniques.

Proteomics in oncology
This activity is mainly aimed at discovering among the proteins we find abnormally secreted by human cancer cell lines new molecules of oncological interest, and in particular novel candidate therapeutic targets or diagnostic/prognostic biomarkers. Ongoing projects include the characterization of the secretomes of cancer cell lines in vitro to identify the most relevant abnormalities. The complex alterations observed in the cancer secretomes are then rationalized and interpreted by using systems biology tools that are able to highlight the functional networks most significantly perturbed.

Inflammation and neurodegeneration induced by environmental agents

We are studying the neurotoxic effects of endotoxin (LPS) and other environmental proinflammatory agents on neuronal cell primary cultures. The mechanisms leading to the activation of the inflammatory reaction are studied by biochemical and immunochemical methods, and proteomic approaches. Novel anti-inflammatory drugs of natural origin are being tested in this model, with the aim of evaluating their neuroprotective effects.

Laboratory of Environmental Chemistry and Toxicology Development and use of analytical methods to evaluate contamination in water bodies, soil, biota, human samples in exposed population
Analytical methods are developed to study environmental pollutants in water ecosystems, landfills, contaminated sites. Qualitative and quantitative analyses of organic pollutants are done by mass spectrometry (GC-MS, LC-MS, LC-MS/MS). Typical analyses include PCDD/F, PCB, PAH, polybrominated diphenylethers, pesticides, endocrine disruptor chemicals, and industrial pollutants.

Studies on environmental, toxicological and ecotoxicological properties of chemicals

Research is carried out on pollutant properties, searching literature data, comparing and evaluating different sources, and mainly developing predictive models to cope with the lack of experimental data. Thus, we develop models starting merely from the chemical structure. The research addresses the different kinds of chemical descriptors and chemical fragments, obtained with different software. Then, we develop models using algorithms such as neural network, fuzzy logic, genetic algorithms, classifiers, multivariate analysis, etc. Different methods are compared and integrated within a structured ensemble. Standardized methods for pesticides were developed and validated according to OECD guidelines.

Risk assessment of pollutants

Studies are aimed at assessing the risk of pollutants for human population and environment. For this we model transport and diffusion of pollutants, to obtain a predicted concentration on given space and time scales. Such an activity is integrated with those above described on chemical analyses and toxicity prediction, to achieve a continuous transfer of data and research.

Research on pollutants emitted in the atmosphere (Unit of Industrial and Environmental Hygiene)
Studies address different aspects of atmospheric pollution. Research deals with: sampling areas

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around the pollution source, chemical analyses, transport modeling depending on meteorological conditions and orography, risk assessment for population and environment. Qualitative and quantitative analyses are done by gas chromatography-mass spectrometry using high resolution for PCDDs/PCDFs, and negative ion-chemical ionization for PCBs.

Laboratory of Mass Spectrometry Particulate air pollution

Epidemiological studies consistently show an association between an increasing number of pathologies, both acute and chronic, and particulate air pollution. This has been shown not only in respiratory, but in cardiovascular diseases as well. Airborne particulate sampling and analysis strategies are developed to characterize both adsorbed compounds and exposition in different activities.

Method development in environmental sciences

Methods, analytical methodologies, instrumentation and software for data acquisition and reduction, are developed for environmental studies. High-sensitivity instrumentation, mainly based on mass spectrometry, is developed for trace and ultra-trace analysis. Also, transportable instrumentation is developed for field studies or continuous monitoring.

Characterization of environmental odor annoyance

Characterization of odors poses several analytical problems because they result from a complex mixture of compounds (odorants) stimulating receptors in the nasal cavity. Most odorants are volatile organic compounds (VOC) generated by bacterial degradation of organic matter. They are often present at trace levels, while numerous sources can contribute to the total odor. Using sampling techniques specifically developed for olfactometry, solid phase microextraction and GC/MS analysis, we can detect traces (low ppb to high ppt) of a wide polarity/volatility range of airborne VOC odorant compounds. With a chemometric approach, we can characterize the sources of emissions, assess odor control methods, and identify emissions that contribute to odors in ambient air.

Laboratory of Food Toxicology Chemical contaminants in food

We are studying human exposure to dietary PCBs and dioxins in Italy. In particular, contaminants were measured in samples of human milk collected from mothers living in highly contaminated areas i Further studies were aimed at measuring PCBs and dioxins in samples of fish caught in Italy and in food items from an Italian area at high risk of contamination. .

Therapeutic and illicit drugs in the environment

Pharmaceuticals are a class of emerging environmental pollutants. We have organized a campaign to detect the presence of pharmaceuticals and their metabolites in Italian rivers and sewage treatment plants and in samples of drinking water, with the aim of characterizing the contamination and assessing related risks. Further ongoing studies are aimed at investigating a possible relationship between antibiotic occurrence and resistance in environmental bacteria. The possible presence of illicit drugs in water samples from sewage treatment plants and rivers was investigated, starting with cocaine and its metabolites. Their levels, used to estimate drug abuse in the local population, revealed that cocaine consumption greatly exceeds official estimates. This approach has been subsequently extended to include other common drugs of

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abuse such as cannabis, opiates (heroin, morphine), and amphetamines (amphetamine, methamphetamine, ecstasy). Our evidence-based method allows monitoring of patterns and trends of drug abuse in local communities, and is able to detect qualitative and quantitative consumption changes in real time. This tool can therefore complement survey methods in more realistically describing the drug abuse phenomenon. Ongoing studies are focussed to assess consumptions at national scale, in collaboration with the National Agency for Drug Policy, at regional scale in collaboration with Regione Lombardia, and locally, in collaboration with Metropolitana Milanese.

Unit of Environmental Pollutants Risk Assessment Toxicological risk assessment

Starting from real cases of contamination, the unit aims to develop methods for the exposure assessment also employing probabilistic approaches, and more refined statistical models. The activities focus on risk assessment related to specific environmental conditions, or human activities, which pose a risk for human health. These studies include risk assessments related to contamination of water or soil, emissions of toxics pollutants from waste management processes, including transfer of these compounds into the food chain. During 2010, studies focused on health effects due to waste disposal into landfills, and on soil treatment with sludge and the potential transfer of persistent organic pollutants across the food chain.

Exposure to environmental pollutants

Research activities also include measurement of contaminants in environmental samples, and assessment of human exposure. Specific research projects focused on the analysis of polychlorinated and polybrominated dioxins and furans (PCDD, PBDD, PCDF and PBDF), polychlorinated biphenyls (PCBs), perfluorooctanoic acid (PFOA), and perfluorooctane sulphonate (PFOS), in aquatic organisms at different levels of the food chain, and farmed fish coming from different areas of the Mediterranean sea. The purpose wass to estimate, for the general Italian population, the exposure to these pollutants trough fish consumption.

Evaluation of toxicological data

Toxicological data, resulting from in vivo sub-chronic studies in rats exposed to individual dioxin-like and non dioxin-like PCBs are evaluated in detail, in order to investigate the doseresponse relationship and the applicability of the benchmark dose approach.

Unit of Analytical Instrumentation Development and application of analytical methods for compounds of biological and environmental interest.
Biological fluids and environmental samples are analysed mainly using solid phase extraction (SPE) and liquid chromatography - mass spectrometry (LC-ESI-MS/MS). Proteins and peptides are also analysed by laser desorption ionization techniques. Tissue samples are directly analyzed by using MALDI or PALDI Imaging (Matrix or nanoParticle Laser Desorption Ionization). Available instruments include liquid chromatographs and mass spectrometers equipped with different analyzers: time of flight (TOF), triple quadrupoles, ion traps and high resolution LTQOrbitrap, with conventional and nanoElectroSpray sources. Substances of interest include proteins, peptides, hormones, pharmaceuticals, drugs of abuse, and other environmental contaminants (pesticides, perfluorinated compounds).

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DEPARTMENT OF NEUROSCIENCE
STAFF Head Gianluigi FORLONI, Biol.Sci.D.

Laboratory of Biology of Neurodegenerative Disorders

Head Gianluigi FORLONI, Biol.Sci.D.

Laboratory of Cell Death and Neuroprotection

Head Tiziana Borselllo, Biol.Sci.D.

Laboratory of Experimental Neurology

Head Annamaria VEZZANI, Biol.Sci.D.

Neuro-glia communications Unit

Head Teresa Ravizza, Biol.Sci.D.

Laboratory of Experimental Psychopharmacology

Head
Luigi CERVO, Ph.D.

Laboratory of Geriatric Neuropsychiatry

Head Ugo LUCCA, MSc

Epidemiology and Social Psychiatry Unit

Head Barbara DAVANZO, Philos.D.

Geriatric Epidemiology Unit

Head Mauro TETTAMANTI, Biol.Sci.D.

Geriatric Pharmacology Unit

Head Emma RIVA, M.D.

Laboratory of Inflammation and Nervous System Diseases

Head Pharmacology of septic shock Head
Maria Grazia DE SIMONI, Biol.Sci.D.

Pia VILLA, Biol. Sci. D

Laboratory of Molecular Neurobiology

Head Caterina BENDOTTI, Farm.D.

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Laboratory of Neurobiology of Prions

Head Roberto CHIESA, Biol. Sci. D

Laboratory of Neurochemistry and Behavior

Head Roberto William INVERNIZZI, Biol. Sci D

Pharmacology of Cognitive Behavior Unit

Head Mirjana CARLI, Ph.D.

Laboratory of Neurological Disorders

Head Ettore BEGHI, M.D.

Laboratory of Quality Assessment of Geriatric Services Unit

Head Alessandro NOBILI, M.D.

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CURRICULA
Gianluigi Forloni, obtained the Degree of Biological Science at the University of Milan in 1985. After two years of post doc at the Department of Neuroscience and Psychiatry at Johns Hopkins University in Baltimore, USA, he came back to the Mario Negri Institute and between 1992 and 1996 he was the head of the Neurobiology of Alzheimer's disease Unit; since 1996 he is the Head of the Biology of Neurodegenerative Diseases Lab and since 2002 also the Head of the Neuroscience Department. His scientific interest is focused on the biological and genetic bases of aging-related disorders in particular Alzheimers disease, Prion-related encephalopathies and Parkinsons disease. He has been member of several European committees for the examination of projects in the neuroscience field. He is now member of the coordination group of the European IMI Consortium PharmaCog. He is President of the Italian Association on Brain Aging Research (AIRIC) and member of the European Academy of Sciences. He is the author of more than 190 peer-reviewed scientific articles and about 30 reviews or book chapters.
Selected publications Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O.,Tagliavini F. Neurotoxicity of a prion protein fragment. Nature 362: 543-546 (1993) Forloni, G., Tagliavini, F.,Bugiani, O. and Salmona, M. Amyloid in Alzheimers disease and prion-related encephalopathies: Studies with synthetic peptides. Progr. Neurobiol. 49: 287- 315 (1996) Forloni G. Iussich, S. Awan T. Colombo L. Angeretti, N. Girola, L. Bertani, I. Poli, G. Caramelli, M. Bruzzone, MG.Farina, L. Limido, L. Rossi, G. Giaccone G. Ironside, JW. Bugiani, O.Salmona M. and Tagliavini, F. Tetracyclines affect prion infectivity Proc. Natl. Acad. Sci . New York 99: 10849-10854 (2002) Fioriti, L. Angeretti, N.. Colombo, L., De Luigi A., Manzoni, C., Colombo A., Morbin, M., Tagliavini, F., Salmona, M. Chiesa, R. Forloni, G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-StrusslerScheinker disease amyloid protein. J. Neurosci. 27: 576-83 (2007) Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron. 60: 598-609 (2008) Albani D, Polito L, Batelli S, De Mauro S, Fracasso C, Martelli G, Colombo L, Manzoni C, Salmona M, Caccia S, Negro A, Forloni G. The SIRT1 activator resveratrol protects SK-N-BE cells from oxidative stress and against toxicity caused by alpha-synuclein or amyloid-beta (1-42) peptide. J Neurochem. 110:1445-56 (2009). Balducci, C., Beeg, M., Stravalaci, M., Bastone, A.,, Sclip, A., Biasini, E., Tapelll., Colombo, L. Canzoni, C., Borsello, T., Chiesa, R., Gobbi, M., Salmona M. Forloni, G., A Synthetic oligomers impair memory independently of cellular prion potein Proc. Natl. Acad. Sci USA, 107: 2295-2300 (2010)

Ettore Beghi graduated in Medicine in 1972 and received his specialty in neurology in 1976 at the University of Milan. He trained in epidemiology with a fellowship at the Department of statistics and Epidemiology of the Mayo Clinic in Rochester, MN (USA). He is Head of the Laboratory of Neurological Disorders at the Mario Negri Institute, Director of the Neurophysiology/Epilepsy Unit and Professor of Neuroepidemiology at the University of Milano-Bicocca, Monza. He is member of the editorial board of the journals Epilepsia, Neuroepidemiology, Inpharma, Drugs in R & D, Clinical Drug Investigation, Neurological Sciences and is a referee of several national and international medical journals. The main areas of interest and research include studies on the descriptive, analytic, and experimental epidemiology in the field of epilepsy, peripheral neuropathies, headache, and amyotrophic lateral sclerosis.
Selected publications E. Beghi, G. Logroscino, A. Chi, O. Hardiman, A. Millul, D. Mitchell, R. Swingler, B.J. Traynor. Amyotrophic lateral sclerosis, physical exercise, trauma and sports: results of a population-based pilot case-control study. Amyotrophic Lateral Sclerosis 2010; 11: 289-292. E. Beghi, A. Carpio, L. Forsgren, D.C. Hesdorffer, K. Malmgren, J.W. Sander, T. Tomson, W.A. Hauser. Recommendation for a definition of acute symptomatic seizure. Epilepsia 2010; 51: 671-675. A. Del Felice, E. Beghi, G. Boero, A. La Neve, G. Bogliun, A. De Palo, L.M. Specchio. Early versus late remission in a cohort of patients with newly diagnosed epilepsy. Epilepsia 2010; 51: 37-42. G. Logroscino, B.J. Traynor, O. Hardiman, A. Chi, D. Mitchell, R.J. Swingler, A. Millul, E. Benn, E. Beghi. Incidence of amyotrophic lateral sclerosis in Europe. J Neurol Neurosurg Psychiatry 2010; 81: 385-390. Leone MA, Solari A, Beghi E, for the FIRST Group. Treatment of the first tonic-clonic seizure does not affect long-term remission of epilepsy. Neurology 2006; 67: 2227-2229

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Van den Broek M, Beghi E, for the RESt-1 Group. Accidents in patients with epilepsy: type and complications. A European Cohort Study. Epilepsia 2004; 45: 667-672.

Caterina Bendotti, got her degree in Pharmacy at the University of Milano in 1984; In 1986 -1988 she was postdoc at the Genetic Developmental Lab, Dept. of Physiology of the Johns Hopkins University, Baltimore, USA. In 1988 -1992 she was research fellow in the laboratory of Neuropharmacology and in the 1992, she became head of the Molecular Neurobiology Unit in Institute, since 1998 she is head of laboratory. The major research interest is the study of pathogenetic mechanisms of familial Amyotrophic Lateral Sclerosis.. Since 2002 she is a member of the editorial board of Journal of Neurochemistry. In 2002-2003 has been Member of Scientific Committees of the International Symposia on ALS held in Milano, 17-19 Novembre,2003. In 2003-2007 has been member of the Italian Ministry of Health Committees for the diagnosis, cure, care and assistance of patients with ALS. Since 2005-2009 she has been member of the Executive Council of the Italian Society of Neuroscience. Since 2006 is member of the Research Advisory Panel of the MND Association, UK. Scientific reviewer of 11 international scientific journals. In 2007 she has co-organised the first international meeting on Mutant SOD1 and familial ALS: from the molecule to man held in Milano (13-16 September). She is author and co-author of 110 articles 100 of which with peer-review. Rapporteur of many communications in national and international meetings.
Selected publications Crippa V, Sau D, Rusmini P, Boncoraglio A, Onesto E, Bolzoni E, Galbiati M, Fontana E, Marino M, Carra S, Bendotti C, De Biasi S, Poletti A. The small heat shock protein B8 (HspB8) promotes autophagic removal of misfolded proteins involved in amyotrophic lateral sclerosis (ALS). Hum Mol Genet. 19(17):3440-56, 2010 Peviani M, Caron I, Pizzasegola C, Gensano F, Tortarolo M, Bendotti C. Unraveling the complexity of amyotrophic lateral sclerosis: Recent advances from the transgenic mutant SOD1 mice. CNS Neurol Disord Drug Targets. 9(4):491503, 2010 Ludolph AC, Bendotti C, Blaugrund E, Chio A, Greensmith L, Loeffler JP, Mead R, Niessen HG, Petri S, Pradat PF, Robberecht W, Ruegg M, Schwalenstcker B, Stiller D, van den Berg L, Vieira F, von Horsten S. Guidelines for preclinical animal research in ALS/MND: A consensus meeting. Amyotroph Lateral Scler.11(1-2):38-45, 2010 Basso M, Samengo G, Nardo G, Massignan T, D'Alessandro G, Tartari S, Cantoni L, Marino M, Cheroni C, De Biasi S, Giordana MT, Strong MJ, Estevez AG, Salmona M, Bendotti C, Bonetto V. Characterization of detergent-insoluble proteins in ALS indicates a causal link between nitrative stress and aggregation in pathogenesis.PLoS One. 4(12):e8130. 2009 Bendotti C, Carr MT. Amyotrophic lateral sclerosis: mechanisms and countermeasures. Antioxid Redox Signal. 11(7):1519-22, 2009 Pizzasegola C, Caron I, Daleno C, Ronchi A, Minoia C, Carr MT, Bendotti C.Treatment with lithium carbonate does not improve disease progression in two different strains of SOD1 mutant mice. Amyotroph Lateral Scler. 10(4):221-8, 2009 Nardo G, Pozzi S, Mantovani S, Garbelli S, Marinou K, Basso M, Mora G, Bendotti C, Bonetto V.Nitroproteomics of peripheral blood mononuclear cells from patients and a rat model of ALS. Antioxid Redox Signal. 11(7):1559-67, 2009 Cheroni C, Marino M, Tortarolo M, Veglianese P, De Biasi S, Fontana E, Zuccarello LV, Maynard CJ, Dantuma NP, Bendotti C.Functional alterations of the ubiquitin-proteasome system in motor neurons of a mouse model of familial amyotrophic lateral sclerosis. Hum Mol Genet. 18(1):82-96, 2009

Tiziana Borsello got her Degree in Biological Science at the University of Torino in 1990 and she then obtained a PhD in Neuroscience at the University of Turin Medical School. She won 1 year fellow of the European Science Foundation scholarship for work at the Netherlands Research Institute of Amsterdam. From 1997 to 1999 she was a Researcher at the Institute of Neurobiology, CNR, Rome Italy. In the period 1999-2003 she was Premier Assistant, Dpartement de Biologie Cellulaire et de Morphologie, Universit de Lausanne, Switzerland, and then became Maitre Assistant and group leader in the same institute. In 2004 joined the Biol. Neurodeg. Disorders Lab at the "Mario Negri Institute. In 2005 won the Prize of the Pfizer Foundation, Neuroscience and Diseases Nervous System. Since 2006 she is the Head of the Unit: Neuronal Death and Neuroprotection. Her main scientific interest is understanding the role of signaling pathways in neuronal death after different stress-stimuli and the neuroprotection. In particular, the present research is focused on the study of the mechanisms of excitotocic stress, ischemia, Traumatic Brain Injury and the cell death pathways in neurodegenerative diseases as Alzheimer, with the challenge to define the neuronal death pathways to design more specific methods of neuroprotection.
Selected pubblications Antoniou X, Falconi M, Di Marino D, Borsello T. JNK3 as a therapeutic target for Neurodegenerative disease. J Alzheimers Dis. 2010 Feb 24. Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, Tapella L, Colombo L, Manzoni C, Borsello T, Chiesa R, Gobbi M, Salmona M, Forloni G. Synthetic Amyloid-Beta Oligomers Impair Long-Term Memory Independently Of Cellular Prion Protein. Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):2295-300 Colombo A, Bastone A, Ploia C, Sclip A, Salmona M, Forloni G, Borsello T. JNK Regulates App Cleavage And

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Degradation In A Model Of Alzheimer's Disease Neurobiology Of Disease, 2009 Mar;33(3):518-25 Borsello T Ed Neuroprotection: Methods In Molecular Biology Published By Humana Press, Usa HYPERLINK "http://www.humanapress.com/"Humana Press, USA, Methods in Molecular Biology, June 2007 Colombo A, Repici M, Pesaresi M, Santambrogio S, Forloni G, Borsello T. The Tat-Jnk Inhibitor Peptide Interferes With Beta Amyloid Protein Stability Cell Death Differ. 2007, 14:1845-8. Borsello T and Forloni G. JNK signalling: a possible target to prevent neurodegeneration. Current Pharmaceutical Design 2007, 13, 1875-1886 Centeno C., Repici M., Chatton J. Y., Riederer B. M., Bonny C., Nicod P., Price M., Clarke P. G., Papa S., Franzoso G. and Borsello T. Role of the JNK pathway in NMDA-mediated excitotoxicity of cortical neurons. Cell Death Differ , 2007, 14: 240-253. Borsello T. and Bonny C.Use of cell-permeable peptides to prevent neuronal degeneration. Trend in Mol. Med. 2004, 10: 239-44 Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nature Med. 2003, 9: 1180-6

Luigi Cervo was awarded the degree of Doctor of Philosophy (Ph.D.) from the Open University, Milton Keynes, U. K. in 2005. Since 2006 he has been the head of the Experimental Psychopharmacology Laboratory. From 1978 to 2001 he has been a research fellow and then Chief of the Behavioural Pharmacology Unit in the laboratory of Neuropharmacology and in 1981 he was awarded the degree in Biochemical Research from the M. Negri Institute. Between 1981 and 1983 he spent two years as a research fellow in the Department of Psychiatry at the Chicago University, Illinois, U.S.A. His main research interests concern Neuropsychopharmacology and the mechanism of action of psychotropic drugs. In particular the role of receptors subtypes for serotonin, dopamine, noradrenaline and glutamate in drug dependence and drug craving, depression, anxiety. Author and co-author of several peer-review articles, author of communications in international meetings, he is scientific reviewer of several international scientific journals. Member of Society for Neuroscience, European Behavioural Pharmacological Society, Italian Society for Neuroscience and Italian Society of Neuropsychopharmacology.
Selected publications Cervo L, Carnovali, F, Stark JA, Mennini T. Cocaine-seeking behavior in response to drug-associated stimuli in rats: involvement of D3 and D2 dopamine receptors. Neuropsychopharmacology 2003; 28: 1150-1159 Grignaschi G, Burbassi S, Zennaro E, Bendotti C, Cervo L. A single high dose of cocaine induces behavioural sensitization and modifies mRNA encoding GluR1 and GAP-43 in rats. Eur J Neurosci 2004; 20:2833-2837 Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi R. Deficits of serotonin synthesis cause resistance to antidepressants, J Neuroscience 2005; 25: 8165-8172 Cervo L, Cocco A, Petrella C, Heidbreder CA. Selective antagonism at dopamine D3 receptors attenuates cocaineseeking behaviour in the rat. Int J Neuropsychopharmacol. 2007; 10: 167-181. Burbassi S, Cervo L. Stimulation of serotonin(2C) receptors influences cocaine-seeking behavior in response to drugassociated stimuli in rats. Psychopharmacology (Berl). 2008; 196: 15-27. Burattini C, Burbassi S, Aicardi G, Cervo L. Effects of naltrexone on cocaine- and sucrose-seeking behaviour in response to associated stimuli in rats. Int J Neuropsychopharmacol. 2008; 11, 103-109. Marchesi F, Piemonti L, Fedele G, Destro A, Roncalli M, Albarello L, Doglioni C, Anselmo A, Doni A, Bianchi P, Laghi L, Malesci A, Cervo L, Malosio M, Reni M, Zerbi A, Di Carlo V, Mantovani A, Allavena P. The chemokine receptor CX3CR1 is involved in the neural tropism and malignant behavior of pancreatic ductal adenocarcinoma. Cancer Res. 2008; 68, 9060-9069. Fumagalli F, Franchi C, Caffino L, Racagni G, Riva MA, Cervo L. Single session of cocaine intravenous selfadministration shapes goal-oriented behaviours and up-regulates Arc mRNA levels in rat medial prefrontal cortex Int J Neuropsychopharmacol. 2009; 12:423-9. Calcagno E, Guzzetti S, Canetta A, Fracasso C, Caccia S, Cervo L, Invernizzi RW. Enhancement of cortical extracellular 5-HT by 5-HT1A and 5-HT2C receptor blockade restores the antidepressant-like effect of citalopram in non-responder mice. Int J Neuropsychopharmacol. 2009 12: 793-803. Watson J, Guzzetti S, Franchi C, Di Clemente A, Burbassi S, Emri Z, Leresche N, Parri HR, Crunelli V, Cervo L. Gamma-hydroxybutyrate does not maintain self-administration but induces conditioned place preference when injected in the ventral tegmental area. Int J Neuropsychopharmacol. 2010, 13:143-53.

Roberto Chiesa graduated in Biological Sciences with major in Genetics at the University of Pavia in 1991, and obtained a Ph.D. in Pharmacology at the Mario Negri Institute for Pharmacological Research of Milan in 1994. From 1996 through 2000 he was Research Associate at the Department of Cell Biology and Physiology of Washington University in St. Louis, MO, USA. In 2001 Dr. Chiesa moved back to the

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Mario Negri Institute where he is currently head of the Prion Neurobiology lab in the Department of Neuroscience. He also holds an Associate Telethon Scientist position (Dulbecco Telethon Institute, Telethon Foundation).
Selected publications Chiesa R, Piccardo P, Ghetti B, Harris DA Neurological illness in transgenic mice expressing a prion protein with an insertional mutation. Neuron. 21:1339-51 (1998) Fioriti L, Dossena S, Stewart LR, Stewart RS, Harris DA, Forloni G, Chiesa R. Cytosolic prion protein (PrP) is not toxic in N2a cells and primary neurons expressing pathogenic PrP mutations. J Biol Chem. 280:11320-8 (2005) Biasini E, Massignan T, Fioriti L, Rossi V, Dossena S, Salmona M, Forloni G, Bonetto V, Chiesa R Analysis of the cerebellar proteome in a transgenic mouse model of inherited prion disease reveals preclinical alteration of calcineurin activity. Proteomics. 6:2823-34 (2006) Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron. 2008, 60:598-609 (2008). Biasini E., Tapella L., Mantovani S., Stravalaci M., Gobbi M., Harris D.A. and Chiesa R. (2009) Immunopurification of pathological prion protein aggregates. PloS ONE, 4(11): e7816 Massignan T. Stewart R.S., Biasini E. Solomon I.H., Bonetto V., Chiesa R. and Harris D.A. (2010) A novel, drug-based, cellular assay for the activity of neurotoxic mutants of the prion protein. J. Biol. Chem. 285: 7752-7765 Balducci C., Beeg M., Stravalaci M., Bastone A., Sclip A., Biasini E., Tapella L., Colombo L., Manzoni C., Borsello T., Chiesa R., Gobbi M., Salmona M., Forloni G. (2010) Ab oligomers impair memory independently of cellular prion protein. Proc. Natl. Acad. Sci. 107: 2295-2300 Massignan T., Biasini E., Lauranzano E., Veglianese P., Pignataro M., Fioriti L., Harris D.A., Salmona M., Chiesa R., and Bonetto V. (2010) Mutant prion protein expression is associated with an alteration of the Rab GDP dissociation inhibitor alpha (GDI)/Rab11 pathway. Mol Cell Proteomics 9: 611-22 Biasini E., Tapella L., Restelli E., Pozzoli M., Massignan T., and Chiesa R. (2010) The hydrophobic core region governs mutant prion protein aggregation and intracellular retention. Biochemical Journal 430: 477-86 Restelli E., Fioriti L., Mantovani S., Airaghi S., Forloni G., and Chiesa R. (2010) Cell type-spcific neuroprotective activity of untranslocated prion protein. PloS ONE, 5(10): e13725

Maria Grazia De Simoni, Doctoral Degree in Biological Sciences in 1977, University of Milano, Italy, 110/110 summa cum laude. 1981: PhD in Neuropharmacology. 1981-1982: European Community fellowship for "Advanced Professional Training", INSERM U 171, Universit Claude Bernard, Lyon, France. Presently: Head of the Laboratory of Inflammation and Nervous System Diseases, Mario Negri Institute Milan, Italy. She developed a long lasting expertise in experimental models of CNS diseases. The main scientific interests presently include the pathogenesis of cerebral ischemia/reperfusion and of traumatic brain injury; the inflammatory response as target of therapeutic strategies; peripheral inflammatory markers in CNS diseases; the protective mechanisms of stem cells. Member of the Scientific Board of Associazione Italiana per la Ricerca sullInvecchiamento Cerebrale (AIRIC) and of the Editorial Board of Stroke, a Journal of Cerebral Circulation. Author of more than 100 scientific papers on peer-reviewed international journals.
Selected publications - Gesuete R, Orsini F, Zanier ER, Deli MA, Albani D, Bazzoni G, De Simoni MG. Glial cells drive preconditioning-induced blood-brain-barrier protection. Stroke, 2011 (in press). - Zanier ER, Brandi G, Peri G, Longhi L, Zoerle T, Tettamanti M, Garlanda C, Sicurt A, Valaperta S, Mantovani A, De Simoni MG, Stocchetti N. Cerebrospinal fluid pentraxin-3 early after subarachnoid hemorrhage is associated with vasospasm. Intensive Care Medicine 2011, in press. - Ortolano F, Maffia P, Dever G, Rodolico G, Millington OR, De Simoni MG, Brewer JM, Bushell T, Garside P, Carswell HV. Advances in imaging of new targets for pharmacological intervention in stroke: real-time tracking of T-cells in the ischemic brain. Br J Pharmacol 2010; 159: 808-811. - Ortolano F, Colombo A, Zanier ER, Sclip A, Longhi L, Perego C, Stocchetti N, Borsello T, De Simoni MG. c-Jun Nterminal kinase pathway activation in human and experimental cerebral contusion. J Neuropathol Exp Neurol 2009; 68: 964-971. - Gesuete R, Storini C, Fantin A, Stravalaci M, Zanier ER, Orsini F, Vietsch H, Mannesse MLM, Ziere B, Gobbi M, De Simoni MG. Recombinant C1-inhibitor in Brain Ischemic Injury. Annals of Neurology 2009; 66: 332-342. - Longhi L, Perego C, Ortolano F, Zanier E R, Bianchi P, Stocchetti N, McIntosh T and De Simoni MG .C1-Inhibitor attenuates neurobehavioral deficits and reduces contusion volume following controlled cortical impact brain injury in mice. Critical Care Medicine 2009; 37: 659-665. - Capone C, Frigerio S, Fumagalli S, Gelati M, Principato M C, Storini C, Montinaro M, Kraftsik R, De Curtis M, Parati E, De Simoni MG. Neurosphere - derived cells exert a neuroprotective action by changing the ischemic microenvironment. PLoS ONE 2007; 2(4): e373.

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- Balosso S, Ravizza T, Perego C, Peschon J, Campbell IL, De Simoni MG, Vezzani A. Tumor necrosis factor- inhibits seizures in mice via p75 receptors. Annals Neurol 2005; 57: 804-812. - Bergamaschini L, Rossi E, Storini C, Pizzimenti S, Distaso M, Perego C, De Luigi A, Vergani C, De Simoni MG. Peripheral treatment with enoxaparin, a low-molecular weight heparin, reduces plaques and -amyloid accumulation in a mouse model of Alzheimers disease. J. Neurosci 2004; 24: 4181-4186.

Roberto W. Invernizzi started his career in the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri in 1976, where, at present, he heads the Laboratory of Neurochemistry and Behavior. In 1986 he got his degree in Biological Sciences at the Universit Statale di Milano and in 1996 he was nominated head of the Intracerebral Microdialysis Unit. Of particular interest to Invernizzis research team is the study of the neurochemical mechanisms and neuronal circuitries involved in the pathology of the main psychiatric diseases, such as depression and schizophrenia and in the mechanism of action of psychotropic drugs. Since 1987 he applied the intracerebral microdialysis technique to study the in vivo release of monoamines. Using this technique, Invernizzis team first contributed to clarifying the role of serotonergic and adrenergic autoreceptors in the effect of antidepressant drugs suggesting new hypotheses on their mechanism of action. Currently, Invernizzis laboratory is involved in two main collaborative projects aimed at clarifying the neurochemical mechanisms involved in the resistance to antidepressant drugs and the role of glutamatergic and serotonergic mechanisms in attentional processes. Reviewer for various international journals in the field of pharmacology and neurochemistry. Author and co-author of more than 60 peer-reviewed articles. Member of the Italian Society of Neuroscience and the Italian Society of Pharmacology.
Selected publications Calcagno E, Invernizzi RW Strain-dependent serotonin neuron feedback control: role of serotonin receptors. J Neurochem 2010; 114: 1701-1710 Calcagno E, Guzzetti S, Canetta A, Fracasso C, Caccia S, Cervo L, Invernizzi RW. Enhancement of cortical extracellular 5-HT by 5HT1A and 5-HT2C receptor blockade restores the antidepressant-like effect of citalopram in non-responder mice. Int J Neuropsychopharmacol 2009 12: 793-803 Calcagno E, Carli M, Baviera M, Invernizzi RW. Endogenous serotonin and serotonin2C receptors are involved in the ability of M100907 to suppress cortical glutamate release induced by NMDA receptor blockade. J Neurochem 2009 108 : 521-532 Balosso S, Ravizza T, Pierucci M, Calcagno E, Invernizzi RW, Di Giovanni G, Esposito E, Vezzani A. Molecular and functional interactions between tumor necrosis factor-alpha receptors and the glutamatergic system in the mouse hippocampus: Implications for seizure susceptibility. Neuroscience 2009 161 : 293-300 Baviera M, Invernizzi RW, Carli M. Haloperidol and clozapine have dissociable effects in a model of attentional performance deficits induced by blockade of NMDA receptors in the mPFC. Psychopharmacology 2008; 196: 269-280. Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi RW Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: Role of serotonin. Eur. J. Pharmacol. 2008; 594: 117-124 Calcagno E, Canetta A, Guzzetti S, Cervo L, Invernizzi RW. Strain differences in basal and post-citalopram extracellular 5-HT in the mouse medial prefrontal cortex and dorsal hippocampus: relation with tryptophan with tryptophan hydroxylase-2 activity. J Neurochem 2007; 103 : 1111-1120 Invernizzi RW, Pierucci M, Calcagno E, Di Giovanni G, Di Matteo V, Benigno A, Esposito E. Selective activation of 5HT2C receptors stimulates GABA-ergic function in the rat substantia nigra pars reticulata: a combined in vivo electrophysiological and neurochemical study. Neuroscience 2007 144 : 1523-1535

Ugo Lucca got his Master of Science, University of Aberdeen - UK, 1999. At the Mario Negri Institute he was investigator from 1986- 1995, head of the "Clinical Evaluation of Antidementia Drugs Unit" (1995-1996) and, since 1996, head of the "Laboratory of Geriatric Neuropsychiatry". The main areas of interests include epidemiology and clinic features of dementia; natural history of dementia; neuropsychiatric disorders of the elderly; instruments for the screening diagnosis and clinical course assessment of dementia; clinical evaluation of anti dementia treatments and CNS active drugs (phase I, II, III, IV and observational studies).
Selected publications Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-L-carnitine treatment in Alzheimer's disease. Neurology 1991; 41:1726-1732 Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimers disease: a prospective study. J Am Geriats Society 1993; 41: 45-49. Lucca U, Tettamanti M, Forloni G, Spagnoli A. Nonsteroidal anti-inflammatory drug use in Alzheimers disease. Biological Psychiatry 1994; 36: 854-856. Imbimbo BP, Martelli P, Troetel WM, Lucchelli F, Lucca U, Thal LJ, and the Eptastigmine Study Group. Efficacy and safety of eptastigmine for the treatment of patients with Alzheimers disease. Neurology 1999; 52: 700-708.

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Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B12 in mild cognitive impairment, Alzheimers disease and Vascular Dementia. Am J Clinical Nutr 2004; 80: 114-122. Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. New England Journal of Medicine 2006; 355: 1390. Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E.Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: the "Health and Anemia" study. PLoS ONE. 3(4):e1920 (2008)

Alessandro Nobili got his degree in Medicine (Milan, 1990). Master in Biotechonological Research, Regione Lombardia, Milan 1988. International School of Pharmacology, 31 Course on: Drug Epidemiology and Post-marketing Surveillance, Erice, September 1990. Course on: Methods in Epidemiological Research, Milan, October 1990. Course: Long Term Clinical Trials, Cogne January 1991. Main areas of interest Methodology of Randomized Clinical Trials; Pharmacoepidemiology and postmarketing surveillance research; Drug utilization studies; Quality assessment of geriatric services; Qualitative studies on caregiver role in the care of patients with dementia; Methodological evaluation of the Special Care Unit for Alzheimer Disease patients; Methodology of drug information. Employment and research experience Chief of the Unit of Quality Assessment of Geriatric Services Chief of the Drug Information Services for the Elderly, Laboratory of Geriatric Neuropsychiatry, Istituto di Ricerche Farmacologiche Mario Negri, Milan. Editorial Board of the MICROMEDEX Inc., Englewood, Colorado 80111-4740 USA. National Expert accredited by Italian Ministry of Health for The Italian (AIFA) and European Agency for the Evaluation of Medicinal Products (EMEA). Head of the Laboratory of the Quality Assessment of Geriatric Services at the Mario Negri Institute since 2007.
Selected publications Nobili A, Riva E, Tettamanti M, et al. The effect of a structured intervention on cergivers of patients with dementia and problem behaviour: a randomized controlled pilot study. Alzheimer Dis Assoc Disord 2004; 18: 75-82. Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M. Alzheimer special care units compared with traditional nursing home for dementia care: are there differences at admission and in clinical outcomes? Alzheimer Dis Assoc Disord. 2008; ;22:352-61. Nobili, L. Pasina, M. Tettamanti, U. Lucca, E. Riva, I. Marzona, L. Monesi, R. Cucchiani, A. Bortolotti, I. Fortino, L. Merlino, G. Walter Locatelli, G. Giuliani. Potentially severe drug interactions in elderly outpatients: results of an observational study of an administrative prescription database. Journal of Clinical Pharmacology and Therapeutics 2009; 34: 377-386. Alessandro Nobili, Luca Pasina, Silvia Trevisan, Emma Riva, Ugo Lucca, Mauro Tettamanti, Marina Matucci, Massimo Tarantola. Use and misuse of antipsychotic drugs in patients with dementia in Alzheimer special care units. Int Clin Psychopharmacol 2009; 24:97-104. Marengoni A, Bonometti F, Nobili A, Tettamanti M, Salerno F, Corrao S, Iorio A, Marcucci M, Mannucci PM; Italian Society of Internal Medicine (SIMI) Investigators. In-hospital death and adverse clinical events in elderly patients according to disease clustering: the REPOSI study. Rejuvenation Res. 2010;13:469-77. Marcucci M, Iorio A, Nobili A, Tettamanti M, Pasina L, Marengoni A, Salerno F, Corrao S, Mannucci PM; REPOSI Investigators. Factors affecting adherence to guidelines for antithrombotic therapy in elderly patients with atrial fibrillation admitted to internal medicine wards. Eur J Intern Med. 2010; 21:516-23.

Annamaria Vezzani got her Degree in Biological Science at the University of Milan in 1978 and she specialized in Neuropharmacology at the Mario Negri Institute in 1982. She spent her post-doctoral period in Baltimore at the University of Maryland in 1983-1984 working on the mechanisms of epileptogenesis in experimental models of epilepsy. She spent additional post-doctoral periods at the University of Stockholm and at the Karolinska Institute between 1985 and 1999. She was on sabbatical at the Albert Einstein College of Medicine in 2002 in the laboratory of Developmental Epilepsy. She is involved in studies on the biochemical and molecular mechanisms involved in the etiopathogenesis of seizures disorders using experimental models of epilepsy. The present research is focused on the functional role of neuroactive peptides and inflammatory mediators in the modulation of neuronal excitability and seizure-related neurodegeneration. Focus of the research is also on the mechanisms of pharmacoresistance. Since 1997 she is the Head of the Laboratory of Experimental Neurology at the Mario Negri Institute. She is member of the Editorial Board of Epilepsy Currents and Neuroscience and Associate Editor for Exp Models of Epilepsia. She is appointed of the Chair of the Commission on Neurobiology of International League Against Epilepsy which is promoting initiatives for improving translational research in epilepsy.

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Selected publications Vezzani A, Conti M, De Luigi A, Ravizza T, Moneta D, Marchesi F, De Simoni MG. Interleukin-1beta immunoreactivity and microglia are enhanced in the rat hippocampus by focal kainate application: functional evidence for enhancement of electrographic seizures.(1999) J Neurosci.19:5054-65. Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S., Lundkvist J., Iverfeldt K. and Bartfai T. Powerful anticonvulsant action of IL-1 receptor antagonist upon intracerebral injection and astrocytic overexpression in mice (2000) Proc Natl Acad Sci USA, 97: 11534 Rizzi M, Caccia S, Guiso G, Richichi C, Gorter JA, Aronica E, Aliprandi M, Bagnati R, Fanelli R, D'Incalci M, Samanin R, Vezzani A. Limbic seizures induce P-glycoprotein in rodent brain: functional implications for pharmacoresistance (2002) J Neurosci, 22: 5833 Balosso S, Ravizza T, Perego C, Peschon J, Campbell I, De Simoni MG, Vezzani A. TNF-alpha inhibits kainic acidinduced seizures in mice via p75 receptors (2005) Ann Neurol, 57: 804-12 Ravizza T, Gagliardi B, No F, Boer K, Aronica E and Vezzani A. Innate and adaptive immunity during epiletogenesis and spontaneous seizures: evidence from experimental models and human temporal lobe epilepsy (2008) Neurobiol Dis, 29: 142 No F, Pool AH, Nissinen J, Gobbi M, Bland R, Rizzi M, Balducci C, Ferraguti F, Sperk G, During MJ, Pitknen A, Vezzani A. Neuropeptide Y gene therapy decreases chronic spontaneous seizures in a rat model of temporal lobe epilepsy (2008) Brain, 131:1506 Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional pathway mediates the proconvulsive effects of interleukin-1beta. (2008) Brain, 131:3256 Maroso M, Balosso S, Ravizza T, Liu J, Aronica E, Iyer A, Rossetti C, Molteni M, Casalgrandi M, Manfredi AA, Bianchi ME and Vezzani A. Toll-Like Receptor 4 (TLR4) and High Mobility Group Box 1 (HMGB1)are involved in ictogenesis and can be targeted to reduce seizures (2010) Nature Medicine, 16:413-9.

Mirjana Carli started his scientific career in the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri Milan in 1977, where, at present, she is head of the Pharmacology of Cognitive Behaviour Unit. She spent a few years in the laboratory of Cognitive Neuroscience, Dept. of Experimental Psychology, University of Cambridge (UK) directed by Prof. Trevor W. Robbins. Here she took interest in the role of brain monoamines in attention, and for this purpose developed several behavioral tests for rats. In 1986 she returned to the laboratory of Neuropharmacology of the Istituto di Ricerche Farmacologiche Mario Negri. Here she devoted her efforts to the study of the role played by neuronal mechanisms in cognitive processes such as memory, attention and executive functions. Her work has improved the knowledge of the role played by some serotonin receptors in cognitive processes.
Selected publications Carli M, Baviera M, Invernizzi R, Balducci C Dissociable contribution of 5-HT1A and 5-HT2A receptors in the medial prefrontal cortex to different aspects of executive control such as impulsivity and compulsive perseveration in rat Neuropsychopharmacology 2006; 31: 757-767 Greco B, Carli M Reduced attention and increased impulsivity in mice lacking NPY Y2 receptors: Relation to anxiolyticlike phenotype Behav Brain Res 2006; 169: 325-334 Carli M, Baviera M, Invernizzi R, Balducci C The serotonin 5-HT2A receptors antagonist MI00907 prevents impairment in attentional performance by NMDA receptor blockade in the rat prefrontal cortex Neuropsychopharmacology 2004; 29: 1637-1647 Balducci C, Nurra M, Pietropoli A, Samanin R, Carli M Reversal of visual attention dysfunction after AMPA lesions of the nucleus basalis magnocellularis (NBM) by the cholinesterase inhibitor donepezil and by a 5-HT(1A) receptor antagonist WAY 100635 Psychopharmacology (Berl) 2003; 167: 28-36 Carli M, Balducci C, Samanin R. Stimulation of 5-HT1A receptors in the dorsal raphe ameliorates the impairment of spatial learning caused by intrahippocampal 7-chloro-kynurenic acid in naive and pretrained rats Psychopharmacology (Berl) 2000; 158: 39-47 Carli M, Samanin R The 5-HT1A receptor agonist 8-OH-DPAT reduces rats' accuracy of attentional performance and enhances impulsive responding in a five-choice serial reaction time task: Role of presynaptic 5-HT1A receptors Psychopharmacology (Berl) 2000; 149: 259-268

Barbara DAvanzo obtained her master in Philosophy in 1989 at the University of Milan. Her main field of interest is epidemiologic research in mental health. She was involved in the analysis of the implementation of the psychiatric reform in Italy and quality evaluation of services and their recent modifications with specific attention to the role of psychiatric residential facilities in the community service networks; evaluation of effectiveness of the most common psychosocial interventions; suicide trend monitoring and study of suicide prevention programs and initiatives. More recently, she is working on issues like recovery-oriented services, consumers empowerment, and methods of participation of

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consumers to evaluation of services, and acknowledgment of the value of the consumers point of view about psychiatric treatments and services. She worked as researcher in the Laboratory of General Epidemiology between 1991 and 1996, and she is Chief of the Unit of Epidemiology and Social Psychiatry since 2002. Member of the Scientific National Board of WAPR Italy and of the World Head Office of the WAPR.
Selected publications Barbato A, Parabiaghi A, Panicali F, Battino N, D'Avanzo B, De Girolamo G, Rucci P, Santone G, PROGRES-Acute Group. Do patients improve after short psychiatric admission? A cohort study in Italy Nord J Psychiatry 2010; E-pub Campi R, Barbato A, D'Avanzo B, Guaiana G, Bonati M Suicide in Italian children and adolescents J Affect Disord 2009; 113:291-295 Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q 2008; 79:121-132 D'Avanzo B, Aliprandini E, Beghi M, Cornaggia C M, Erlicher A, Frova M, Mascarini A, Miragoli P, Righi A. Strutture residenziali e semiresidenziali nei servizi di salute mentale. Dove sta la differenza? Epidemiologia e Psichiatria Sociale 2008; 17:57-64 Barbato A, D'Avanzo B. Marital therapy for depression. Cochrane Database Systematic Reviews 2006; Issue 2. Parabiaghi A, Barbato A, D'Avanzo B, Erlicher A, Lora A. Assessing reliable and clinically significant change on Health of the Nation Outcome Scales: method for displaying longitudinal data. Aust N Z J Psychiatry 2005; 39: 719-725. Barbato A, D'Avanzo B. Involuntary placement in Italy. Br J Psychiatry 2005; 186: 542-543. Guaiana G, Andretta M, Corbari L, Mirandola M, Sorio A, D'Avanzo B, Barbui C. Antidepressant drug consumption and public health indicators in Italy, 1955-2000. J Clinical Psychiatry 2005; 66: 750-755. D'Avanzo B, Battino R N, Gallus S, Barbato A. Factors predicting discharge of patients from community residential facilities: A longitudinal study from Italy. Aust N Z J Psychiatry 2004; 38: 619-628. D'Avanzo B, Barbato A, Barbui C, Battino N, Civenti G, Frattura L. Discharges of patients from public psychiatric hospitals in Italy between 1994 and 2000. Int J Social Psychiatry 2003; 49: 27-3

Emma Riva, Medical Doctor degree in 1984 University of Milan, PhD in 1990 in Cardiovascular Pathophysiology at the University of London (UK) Training: Research Assistant, Department of Pharmacology, Medical School, University of Ottawa, Canada; Internship in Internal Medicine, Ospedale Luigi Sacco, Milan; Cardiac Fellow, St Thomas' Hospital, London, UK. Field of interest: Prevalence and effects of anemia on cognitive, functional and clinical variables in the elderly; Problem behaviors in dementia; Burden for care-givers of Alzheimer Disease patients; End of life care. Present and past roles in Institute Head of the Geriatric Pharmacology Unit, Istituto "Mario Negri", Milan; Scientific Director of the hospice Via di Natale Franco Gallini, Aviano, Italy; Consultant Istituto Geriatrico Pio Albergo Trivulzio, Milan: Project member of PREDICT (Policy Review and Evaluation of Dementia and Institutional Care Trends): a Transnational Comparison.
Selected publications Riva E, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Lucca U. Association of mild anemia with hospitalization and mortality in the Elderly: The Health and Anaemia Population-based Study. Haematologica 2009; 94:22-28 Nobil i A, Franchi C, Pasina L, Tettamanti M, Baviera M, Monesi L, Roncaglioni C, Riva E, Lucca U, Bortolotti A, Fortino, I Merlino L. Drug utilization and polypharmacy in an Italian elderly population: the EPIFARM-Elderly Project. Pharmacoepidemiology and Drug Safety, 2011 DOI: 10.1002/pds.2108 Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta MG, Riva E. Prevalence, incidence and types of mild anemia in the elderly: the "Health and Anemia" population-based study. Haematologica. 2010;95(11):1849-1956 Pasina L, Nobili A, M. Tettamanti M, Riva E, Lucca U, Piccinelli R, Defendi L, Perego L, Lucifora S, Bulla C. Coprescription of gastroprotective agents in patients taking non-selective NSAIDs or COX-2 selective inhibitors: analysis of prescriptions. Int J Clini Pharmacol and Ther 2010;48:735-743 Avanzini F, Marelli G, Donzelli W, Sorbara L, Palazzo E, Bellato L, Colombo EL, Roncaglioni MC, Riva E, De Martini. Hyperglycemia during acute coronary syndrome: a nurse-managed insulin infusion protocol for stricter and safer control. Eur J Cardiovasc Nursig 2009;8:182-189 Nobili A, Pasina L, Tettamanti M, Lucca U, Riva E, Marzoni I, Monesi L, Cucchiani R, Bortolotti A, Fortino I, Merlino L, Locatelli W. Giuliani G. Potentially severe drug interactions in elderly outpatients: results of an observational study of an administrative prescription database. J Clin Pharm Ther 2009;34:1-10 Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E. Association of Mild Anemia with Cognitive, Functional, Mood and Quality of Life Outcomes in the Elderly: The Health and Anemia Study. Plos ONE 2008;3(4):e1920 Tettamanti M, Garr MT, Nobili A, Riva E, Lucca U. Low folate and risk of cognitive and functional deficits in the very old: The Monzino 80-plus study. J Am Coll Nutr 2006;25:502-508

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Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol 2006;63:154-155

Mauro Tettamanti got his Biology Degree at the Universit degli Studi di Milano in 1986, and the specialisation in Epidemiology and Medical Statistics in 1993, at the Universit degli Studi di Pavia. Teaching experience Introduction course to statistics, Master in Ergonomy, Politecnico di Milano, years 2001-2004 Areas of interest: Planning, conduction and analysis of clinical trials and epidemiologic researches in the geriatric field: Phase I, II, III and observational studies on the efficacy of drugs on neurologic disorders, with special emphasis on dementia; Effects of multi-disciplinary interventions on geriatric/dementia patients; Epidemiology and risk factors of dementia; Care of patients with terminal illness; Association of anemia with prevalence of diseases and cognitive problems Scholarship between 1989 and 1998, Senior Researcher since 1999 and Head of the Unit of Geriatric Epidemiology at the Mario Negri Institute since 2001.
Selected publications

Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-L-carnitine treatment in Alzheimer's disease. Neurology 1991; 41:1726-1732. Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimer's disease: A prospective study. J Am Geriatr Soc 1993; 41:45-49 Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B12 in mild cognitive impairment, Alzheimer disease, and vascular dementia. Am J Clin Nutr 2004; 80: 114-122 Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol 2006; 63:154-155 Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. N Engl J Med 2006; 355:1390 Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U. Low folate and the risk of cognitive and functional deficits in the very old: The Monzino 80-plus study. J Am Coll Nutr 2006; 25: 502-508 Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M.Alzheimer special care units compared with traditional nursing home for dementia care: are there differences at admission and in clinical outcomes? Alzheimer Dis Assoc Disord. 22:352-6 (2008).

ACTIVITIES
The Department of Neuroscience is formed by ten Laboratories; the activities of research are devoted to the study of neurological and psychiatric diseases, evaluated by the biological point of view, clinical and epidemiological aspects and the quality of care. Together with these activities, in the Department other more general expertise are present. Drug information service and preparation of protocols for clinical trial and epidemiological studies are activities in charge of the Neuroscience Department. Traditionally part of the Department was devoted to the creation of experimental models for the pharmacological, neurochemical and pathogenetic studies in Alzheimer or prion's diseases, epilepsy, depression and cognitive impairment. More recently, consolidated expertise were created in the pathogenesis of amyotrophic lateral sclerosis (ALS), cerebral stroke and drug abuse. Some of these disorders, like epilepsy, ALS and Alzheimer's disease are investigated from the clinical and epidemiological points of view for the evaluation of drug and care efficacy. The activities of the Department are aimed to an integration of the different expertise to develop multidisciplinary approaches. The purpose is to address at different levels, knowledge, therapy and clinical practice to the numerous questions, largely unresolved, proposed by the disorders of nervoussystem.

MAIN FINDINGS
The intracerebral application of synthetic amyloid 1-40 e 1-42 in oligomeric form is associated with a cognitive damage that does not occur when the pepetides are applied in

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monomeric form of fibrils species. In contrast with other evidence, the effect of oligomers is reversible and idependent from the presence of cellular prion protein. Analysis by Magntic Resonance Image (MRI) of Alzheimers disease (AD) experimental models showed structural alterations aging associated reminiscent of the brain structural changes reported in AD patients. The exposure of cultured hippocampal cells to amyloid 1-42 in oligomeric form induces an alteration of dendritic spines. This effect was evident also when the oligomers did not induce cell death. A chronic treatment with doxycycline, a tetracycline that pass the blood brain barrier, with antiamyloidogenic activity, reduces the aggregates of amyloid and antagonize the dysfunction indeuced by oligomers By performing the first in vivo chronic-treatment with the specific cell-penetrating JNK inhibitor peptide D-JNKI-1 we demonstrated that JNK is regulating the main pathogenic mechanisms of AD and might hold promise as an innovative and therapeutic target against it. We obtained a complete rescue in AD mouse model of both long-term potentiation and longterm recognition memory impairments in D-JKNI1 treated mice. The functional recovery shows a strong relationship with the JNK signaling pathway and reduction of Ab oligomers, derived by APP cleavage. The overexpression of SUMO-1 neuroblastoma cells lines reduces phosphorylation of JNKs and c-Jun in H2O2 stress conditions and decreases JNKs expression level. SUMO-1 has a possible dual role in modulating JNKs stress response: contrasting JNKs phosphorylation results in inhibition of cell death and controlling JNKs degradation. Design and synthesis of the new cell permeable inhibitor peptides MKK7 inhibitor: The in vitro inhibitory tests showed no toxicity of these peptides and a high neuroprotective effect against excitotoxicity The resveratrol, a well-known antioxidant induces its neuroprotective effect through the activation of SIRT.1 In the transgenic mice overexpressing a mutated form of human prion protein associated with CJD generated in the Institute has been found a significant alteration of endoplasmic reticulum associated with the presence of mutated prion protein. It has been developed a cellular assay to evaluate the neurotoxicity of mutated prion proteins in immoratilized cell lines. The assay can be used to understand the molecular basis of PrP neurotoxicity and to evaluate potential therapeutic approaches. Proteomic analysis in cellular model of fatal familial insomnia it has been show that alterations of intracellular protein transport might play a role in the pathogenesis of grnetic prions diseases Polymorphisms in gene encoded for serotonin transporter protein influenced the risk to develop Parkinsons disease

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In a prospective population-based study in the very old (Monzino 80-plus Study), the the risk associated with a family history of deementia was significantly high also among the oldes old, but, at variance with the prevalence of dementia, it decreases with age and is not significant after 90. The same prospective population-based study (Monzino 80-plus Study) failed to demonstrate a significant association between diabetes or glucose concentration and dementia in the very old. Though increasing with age, apathy is pervasive among dementia sufferers and its prevalence and severity increases along the disease course. Together with depression and possibly alone, apathy could be a prodromic manifestation of dementia ( Monzino 80-plus Study). In a prospective ambulatory population of cognitively normal or mildly cognitively impaired elderly, dissecting mild cognitive impairment into severity levels was found to increase the accuracy of progression predictions to dementia. More than one out of 10 elderly persons (65 years or older) were anemic and most of the cases had a mild grade anemia. Hemoglobin concentrations decreased and prevalence of (mild) anemia increased with increasing age. After controlling for many potential confounders, mild anemia was found to be associated with poorer health conditions and with increased risk of clinically relevant outcome such as hospitalization and mortality (Health and Anemia Study). In Trentino, where the Form for Hospital Assessment of Self-Harm and Suicide Attempts has been in use since June 2009, was found a rate of 37/100,000 cases of self-harm and suicide attempts seen in the Emergency Departments of the province, with women showing higher rates. The quality assessment of the network of mental health services indicated that a few areas needed to be improved: pharmacotherapy, self-help, facilities of the psychiatric ward, information on illness and the drugs, possibility to choose the profesional to be cared by, wait time for the appointments and at the phone. According to the GiSAS survey questionnaire, the majority of psychiatrists (63%) trusted in the superiority of second-generation antipsychotics. Those who indicated pharmaceutical representatives as their main source of knowledge of antipsychotics (n=103) all preferred the other SGAs over clozapine (OR 2.1, CI 1.13.9; p=0.01). The influence of industry-mediated information might have affected opinions on SGAs and the lack of uncertainty about antipsychotics attitudes may have hindered trial participation. In years 2000-2006 the prescriptions of fluoxetine, paroxetine and mirtazapine increased. On the contrary, the use of reboxetine progressively declined and was associated with higher discontinuation rates. These findings are consistent with recent experimental evidence. Both treated incidence and prevalence of mental disorders in Lombardy increased between 1999 and 2009. Incidence of schizophrenic and personality disorders decreased and that of affective and neurotic disorders increased, while increasing prevalence concerned all diagnostic groups. The majority of patients, even those with schizophrenia, were cared at outpatient level through clinical and community packages.

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Patients with dementia resident in Alzheimers special care units (ASCU) had a lower rate of hospitalisation and use of physical restraints than those in traditional nursing homes. In ASCU 60% of patients with dementia were taking at least one antipsychotic, 49% typical and 51% atypical. More than 50% of patients exposed to antipsychotics at baseline, were still taking the drug after 18 months of follow-up. The use of antipsychotic agents was strongly related to the presence of agitation, irritability, delusions, anxiety, night-time behaviour and aberrant motor behaviour. In the Lecco Local Health Authority 16% of elderly patients were exposed to potential severe drug-drug interactions; age and number of chronic drugs were associated with an increasing risk of DDIs. Since physicians still have some difficulty in managing this topic, it is essential to provide them with adequate information on which factors raise the risk of DDIs. Age, local health unit (LHU) of residence, number of drugs and co-prescribed PIDs were predictors of hospitalization for hemorrhage. During 2005 in Lombardy Region, 76% of the elderly aged 65 years ore more (76% women and 75% men) received at least one chronic drug, 46% were exposed to polypharmacy (46% women and 45% men) and 20% to chronic polypharmacy (18% women and 22% men). Elderly in the age groups of 75-79, 80-84 and 85-89 years had the highest risk to be exposed to chronic polypharmacy (OR 2.25; 95%CI: 2.23-2.27, OR 2.68; 95%CI: 2.65-2.71, and OR 2.84; 95%CI: 2.79-2.89 respectively). During 2005, 34 % of the population living in Lombardy Region received at least one antibiotic drug prescription. The highest prescription prevalence was observed in the 0-17 and 80 or more year age ranges (41.6% and 41.9%, respectively). Patients aged <18 years (OR= 1.73; 95% CI 1.73, 1.74), aged 65 or older (OR= 1.64; 95% CI 1.63, 1.65), and those that live in Brescia (OR 1.66, 95% CI 1.65, 1.66) had a statistically significant higher risk of antibiotic drug exposure. In a large population sample of subject living in Lomabrdy Region, the use of paroxetine and fluoxetine peaked in 2002 and then decreased. The prescripition rates of mirtazapine gradually increased all through the study period: from 0.07% in 2000 to 0.13% in 2006. On the contrary, the prescription rates of reboxetine showed a different trend and progressively decreased from 0.20 in 2000 to 0.04 in 2006. In a sample of 38 internal medicine and geriatric wards, at hospital admission 52% of 1332 elderly patients aged 65 years or older taken five or more different drugs (polypharmacy) and were in the ward for a mean of 11 days. At hospital discharge there was an increase in the rate of patient with polypharamacy (+13%) and with multiple disease (+16%). Among elderly patients admitted with a diagnosis of AFF to internal medicine wards, an appropriate antithrombotic prophylaxis was taken by less than 50%, with an underuse of VKAs prescription independently of the level of cardio-embolic risk. Hospitalization did not improve the adherence to guidelines. After multiadjustment, the diagnosis of dementia was associated with in-hospital death (OR = 2.1; 95% CI = 1.0 - 4.5). Having dementia and at least one adverse clinical event during hospitalization showed an additive effect on in-hospital mortality (OR = 20.7 ;95% CI = 6.9 61.9).

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There is a direct correlation between ALS and mechanical trauma as a result of the following observations: The risk of ALS increases with the number of traumatic events and the severity of injuries. There is an inverse correlation between ALS and coffee intake. The treatment of the first unprovoked seizure does not affect short and long-term mortality in patients with epilepsy. The prevalence of some neurological disorders in Albania differs from that of other European countries. Differences can be explained by the study methodology and by the diverse distribution of genetic and environmental risk factors. The incidence of acute symptomatic seizures in patients with a firstly diagnosed stroke followed prospectively is fairly low. Cerebral hemorrhage and cortical lesions are the only independent predictors of acute symptomatic seizures while hypercholesterolemia reduces the risk of hemorrhagic stroke Patients with epilepsy-headache comorbidity differ from patients with epilepsy or headache alone in terms of family history and severity of the clinical picture. About one-fourth of patients with epilepsy starting or changing an antiepileptic drug are unsatisfied with treatment. Poor satisfaction with treatment is most frequent in patients with long-lasting disease, adverse drug reactions, and with parents/caregivers declaring poor quality of life. We have demonstrated the crucial involvement of some pro- and anti-inflammatory cytokines in seizures using experimental models of epilepsy in rodents, thus describing a new etiopathological mechanism which may be relevant for human epilepsy. We have demonstrated that membrane-bound drug transport proteins are functionally activated by seizures and have a significant role in decreasing the brain concentrations of antiepileptic drugs in experimental models. Pharmacological intervention to block the activity of these proteins may contribute to reverse multidrug resistance in epilepsy. Gene therapy studies highlight the possibility to significantly reduced spontaneous seizure that are refractory to anticonvulsant drugs opening the perspective of using gene therapy in pharmacoresistant forms of epilepsy. Mannose-binding lectin (MBL), a key protein in the complement lectin pathway, is a novel target for stroke treatment Microglia can favour protective actions in the ischemic environment Stem cells from umbilical cord blood decrease post-traumatic brain functional deficts and lesion in injuried mice In some motor neurons of mice with mutant SOD1 is evident an early induction of the protein, HspB8, which promotes the transport of the altered proteins to the autophagy system of the cell for being destroyed and thus preventing their accumulation. The increase of HspB8 is also evident in the motor neurons that survive in the terminal stages of disease suggesting a neuroprotective role of this protein. This discovery opens the way for a possible therapeutic strategy, based on the identification and application of substances that

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promote the activation of HspB8 in the entire population of motor neurons in order to protect them from the accumulation of protein aggregates and from death Differences in feedback control of serotonergic transmission influence the efficacy of SSRIs Co-treatment with 5-HT2C receptor antagonists enhance the effects of SSRI on serotonergic transmission and restore their antidepressant-like effect in non-responder mice The blockade of NMDA receptors of the rat prefrontal cortex induces an increase of glutamate release, activates the transcription factor CREB in the dorsal striatum and is deleterious for prefrontal cortex-dependent cognitive functions 5-HT2A receptor antagonists and 5-HT1A and 5-HT2C receptor agonists prevent the increase of glutamate release and attentional deficits caused by NMDA receptors blockade suggesting that some serotonin receptor subtypes might constitute a molecular target for the development of drugs for the treatment of cognitive deficits of schizophrenia We show that in a glutamate NMDA model of cognitive deficit of schizophrenia antipsychotics may be differentiated by a selective effect of typical antipsychotics on compulsive perseveration, and atypical antipsychotics on impulsivity. A single session of cocaine self-administration is sufficient to shape rat behavior towards goaldirected behaviors and selectively up-regulate Arc expression in mPFC. This is the first evidence that the mPFC's function is already profoundly influenced by the first voluntary cocaine exposure. Gamma-hydroxybutyric acid (GHB) does not maintain self-administration but induces conditioned place preference when injected in the ventral tegmental area. Environmental stimuli associated to drug self-administration induce drug seeking behavior when present to rodents after a long period of abstinence. Bifeprunox, a partial agonist at dopamine D2 and serotonin1A receptors, influences nicotineseeking behaviour in response to drug associated stimuli in rats Genetic differences in serotonin synthesis may contribute to the efficacy of SSRIs in a murine model predictive of the antidepressant activity. In non-responder mice, 5-HT1A and 5-HT2C receptor antagonists restore the antidepressant-like to the SSRI.

NATIONAL COLLABORATIONS
Associazione Familiari Insonnia Familiare Fatale malattie da prioni, Treviso Associazione Italiana GIST A.I.G. Associazione per la Ricerca Neurogenetica, Lamezia Terme (CS) e ASL 6, Regione Calabria Agenzia di Sanit Pubblica del Lazio, Regione Lazio Assessorato alla Salute, Comune di Milano

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Azienda Ospedaliera Ospedali Riuniti di Bergamo Azienda Sanitaria Locale di Bergamo Azienda ULS TO2, Torino CEND, Centro Eccellenza per le Malattie Neurodegenerative, Universit di Milano Centro di Terapie per lAdolescenza, Milano Centro Fatebenefratelli San Giovanni di Dio, Cernusco sul Naviglio Centro di Neurofarmacologia, Dipartimento di Scienze Farmacologiche, Universit di Milano Centro Studi in Psichiatra, ASL 2, Torino Centro Parkinson-Istituti Clinici di Perfezionamento Clinica IRCSS S. Maria Nascente, Milano Clinica Neurologica III Universit di Milano, Azienda Ospedaliera S. Paolo, Milano Clinica Psichiatrica, Universit Milano Bicocca Consorzio Ricerche Luigi Amaducci, CRIC, Arcugnano (Vc) Consorzio MIA, Milano DIBIT, San Raffaele Scientific Insitute, Milano. Dipartimento di Chimica Biologica, Universit di Padova Dipartimento di Chimica, Universit egli Studi di Firenze Dipartimento di Chirurgia "P. Valdoni" - Lab., Ricerca Center for Research in Neurobiology "Daniel Bovet" (CRiN), "Sapienza" Universit di Roma Dipartimento Endicronologia, Universit di Milano Dipartimento Farmaco Chimico Tecnologico, Universit di Siena Dipartimento di Farmacologia Medica, Universit di Milano Dipartimento di Fisiologia Umana, Facolt di Medicina, Universit di Milano Dipartimento di Medicina e Sanit Pubblica, Sezione di Psichiatria e Psicologia Clinica, Universit di Verona Dip. di Morfofisiologia, Scuola di medicina Veterinaria, Universit di Torino, Grugliasco (TO). Dip. Neurologia, IRCCS Fondazione Maugeri, Pavia Dipartimento Neurologia, Ospedale Molinette, Torino Dipartimento di Neurologia Universit di Milano, Ospedale Luigi Sacco. Dipartimento di Neuroscienze, Universit di Parma, Parma Dipartimento di Salute Mentale di Niguarda, Milano Dipartimento di Salute Mentale ASL 3 Genovese, Genova Dipartimento di Salute Mentale San Carlo, Milano Dipartimento di Salute Mentale della Ulss 5 Ovest Vicentino Dip. di Scienze Biomolecolari e Biotecnologie, Universit di Milano Dipartimento di Scienze Fisiologiche Universit di Pavia, Pavia Dipartimento Scienze Neurologiche, Universit di Genova, Genova Dipartimento Scienze Neurologiche, Ospedale Maggiore Policlinico di Milano Direzione Generale Famiglia e Solidariet Sociale, Regione Lombardia, Milano Direzione Generale Sanit, Regione Lombardia, Milano Direzione Regionale Sanit e Servizi Sociali, Regione Umbria Divisione di Ematologia, Universit di Pavia e Fondazione IRCCS Policlinico S. Matteo, Pavia Divisione Neurologica, Universit di Bologna EPAPSY, Scientific Association for Regional Development and Mental Health, Athens, Greece Evidentia Medica, Grottaferrata, Roma Federazione Alzheimer Italia, Milano Franco Calori Cell Factory, Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano Fondazione Clelio Angelino Fondazione Cecchini Pace, Milano Fondo Edo Tempia Hospice Franco Gallini, Aviano (PN)

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IRCSS "Casa Sollievo della Sofferenza", San Giovanni Rotondo IRCCS Istituto Auxologico Italiano, Milano Istituto di Ricovero e Cura a Carattere Scientifico IRCCS (I.N.R.C.A.), Ancona IRCSS Fatebenefratelli di Brescia IRCSS "San Raffaele", Milano Istituto Europeo di Oncologia, IRCCS, Milano Istituto di Farmacologia e Farmacognosia, Universit di Urbino Istituto di Farmacologia, Universit di Milano Istituto di Fisiologia Umana II Universit degli Studi di Milano, Milano Istituto G. Ronzoni, Milano Istituto Nazionale Neurologico Carlo Besta, Milano Istituto Scientifico Humanitas Istituto di Scienze e Tecnologie della Cognizione, CNR, Roma Istituto "Stella Maris", IRCCS, Calambrone (PI) Istituto Superiore di Sanit, Roma Istituto Zooprofilattico Piemonte Liguria Val D'Aosta,Torino Laboratorio di Immunopatologia Renale, Ospedale San Carlo, Milano Laboratorio di Neuroscienze, Centro Dino Ferrari, Universit di Milano Lega Italiana per la Lotta contro i Tumori Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genova Ospedale del Bambin Gesu, Roma Ospedale Regionale Ca Fondello, Treviso Ospedale "Molinette", Torino Polo Oncologico, ASL 12, Biella Polo Tecnologico, IRCCS S. Maria Nascente, Fondazione Don Carlo Gnocchi Onlus, Milano Provincia Lombardo-Veneta Ordine Ospedaliero San Giovanni di Dio, Fatebenefratelli di Cernusco sul Naviglio Progetto Itaca, associazione Volontari per la Salute Mentale ONLUS, Milano Scuola di Specializzazione in Psicoterapia IRIS-Insegnamento e Ricerca Individuo e Sistemi, Milano Scuola di Terapia Cognitiva Studi Cognitivi, Milano Societ Italiana Medicina Interna, Roma Unione Nazionale delle Associazioni per la Salute Mentale (UNASAM), Milano Unit di Geriatria, Ospedale Maggiore IRCCS, Universit di Milano Unit Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano Unit Operativa di Psichiatria, Azienda Ospedaliera San Gerardo di Monza, Monza, Unit Operativa di Psichiatria di Garbagnate, Azienda Ospedaliere Salvini di Garbagnate, Garbagnate Milanese Unit Operativa di Psichiatria, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena di Milano, Milano Universit degli Studi di Foggia Universit Cattolica del Sacro Cuore di Roma Universit dellInsubria, Varese Universit del Piemonte Orientale, Novara Universit di Milano, IRCCS Ospedale Maggiore, Milano Universit Milano-Bicocca, Monza Universit La Sapienza, Roma U.O. Neurologia, Clinica S. Maria, IRCCS, Castellanza (VA). UNASAM, Unione Nazionale delle Associazioni per la Salute Mentale Unit Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano Web Medica, Grottaferrata, Roma

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INTERNATIONAL COLLABORATIONS
Albert Einstein College of Medicine, Bronx, NY, USA Atomic Energy Commission, Service de Neurovirologie, Fontenay-aux-Roses, France Beaumont Hospital, Dublin, Ireland Brain Repair Centre, University of Cambridge, Cambridge, UK Cambridge Centre for Brain Repair, University of Cambridge, UK Centre for Neuroscience Research and Division of Biomolecular Sciences, GKT School, Kings College, London, UK Centre National de la Recherche Scientifique, Paris. France Chorley & South Ribble General Hospital, Chorley, Cochrane Schizophrenia Group, Universit di Nottingham, Nottingham, UK Columbia Univ, Haverstraw, NY, USA Department of Anatomy and Physiology, Laval University, Quebec Department of Biochemistry, Boston University, Boston USA Department of Cell Biology, Washington University, St Louis, USA Department of Chemistry, The Australian National University, Canberra City, Australia Department of Experimental Psychology, University of Cambridge, UK Department of Neuroscience, Physiology & Pharmacology University College London, , UK Department of Pathology and Infectious Diseases Royal Veterinary College, Herts, UK Department of Psychiatry, Medical Center University of Mississippi, Jackson, USA Directorate General for the Health and Consumer Protection, European Commission, Luxembourg Division of Medical Genetics, CHUV Lausanne, Switzerland Divisione di Geriatria, Ospedali Regionali di Lugano e Mendrisio, Svizzera EPAPSY, Scientific Association for Regional Development and Mental Health, Athens, Greece European Union of Family Associations of People with Mental Illness (EUFAMI) Geriatric Division and Department of Metabolic Diseases, Ospedali Regionali of Lugano and Mendrisio, Switzerland HSPH Harvard University, Boston, USA IBCM, University of Lausanne, Lausanne, Switzerland INSERM U 751, Marseille, France Institut de Gntique Humaine du CNRS, Montpellier, France Jefferson Med Coll, Philadelphia, USA Karolinska Institutet, Stockholm, Sweden Kings College Hospital, London, UK Lancaster University, Lancaster, UK. Lexicon Pharmaceuticals Texas, USA Max-Delbrck-Center for Molecular Medicine, Berlin, Germany MPRC, Univ Baltimore, Baltimore, MD, USA National Insitute on Aging, NIH, Baltimore, USA Neuroprion, Network of Excellence, WP VI, EC Neurological Department of the University of Tirana, Albania Neurology, GlaxoSmithKline, New Frontiers Science Park North, Harlow UK Ninewells Hospital and Medical School, Dundee, Scotland UK Northern Illinois University, DeKalb, IL, USA Novartis Pharma, Basel, Switzerland NYU, NY, USA Observatoire National Sant mentale et Prcarit, Rgion Rhne-Alpes, Lione, France Ohio State Univ, Columbus, Ohio, USA Robarts Research Institute, London, Ontario, Canada Royal Manchester Children's Hospital, Manchester, UK

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Royal Preston Hospital, Preston, UK Sergievsky Center, Columbia University, New York, NY, USA Servizio di Geriatria, Ospedale della Beata Vergine, Mendrisio, Switzerland The Scripps Research Institute, Jupiter, Florida, USA Technology Park of Bizkaia, Bizkaia, Spain Toxicology Unit MRC, Leicester, UK University of Alberta, Canada University of Bristol, Frenchay Hospital, Frenchay, Bristol, UK. University of Bristol, School of Medical Sciences, UK Univ of California at Irvine, Irvine, CA, USA University of Cardiff, United Kingdom University of Chicago, Chicago, IL , USA. Univ of Colorado, Denver, USA University Hospital, London, ON, Canada Univ of Innsbruck, Innsbruck, Austria University of Lausanne, Lausanne Switzerland Univ of Maryland, Baltimore, USA University of Maastricht, the Netherlands University of Rijeka Medical School, Rijeka, Croatia University of Szeged, Hungary Universit de la Mditerrane -Hpital de la Timone Marseille, France Universit Victor Segalen, Bordeaux, France Unit of Molecular Genetics, CHUV Lausanne, Switzerland Virtanen Institute for Molecular Sciences, University of Kuopio, Finland Vrije Universiteit Medical Center, Amsterdam, The Netherlands Walton Hospital, Liverpool, UK WAPR (World Association for Psychosocial Rehabilitation) Washington University, St Louis, MI,USA Weill Cornell Medical College, New York, USA World Mental Health, Department of Mental Health and Substance Abuse, Geneva, Switzerland World Association for Psychosocial Rehabilitation World Health Organization, Disability and Rehabilitation Team

EDITORIAL BOARD MEMBERSHIP

Annals Pharmacotherapy (Nobili) Biochemical Journal (Chiesa) Brain Aging (Forloni) Clinical Drug Investigation (Beghi) Clinical Neurology and Neurosurgery (Beghi) Cochrane Collaboration, Epilessia (Beghi) Dialogo sui Farmaci (Nobili) Drugs in the R&D (Beghi) Early Intervention in Psychiatry (Barbato) Epidemiologia e Prevenzione (Lucca) Epilepsia (Beghi, Vezzani, assistant editor) Epilepsy Current (Vezzani) Epilepsy Research (Vezzani) Inpharma (Beghi) International Journal of Mental Health (Barbato)

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International Journal of Molecular Epidemiology and Genetics (Forloni, senior, Albani associate) Journal of Alzhiemers disease (Albani, Forloni) Journal of Neurochemistry (Bendotti) Journal of Neuroscience Online (Forloni) Neurological Sciences (Beghi) Neuroepidemiology (Beghi) Neuroscience (Vezzani) Open Aging Journal (Forloni) Open Geriatric Medicine Journal (Forloni) Psichiatria di Comunit (Barbato) Quality of Life Research (Barbato) Ricerca & Pratica (Nobili) Stroke (De Simoni, Associate editor)

PEER REVIEW ACTIVITIES

Acta Neurologica Scandinavica Acta Psychiatrica Scandinava Alzheimer's & Dementia Alzheimer Disease and Associated Disorders American Journal of Clinical Nutrition American Journal of Hematology American Journal of Human Genetics American Journal of Pathology American Journal of Physiology Annals of Neurology Annals of Pharmacotherapy Archives of Internal Medicine Arthritis Research & Therapy Behavioural Brain Research Behavioural Neuroscience Biochimica et Biophysica Acta Biochemical Journal Biochemistry BioMed Central Neurology Biological Psychiatry BMC Psychiatry BMC Public Health Brain Research Brain Research Bulletin Brain Research Review Clinical Drug Investigation Clinical Neurology and Neurosurgery Clinical Pharmacokinetics Clin Pharm Therapy CNS Drugs Dialogo sui farmaci Drugs

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Epidemiologia e Psichiatria Sociale Epilepsia Epilepsy & Behavior European Journal of Immunology European Journal of Neuroscience European Journal of Pharmacology European Journal of Public Health Experimental Neurology European Neuropsychopharmacology Expert Opinion on Pharmacotherapy FASEB Journal FEBS letters Fundamental Clinical Psychopharmacology Future Drugs Giornale di Neuropsichiatria dellEt Evolutiva Glia International Journal of Neuropsychopharmacology Journal of Alzhiemers disease JAMA Journal of the American Board of Family Practice Journal of Biological Chemistry Journal of Cell. Biology Journal of Cell Physiology Journal of Cerebral Blood Flow and Metabolism Journal of Chemical Neuroanatomy Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Science Journal of Gerontology Journal of Headache and Pain Journal of Histochemistry and Cytochemistry Journal of Immunology Journal of Internal Medicine Journal of Neurochemistry Journal of Neuroimmunology Journal of Neurology, Neurosurgery and Psychiatry Journal of Neuroscience Journal of Pharmacology and Experimental Therapeutics Journal of Pharmacy and Pharmacology Journal of Psychopharmacology Journal of Psychosomatic Research Journal of Structural Biology Journal of Virology Life Sciences Lancet Lancet Neurology Molecular Brain Research Molecular and Cellular Neuroscience Molecular Therapy Nature Neuroscience Neuroepidemiology Neurology Neurological Sciences

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Neurobiology of Learning and Memory Nerobiology of Aging Neurobiology of Diseases Neuropharmacology Neuropsychopharmacology Neuroscience Neuroscience Letters N.S. Archives Pharmacology Parkinsonism & Related Disorders Pharmacological Research Pharmacoepidemiology and Drug Safety Pharmacology Biochemistry & Behavior PlosONE Proc Natl Acad Sci, USA Progress in Neuro-Psychopharmacology & Biological Research Psychopharmacology Schizophrenia Research Social Psychiatry and Psychiatric Epidemiology Synapse Trends Molecular Medicine Vaccine

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Agenzia Europea di Valutazione dei Medicinali (EMEA) Agenzia Italiana per il Farmaco (AIFA) Associazione Italiana per la Ricerca sullInvecchiamento Cerebrale (AIRIC, Presidenza) Board of "Master in Advanced Technologies for the Study of Neurodegenerative Diseases", Milan University Comitato di coordinamento internazionale del progetto europeoQuelles professionnalits en sant mentale. Perspectives croises, usagers, lus professionnels. Commissione sulla Health Care Policy della Lega Internazionale contro lEpilessia (ILAE) Comitato Ordinatore del Master in "Tecnologie Avanzate Applicate alle Patologie Neurodegenerative", Universit di Milano Committee for Proprietary Medicinal Products (CPMP) presso LEMEA Consiglio Direttivo AIRIC Coordination Group IMI-PharmaCog project Direttivo della Lega Italiana contro lEpilessia (LICE) Editorial Committee, Guidelines of community based rehabilitation, World Health Organization. Esperto Nazionale, accreditato dallAIFA (Agenzia Italiana del Farmaco), per lEMEA (Esperto per il Medical Research Council (MRC), UK Gruppo di Approfondimento Tecnico per lo sviluppo dellarea Promozione della salute mentale, Regione Lombardia Gruppo di lavoro sull'epilessia dell'Organizzazione Mondiale della Sanit Gruppo di Studio sullEpilessia della Societ Italiana di Neurologia (SIN) Gruppo di Studio sulla Qualit della Vita della Societ Italiana di Neurologia (SIN) Gruppo di Studio sulla Sclerosi Laterale Amiotrofica della Societ Italiana di Neurologia (SIN) Medical Research Council Strategic Grant Application, UK Mental Health Working Party, gruppo di lavoro nominato dal Direttorato Generale per la

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Protezione del Consumatore della Commissione Europea (DG-SANCO), Bruxelles. Gruppo di coordinamento Neuroprion NoE, EU International Committee su Epilepsy and the Law International Organizing Committee e coordinator della segreteria al Global Forum for Community Mental Health, istituito dal Department of Mental Health della World Health Organization. International Subcommittee della American Academy of Neurology International Steering Committee dellEuropean Network on mental health promotion and mental disorder prevention (EMHPA). International Subcommittee dellAmerican Academy of Neurology National Institutes of Health of the USA and World Health Organization supported project on The Future of Psychiatric Diagnosis: Refining the Research Agenda. Neurobiology Commission of the International League Against Epilepsy Neuroepidemiology Section of the American Academy of Neurology (Chair uscente) Research Advisory Panel, MND Association, UK Task Force sullepidemiologia dellepilessia della ILAE Scientific Advisory Board of Sheffield Institute Foundation for MND Scientific Advisory Board del Thierry Latran Foundation, Francia Working Group on Epilepsy della World Health Organization (WHO)

EVENT ORGANIZATION
8a Giornata di studio sulla malattia di Alzheimer Polipatologie e politerapie nel paziente con demenza Stili di vita e insorgenza di demenza 13 March 2010, Ateneo Veneto, Venezia Third GiSAS Investigators Meeting Salerno 28 January 2010 Fourth GiSAS Investigators Meeting Milano 9 November 2010

GRANTS AND CONTRACTS

Abbott GmbH & Co. KG Agenzia di Sanit Pubblica del Lazio Amgen, Milano ASL 2 Piemonte. ASL TO1 Torino Assessorato alla Salute, Comune di Milano Association pour la recherch sur la SLA, France Azienda USL 3 Pistoia e Valdinievole Bristol-Myers Squibb Boehringer Ingelheim Centro Studi in Psichiatria ASL TO2, Torino CURE EISAI Epilepsy

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Dana Foundation Dipartimento di Salute Mentale, Azienda Ospedaliera Niguarda Ca Granda, Milano Dipartimento di Salute Mentale di Pistoia e Valdinievole Evidentia Medica, Grottaferrata (Roma) Fondazione Cariplo, Milano Fondazione Golgi-Cenci, Abbiategrasso Fondazione Mariani, Milano Fondazione Italo Monzino, Milano Fondazione Vialli e Mauro per la Ricerca FP6, European Union Glaxo-SmithKline, Italy Hospice "via di Natale Franco Gallini", Aviano (PN) Human Frontiers Scientific Programme IMI-PharmaCog IMPHA II, DG-SANCO, Public Health and Consumers' Protection (Directorate General Istituto Comprensivo Statale "G.D. Romagnosi", Carate Brianza (MI) Istituto Regionale Lombardo di Formazione per lAmministrazione Pubblica IREF I.R.I.S Istituto Superiore di Sanit Janssen-Cilag H. Lundbeck A/S, Danimark Hoffmann-La Roche AG, Svizzera Ministero della Ricerca Scientifica Ministero della Salute MND Association, UK Newron Ospedale Casa Sollievo di San Giovanni Rotondo Pharming Provincia Autonoma di Trento Progetto Itaca, Milano Regione Lombardia, Assessorato alla Famiglia e Solidariet Sociale e Assessorato alla Sanit, Milano Rimoldi e Bergamini Rotary Clubs Gruppo 1, Milano Rotary Clubs Milano Naviglio Grande San Carlo, Milano Scala, Inner Wheel Milano San Carlo Sanofi-Aventis SELECTA MEDICA, Pavia Servier Laboratories, Parigi Sigma-Tau Telethon Unione Nazionale Associazioni per la Salute Mentale UNASAM Vertex WebMedica, Grottaferrata (Roma). World Health Organisation

SCIENTIFIC PUBLICATIONS (2010)

Aguglia U, Beghi E, Labate A, Condino F, Cianci V, Mumoli L, Gasparini S, Quattrone A, Gambardella A.Age at onset predicts good seizure outcome in sporadic non-lesional and mesial

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temporal sclerosis based temporal lobe epilepsy. J Neurol Neurosurg Psychiatry. 2010 Oct 22. [Epub ahead of print] Albani D, Polito L, Signorini A, Forloni G. Neuroprotective properties of resveratrol in different neurodegenerative disorders. Biofactors. 2010; 36:370-6 Albani D, Polito L, Forloni G. Sirtuins as novel targets for Alzheimer's disease and other neurodegenerative disorders: experimental and genetic evidence. J Alzheimers Dis. 2010; 19:11-26.. Antoniou X and Borsello T., Cell permeable peptides: A promising tool to deliver neuroprotective agents in the brain Pharmaceuticals 2010; 3: 379-392 Antoniou X, Falconi M, Di Marino D, Borsello T.,JNK3 as a Therapeutic Target for Neurodegenerative Diseases. J Alzheimers Dis. 2010 Feb 24. [Epub ahead of print] Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, Tapella L, Colombo L, Manzoni C, Borsello T, Chiesa R, Gobbi M, Salmona M, Forloni G.Synthetic amyloid-beta oligomers impair long-term memory independently of cellular prion protein. Proc Natl Acad Sci U S A. 2010; 107: 2295-300 Balducci C, Tonini R, Zianni E, Nazzaro C, Fiordaliso F, Salio M, Vismara L, Gardoni F, Di Luca M, Carli M, Forloni G. Cognitive Deficits Associated with Alteration of Synaptic Metaplasticity Precede Plaque Deposition in AbetaPP23 Transgenic Mice. J Alzheimers Dis. 2010 21:1367-1381 Barbato A, Parabiaghi A, Panicali F, Battino N, D'Avanzo B, De Girolamo G, Rucci P, Santone G PROGRES-Acute Group. Do patients improve after short psychiatric admission? A cohort study in Italy. Nord J Psychiatry 2010 E-pub. Beghi E.Treating epilepsy across its different stages. Ther Adv Neurol Disord. 2010; 3:85-92 Beghi E, Carpio A, Forsgren L, Hesdorffer DC, Malmgren K, Sander JW, Tomson T, Hauser WA. Recommendation for a definition of acute symptomatic seizure. Epilepsia. 2010; 51:671-5. Beghi E, Bussone G, D'Amico D, Cortelli P, Cevoli S, Manzoni GC, Torelli P, Tonini MC, Allais G, De Simone R, D'Onofrio F, Genco S, Moschiano F, Beghi M, Salvi S. Headache, anxiety and depressive disorders: the HADAS study. J Headache Pain. 2010; 11:141-50. Beghi E, Logroscino G, Chi A, Hardiman O, Millul A, Mitchell D, Swingler R, Traynor BJ. Amyotrophic lateral sclerosis, physical exercise, trauma and sports: results of a populationbased pilot case-control study. Amyotroph Lateral Scler. 2010; 11:289-92. Biasini E., Tapella L., Restelli E., Pozzoli M., Massignan T., and Chiesa R. The hydrophobic core region governs mutant prion protein aggregation and intracellular retention. Biochemical Journal 2010; 430: 477-86 Bruni AC, Bernardi L, Colao R, Rubino E, Smirne N, Frangipane F, Terni B, Curcio SA, Mirabelli M, Clodomiro A, Di Lorenzo R, Maletta R, Anfossi M, Gallo M, Geracitano S,

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Tomaino C, Muraca MG, Leotta A, Lio SG, Pinessi L, Rainero I, Sorbi S, Nee L, Milan G, Pappat S, Postiglione A, Abbamondi N, Forloni G, St George Hyslop P, Rogaeva E, Bugiani O, Giaccone G, Foncin JF, Spillantini MG, Puccio G.Worldwide distribution of PSEN1 Met146Leu mutation: a large variability for a founder mutation. Neurology. 2010; 74:798-806. Caccia S, Pasina L, Nobili A. New atypical antipsychotics for schizophrenia: iloperidone Drug Des Devel Ther 2010; 4: 33-48 Calcagno E, Invernizzi RW. Strain-dependent serotonin neuron feedback control role of serotonin 2C receptors. J Neurochem. 2010; 114:1701-10. Carli M, Calcagno E, Mainolfi P, Mainini E, Invernizzi RW. Effects ofaripiprazole, olanzapine, and haloperidol in a model of cognitive deficit of schizophrenia in rats: relationship with glutamate release in the medial prefrontal cortex. Psychopharmacology (Berl). 2010 Nov 4. [Epub ahead of print] Carli M, Calcagno E, Mainini E, Arnt J, Invernizzi RW. Sertindole restore attentional performance and suppresses glutamate release induced by the NMDA receptor antagonist CPP. Psychopharmacology (Berl). 2010 Nov 4. [Epub ahead of print] Chiappedi M, Beghi E, Ferrari-Ginevra O, Ghezzo A, Maggioni E, Mattana F, Spelta P, Stefanini MC, Biserni P, Tonali P.Response to rehabilitation of children and adolescents with epilepsy. Epilepsy Behav. 2011 20: 79-82. Chi A, Borghero G, Calvo A, Capasso M, Caponnetto C, Corbo M, Giannini F, Logroscino G, Mandrioli J, Marcello N, Mazzini L, Moglia C, Monsurr MR, Mora G, Patti F, Perini M, Pietrini V, Pisano F, Pupillo E, Sabatelli M, Salvi F, Silani V, Simone IL, Sorar G, Tola MR, Volanti P, Beghi E; LITALS Study Group. Lithium carbonate in amyotrophic lateral sclerosis: lack of efficacy in a dose-finding trial. Neurology. 2010; 75: 619-25 Congedo M, Causarano RI, Alberti F, Bonito V, Borghi L, Colombi L, Defanti CA, Marcello N, Porteri C, Pucci E, Tarquini D, Tettamanti M, Tiezzi A, Tiraboschi P, Gasparini M; Bioethics and Palliative Care in Neurology Study Group of Italian Society of Neurology. Ethical issues in end of life treatments for patients with dementia. Eur J Neurol 2010; 17: 774779. Crippa V, Carra S, Rusmini P, Sau D, Bolzoni E, Bendotti C, De Biasi S, Poletti A. A role of small heat shock protein B8 (HspB8) in the autophagic removal of misfolded proteins responsible for neurodegenerative diseases. 2010; Autophagy. 6: 958-60, Crippa V, Sau D, Rusmini P, Boncoraglio A, Onesto E, Bolzoni E, Galbiati M, Fontana E, Marino M, Carra S, Bendotti C, De Biasi S, Poletti A.The small heat shock protein B8 (HspB8) promotes autophagic removal of misfolded proteins involved in amyotrophic lateral sclerosis (ALS). Hum Mol Genet. 2010 19:3440-56, Dub CM, Ravizza T, Hamamura M, Zha Q, Keebaugh A, Fok K, Andres AL, Nalcioglu O, Obenaus A, Vezzani A, Baram TZ. Epileptogenesis provoked by prolonged experimental febrile seizures: mechanisms and biomarkers. J Neurosci. 2010; 30: 7484-94.

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Epis R, Marcello E, Gardoni F, Vastagh C, Malinverno M, Balducci C, Colombo A, Borroni B, Vara H, Dell'Agli M, Cattabeni F, Giustetto M, Borsello T, Forloni G, Padovani A, Di Luca M. Blocking ADAM10 synaptic trafficking generates a model of sporadic Alzheimer's disease. Brain. 2010; 133:3323-35 Fumagalli S, Coles JA, Ejlerskov P, Ortolano F, Bushell T, Brewer JM, De Simoni MG, Dever G, Garside P, Maffia P, Carswell HV. In vivo real time multiphoton imaging of T lymphocytes in mouse brain after experimental stroke. Stroke, 2010. E-pub Gallucci M, Ongaro F, Meggiolaro S, Antuono P, Gustafson DR, Forloni GL, Albani D, Gajo GB, Durante E, Caberlotto L, Zanardo A, Siculi M, Muffato G, Regini C. Factors related to disability: Evidence from the "Treviso Longeva (TRELONG) Study" Arch Gerontol Geriatr. 2010 Jun 8. [Epub ahead of print] Gesuete R, Orsini F, Zanier ER, Deli MA, Albani D, Bazzoni G, De Simoni MG. Glial cells drive preconditioning-induced blood-brain-barrier protection. Stroke, 2010. E-pub. Gironi M, Saresella M, Marventano I, Guerini FR, Gatti A, Antonini G, Ceresa L, Morino S, Beghi E, Angelici A, Mariani E, Nemni R, Clerici M. Distinct cytokine patterns associated with different forms of chronic dysimmune neuropathy. Muscle & Nerve. 2010; 42: 864-70 Lescai F, Pirazzini C, D'Agostino G, Santoro A, Ghidoni R, Benussi L, Galimberti D, Federica E, Marchegiani F, Cardelli M, Olivieri F, Nacmias B, Sorbi S, Bagnoli S, Tagliavini F, Albani D, Boneschi FM, Binetti G, Forloni G, Quadri P, Scarpini E, Franceschi C. Failure to replicate an association of rs5984894 SNP in the PCDH11X gene in a collection of 1,222 Alzheimer's disease affected patients. J Alzheimers Dis. 2010; 21: 385-8. Logroscino G, Traynor BJ, Hardiman O, Chi A, Mitchell D, Swingler RJ, Millul A, Benn E, Beghi E; EURALS. Incidence of amyotrophic lateral sclerosis in Europe. J Neurol Neurosurg Psychiatry. 2010; 81: 385-9 Ludolph AC, Bendotti C, Blaugrund E, Chio A, Greensmith L, Loeffler JP, Mead R, Niessen HG, Petri S, Pradat PF, Robberecht W, Ruegg M, Schwalenstcker B, Stiller D, van den Berg L, Vieira F, von Horsten S. Guidelines for preclinical animal research in ALS/MND: A consensus meeting. Amyotroph Lateral Scler. 2010; 11: 38-45, Marcucci M, Iorio A, Nobili A, Tettamanti M, Pasina L, Marengoni A, Salerno F, Corrao S, Mannucci PM; REPOSI Investigators. Factors affecting adherence to guidelines for antithrombotic therapy in elderly patients with atrial fibrillation admitted to internal medicine wards. Eur J Intern Med. 2010; 21:516-23. Marengoni A, Bonometti F, Nobili A, Tettamanti M, Salerno F, Corrao S, Iorio A, Marcucci M, Mannucci PM; Italian Society of Internal Medicine (SIMI) Investigators. In-hospital death and adverse clinical events in elderly patients according to disease clustering: the REPOSI study. Rejuvenation Res. 2010; 13:469-77. Marengoni A, Corrao S, Nobili A, Tettamanti M, Pasina L, Salerno F, Iorio A, Marcucci M, Bonometti F, Mannucci PM; on behalf of SIMI Investigators SIMI denotes the Italian Society of Internal Medicine. The participating units and co authors are listed in the Appendix. In-hospital death according to dementia diagnosis in acutely ill elderly patients: the REPOSI study. Int J

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Geriatr Psychiatry. 2010 Nov 9. Maroso M, Balosso S, Ravizza T, Liu J, Aronica E, Iyer AM, Rossetti C, Molteni M, Casalgrandi M, Manfredi AA, Bianchi ME and Vezzani A. Toll-Like Receptor 4 (TLR4) and High Mobility Group Box 1 (HMGB1) are involved in ictogenesis and can be targeted to reduce seizures Nature Medicine, 2010; 16: 413-9. Massignan T., Biasini E., Lauranzano E., Veglianese P., Pignataro M., Fioriti L., Harris D.A., Salmona M., Chiesa R., and Bonetto V. Mutant prion protein expression is associated with an alteration of the Rab GDP dissociation inhibitor alpha (GDI)/Rab11 pathway. Mol Cell Proteomics 2010; 9: 611-22 Massignan T., Stewart R.S., Biasini E., Solomon I.H., Bonetto V., Chiesa R. and Harris D.A. A novel, drug-based, cellular assay for the activity of neurotoxic mutants of the prion protein. J. Biol. Chem. 2010: 285: 7752-7765 Mecarelli O, Capovilla G, Romeo A, Rubboli G, Tinuper P, Beghi E.Past and present public knowledge and attitudes toward epilepsy in Italy. Epilepsy Behav. 2010; 18: 110-5 Noe F, Vaghi V, Balducci C, Fitzsimons H, Bland R, Zardoni D, Sperk G, Carli M, During MJ, Vezzani A Anticonvulsant effects and behavioural outcomes of rAAV serotype 1 vectormediated neuropeptide Y overexpression in rat hippocampus. Gene Ther. 2010; 17: 643-52 Ortolano F, Maffia P, Dever G, Rodolico G, Millington OR, De Simoni MG, Brewer JM, Bushell T, Garside P, Carswell HV. Advances in imaging of new targets for pharmacological intervention in stroke: real-time tracking of T-cells in the ischemic brain. Br J Pharmacol 2010; 159: 808-811. Pasina L, Nobili A, Tettamanti M, Riva E, Lucca U, Piccinelli R, Defendi L, Perego L, Lucifora S, Bulla C. Co-prescription of gastroprotective agents in patients taking non-selective NSAIDs or COX-2 selective inhibitors: analysis of prescriptions. Int J Clin Pharmacol Ther. 2010, 48:735-43 Peviani M, Caron I, Pizzasegola C, Gensano F, Tortarolo M, Bendotti C. Unraveling the complexity of amyotrophic lateral sclerosis: Recent advances from the transgenic mutant SOD1 mice. CNS Neurol Disord Drug Targets. 2010, 9: 491-503, Piccinelli P, Beghi E, Borgatti R, Ferri M, Giordano L, Romeo A, Termine C, Viri M, Zucca C, Balottin U.Neuropsychological and behavioural aspects in children and adolescents with idiopathic epilepsy at diagnosis and after 12 months of treatment. Seizure. 2010; 19:540-6. Politis A, Olgiati P, Malitas P, Albani D, Signorini A, Polito L, De Mauro S, Zisaki A, Piperi C, Stamouli E, Mailis A, Batelli S, Forloni G, De Ronchi D, Kalofoutis A, Liappas I, Serretti A.Vitamin B12 levels in Alzheimer's disease: association with clinical features and cytokine production. J Alzheimers Dis. 2010; 19: 481-8. Pous MF, Camporese M, Nobili A, Frau S, Del Zotti F, Conforti A, Zimol R, Giustetto G, Zermiani G, Lombardo G, Mezzalira L. Quality assessment of information about medications in primary care electronic patient record (EPR) systems. Inform Prim Care. 2010;18:109-16

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Pozzi L, Greco B, Sacchetti G, Leoni G, Invernizzi RW, Carli M. Blockade o serotonin 2A receptors prevents PCP-induced attentional performance deficit and CREB phosphorylation in the dorsal striatum of DBA/2 mice. Psychopharmacology (Berl). 2010; 208: 387-99. Repici M, Wehrl R, Antoniou X, Borsello T, Dusart I c-Jun N-Terminal Kinase (JNK) and p38 Play Different Roles in Age-Related Purkinje Cell Death in Murine Organotypic Culture. Cerebellum. 2010 Dec 30. [Epub ahead of print] Restelli E, Fioriti L, Mantovani S, Airaghi S, Forloni G, Chiesa R Cell type-specific neuroprotective activity of untranslocated prion protein. PLoS One. 2010; 5:e13725. Rischio and Prevenzione. Investigators.Efficacy of n-3 polyunsaturated fatty acids and feasibility of optimizing preventive strategies in patients at high cardiovascular risk: rationale, design and baseline characteristics of the Rischio and Prevenzione study, a large randomised trial in general practice. Trials. 2010; 28;11:68. Termine C, Ferri M, Livetti G, Beghi E, Salini S, Mongelli A, Blangiardo R, Luoni C, Lanzi G, Balottin U. Migraine with aura with onset in childhood and adolescence: long-term natural history and prognostic factors. Cephalalgia. 2010; 30: 674-8 Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta MG, Riva E. Prevalence, incidence and types of mild anemia in the elderly: the "Health and Anemia" population-based study. Haematologica. 2010; 95:1849-56. Urru SA, Veglianese P, De Luigi A, Fumagalli E, Erba E, Gonella Diaza R, Carr A, Davoli E, Borsello T, Forloni G, Pengo N, Monzani E, Cascio P, Cenci S, Sitia R, Salmona M. A new fluorogenic peptide determines proteasome activity in single cells. J Med Chem. 2010; 53:7452-60. Valerio A, Bertolotti P, Delbarba A, Perego C, Dossena M, Ragni M, Spano PF, Carruba MO, De Simoni MG, Nisoli E. Glycogen synthase kinase-3 inhibition reduces ischemic cerebral damage, restores impaired mitochondrial biogenesis and prevents ROS production. J Neurochem, 2010. E-pub Vezzani A, Balosso S, Maroso M, Zardoni D, No F, Ravizza T. ICE/caspase 1 inhibitors and IL-1beta receptor antagonists as potential therapeutics in epilepsy. Curr Opin Investig Drugs. 2010; 11: 43-50 Watson J, Guzzetti S, Franchi C, Di Clemente A, Burbassi S, Emri Z, Leresche N, Parri HR, Crunelli V, Cervo L.Gamma-hydroxybutyrate does not maintain self-administration but induces conditioned place preference when injected in the ventral tegmental area. Int J Neuropsychopharmacol. 2010; 13:143-53. Zanier ER, Brandi G, Peri G, Longhi L, Zoerle T, Tettamanti M, Garlanda C, Sicurt A, Valaperta S, Mantovani A, De Simoni MG, Stocchetti N. Cerebrospinal fluid pentraxin-3 early after subarachnoid hemorrhage is associated with vasospasm. Intensive Care Medicine, 2010. E-

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Zanier E R, Refai D, Zipfel G J, Zoerle T, Longhi L, Esparza T J, Spinner M L, Bateman R J, Brody D L, Stocchetti N. Neurofilament light chain levels in ventricular cerebrospinal fluid after acute aneurysmal subarachnoid haemorrhage. J Neurol Neurosurg Psychiatry 2010, E-pub

LAY PRESS SELECTION (2010)

Clavenna A, Bonati M, Sequi M, Nobili A, Franchi C, Tettamanti M, Bortolotti A, Merlino L, Fortino I. La prescrizione di farmaci per i bambini e gli anziani in Regione Lombardia. Ricerca & Pratica 2010; 26: 9-18. Forloni G. Ricerca traslazionale o semplicemente razionale, la ricerca per la malattia di Alzheimer, Ricerca & Pratica 2010; 26: 183-192 Franchi, C. Arosio, F, Salvini Porro, G., Colombo, A., Nobili, Studio della qualit delle Unit di Valutazione Alzheimer in Lombardia Giornale di Gerontologia, 2010 58:278-282 Nobili A. A proposito di anziani e farmaci. Ricerca & Pratica 2010; 26: 31-3. Nobili A, Pasina L, Garattini S. Il caso del clopidogrel e degli inibitori di pompa protonica. Aggiornamento Medico 2010; 34: 56-59. Nobili A, Pasina L, Garattini S. Farmacogenetica: lesempio della warfarina. Aggiornamento Medico 2010; 34: 84-88.

RESEARCH ACTIVITIES Laboratory of Biology of Neurodegenerative Disorders Alzheimer's disease: genetic studies and clinical investigations
In collaboration with different neurological centers and the laboratory of Geriatric Neuropsychiatry it has been created a bank of blood samples for DNA of patients with Alzheimers disease (AD), in familial (FAD) or sporadic form (SAD), and patients with vascular dementia (VD). In all subjects the diagnosis of dementia is performed according to the international guidelines. Since 2005 we also started the collection of blood samples from subjects with front-temporal dementia. The genetic studies are aimed at identifying causal factors in FAD and risk factors in SAD. Mutations on genes encoding proteins involved in the physiopathology of AD were investigated. The pathogenic role of these mutations is under investigation using fibroblasts obtained from skin biopsy. Furthermore, we continued the screening of FAD samples for the genes encoding for presenilin 1 and 2 (PS-1 and PS-2) and APP, missense mutations in these three genes were associated with AD.

Alzheimer's disease: preclinical studies

The formation of amyloid (A) deposits in brain parenchyma and on the wall of cerebral blood vessels is an early event in AD and there are now numerous genetic, biochemical and neuropathological studies pointing to a causal role of A in the pathogenesis of AD. Thus, prevention the formation of A aggregates or their elimination once formed is a potential therapeutic approach to the disease. This aim is strongly persecuted with different strategies

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including the regulation of enzymes responsible of the synthesis and degradation of A and the enzymes influencing the metabolism of amyloid precursor protein (APP). In the lab, we developed the idea to interfere directly with the A deposits formation using anti-amyloidogenic drugs. The experimental studies have shown the potential therapeutic activity of these drugs in AD, and now they will be tested in a clinical setting.

Alzheimers disease: Translational studies

In the frame of the international study IMI-PharmaCog several protocols have been set up for the MRI analysis in various transgenic mice models of Alzheimers disease (AD). The PharmaCog project focused on the optimization of the translational studies to facilitate the therapeutic approaches considering in experimental models and in the clinical studies the same parameters, behaviorally, biochemically and of imaging. In this contest it will be analyzed longitudinally in single, carrying human amyloid precursor protein mutated (APP) associated to AD, double carrying APP and mutated PS1 transgene, and triple transgenic mice carrying APP, PS2 and mutated tau transgene. We planned to peform the MRI analysis in the same animals at 4, 8, 12, 18 and 24 months, the analysis will be structural, functional and spettroscopical. The strumental parameters (ROI, T2, DTI) have been harmonized with the partners developing similar approaches in humans. The first results seem to confirm the presence of a hippocampal atropy in double transgenic mice similar to that shown in AD.

The role of oligomers in the Alzheimer pathogenesis

Recent data have shown the essential role played by oligomers, small and soluble aggregates of A, in the Alzheimer pathogenesis and in particular in the cognitive decline associated to the disease. In collaboration with the Department of Biochemistry and Molecular Pharamacology we developed some in vivo models to analyze the neuronal dysfunction induced by A 142 but not in monomeric or fibrillar species. The intracerebral application of these different forms confirmed that A oligomers induced behavioral impairment while monomeric or fibrillar forms of A did not affect the cognitive behavior. The intracellular signaling pathways, by which A oligomers induce synaptic failure and consequently neuronal degeneration are poorly understood. Nevertheless increasing evidence indicate the involvement of kinases-dependent signaling pathways, and more specifically the JNK signaling pathway in these early degenerative events. The JNK kinase phosphorylates APP (amyloid precursor protein) and its relevance in both neuronal death and brain plasticity is well established. We recently demonstrated, by using the specific cell penetrating JNK inhibitor peptide (D-JNKI1) characterized by dr. Borsello, that JNK is responsible for APP phosphorylation at Thr668, and that its specific inhibition reduced the APPs and A fragments production in primary cortical neurons. In addition, we could show that JNK inhibition leads to a shift from the amyloidogenic to the non-amyloidogenic pathway, a result with potentially important therapeutic implications.

Sirtuins and aging

The sirtuins are a family of conserved proteins with de-acetylation activity. In human beings the sirtuins are coded by 7 different genes and are localized in the citosol, within the nuclei and in the cellular mitochondria. SIRT-1, the better known sirtuin, is involved in the aging physiology and energetic metabolism, its activation induced beneficial effects in Alzheimer and Parkinson experimental models. We studied sirtuins from different points of view, genetic, cellular and behaviorally. The genetic studies are devoted to identify alterations associated to AD in Italian populations. During the screening of all sirtuin genes, we found several single nucleic polymorphisms that now are investigated in a larger population (560 AD subjects). The cellular studies are focused on the role of SIRT-1 and SIRT-2 in the cell death mechanisms and oxidative stress in cellular models of AD. Since sirtuins have been involved in the energetic

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metabolism, and mental as well as physical exercise exert protective effect in AD, we are evaluating in AD animal models if sirtuins are able to mediate the beneficial effects of physical exercise and environmental stimulation.

Genetics of aging
In collaboration with the Geriatric Neuropsychiatry Lab for the Monzino 80-plus study and with dr. Maurizio Gallucci from the ARGel Association in Treviso for Trelong study we collected a large number of blood samples from subjects over seventy. In these samples we are performing a genetic analysis to identify genetic profiles associated to the longevity and /or to the agingassociated pathologies with specific attention to the dementias. The aim is to cross the genotype/phenotype profile with pathologies and environmental aspects including lifestyle, diet and economical conditions to identify risks and protective factors. Initially the subjects were genotypized for ApoE, whom allele E4 is a well-known risk factor for Alzheimers disease and several other disorders and sirt-1 a gene codified for protein member of a enzymatic family of sirtuins associated to the longevity in several experimental models. The results are interesting but before drawing any conclusion we need to consider the numerous other parameters collected in our database.

Sirtuins and aging

The sirtuins are a family of conserved proteins with de-acetylation activity. In human the sirtuins are coded by 7 different genes and are localized in the citosol, within the nuclei and in the cellular mitochondria. SIRT-1, the better known sirtuin, is involved in the aging physiology and energetic metabolism, its activation induced beneficial effects in Alzheimer and Parkinson experimental models. We studied sirtuins from different points of view, genetic, cellular and behaviorally. The genetic studies are devoted to identify alterations associated to AD in Italian populations. During the screening of all sirtuin genes, we found several single nucleic polymorphisms that now are investigated in larger population (560 AD subjects). The cellular studies are focused on the role of SIRT-1 and SIRT-2 in the cell death mechanisms and oxidative stress in cellular models of AD. Since sirtuins have been involved in the energetic metabolism, and mental as well as physical exercise exert protective effect in AD, we are evaluating in AD animal models if sirtuins are able to mediate the beneficial effects of physical exercise and environmental stimulation.

Genetics of aging
In collaboration with Geriatric Neuropsychiatry Lab for the Monzino 80-plus study and with dr. Maurizio Gallucci from the ARGel Association in Treviso for Trelong study we collected a large number of blood samples from subjects over seventy. In these samples we are performing a genetic analysis to identify genetic profiles associate to the longevity and /or to the agingassociated pathologies with specific attention to the dementias. The aim is to cross the genotype/phenotype profile with pathologies and environmental aspects including lifestyle, diet and economical conditions to identify risks and protective factors. Initially the subjects were genotypized for ApoE, whom allele E4 is a well-known risk factor for Alzheimers disease and several other disorders and sirt-1 a gene codified for protein member of an enzymatic family of sirtuins associated to the longevity in several experimental models. The results are interesting but before drawing any conclusion we need to consider the numerous other parameters collected in our database.

Parkinsons Disease: genetic studies

Parkinsons disease (PD) is the second more diffuse neurodegenerative disorder with an unknown pathogenesis, however for PD several therapies are available and, although at the symptomatic level, their efficacies is well-established. In the etiological studies on PD the

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genetic component has been traditionally considered with scarce interest whereas the environmental causes were carefully evaluated. This orientation was based on the evidence that the exposure to several toxins can mimic the PD pathology. However the genetic studies in the last few years have completely changed the perspective with the identification of mutations on two genes, encoding for alpha-synuclein and parkin, associated to the juvenile forms of the disease. A mutation on alpha synuclein gene is an extremely rare event, only three mutations identified until now, the parkin mutations are numerous either in puntiform or in deletion form. The mutations on alpha-synuclein gene are dominant while the parkin mutations are associated with PD in recessive form. We collected, in collaboration with several neurological centers, blood samples from PD subjects and the screening of the samples involved genes like alphasynuclein, parkin, DJ-1 and other factors potentially involved in PD. Recently, an association between polymorphisms occurring in gene for serotonin transporter and the appearance of depression in PD subjects has been investigated. The results indicate no association between the serotonin transporter gene polymorphisms and depression in PD, but a direct association between these polymorphisms and PD itself. This indicates a more relevant involvement o serotonergic system in PD pathogenesis compared to whom is generally considered.

Parkinsons disease: studies in vitro

The identification of the mutations associated to Parkinsons disease (PD) gave a substantial contribute to understand the disease and allowed the development of cellular models to investigate the pathogenesis of the disease. In the past we showed the potential neurotoxic activity of alpha-sinuclein using the synthetic peptide homologous to the fibrillogenic fragment 61-95 (NAC) of the protein. Successively with the help of dr. Negro at the Department of Biochemistry at the University of Padova we prepared cDNA vectors including the sequence of wild type and mutated alpha-synuclein. Their transfection to the PC12 cells induced in specific conditions a cellular damage. More recently alpha-synuclein was associated to a TAT sequence capable to transport inside the cells the protein. With this method the intracellular concentration of alpha-sinuclein was better controlled. In a micromolar range alpha-synuclein was toxic, but in nanomolar range, it exerted neuroprotective effect against oxidative stress induced by hydrogen peroxide. This double effect dose-dependent was confirmed in an inducible model. More recently again in collaboration with Dr. Negro (Padua University), we obtained the recombinant form of DJ-1 associated with TAT (TAT-DJ-1). This protein is similar to alphasynuclein, mutations of its sequence has been associated to PD. TAT-DJ-1 silencing by small interference RNA (siRNAi) were used to study the interaction between DJ-1 and alpha synuclein..

Synaptic Dysfunction
Nowadays it is assumed that AD is a synapse-related pathology leading to synaptic dysfunction and loss, a phenomenon that precedes extensive amyloid deposition in the brain. At the same time, soluble diffusible forms of A can perturb in an early stage of the disease the synaptic function causing a reduction of dendritic spines density in the cortex and hippocampus, an acute inhibition of long term potentiation (LTP) and the loss of critical spine proteins (e.g. membrane expression of NMDA receptors). However, the relationship between A and synapses loss remains unclear and more efforts are necessary to better understand the mechanisms underlying A synaptic toxicity. Our aim is to study the effects of A oligomers on MAPKs pathways and elucidate the link between synaptopathies and the activation/inhibition of the JNK, p38 and ERK cascades. This study can potentially be a breakthrough in the comprehension of AD pathogenesis: understanding the cellular and molecular alterations that lead to AD will help in developing effective and preventive therapeutic strategies in order to counteract or nullify the degenerative processes activated by A.

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MAPKs (ERK, p38 and JNK) are also implicated in the regulation of the immediate early signaling events that modulate synaptic plasticity by controlling LTP, LTD and the recycling of glutamate receptors (NMDAR and AMPAR) as well as their expression. Nevertheless, MAPKs involvement in the regulation of synaptic function and dysfunction and the mechanisms by which they trigger the synaptic loss induced by A oligomers are largely unknown.

The cargo strategy as a key tool in neuroprotection

The possibility to target protein complexes and enzymes involved in intracellular signaling pathways by means of cell permeable peptide (CPP) conjugates represents a novel, versatile and extremely powerful way of blocking the propagation of intracellular signaling events or intracellular processes, with an unprecedented specificity allowing for reduction of side effects. D-JNKI1 is a cell-permeable, biologically-active peptide consisting of a carboxyl terminal sequence derived from the JNK binding domain of the scaffold protein JIP-1/IB1 (JBD20), and an amino terminal portion containing the HIV-TAT 48-57 transporter sequence. D-JNKI-1 has been designed to block the interaction, mediated by JBD domain, between JNK and its targets. D-JNKI1 afforded powerful protection against NMDA excitotoxicity in cortical neurons and against cerebral ischemia in vivo, as well as in two other neurodegenerative models (hair-cell loss in animal models of sudden deafness and retinal ganglion cell loss following optic nerve crush in vivo. The peptide progressed in clinical trails for preventing brain ischemia/stroke (see CHUV/Xigenpharma Lausanne web-link). Among the possible targets for neuroprotection there is MKK7, that is activated, unlikely MKK4, during NMDA-stress,. To inactivate MKK7 we use lentiviruses because of the particular capability of integrating genetic material into the genome of non-dividing cells stably. Our lentiviral vector will carry a single si-RNA duplex of MKK7. A second approach is to test in vitro the specific inhibitor: Gadd45, a molecule active on MKK7. Gadd45 binds to MKK7 directly and blocks its catalytic activity, the binding between Gadd45/MKK7 being tighter than JIP1/MKK7. The endogenous Gadd45 interacts to MKK7 through direct, high-affinity contact but not with the other JNK upstream kinase, MKK4. For this study we initially plan to use a viral system that will allow us to observe the role of this pathway in excitotoxicity, with a final goal of producing a cell-permeable peptide with a more specific effect in the prevention of neuronal death.

SUMOlization in acute and chronic diseases

Small ubiquitin-like modifier (SUMO) is a group of proteins responsible for post translational modifications influencing protein function, localization and stability. Recently, protein Sumoylation has attracted neuroscientists since it is implicated in the altered protein dynamics that are associated with various aspects of neurodegenerative disease, including stroke and Alzheimer's Disease (AD). Our hypothesis is that SUMOylation confers neuroprotection against stressful stimuli through regulation of important stress signaling pathways. The aim of this study is to investigate the role of SUMOylation in models of acute (ischemia) and chronic brain diseases (AD). At present we are characterising the changes in expression and localisation of SUMO1-2/3 in an in vitro model of ischemia (NMDA application) as well as in a model of AD (oligomers application).

Laboratory of Neurological Disorders Epidemiological studies on amyotrophic lateral sclerosis (ALS)

Included are studies on the incidence, risk factors and mortality of ALS. The data are obtained from a regional registry of the disease activated in 1998 and including all patients with newly

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diagnosed ALS identified in the Lombardy region. Using similar study protocols, the same data are collected in two additional regional registries (from Piemonte and Puglia) included in a network with the Lombard registry. Information obtained from patients enrolled in the Lombard registry and from cases examined by members of the Italian ALS Study Group has been used to assess the validity and reliability of diagnostic criteria for ALS and selected disability scales. Based on the data recorded, the annual incidence of ALS is comparable to that obtained in other Western countries where ALS registries have been activated, and is among the highest ever published (1.9 per 100,000). Mortality of ALS has been found to be comparable to that of studies from similar populations studied with the same protocol. The study on the validation of the current diagnostic criteria for ALS (the El Escorial criteria) showed that to be considered valid and reliable, the criteria should be used after proper training of the investigators. In October 2004, the Laboratory of Neurological Disorders has started a European collaborative group for the ALS registries (EURALS) with the intent to create a common database (completed in the year 2005) with the participation of the existing regional and national disease registries. With the collaboration of the UK and Irish groups participating in the EURALS collaboration, a scientific report has been published on a meta-analysis of the incidence of ALS, performed by pooling data from the 1998-99 cohorts of patients enrolled in the populationbased registries. Two studies have been recently concluded: 1. A case-control study on trauma and risk of ALS (in collaboration with the Italian registries); 2. A survey of the prevalence of cognitive impairment and extrapyramidal signs in patients with newly diagnosed ALS (Italian registries). Other studies are ongoing under the coordination of the Laboratory of Neurological Disorders: 1. A case-control study on ALS, physical exercise and sport (in collaboration with the EURALS Consortium). 2. A study of the mortality of ALS in the 1998-99 cohort of patients from the European population-based registries (EURALS Consortium).

Treatment of the first epileptic seizure and short and long-term mortality
A cohort of 419 patients with a first unprovoked seizure, randomized to immediate treatment of to treatment at the time of relapse, were followed for up to 20 years in search of short-term and long-term mortality. The mortality in this cohort was compared to that of the Italian population and measured with the Standardized Mortality Ratio (SMR). The SMR was not significant (1.2; 95% CI 0.9-1.7) and was not changed for patients in whom the treatment of the first seizure was withhold.

Innovative therapeutic strategies in patients with epilepsy

A cohort of patients with a first unprovoked seizure, randomised since 1988 by several Italian centers to immediate treatment or to treatment only at the time of a seizure relapse, was followed to verify the impact of the two therapeutic strategies on the long-term prognosis of epilepsy, measured by the chance of achieving 5-year remission. To provide a pragmatic definition of drug resistance in childhood epilepsy, children refractory to two antiepileptic drugs (in sequence or in combination) were randomised to the use of a third drug or to the optimization of the existing treatment and followed for up to three years. Therapeutic response was measured by the achievement of a six-month period of remission. The study has been conducted in collaboration with the IRCCS Stella Maris of Calambrone (PI). The study has been concluded prior to completing patient recruitment because the eligible patients could not be easily traced. The available data have processed and analyzed and a scientific manuscript is in preparation Epidemiology of neurological disorders in Albania With the collaboration of the Fondazione Mariani and the Neurological Department of the University of Tirana, an epidemiological survey has been completed to assess the prevalence

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and incidence of several neurological conditions (stroke, epilepsy, headache, dementia, peripheral neuropathy, multiple sclerosis) comparing an urban and a rural community (Tirana and Saranda). In 2005, a study on the validation of the diagnostic criteria was conducted. A total of 9869 persons (Tirana 4953; Saranda 4916) were screened. The prevalence rates of the clinical conditions (expressed as number per 1000) are, in decreasing order, 258 (headache), 36 (polyneuropathy), 15 (epilepsy), 13 (stroke), 11 (dementia), 9 (parkinsonisms), 5 (cerebral palsy), and 0.3 (multiple sclerosis).

Headache and comorbidity

Headache is frequently associated with other comorbid disorders. Epilepsy is one of the commonest associated clinical conditions. On this background, an observational study has been recently completed. The aim of the study was to show any differences between patients with epilepsy-headache comorbidity and those with epilepsy or headache alone. A total of 1167 patients were examined (156 with epilepsy-headache comorbidity, 675 with headache alone, and 336 with epilepsy alone). Differences between the three groups were found for family history of epilepsy (prevailing in patients with epilepsy-headache comorbidity) or headache (prevailing in patients with headache alone) and for clinical features (less severe in patients with epilepsy-headache comorbidity).

Cerebrovascular disorders and risk of epilepsy

Epilepsy is a frequent complication of stroke. Acute symptomatic seizures (i.e. seizures occurring in the seven days after stroke) can occur in up to two-thirds of cases and epilepsy (i.e. repeated unprovoked seizures) in 2-4%. There are no consistent findings on the risk factors for acute symptomatic seizures, unprovoked seizures and epilepsy in patients with stroke. For these reasons, in 2007 a multicenter national prospective survey has been started to assess the risk of seizures and epilepsy (and the main risk factors) in a cohort of patients with a first ischemic or hemorrhagic stroke followed for a maximum period of 24 months. The study was also implemented to assess the feasibility of a pragmatic therapeutic trial on the prophylaxis of seizures and epilepsy in stroke. Based on the analysis of 714 patients, the incidence of acute symptomatic seizures was 6.7%. Cerebral hemorrhage and a cortical lesion increased overall seizure risk while hypercholesterolemia decreased the risk of hemorrhagic stroke .

Therapeutic trials in neurological disorders

During the year 2010 six therapeutic trials sponsored by the Italian Drug Agency (AIFA) and a therapeutic trial sponsored by the Italian Ministry of Health were ongoing. Included are: 1. A randomized double-blind parallel-group placebo-controlled trial on the efficacy and tolerability of L-acetylcarnitine in ALS; 2. A randomized open-label parallel-group trial comparing Erythropoietin to Methyl-prednisolone in patients with acute spinal cord injury; 3. A randomized double-blind parallel-group placebo-controlled trial on the efficacy and safety of valproate in medication-overuse headache; 4. A randomized open-label trial of the efficacy of a comprehensive rehabilitation program for the prevention of falls in Parkinsons disease; 5. A randomized open-label trial on the efficacy of an active monitoring of the adverse effects of antiepileptic drugs and of relevant drug interactions; 6. A randomized open-label trial on the efficacy of an educational program for physicians working in nursing homes.. The first trial aims at finding a potentially effective drug in a clinical condition for which there is only one product (Riluzole) with at best modest efficacy on survival. L-acetylcarnitine has been found to improve survival in experimental models of motor neuron disease. The second trial intends to verify the efficacy of erythropoietin, a drug shown to mitigate the effects of traumatic spinal shock and accelerate recovery in experimental animals. The drug chosen for comparison (Methylprednisolone at high doses) has been selected for being the present gold standard in

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clinical practice. The third trial aims at verifying whether valproate (a drug commonly used for the prophylaxis of migraine) abates symptoms occurring in drug-overuse headache, a common and frequently invalidating variety of chronic idiopathic headache. The fourth trial aims at assessing whether a comprehensive rehabilitation program compared to usual care is followed by a reduction in the incidence of falls in patients with Parkinsons disease at risk of falls. The fifth trial aims at verifying the added value of an active monitoring of adverse drug interactions compared to usual care in patients receiving antiepileptic drugs associated to other compounds. The sixth trial aims to verify the added value of a web-based educational program in reducing the number of inappropriate prescriptions compared to usual care. The laboratory of neurological disorders is the coordinator of the first trial and a partner in the other trials, where the main tasks include protocol and CRF preparation, statistical analysis, and preparation of the final scientific report.

Satisfaction with antiepileptic treatment in children and adolescents with epilepsy

Epilepsy is a disease requiring chronic use of drugs having adverse effects which may affect satisfaction with treatment. Poor satisfaction, particularly in children and adolescents, may affect compliance and quality of life of patients and their parents and caregivers. Two hundred and 93 children and adolescents with epilepsy aged 3 through 17 years were included in a nationwide prospective observational study at treatment onset or at time of treatment modification and reassessed at one and three months. At the end of the first month, dissatisfaction with treatment was manifested by 29.4% of cases. The percentage fell to 24.2% at three months. Patients with newly diagnosed epilepsy were better satisfied than patients diagnosed in the past. The variables associated with poor satisfaction with treatment included adverse drug reactions, disease duration and poor quality of life of parents/caregivers.

Laboratory of Experimental Neurology Role of inflammatory molecules in ictogenesis and epileptogenesis

We are studying the role of IL-1beta and HMGB1 systems in the genesis and propagation of seizures and in the associated neurodegenerative phenomena. We have demonstrated that epileptic activity induces the synthesis of these pro-inflammatory molecules, danger signals and their specific receptors. In particular, IL-1beta and HMGB1 have proconvulsant actions while their receptor antagonist (IL-1Ra, BOX-A, Toll-like receptors inhibitors) or IL-1beta synthesis inhibitors, have anticonvulsant activities. We are actively studying the role of these molecules in epilepsy models with the intent of promoting their clinical applications in drug-resistant epileptic patients. This possibility is encouraged by the clinical use of some of these molecules (e.g. anakinra, the IL-1R antagonist) in chronic inflammatory and autoimmune diseases in humans. We are studying pharmacological approaches to block IL-1beta- and HMGB1-signaling involved in the proconvulsant effects of these molecules

Epilepsy and postnatal development

Seizure susceptibility is higher in early infancy although the immature brain appears to be less susceptible to epileptogenesis. Using experimental models of seizures induced during pos-tnatal development in rodents, we are studying the mechanisms involved in age-dependent seizure susceptibility and the associated neuronal injury. Our studies are primarily focused on inflammatory pathways, angiogenic processes and blood-brain barrier damage.

Blood-brain barrier and epileptogenesis

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We are studying BBB permeability and microvasculature changes induced in the brain by seizures or by neurotrauma or infection and how these modifications may affect the process of epileptogenesis. Experimental models of symptomatic epilepsy are used.

New therapeutic approaches of In vivo gene transfer

This study concerns the use of adeno-associated viral vectors to introduce genes with therapeutic potential in the brain, thus increasing the synthesis of specific proteins to produce long-lasting anticonvulsant effects. We have demonstrated that adeno-associated viral vector carrying the human neuropeptide Y gene, significantly increases the brain concentration of this peptide after its intrahippocampal injection for a prolonged time (at least up to 5 months after a single intracerebral injection). The rats overexpressing this peptide are less susceptible to seizures. Future development of this study concerns the optimization of the transgene transfer technology to envisage a possible clinical application

In vivo MRI/MRS to determine glia activation and blood-brain barrier damage

This study is focused on in vivo magnetic resonance imaging (MRI) and spectroscopy (MRS) techniques to evaluate the role of glia activation and blood-brain barrier damage in the epileptic process. Our intention is to explore whether these two phenomena can be used as biomarkes of epileptogenesis. This information may provide a clinically applicable method for predicting the development of spontaneous seizures in individual at risk, thus permitting to envisage prevention strategies.

Laboratory of Geriatric Neuropsychiatry Population study on the prevalence of dementias in the older-old
Parallel to the progressive increase of individuals aged 80 years or older within the elderly population (65+), the number of demented patients of 80 years or older makes up an ever increasing fraction of the total population affected by dementia. As very often happens, the exclusion from studies of subjects in the oldest age classes tends to inevitably underestimate the total number of individuals affected by dementia present in the population. To fill this gap, a door-to-door population study on the prevalence, incidence, risk factors and evolution of dementias and age-associated cognitive deficits has been set up in an elderly population aged 80 years or older living in eight small towns of Varese Province. The study is funded by a grant from the Fondazione Italo Monzino, Milano.

Health and Anemia in the elderly population

A large survey in old residents of Biella (65 years or older) has been conducted in collaboration with the Local Health Authority of Biella (ASL 12) to estimate the prevalence and incidence of anemia (mild, moderate and severe) in the elderly population and to investigate the association of mild anemia with hospitalization and mortality. In a large subgroup of eldely subjects also the effects of mild grade anemia on cognitive, functional, mood, and quality of life variables have been investigated.

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Evaluating risk profiles in hospitalised elderly subjects

In collaboration with the Geriatric Division of the Ospedali Regionali of Lugano and Mendrisio, Switzerland, hospitalized and ambulatory patients are evaluated from a neuropsychological, functional and mobility point of view to estimate the impact of these factors on heath-related outcomes and disease progression (Canton Ticino Study).

Longitudinal follow-up of individuals with mild cognitive impairment (MCI)

In collaboration with the Geriatric Unit of Ospedali Regionali of Lugano and Mendrisio, Switzerland, the follow-up study of all Mild Cognitive Impairment or Questionable Dementia (CDR 0.5) patients seen at the Memory Clinic of the Hospitals is continuing to estimate the rate of conversion to dementia and to evaluate the possible risk factors associated with conversion (Canton Ticino Study).

Quality of care of terminally ill oncological subjects

In 1999 we started a collaborative programme with the hospice via di Natale Franco Gallini in Aviano (PN). The present aim of the collaborative research project is to assess the hospice activities after its opening and the analysis of the data collected in the first decade of activity (2000-2009).

Randomised controlled trial of the Italian Group for the Study of the Second Generation Antipsychotics GiSAS
The study aims at evaluating efficacy and safety of three antipsychotic drugs - aripiprazole, olanzapine and haloperidol by a pragmatic design involving a large sample of patients with schizophrenia treated in community psychiatric services across Italy. This is the first large multicentre trial on this subject ever realized in Italy. The target is to recruit 260 patients. Of the over 50 centers involved, 43 are currently actively participating. These centers have recruited 243 patients, among whom 129 completed the one-year follow-up.

GiSAS survey
This study examined the opinions of psychiatrists from centers participating in the GiSAS trial, a pragmatic RCT of antipsychotic drugs in schizophrenia in order to explain and possibly improve the limited Patient recruitment in randomized clinical trials in general and in the GiSAS study. A survey of all clinicians working in the trial recruiting centers was performed exploring four critical areas: reasons for trial participation, recruitment barriers, antipsychotic preferences and sources of information about antipsychotics.

Monitoring of self-harm and suicide attempts

In the framework of a wider project financed by the Ministry of Health (Call 2006) and led by the Public Health Agency of the Lazio Region, the Unit of Epidemiology and Social Psychiatry has enlarged the network of services where the Form for Hospital Assessment of Self-Harm and Suicide Attempts is used. Data collection has started in various centres, covering a total of more than 1 million inhabitants in various areas of North Italy. In the Province of Trentino, services for adolescents have adopted the schedule after the necessary modifications. The Unit is also involved on a project financed by the Italian Centre for Disease Control, where the form is

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further introduced in areas with high incidence of suicide and family practitioners will be trained about depression identification and suicide risk detection.

European Network of Bipolar Research Export Centres - ENBREC

ENBREC is designed to build an EU-wide network of expert centres specialised in research and care on bipolar disorders, in order to integrate research efforts on the mechanism of disease, and optimized diagnostic and treatment. Common tools and practice, training and information will help structuring the European bipolar disorders research community and translate research outcome into healthcare. Epidemiology and Social Psychiatry Unit reviewed research findings on effective psychosocial interventions and planned, in collaboration with the Department of Mental Health of San Carlo Hospital in Milan, an investigation on use of psychosocial intverventions for bipolar disorders in routine clinical practice. A psychoeducation intervention, integrated with self-help and expression of the direct experience of patients was designed and proposed to the patients in charge. A total of 35 patients were included in three intervention groups.

Quality Evaluation of a Department of Mental Health with the participation of users

The study investigates the quality of assistance offered to patients with severe mental illnesses and intensively cared by the mental health services, through an instrument developed with a substantial contribution of users and distributed by them. Of the 290 patients identified to be interviewed, 204 gave valid questionnaires.

HoNOS-5 Study: towards the development of a clinically oriented informative system

The study was approved by the Italian Health Department (Call 2008) and adopted as outcome measure the Health of the Nation Outcome Scale (HoNOS). Its aim is to evaluate the clinical outcomes of a cohort of subjects referring to a group of 25 Italian mental health services and to create a dataset containing all available information drawing on already operating informative flows, thus developing a clinically oriented informative system. More than 6,500 patients were recruited and evaluated through the HoNOS scale and subsequently followed-up and reassessed in July and November. The last follow-up is scheduled for March 2011.

Michaels game: a table game for group cognitive-behavioural therapy of psychotic symptoms.
Cognitive therapies (CBT) of psychotic symptoms have shown to be effective in the treatment of psychoses. Their spreading within naturalistic settings is, however, limited. Michaels Game, a training module for hypothetical reasoning, coming in the form of a game of cards, is a treatment inspired by CBT approach. It was conceived as a tool to promote the spreading of CBT in natural clinical settings. The Unit coordinates the participation of two centers in Italy to a multi-center international non-profit clinical trial evaluating the effectiveness of Michaels Game program on adult patients who show residual psychotic symptoms. In the two center of Brescia, 10 patients were followed up to nine months and the study was concluded. In the center of Senigallia, 11 subjects were included and the experimental phase was concluded.

Use of the Regional Database on Drug Prescriptions

Although reboxetine has been prescribed for many years in Europe for the treatment of depression, a recent meta-analysis concluded that it is ineffective and potentially harmful. We performed a population-based prescription study comparing use of reboxetine, fluoxetine, paroxetine and mirtazapine in a large sample of adults in Lombardy.

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Use of the Regional Psychiatric Information System to monitor access to mental health services and patterns of care
Data on mental health services in Lombardy were collected through rhe regional information system and analyzed in terms of treated prevalence, treated incidence, contnuity of care and packages of care for the period 1999-2009 and involving 120.000 patients corresponding to a rate of 146/10,000 population aged 15 and over.

Laboratory of Inflammation and Nervous System Diseases The complement system in stroke and traumatic brain injury experimental models
Previous studies of ours have indicated that complement and related inflammatory systems such as contact/kinin and fibrinolytic systems may represent novel targets for reducing ischemia/reperfusion injury. We have shown that C1-INH, a serine-protease inhibitor that acts as a major regulator of both complement and kinin systems, remarkably improves the functional and neuropathological consequences of focal transient as well as permanent ischemia induced by middle cerebral artery occlusion in mice. In particular we have shown that plasmaderived(pd)C1-INH effectively and markedly reduces brain ischemia/reperfusion injury, inducing a decrease of the 90% of the ischemic lesion and that its actions lead to inhibition of cell recruitment, inflammation and apoptosis. In addition we could show that pdC1-INH protective effects are present also in traumatic brain injury models. More recently we demonstrated that the recently available recombinant human(rh) C1-INH is as effective as pdC1-INH in reducing the ischemic lesion but it has a much wider therapeutic window, being protective whan adiminstered up to 18 h after transient ischemia and up to 6 h after permanent ischemia. We could show that a major target of rhC1-INH neuroprotection is mannose-binding lectin (MBL), a key protein in the complement lectin pathway. Thus C1-INH, which is presently used as replacement therapy in patients with C1-INH deficiency, possesses potent, multi-faceted neuroprotective actions that may be beneficial in acute brain injury. Ongoing studies are focussed on MBL as a novel target for stroke and traumatic brain injury.).

Stem cells as a therapeutic approach in stroke and traumatic brain injury

We have previously demontrated a beneficial effect of neural stem cells after transient brain ischemia Ethical issues involved in stem cell research and the limited availability of most adult stem cells outline the need to look for other cell populations. We are presently focussing on stem cells obtained from human umbilical cord blood that are an easily available source of progenitors with multilineage capacity and could represent an ideal candidate for cell-based therapy after acute brain injury. The aim of the research is to verify the conditions for the effectivenss of Human Umbilical Cord Blood Mesenchymal Stem Cells (CB-MSC, provided by Cell Factory, Ospedale Maggiore Policlinico di Milano) in reducing the ischemic and traumatic brain injury (TBI) and to investigate the mechanisms triggered by their infusion in the injured brain. CB-MSC are infused into the brain ventricle of injured mice. At different time points and up to 3 months, several parameters are being evaluated: distribution and phenotype of injected cells, neurodegeneration, behavioral deficits, cytokine and trophic factor gene expression, microglia activation. In our TBI model the results obtained show that CB-MSC: 1) migrate towards the contused tissue and

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survive in the injured brain up to 5 weeks postinjury; 2) induce an early and persistent attenuation of functional impairments related to sensory/motor activity and cognitive functions up to 6 weeks post-TBI; 3) significantly reduce the contusion volume as assessed one month after trauma; 4) modify the brain environment by modulating the inflammatory response and stimulating the production of trofic factors. These data indicate that this type of stem cells may represent a useful therapeutic tool in TBI patients. In our focal ischemia model the results obtained up to now indicate a similar protective effect on sensorymotor functions. Ongoing studies include: 1) definition of the long term effects on behavioural and anatomical damage after ischemia; 2) evaluation of the trophic effects of CB-MSC and of the reciprocal interaction between CB-MSC and injured environment with specific analysis of the protective/toxic role of microglia; 2) definition of the mechanisms of homing of stem cells in the injured brain; 3) evaluation of the ability of CB-MSC to differentiate into functional neural progeny 3 months after transplantation.

Ischemic preconditioning and the blood-brain barrier

Recent studies indicate that cell death resulting from ischemic injury can be reduced when a sublethal injurious stimulus or ischemic episode occurs hours or days before a severe ischemic insult. This phenomenon is known as ischemic pre-conditioning (IPC). Elucidating the mechanisms responsible of ischemic preconditioning provides an opportunity to identify the putative candidates that can confer neuroprotection against acute brain injury. A major goal is to identify underlying endogenous protective signals, with the long-term goal to allow therapeutic augmentation of the endogenous protective mechanisms in cerebral ischemia. Although most attention has been focused on the neuronal effects of IPC, recent studies have shown that IPC reduces ischemia-induced cerebrovascular damage. We have specifically investigated the role of BBB in cerebral ischemia and preconditioning. To this purpose we have established a bidimensional BBB model by means of co-cultures of mouse brain endothelial cells and glial cells. BBB has been exposed to oxygen-glucose deprivation (OGD) to mimic ischemic injury. We have specifically: 1) elucidated BBB involvement in cerebral ischemia and IPC in in vivo and in vitro mouse models; 2) identified mediators and cell types involved in activation and maintenance of ischemic tolerance in the cerebrovascular unit. The results obtained have shown that BBB can be directly preconditioned and that astrocytes are the driving gear of this protective mechanism.

In vivo real time imaging in ischemic mouse brain by two-photon microscopy

Ischemic stroke triggers vascular responses including blood flow rearrangements, blood brain barrier disruption and expression of adhesion molecules that stimulate immune cell infiltration. These mechanisms contribute to the progression of the ischemic damage after the acute event and represent potential therapeutic targets. In vivo imaging of the brain at cellular resolution in 3D provides an ideal tool to get an insight in these dynamic events. We have recently established an original approach by means of two-photon microscopy that allows the visualisation and measurement of dynamic events taking place in the brain. Two-photon microscope benefits from high-energy electronic transition in a fluorescent molecule due to the cooperation of two low-energy photons, thus enabling imaging over long periods in living animals. On-going projects (carried on in collaboration with the Centre For Biophotonics at the Strathclyde University of Glasgow) focus on the blood flow dynamics following focal brain ischemia and on the tracking of lymphocytes infiltrated in the ischemic territory. We obtained highly detailed imaging and quantification of vascular dynamics and moving lymphocytes in the brain in vivo after stroke. In particular we: 1) measured the massive vascular rearrangement

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occurring during and after ischemia, such as blood flow speed variations and temporal dynamics of extravasation appearance; 2) collected data as number of infiltrated lymphocytes, their track velocity, displacement rate and meandering index thus providing a comprehensive description of lymphocyte behaviour in the brain. The final aim of the project will be to elucidate dynamic events associated with the evolution of ischemic damage over time thus providing the bases for a rational manipulation of blood supply and immune responses for therapeutic intervention in stroke.

Laboratory of Molecular Neurobiology Study on pathogenic mechanisms of Amyotrophic Lateral Sclerosis

Role of protein aggregation One of the pathological features of ALS is the accumulation of protein aggregates in cell bodies and axons of motor neurons. Having shown that the alteration of the ubiquitin / proteasome contributes to the progression rather than the pathogenesis of the disease (see activity report 2009) in SOD1G93A mice, a model of familial ALS, this year we focused on another protein degradation system, the lysosome-autophagy. We observed that in the spinal cord of SOD1G93A mice at the end stage of disease there is an increase in the levels of LC3II which is a marker for the formation of autophagosomes. This indicates that the activation of autophagy could be an alternative defense mechanism of the cell aimed to eliminate the abnormal protein aggregates when the proteasome is partially inhibited as we demonstrated in the previous study. Furthermore, in collaboration with the Department of Endocrinology, Applied Biology and Pathophysiology of the University of Milan, we have shown that in motor neurons of SOD1G93A mice there is an increased expression of a chaperone protein, HspB8, already at the presymptomatic stage and this phenomenon persists in the motor neurons that survive at the terminal stages of disease. In vitro studies on motor neuron cell lines have shown that HspB8 by interacting with a complex of other proteins such as Bag3/Hsc70/CHIP, is able to reduce aggregation and increase the solubility of SOD through the activation of the autophagic degradation pathway. The results of this work were published in Human Molecular Genetics and Autophagy. Comparative analysis of gene expression and proteome profiling of two mouse models of familial ALS with phenotypic differences of disease. (MNDA UK Project, No. 6054 and EUROMOTOR FP7 program) Recently, we observed that two genetically distinct strains of mice (C57 and 129Sv), but carriers of the same number of copies of the transgene for human SOD1 with G93A mutation, develop a different phenotype of ALS as regard the age of onset and illness duration. The genetic variability is probably a reason for differences in age of onset and severity of disease in patients with both familial and sporadic ALS. Hence our interest was focused on the study of two strains of mice and patients C57/G93A and 129Sv/G93A the behavioral, biochemical and histopathological order to understand the mechanisms responsible for the different development of the disease. The study funded by the Motor Neuron Disease Association is done in collaboration with the group of Professor Pamela Shaw, University of Sheffield (UK). The laboratory of Sheffield isolated the spinal motor neurons of the two strains of mice at different stages of the disease by laser-disector and performed the comparative transcriptome analysis between the two models. Numerous data have emerged showing a profile of gene expression with some similarities and many differences between the two strains of mice. The data are now being analyzed by the Gene Ontology Classification to identify specific pathways involved in the different phenotype. In parallel we have demonstrated histopathological and biochemical differences between the two

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SOD1G93A mouse models especially regarding the accumulation of insoluble proteins and the formation of protein inclusions. Mice with a more aggressive disease show a greater amount of aggregates at the early stages of the disease which is associated with a reduced expression of the proteasome. We also noted that mice with more severe disease have levels of alpha beta crystallin, a chaperone involved in the correct folding of proteins, 10 times lower than those of mice with slower disease suggesting a protective role of this protein. We are currently planning to examine whether an increase in expression and function of this protein is able to protect motor neurons and slow the progression of the disease. In vitro studies on neuron-glia interaction To investigate further the role of inflammatory mechanisms, we have set up in vitro co-coltures of spinal neurons and astrocytes derived from SOD1G93A mice embryos. This cellular system reproduces the selective motor neuron degeneration induced by the expression of mutant SOD1 and allows to investigate the role of TNFalpha and associated pathways. Our results firstly demonstrated the protective effect of p38MAPK inhibitors in this model, confirming the hypothesis derived by the in vivo studies, and the involvement of TNFalpha and its receptors type 2 in the motor neuron death. These findings suggest a possible new toxic pathway that can represent an innovative target for curative intervention. Further studies are in progress to validate this hypothesis. Altered axonuclear communication in motor neurons of a mouse model of familial amyotrophic lateral sclerosis (European collaborative project FP6 program EU-NES AXON SUPPORT))

The objective of this study is to test the hypothesis that a defect in the communication of specific signals between the axonal compartment and peripheral nuclei of motor neurons is responsible for the death of motor neurons in models of familial ALS. In particular, we focused our attention to the study of two proteins whose interaction fosters a regenerative response following acute axonal injury: vimentin and importin beta. We observed that in pure motor nerves (nerve quadriceps) but not in sensory nerves (saphenous nerve) of SOD1G93A rats, there is an increase of importin beta which is not associated with an increase in vimentin, at variance with what happens during the axon regeneration after an acute nerve insult. Hence we hypothesize that the non-regenerative response of axons in models of ALS is due to the lack of an increase in vimentin. To verify this hypothesis, we developed SOD1G93A mice that express a green fluorescent protein only in motor neurons (Hb9-SOD1G93A mice) and therefore allows to distinguish specifically the sensory from the motor axons under the structural and biochemical profile. The characterization of these mice is currently under study. Understanding the role of vimentin in the disease progression is important to determine whether this protein could be a therapeutic target for amyotrophic lateral sclerosis. Therapeutical interventions in mouse model of ALS
Induction and mobilization of bone marrow hematopoietic stem cells (HSC) by GCSF (granulocyte colony stimulating factor) in SOD1G93A mice. (Project supported by Thierry Latran Foundation) We evaluated the effect of GCSF on the distribution and cellular differentiation of the HSC in the nervous tissue of SOD1G93A transgenic mice and verified the effectiveness of this treatment on the progression of the disease and on neuropathological changes. We have seen that the cell proliferation assessed by incorporation of a fluorescent molecule (EDU) in DNA was increased in the spinal cord of SOD1G93A mice but this effect was only partially increased by treatment with GCSF. While the preliminary results showed a tendency to ameliorate the

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motor deficit of SOD1G93A mice after treatment with GCSF a subsequent study did not confirm this result. We are now considering other experimental conditions of treatment with GCSF and other compounds very active in the mobilization of bone marrow in order to find the ideal conditions for producing a more efficient infiltration of stem cells in the CNS. Studies on the effects of the long-chain omega-3 polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid in the G93A SOD1 mouse This is part of a collaborative project with Dr. A. Michael-Titus of Queen Mary University of London supported by the MND Association) This study is based on the observation by Dr. Michael-Titus of a neuroprotective effect of a diet enriched with fatty acids eicosapentaenoic acid (EPA), acid docosahexaenoico (DHA) on motor neurons in a rat model of spinal trauma. After a preliminary study in which we observed a trend toward improvement in the course of the disease in SOD1G93A mice treated with EPA but not with DHA or with the combination of the two compounds, we evaluated the effect of three doses of EPA alone. Unfortunately, none of the 3 doses improved the progression of the disease although there was a significant decrease in the levels of nitrotyrosine, a marker of oxidative stress in the spinal cord of mice treated with EPA.

Studies aimed at identifying biomarkers for the diagnosis and progression of the disease in ALS patients
In collaboration with the Translational Proteomics Laboratory of the Department of Biochemistry, directed by Dr. Valentina Bonetto we performed a proteomic investigation on peripheral blood cells (PBMC) of ALS patients provided by the department of neurology of the Fondazione Salvatore Maugeri, IRCCS, Milan and Centro Nemo from Niguarda Hospital in Milan. We obtained a panel of protein markers that are closely associated with ALS and can discriminate with high significance and specificity patients with the ALS from control patients with other diseases that may show a phenotype similar to ALS at the disease onset(eg, neuropathies , multiple sclerosis). In addition, some markers are able to discriminate between two levels of disease severity. The results are collected in a manuscript that is being submitted to the journal Annals of Neurology.

Study of disease progression by magnetic resonance imaging (MRI) in animal models of ALS
The purpose of this project is to verify whether the analysis of magnetic resonance imaging may be a useful tool for monitoring the progression of ALS in vivo in mice and rats bearing SOD1 G93A mutation, models of the disease. The results obtained so far show that this technique can detect the degeneration of the major cranial motor nuclei, trigeminal nucleus and facial nucleus, before the onset of disease symptoms. This is manifested by an increased signal in T2 at these nuclei, probably related to the presence of vacuoles in damaged motor neurons. In parallel we are conducting histopathological analysis at different times during the course of the disease in order to correlate the degree of degeneration of motor neurons with the modification of the MRI signal. With this study we will determine whether MRI can be a useful means of investigation to check the efficacy of a potential therapy on motor neuron degeneration without sacrificing the animal. Through the use of MRI, and in particular with the technique called Fiber Tracking we were also able to analyze the axonal degeneration in the lumbar spinal cord in ex-vivo isolated spinal cord of SOD1G93A mice. We are now trying to replicate these data in viable mice, during the progression of the disease. This will allow us to assess more accurately and at different levels (somatic and axonal) the degree of degeneration of motor neurons. The use of the technique of 'MRI is very important because it can be directly translated to the clinic as a marker of disease progression in patients.

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Laboratory of Prion Neurobiology Development of a cellular assay for the activity of neurotoxic PrP mutants Several mutated forms of PrP induce neurodegeneration when expressed in transgenic mice; however, they are not toxic when expressed in cultured cells. We found that some PrP mutants sensitize cultured cells to the toxic activity of two classes of antibiotics. Based on this result, we developed a new assay allowing the screening of the cytotoxic activity of mutant PrP in immortalized cell lines. This assay could be useful for dissecting the molecular mechanisms of PrP toxicity, and to test the efficacy of potential therapeutic agents. Proteomic analysis of a cellular model of fatal familial insonnia The PrP mutation D178N/M129 is linked to fatal familial insomnia, characterized by severe sleep abnormalities and autonomic dysfunction. We showed by immuno-electron microscopy that this mutant PrP accumulates abnormally in the endoplasmic reticulum and Golgi of transfected neuroblastoma N2a cells. To investigate the impact of intracellular PrP accumulation on cellular homeostasis, we did a differential proteomic analysis in collaboration with the Translational Proteomics Lab of the Mario Negri Institute. We found changes in proteins involved in energy metabolism, redox regulation and vesicular transport. Rab GDP dissociation inhibitor alpha (GDI) was one of the proteins that changed most. GDI regulates vesicular trafficking by acting on the activity of several Rab proteins. We found a specific reduction in the level of functional Rab11 in mutant PrP expressing cells, associated with impaired post-Golgi trafficking. These data indicate that mutant PrP expression alters the cellular mechanisms governing intracellular membrane traffic. Role of PrP in mediating the neurotoxic activity of amyloid beta oligomers There is evidence that the amyloid- (A) peptide plays a key role in the pathogenesis of Alzheimers disease. In collaboration with the Laboratory of Biology of Neurodegenerative Disorders and the Department of Biochemistry of Mario Negri Institute, we found that acute intracerebroventricular injections of synthetic A oligomers impaired consolidation of the long-term recognition memory, whereas mature A fibrils and freshly dissolved peptide did not. The deficit induced by oligomers was reversible and was prevented by an anti-A antibody. It has been suggested that PrP mediates the impairment of synaptic plasticity induced by A. We confirmed that A oligomers interact with PrP, with nanomolar affinity. However, PrP-expressing and PrP knock-out mice were equally susceptible to this impairment. These data suggest that A oligomers are responsible for cognitive impairment in AD and that PrP is not required. Role of the hydrophobic core of PrP in misfolding It is not clear how PrP mutations cause disease, but structural changes of the mutant protein and aggregation inside the cells may contribute to the brain damage. We modified the central region of mutant PrP, and found that the modified molecules folded properly and reached the cell surface, where non-mutated PrP normally resides, more efficiently. These findings indicate that the central region of PrP plays a pivotal role in governing the folding and cellular fate of the mutant molecules, and may help in devising rational therapeutic approaches for inherited prion diseases.

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Neuroprotective activity of cytosolic PrP A key pathogenic role in prion diseases was proposed for a cytosolic form of PrP. However, it is not clear how cytosolic PrP localization influences neuronal viability, with either cytotoxic or anti-apoptotic effects reported in different studies. The cellular mechanism by which PrP is delivered to the cytosol of neurons is also debated, and either retrograde transport from the endoplasmic reticulum or inefficient translocation during biosynthesis has been proposed. We investigated cytosolic PrP biogenesis and effect on cell viability in primary neuronal cultures. We found that an untraslocated form of cytosolic PrP was produced in certain neurons but not in others, and that this form boosted the resistance of cortical and hippocampal neurons to apoptosis. These results indicate cell type-dependent differences in the efficiency of PrP translocation, and argue that cytosolic PrP targeting might serve a physiological neuroprotective function. Laboratory of Experimental Psychopharmacology Drug Abuse. Neural basis of drug self-administration
To separate the direct pharmacological effects of cocaine from those associated with active drug self-administration we employed a yoked control-operant paradigm and investigated the expression of well established markers of the rapid action of cocaine, i.e. the inducible early genes and trophic factors, in rats after a single intravenous (i.v.) cocaine self-administration session. Animals self-administering cocaine did more active lever-presses than yoked-cocaine (YC) and yoked-vehicle (YV) animals. This goal-oriented behaviour was accompanied by a selective increase in Arc mRNA levels in the medial prefrontal cortex (mPFC). These findings demonstrate that a single session of cocaine i.v. self-administration is sufficient to shape rat behaviour towards goal-directed behaviours and selectively up-regulate Arc expression in mPFC (of SA animals), providing the first evidence that the mPFC's function is already profoundly influenced by the first voluntary cocaine exposure. Ongoing studies are evaluating whether this effect is peculiar to cocaine or common to other drugs of abuse.

GHB mechanism of action

Gamma-hydroxybutyric acid (GHB) is an endogenous brain substance that has diverse neuropharmacological actions, including rewarding properties in different animal species and in humans. As other drugs of abuse, GHB affects the firing of ventral tegmental neurons (VTA) in anaesthetized animals and hyperpolarizes dopaminergic neurons in VTA slices. We investigated the effects of various doses of intravenous GHB in maintaining self-administration and its ability to induce conditioned place preference (CPP) in rats when given orally or injected directly either in the VTA or NAc. Our results indicate that GHB did not maintained selfadministration, while given orally induced CPP. CPP was also observed when GHB was injected in the VTA but not in the NAc. Together with recent in-vitro findings, these results suggest that the rewarding properties of GHB mainly occur via disinhibition of VTA dopaminergic neurons.

Neural basis of drug craving and relapse in the drug abuse assumption
Drug craving, defined as the desire to experience the effect(s) of a previously experienced psychoactive substance is a cardinal feature of drug addiction and is clinically significant

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because of its potential link to relapse. To provide useful indications to the development of novel therapeutic approaches to prevent the use and abuse and the relapse of drug assumption following the outcome of craving, we elaborated experimental models of self-administration and relapse induced by cocaine, nicotine and alcohol-associated cues, after a period of abstinence. Ongoing studies are evaluating the role of several neurochemical mechanisms potentially involved in the drug-seeking behavior.

Search for pharmacological agents capable of modulating the craving and relapse in the consumption of psychotropic substances of abuse
Environmental stimuli associated with the intake of psychotropic substances of abuse may take the ability to induce craving that often preludes to relapse in formally detoxified patients. Studying nicotine in an experimental model of extinction-reinstatement induced by the presentation of environmental stimuli associated with self-administration of psychotropic substance of abuse, it was found that bifeprunox, a high-affinity partial agonist of dopamine D2 receptors and serotonin1A receptors, preferentially reduced nicotine-seeking behaviour in response to drug-associated stimuli in rats after a long period of abstinence.

Resistance to antidepressant drugs: experimental and clinical studies

This project arises from a collaboration between the laboratories of Neurochemistry and Behavior (R.W. Invernizzi), Drug Metabolism (Silvio Caccia), Biology of Neurodegenerative Disorders (GianLuigi Forloni) and focus on behavioural and biochemical characterization of an experimental model of resistance to the antidepressant drugs. Using an animal model which is predictive of the antidepressant activity, the effects of selective serotonin reuptake blockers (SSRI) was evaluated in several mice strains. It was found that DBA/2J and BALB/c do not respond to the antidepressant-like activity of the SSRI. The lack of effect was attributed to genotype-dependent impairment of 5-HT synthesis since DBA/2J and BALB/c carring a single nucleotide polymorphism (C1473G mice) in the gene for the brainspecific isoform of tryptophan hydroxylase-2, the rate-limiting enzyme in the synthesis of serotonin are characterized by a decreased serotonin synthesis. This hypothesis seems to be supported by the observation that DBA/2J and BALB/c mice had less dialysate 5-HT in the medial prefrontal cortex and dorsal hippocampus than C57BL/6J mice. Moreover, in DBA/2J and BALB/c the SSRI raised significantly less extracellular 5-HT when compared to C57BL/6J mice. More recently it was found that 5-HT1A and 5-HT2C receptor antagonists restored the SSRIs effect on either the antidepressant-like activity and the extracellular 5-HT.

Laboratory of Neuronal Death and Neuroprotection The role of oligomers in the pathogenesis of Alzheimers disease
Recent data have shown the essential role played by oligomers, small and soluble aggregates of A, in the pathogenesis of Alzheimer and in particular in the cognitive decline associated to the disease. In collaboration with the Department of Biochemistry and Molecular Pharamacology we developed some in vivo models to analyze the neuronal dysfunction induced by A 140 and 142 structured in oligomers but not in monomeric or fibrillar species. The intracellular signalling pathways, by which A oligomers induce synaptic failure and consequently neuronal degeneration are poorly understood. Nevertheless increasing evidence indicates the involvement of kinases-dependent signalling pathways, and more specifically the JNK signalling pathway in these early degenerative events. The JNK kinase phosphorylates APP (amyloid precursor protein) and its relevance in both

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neuronal death and brain plasticity is well established. We recently demonstrated, by using the specific cell penetrating JNK inhibitor peptide (D-JNKI1) characterized by Dr. Borsello, that JNK is responsible for APP phosphorylation at Thr668, and that its specific inhibition reduces the APPs and A fragments production in primary cortical neurons. We now demonstrated that JNK inhibition by D-JNKI1 gave total functional recovery of both long-term potentiation and long-term recognition memory impairments in TgCRND8 mouse model. The functional recovery shows a strong relationship with a reduction of Ab oligomers, derived by APP cleavage, in brain parenchyma. Our findings demonstrate that JNK is regulating the main pathogenic mechanisms of AD and might hold promise as an innovative and therapeutic target against it.

Synaptic Dysfunction
Nowadays it is assumed that AD is a synapse-related pathology leading to synaptic dysfunction and loss, a phenomenon that precedes extensive amyloid deposition in the brain. At the same time, soluble diffusible forms of A can perturb in an early stage of the disease the synaptic function causing a reduction of dendritic spines density in the cortex and hippocampus, an acute inhibition of long term potentiation (LTP) and the loss of critical spine proteins (e.g. membrane expression of NMDA receptors). However, the relationship between A and synapses loss remains unclear and more efforts are necessary to better understand the mechanisms underlying A synaptic toxicity. Our aim is to study the effects of A oligomers on MAPKs pathways and elucidate the link between synaptopathies and the activation/inhibition of the JNK, p38 and ERK cascades. This study can potentially be a breakthrough in the comprehension of AD pathogenesis: understanding the cellular and molecular alterations that lead to AD will help in developing effective and preventive therapeutic strategies in order to counteract or nullify the degenerative processes activated by A. MAPKs (ERK, p38 and JNK) are also implicated in the regulation of the immediate early signalling events that modulate synaptic plasticity by controlling LTP, LTD and the recycling of glutamate receptors (NMDAR and AMPAR) as well as their expression. Nevertheless, MAPKs involvement in the regulation of synaptic function and dysfunction and the mechanisms by which they trigger the synaptic loss induced by A oligomers are largely unknown.

The cargo strategy as a key tool in neuroprotection

The possibility to target protein complexes and enzymes involved in intracellular signalling pathways by means of cell permeable peptide (CPP) conjugates represents a novel, versatile and extremely powerful way of blocking the propagation of intracellular signalling events or intracellular processes, with an unprecedented specificity allowing for reduction of side effects. D-JNKI1 is a cell-permeable, biologically-active peptide consisting of a carboxyl terminal sequence derived from the JNK binding domain of the scaffold protein JIP-1/IB1 (JBD20), and an amino terminal portion containing the HIV-TAT 48-57 transporter sequence. D-JNKI-1 has been designed to block the interaction, mediated by JBD domain, between JNK and its targets. D-JNKI1 afforded powerful protection against NMDA excitotoxicity in cortical neurons and against cerebral ischemia in vivo, as well as in two other neurodegenerative models (hair-cell loss in animal models of sudden deafness and retinal ganglion cell loss following optic nerve crush in vivo. The peptide progressed in clinical trails for preventing brain ischemia/stroke (see CHUV/Xigenpharma Lausanne web-link). Among the possible targets for neuroprotection there is MKK7, that, unlike MKK4, is activated during NMDA-stress. To inactivate MKK7 we use lentiviruses because of the particular capability of integrating genetic material into the genome of non-dividing cells stably. Our lentiviral vector will carry a single si-RNA duplex of MKK7. A second approach is to test in vitro the specific inhibitor: Gadd45, a molecule that acts on MKK7. Gadd45 binds to MKK7

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directly and blocks its catalytic activity, the binding between Gadd45/MKK7 being tighter than JIP1/MKK7. The endogenous Gadd45 interacts with MKK7 through direct, high-affinity contact but not with the other JNK upstream kinase, MKK4. Using this knowledge we synthesized the first 2 peptides: GADD45 (69-86) by XEGEN, containing the MKK7 binding region only and GADD45 (60-86), by Dr. Mario Salmona (Mario Negri) that contains the MKK7 docking region as well as a region that is essential for MKK7 inactivation. Application of two peptides did not exert toxicity in our in vitro neuronal cultures (12 days in vitro) for concentrations as high as 20uM and for the duration tested. Notably application of both peptides 30 before NMDA application resulted in significant neuroprotection against NMDA toxicity both at 5h and 12h. Our data show convincingly that application of GADD45 (69-86) at a concentration of 20uM leads to complete abolishment of MKK7 phosphorylation, without affecting MKK4 phosphorylation. In a similar way, Western Blot analysis confirmed that application of GADD45 (69-86) completely inhibit MKK7 phosphorylation. Our preliminary data indicate that the GADD45 peptides exert neuroprotective functions, through direct regulation of the MKK7 signalling pathway. Additionally we have performed viral infection experiments with fusion proteins of enhanced green fluorescent protein (eGFP) and GADD45 and confirmed the role of GADD45 in modulation of the MKK7. We are positive on the fact that we will soon be able to test the effect of the GADD45 peptide in an in vivo ischemic model.

SUMOylation in acute and chronic diseases

Small ubiquitin-like modifier (SUMO) is a group of proteins responsible for post translational modifications influencing protein function, localization and stability. Recently, protein SUMOylation has attracted neuroscientists since it is implicated in the altered protein dynamics that are associated with various aspects of neurodegenerative disease, including stroke and Alzheimer's Disease (AD). Our hypothesis is that SUMOylation confers neuroprotection against stressful stimuli through regulation of important stress signalling pathways. The aim of this study is to investigate the role of SUMOylation in models of acute (ischemia) and chronic brain diseases (AD). Studies on neuron are starting using lentiviral technique for SUMO-1 over-expression to confirm cell lines results. JNK activity inhibition regulates APP cleavage and degradation. Since the observed modulation on JNKs activation we are testing if over-expression of SUMO-1 in H4-APPsw (cell line model of Alzheimer disease) can reduce b-amiloid fragments as cellautonomous mechanismSUMOylation of JNK3 has been successfully tested in bacterial assay, cell lines (COS7 and SH-SY5Y) and we are testing in neurons, using SUMO-1 lentivirus with promising results. Preliminary results showed that SUMO-1 regulates JNK3 expression and phosphorylation. Other tests are currently on going on neuronal culture to confirm the results obtained in cell line and bacterial. We think that SUMOylation could be a modulatory mechanism of neuronal death and these findings could be bases for new therapeutical studies against acute and chronic brain pathologies

Laboratory of Neurochemistry and Behavior Resistance to antidepressant drugs

Biological mechanisms underlying the response to antidepressant drugs were studied in strains of mice responder and non-responder to selective serotonin reuptake inhibitors (SSRI) in the forced swimming test (FST), a behavioural procedure widely used to screen potential antidepressant compounds. In 2010 we found that in non-responder mice serotonergic autoregulatory feedback mechanisms are overactive because of an increase in the sensitivity of

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5-HT2C receptors controlling the release of GABA from inhibitory neurons in the midbrain dorsal raph nucleus. Blockade of 5-HT2C receptors with a selective antagonist restored the effects of SSRI in the FST and abolished the excessive inhibitory GABAergic tone. These results suggest that overactive feedback control suppresses the effects of SSRI and identify pharmacological strategies that may enhance the response in treatment-resistant depressed patients

Animal model of cognitive deficit of schizophrenia; typical and atypical antipsychotics

The cognitive deficit is a core symptom of schizophrenia, which has been linked to functional outcome and is relatively independent of psychotic symptoms. The antipsychotics, either typical or atypical, are able to control positive symptoms such as delirium, hallucinations and paranoia. However, the currently available atypical antipsychotics when compared to conventional antipsychotics show somewhat superior efficacy for the management of cognitive deficits in patients with schizophrenia. The cognitive deficit of schizophrenia was modelled in rats and mice, by using a test of attention such as the 5-choice serial reaction time task (5-CSRTT) and injections of glutamate NMDA receptor antagonists into the medial prefrontal cortex (mPFC). This model makes clear links with psychopathology as dysfunctional glutamate neurotransmission in the mPFC has been implicated in cognitive deficits of schizophrenia and the 5-CSRTT is the rat analogue of the continuous performance test used to assess attention and vigilance in schizophrenic patients. Our studies compared the effects of conventional and atypical antipsychotics, including haloperidol, olanzapine, clozapine, sertindole and aripiprazole in this model of cognitive deficit of schizophrenia. The results show that antipsychotics may be differentiated by a preferential effect of typical antipsychotics on compulsive perseveration, and atypical antipsychotics on impulsivity. Biochemical studies show that attention deficits induced by NMDA receptor antagonists are associated with excessive glutamate in the medial prefrontal cortex of the rat, increased phosphorylation of the transcription factor CREB in the frontal cortex and decresed phosphorylation of the same transcription factor in the dorsal striatum.

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DEPARTMENT OF CARDIOVASCULAR RESEARCH

STAFF Head Maria Grazia FRANZOSI, Biol.Sci.D.

Laboratory of Cardiovascular Clinical Pharmacology

Head Roberto LATINI, M.D.

Bio-imaging Unit
Head Fabio FIORDALISO, Biol.Sci.D.

Cardiovascular Endocrine Unit

Head Serge MASSON, Ph.D.

Tissue Culture Unit

Head Giovanna BALCONI, BSc.

Laboratory of Clinical Drug Evaluation

Head Maria Grazia FRANZOSI, Biol.Sci.D.

Bioinformatics Unit
Head Enrico NICOLIS

Laboratory of General Practice Research

Head Maria Carla RONCAGLIONI, Biol.Sci.D.

Laboratory of Medical Statistics

Head Simona BARLERA, Dr.Sci.Pol., MSc.

Laboratory of Clinical Pharmacology

Head Gianni TOGNONI, M.D.

Nursing Research Unit

Head Paola DI GIULIO, R.N., MSc

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CURRICULA
Maria Grazia Franzosi got her Biological Science degree in 1972 at the University of Milan.

Education
1972 1978 Doctoral degree in Biological Sciences, University of Milan, Italy Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche "Mario Negri di Milano, Italy

Main fields of activity

Coordination of multicentric randomised clinical trials. Relationship between genetic and environmental risk factors in coronary events. Pharmacogenetics. Cardiovascular genetic epidemiology. Pharmacoeconomics. Drug Epidemiology and Post-Marketing Surveillance.

Position
from 2002 from 2005 from 2004 from 2001 from 1998 from 1997 from 1996 Director of the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Member of the Coordinating Committee of Master course in Clinical Research University of Milano Member of Steering Committee, Studio GISSI-AF Study, Milano, Italy Member of Steering Committee, Studio GISSI-HF Study, Milano, Italy Member of Steering Committee of the PROCARDIS Research Programme - A genome-wide strategy to identify susceptibility loci in precocious coronary artery disease - University of Oxford, UK Member of Antithrombotic Trialists Collaboration, Oxford, UK

Member of the Steering Committee e National Coordinator for Italy of the Organization to Assess Strategies for Ischemic Syndromes (OASIS-2, OASIS-4 CURE, Michelangelo OASIS-5 e OASIS 6, , CURRENT OASIS-7, FUTURA OASIS-8), of the INTER-HEART study and of the ACTIVE, RELY and AVERROES studies, Population Health Research Institute, McMaster University, Hamilton, Canada
Director of European Coordinating Centre and Member of Steering Committee, Collaborative Organization for RheothRx Evaluation (CORE), McMaster University, Hamilton, Canada Member of Steering Committee, Studio GISSI-Prevenzione, Milano, Italy Member of Fibrinolytic Therapy Trialistss Collaboration, Oxford, UK e del Collaborative Group on Angiotensin Converting Enzyme Inhibitors Trials, National Institutes of Health, Bethesda, Washington, USA Head of the Laboratory of Clinical Drug Evaluation, Istituto di Ricerche Farmacologiche "Mario Negri" Head of the Clinic Drug Evaluation Unit of the Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri" Member of the Scientific and Organising Secretariat, Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardico (GISSI-1, GISSI-2, GISSI-3 studies) Milano, Italy Researcher at the Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri" and at the Regional Center for Drug Information of the Lombardy Region

1994-1996 from 1993 from 2002 1989-2001 1985-1988 from 1984 1975-1984

Selected publications Broadbent HM, Peden JF, Lorkowski S, Goel A, Ongen H, Green F, Clarke R, Collins R, Franzosi MG, Tognoni G, Seedorf U, Rust S, Hamsten A, Farrall M, Watkins H, for the PROCARDIS Consortium. Susceptibility to coronary artery disease and diabetes is encoded by distinct, tightly linked, SNPs in the ANRIL locus on chromosome 9p. Hum Mol Genet 2008; 17: 806-814 GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230 GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1231-1239 Clarke R, Peden JF, Hopewell JC, Kyriakou T, Goel A, Heath SC, Parish S, Barlera S, Franzosi MG, Rust S, Bennett D, Silveira A, Malarstig A , Green FR, Lathrop M, Gigante B, Leander K, de Faire U, Seedorf U, Hamsten A, Collins R, Watkins H, Farrall M, for the PROCARDIS Consortium. Genetic variants associated with Lp(a) lipoprotein level and coronary disease. N Engl J Med 2009; 361: 2518-2528 GISSI-AF Investigators (Writing Committee: Disertori M, Latini R, Barlera S, Franzosi MG, Staszewsky L, Maggioni AP, Lucci D, Di Pasquale G, Tognoni G). Valsartan for prevention of recurrent atrial fibrillation. N Engl J Med 2009; 360: 1606-1617

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The FUTURA/OASIS-8 Trial Group. Low-Dose vs standard-dose unfractionated heparin for percutaneous coronary intervention in acute coronary syndromes treated with fondaparinux: The FUTURA/OASIS-8 Randomized Trial. JAMA 2010; 304: 1339-1349 Wallentin L, Yusuf S, Ezekowitz MD, Alings M, Flather M, Franzosi MG, Pais P, Dans A, Eikelboom J, Oldgren J, Pogue J, Reilly PA, Yang S, Connolly SJ, on behalf of the RE-LY investigators. Efficacy and safety of dabigatran compared with warfarin at different levels of international normalised ratio control for stroke prevention in atrial fibrillation: an analysis of the RE-LY trial. Lancet 2010; 376: 975-983

Simona Barlera got her degree in Political Science, area Statistics at the Universit degli Studi di Milano in Milano in 1992, followed by a master in Medical Statistics at the London School of Hygiene and Tropical Medicine, University of London in 1998. Education and training 1987-1992 Degree in Political Science, course of studies Statistics, Universit degli Studi di Milano, Milano (Italy) 1993-1995 Post-degree Specialization in Pharmacological Research. School of Specialization in Pharmacological Research Of Lombardia Region, Milan 1997-1998 Master of Science in Medical Statistics at the London School of Hygiene and Tropical Medicine, University of London, London. 1998-1999 Visiting Scientist in the Department of Statistical Genetics, Wellcome Trust Centre for Human Genetics, University of Oxford (UK). Main fields of activity Methodology of Clinical Trials in the cardiovascular field. Preparation and viewing of research protocols, planning and conduct of statistical analyses and the reporting of findings on scientific journals. Genetic epidemiology: genome-wide strategies (linkage analysis) to identify susceptibility genes in coronary artery disease; case-control studies in order to identify candidate genes involved in the cardiovascular pathology. Position Held from Oct 2006 1999 -2006 1992-1997 Head of the Laboratory of Medical Statistics, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Head of the Medical Statistics Unit, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Researcher in the Unit of Applied Statistics and Information Technology, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy

Selected publications Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: 221-227 GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230 GISSI-HF Investigators (Writing Commitee:Tavazzi L, Maggioni A P, Marchioli R, Barlera S, Franzosi M G, Latini R, Lucci D, Nicolosi G L, Porcu M, Tognoni G) Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008 372: 1231-1239 Clarke R, Peden JF, Hopewell JC, Kyriakou T, Goel A, Heath SC, Parish S, Barlera S, Franzosi MG, Rust S, Bennett D, Silveira A, Malarstig A , Green FR, Lathrop M, Gigante B, Leander K, de Faire U, Seedorf U, Hamsten A, Collins R, Watkins H, Farrall M, for the PROCARDIS Consortium. Genetic variants associated with Lp(a) Lipoprotein Level and Coronary Disease. N Engl J Med 2009; 361: 2518-2528 GISSI-AF Investigators (Writing Committee: Disertori M, Latini R, Barlera S, Franzosi MG, Staszewsky L, Maggioni AP, Lucci D, Di Pasquale G, Tognoni G). Valsartan for prevention of recurrent atrial fibrillation. N Engl J Med 2009; 360: 1606-1617 Gori F, Specchia C, Pietri S, Crociati L, Barlera S, Franciosi M, Nicolucci A, Signorini S, Brambilla P, Franzosi MG, for GISSI Prevenzione Investigators and SIBioC-GISSI Prevenzione Group. Common genetic variants chromosome 9p21are associated with myocardial infarction and type 2 diabetes in an Italian population. BMC Med Genet 2010; 11: 60

Roberto Latini got his Medical Doctor degree in 1978 at the University of Milan. Education 1970-1978 University of Milan School of Medicine, degree in Medicine

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1981-1983 Merck Sharp & Dohme International Fellow in Clinical Pharmacology Main fields of activity Mechanisms of cardiac damage following ischemia, with focus on eurohumoral activation. Use of stem cells for cardiac repair. Biohumoral investigations within large scale clinical trials in heart failure and atrial fibrillation. Positions from 1990 Head of the Cardiovascular Clinical Pharmacology Laboratory (Cardiovascular Research Department) Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy from 2001 Member of the GISSI-HF Steering Committee from 2004 Member of the GISSI-AF Steering Committee from 2005 Member of the CandHeart Steering Committee 1999-2009 Visiting Professor Dept of Medicine, New York Medical College, Valhalla, NY, USA 1981-1983 Cardiology Fellow (Dr. R. E. Kates, Laboratory) Stanford University Medical Center, CA, USA 1976-1981 Member of the Sub-Group RMs for Drugs (Community Bureau of Reference, Commission of the European Communities) 1973-1990 Fellow at the Laboratory of Clinical Pharmacology of the Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy
Selected publications Galvez BG, Covarello D, Tolorenzi R, Brunelli S, Dellavalle A, Crippa S, Mohammed SAA, Scialla L, Cuccovillo I, Molla F, Staszewsky L, Maisano F, Sampaolesi M, Latini R, Cossu G. Human cardiac mesoangioblasts isolated from hypertrophic cardiomyopathies are greatly reduced in proliferation and differentiation potency. Cardiovasc Res 2009, 83: 707-716 GISSI-AF Investigators (Writing Committee: Disertori M, Latini R, Barlera S, Franzosi MG, Staszewsky L, Maggioni AP, Lucci D, Di Pasquale G, Tognoni G), Valsartan for prevention of recurrent atrial fibrillation. N Engl J Med 2009; 360: 1606-1617 Taccone P, Pesenti A, Latini R, Polli F, Vagginelli F, Mietto C, Caspani L, Raimondi F, Bordone G, Iapichino G, Mancebo J, Guerin C, Ayzac L, Blanch L, Fumagalli R, Tognoni G, Gattinoni L, for the Prone-Supine II Study Group. Prone positioning in patients with moderate and severe acute respiratory distress syndrome. A randomized controlled trial. JAMA 2009; 302: 1977-1984

Masson S, Latini R, Anand IS. An update on cardiac troponins as circulating biomarkers in heart failure. Curr Heart Fail Rep 2010; 7: 15-21
Masson S, Solomon S, Angelici L, Latini R, Anand IS, Prescott M, Maggioni AP, Tognoni G, Cohn JN, on behalf of the Val-HeFT Investigators. Elevated plasma renin activity predicts adverse outcome in chronic heart failure, independently of pharmacologic therapy: Data from the Valsartan Heart Failure trial (Val-HeFT). J Card Fail 2010; 16: 964-970 Rysland R, Masson S, Omland T, Milani V, Bjerre M, Flyvbjerg A, Di Tano G, Misuraca G, Maggioni AP, Tognoni G, Tavazzi L, Latini R, on behalf of the GISSI-HF Investigators. Prognostic value of osteoprotegerin in chronic heart failure: The GISSI-HF trial. Am Heart J 2010; 160: 286-293

Maria Carla Roncaglioni got her Biological Science degree in 1987 at the University of Milan. Education 1987 Doctoral degree in Biological Sciences, University of Milan, Italy 1982-1983 Research Fellow at the Dept. of Biochemistry, Faculty of Medicine, Rijksuniversiteit of Limburg, Maastricht , The Netherland (Prof. C.Hemker); 1998-1999 Visiting Scientist at the Cardiovascular Research Unit, Hammersmith Hospital, London, UK (Prof. A. Maseri) Main fields of activity Coordination of multicenter clinical trials and observational studies in different cardiovascular areas (neurological, angiological, cardiological). Coordination of a network of more than 1000 GPs actively involved in epidemiological and experimental studies in the prevention of cardiovascular diseases. Position from 2001 Head of the Laboratory for General Practice Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1989 Senior Researcher in the Clinical Pharmacology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1974 Researcher in the Laboratory for the Study of Haemostasis and Thrombosis, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy

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Selected publications Tognoni G, Avanzini F, Pangrazzi J, Roncaglioni M C, Bertele V, de Gaetano G, Caimi V, Tombesi M, Colombo Fabio, Barlera S, PPP - Primary Prevention Project. Low-dose aspirin and vitamin E in people at cardiovascular risk: A randomized trial in general practice. Lancet 2001; 357: 89-95 Sacco M, Pellegrini F, Roncaglioni MC, Avanzini F, Tognoni G, Nicolucci A, PPP - Primary Prevention Project. Primary prevention of cardiovascular events with low-dose aspirin and vitamin E in type 2 diabetic patients. Results of the Primary Prevention Project (PPP) trial. Diabetes Care 2003; 26: 3264-3272 Berger JS, Roncaglioni MC, Avanzini F, Pangrazzi J, Tognoni G, Brown DL. Aspirin for the primary prevention of cardiovascular events in women and men: A sex-specific meta-analysis of randomized controlled trials. JAMA 2006; 295: 306-313 Montalvo G, Avanzini F, Anselmi M, Prandi R, Ibarra S, Marquez M, Armani D, Moreira J M, Caicedo C, Roncaglioni MC, Colombo Fabio, Camisasca P, Milani V, Quimi' S, Gonzabay F, Tognoni G. Diagnostic evaluation of people with hypertension in low income country: cohort study of &quot;essential&quot; method of risk stratification. BMJ 2008; 337: a1387 Antithrombotic Trialists' (ATT) Collaboration. Aspirin in the primary and secondary prevention of vascular disease: collaborative meta-analysis of individual participant data from randomised trials. Lancet 2009; 373: 1849-1860 Rischio and Prevenzione Investigators. Efficacy of n-3 polyunsaturated fatty acids and feasibility of optimizing preventive strategies in patients at high cardiovascular risk: rationale, design and baseline characteristics of the Rischio and Prevenzione study, a large randomised trial in general practice. Trials 2010; 11: 68

Gianni Tognoni got his Medical Doctor degree in 1970, University of Milan. Main areas of methodology Randomized clinical trials; outcomes studies; pharmacoepidemiology; pharmacoeconomics; epidemiological monitoring and assessment of health care systems, drug policy; genetic epidemiology; community epidemiology; transfer of technology; health and human rights. Main clinical areas Acute and chronic CV diseases; psychiatry; aging; intensive care; neurodegenerative disordes; hematooncology. Position from 2004 Member, Commission of Human Experimentation of the Italian Drug Agency (AIFA) 2001-2003 Member, Commissione Unica del Farmaco (CUF), Ministry of Health from 2002 Director, Consorzio Mario Negri Sud, S. Maria Imbaro, Chieti. 1996-2002 Coordinator, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano from 1990 Co-Director, Scuola Superiore di Ricerca in Medicina Generale (CSeRMEG) from 1976 Founding member of the International Society of Drug Bulletins (ISDB) Coordinator, Commission of Human Experimentation, Regione Lombardia from 1983 Founder and in the Editorial Board of the nursing research Journal Rivista dell'Infermiere/Assistenza Infermieristica e Ricerca from 1977 Consultant to WHO and other UN agencies for drug selection and policy; training in methods of clinical and epidemiological research in developing countries mainly in Latin America and Africa 1976-1999 Head, Laboratory of Clinical Pharmacology of the Istituto di Ricerche Farmacologiche "Mario Negri", Milano from 1975 Head, Regional Centre for Drug Information (CRIF), Regione Lombardia, Istituto di Ricerche Farmacologiche "Mario Negri", Milano 1969-1974 Research Assistant, Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche "Mario Negri", Milano
Selected publications GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R, Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230 GISSI-AF Investigators (Writing Committee: Disertori M, Latini R, Barlera S, Franzosi MG, Staszewsky L, Maggioni AP, Lucci D, Di Pasquale G, Tognoni G). Valsartan for prevention of recurrent atrial fibrillation. N Engl J Med. 2009; 360: 1606-1617 Palmer SC, Navaneethan SD, Craig JC, Johnson DW, Tonelli M, Garg AX, Pellegrini F, Ravani P, Jardine M, Perkovic V, Graziano G, McGee R, Nicolucci A, Tognoni G, Strippoli GF. Meta-analysis: erythropoiesis-stimulating agents in patients with chronic kidney disease. Ann Intern Med 2010; 153: 23-33 Sattar N, Preiss D, Murray HM, Welsh P, Buckley BM, de Craen AJ, Seshasai SRK, McMurray JJ, Freeman DJ, Jukema JW, Macfarlane PW, Packard CJ, Stott DJ, Westendorp RG, Shepherd J, Davis BR, Pressel SL, Marchioli R, Marfisi RM, Maggioni AP, Tavazzi L, Tognoni G, Kjekshus J, Pedersen TR, Cook TJ, Gotto AM, Clearfield MB, Downs JR, Nakamura H, Ohashi Y, Mizuno K, Ray KK, Ford I. Statins and risk of incident diabetes: a collaborative meta-analysis of

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randomised statin trials. Lancet 2010; 375: 735-742 Sud S, Friedrich JO, Taccone P, Polli F, Adhikari NKJ, Latini R, Pesenti A, Gurin C, Mancebo J, Curley MAQ, Fernandez R, Chan M-C, Beuret P, Voggenreiter G, Sud M, Tognoni G, Gattinoni L. Prone ventilation reduces mortality in patients with acute respiratory failure and severe hypoxemia: systematic review and meta-analysis. Intensive Care Med 2010; 36: 585-599 The NAVIGATOR Study Group. Effect of nateglinide on the incidence of diabetes and cardiovascular events. N Engl J Med 2010; 362: 1463-1476

Giovanna Balconi got her degree at the School for Technicians of Biomedical Institutes of the University of Milan, with a specialisation in Histology in the Pathological Anatomy Laboratory of the same University (1968). Main fields of interest Isolation, culture and characterization of peripheral blood circulating progenitor cells of patients with heart failure. Isolation, culture and characterization of peripheral blood circulating progenitor cells in rodents affected by diabetic cardiomyopathy. In vitro culture and characterization of stem cells for repair of myocardial infarction in experimental animal models. Positions from July 2005 Head of Tissue Culture Unit, Cardiovascular Clinical Pharmacology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Oct 1995 - June 2005 Head of Tissue Culture Unit, Vascular Biology Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Dec 1983 - Oct 1995 Head of Tissue Culture Unit, Anticancer Chemotherapy Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Oct 1968 - Nov 1983 Researcher, Anticancer Chemotherapy Laboratory, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy
Selected publications Cattelino A, Liebner S, Gallini R, Zanetti A, Balconi G, Corsi A, Bianco P, Wolburg H, Moore R, Oreda B, Kemler R, Dejana E. The conditional inactivation of the beta-catenin gene in endothelial cells causes a defective vascular pattern and increased vascular fragility. J Cell Biol 2003; 162: 1111-1122 Cusella De Angelis MG, Balconi G, Bernasconi S, Zanetta L, Boratto R, Galli D, Dejana E, Cossu G. Skeletal myogenic progenitors in the endothelium of lung and yolk sac. Exp Cell Res 2003; 290: 207-216 Galli D, Innocenzi A, Staszewsky L, Zanetta L, Sampaolesi M, Bai A, Martinoli E, Carlo E, Balconi G, Fiordaliso F, Chimenti S, Cusella G, Dejana E, Cossu G, Latini R. Mesoangioblasts, vessel-associated multipotent stem cells, repair the infarcted heart by multiple cellular mechanisms. A comparison with bone marrow progenitors, fibroblasts, and endothelial cells. Arterioscler Thromb Vasc Biol 2005; 25: 692-697 Sarto P, Balducci E, Balconi G, Fiordaliso F, Merlo L, Tuzzato G, Pappagallo GL, Frigato N, Zanocco A, Forestieri C, Azzarello G, Mazzucco A, Valenti M T, Alborino F, Noventa D, Vinante O, Pascotto P, Sartore S, Dejana E, Latini R. Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure. J Card Fail 2007; 13: 701-708 Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G. Cardiac mesoangioblasts are committed, self-renewable progenitors, associated with small vessels of juvenile mouse ventricle. Cell Death Differ 2008; 15: 1417-1428 Balconi G, Lehmann R, Fiordaliso F, Assmus B, Dimmeler S, Sarto P, Carbonieri E, Gualco A, Campana C, Angelici L, Masson S, Mohammed SAA, Dejana E, Gorini M, Zeiher AM, Latini R, GISSI-HF Investigators. Levels of circulating pro-angiogenic cells predict cardiovascular outcomes in patients with chronic heart failure. J Cardiac Fail 2009; 15: 747755

Paola Di Giulio got her Nursing Diploma at the Nursing School of Istituto Nazionale dei Tumori in Milano and her Master in Oncology Nursing at Guildford University (UK) in 1995. Main fields of activity Coordination of multicentre and observational studies in cardiology and palliative care. Coordination of nursing networks. Position from March 2001 Associated professor at the Turin University. from 1997 Responsible of the Nursing Research Unit from 1995 Senior researcher of the Cardiovascular Research Department from 1989 Consultant of the Clinical Phrmacology Laboratory since 1998 Coordinator of the Editorial Board of Assistenza Infermieristica e Ricerca

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Selected publications Saiani L, Di Giulio P, Gruppo PARI-FV (Percorsi Assistenziali e Ricerca Infermieristica-Farmaco Vigilanza). Epidemiologia dei problemi assistenziali legati a farmaci e presidi in RSA e distretto. Assistenza Infermieristica e Ricerca 2007; 26: 123-164 Lepore V, Cecchetto G, Di Giulio P, Saiani L, Samarelli V, Saugo M, Romero M, Scurti V, Tognoni G, Valerio M. Et anziana-molto-anziana, e aspettativa di vita? Assistenza Infermieristica e Ricerca 2007; 26: 234-242 Di Giulio P, Toscani F, Villani D, Brunelli C, Gentile S, Spadin P. Dying with advanced dementia in long-term care geriatric institutions: a retrospective study. J Palliat Med 2008; 11: 1023-1028 Amodeo R, De Ponti A, Sorbara L, Avanzini F, Di Giulio P, De Martini M. Come aumentare le conoscenze dei pazienti con cardiopatia ischemica sulla loro malattia? Utilit di un incontro educazionale tenuto da infermieri. G Ital Cardiol 2009; 10: 249-255 Di Giulio P, Pera C, Scarano M, Ferri B, Lepore V, Miani D, Tognoni G. Rapporto finale dello studio QDF (Qualit di vita, Depressione e Funzioni cognitive) nei pazienti con scompenso cardiaco. Assistenza Infermieristica e Ricerca 2009; 28: 5-38 Gouchon S, Gregori D, Picotto A, Patrucco G, Nangeroni M, Di Giulio P. Skin-to-Skin contact after cesarean delivery: an experimental study. Nurs Res 2010; 59: 78-84

Fabio Fiordaliso got his Biological Science degree in 1995 at the University of Milan. Education 1998 Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy 1995 Doctoral degree in Biological Sciences, University of Milan, Italy Main fields of activity Therapeutical potential of stem cell and antioxidant treatments in experimental model of diabetic cardiomyopathy and in primary myocyte cultures exposed to hyperglycemia. Morphological and structrural analysis of cells and tissue by optical, confocal and electron microscopy. Positions from 2007 Head of Bio-imaging Unit, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche Mario Negri, Milan from 2006 Member of the Heart Failure Association (HFA) of the European Society of Cardiology from 2005 Member of the Working group on myocardial function (WG 4) of the European Society of Cardiology from 2005 Member of the steering committee of the Consorzio of Microscopy and Image Analysis (MIA) from 2001 Senior Research Scientist, Laboratory of Cardiovascular Clinical Pharmacology (Department of Cardiovascular Research), Istituto di Ricerche Farmacologiche Mario Negri, Milan 1997-2001 Post-Doctoral Research Fellow at Cardiovascular Research Institute (Department of Medicine), New York Medical College, Valhalla, New York 1994-1997 Research Fellow, Laboratory of Cardiovascular Clinical Pharmacology (Department of Cardiovascular Research), Istituto di Ricerche Farmacologiche Mario Negri, Milan 1992-1994 Research training, Institute of General Pathology, University of Milan (Italy)
Selected publications Fiordaliso F, Cuccovillo I, Bianchi R, Bai A, Doni M, Salio M, De Angelis N, Ghezzi P, Latini R, Masson S. Cardiovascular oxidative stress is reduced by an ACE inhibitor in a rat model of streptozotocin-induced diabetes. Life Sci 2006; 79: 121-129 Fiordaliso F, De Angelis N, Cuccovillo I, Bai A, Salio M, Serra DM, Bianchi R, Razzetti R, Latini R, Masson S. Effect of -adrenergic and renin-angiotensin system blockade on myocyte apoptosis and oxidative stress in diabetic hypertensive rats. Life Sci 2007; 81: 951-959 Latini R, Brines M, Fiordaliso F. Do non-hemopoietic effects of erythropoietin play a beneficial role in heart failure? Heart Fail Rev 2008; 13: 415-423 Balconi G, Lehmann R, Fiordaliso F, Assmus B, Dimmeler S, Sarto P, Carbonieri E, Gualco A, Campana C, Angelici L, Masson S, Mohammed SAA, Dejana E, Gorini M, Zeiher AM, Latini R, GISSI-HF Investigators. Levels of circulating pro-angiogenic cells predict cardiovascular outcomes in patients with chronic heart failure. J Cardiac Fail 2009; 15: 747755 Remuzzi A, Cornolti R, Bianchi R, Figliuzzi M, Porretta-Serapiglia C, Oggioni N, Carozzi V, Crippa L, Avezza F, Fiordaliso F, Salio M, Lauria G, Lombardi R, Cavaletti G. Regression of diabetic complications by islet transplantation in the rat. Diabetologia 2009; 52: 2653-2661 Diomede L, Cassata G, Fiordaliso F, Salio M, Ami D, Natalello A, Doglia SM, De Luigi A, Salmona M. Tetracycline and its analogues protect Caenorhabditis elegans from amyloid-induced toxicity by targeting oligomers. Neurobiol Dis 2010; 40: 424-31

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Serge Masson obtained his doctorate (PhD) in Biochemistry and Cellular Biology in 1990 at the University of Marseilles (France), followed by a postdoctoral stay at the Panum Institute in Copenhagen (Denmark). Education 1988-1990 Doctorate fellow, Faculty of Medicine, University of Aix-Marseilles, France 1990-1993 Post-doctoral Researcher, Panum Institute and Assistant Lecturer, University of Copenhagen, Denmark 1993 Research Scientist, NMR Laboratory, Hospital San Raffaele, Milan, Italy from 1994 Research Scientist, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Main fields of activity Physiopathology, diagnostic and prognostic role of the activation of neuroendocrine systems in cardiovascular disease Position from 2002 Head of the Cardiovascular Endocrine Unit, responsible for Quality Assurance for the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 2002 Tutor of fellows of the School of Specialists in Pharmacological Research, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 2002 Fellows of the American Heart Association (Basic Council) and the Working Group on Myocardial Function of the European Society of Cardiology
Selected publications Latini R, Masson S, Anand I S, Missov E, Carlson M, Vago T, Angelici L, Barlera S, Parrinello G, Maggioni AP, Tognoni G, Cohn J N, Val-HeFT Investigators. Prognostic value of very low plasma concentrations of troponin T in patients with stable chronic heart failure. Circulation 2007; 116: 1242-1249 Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators. Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial). J Am Coll Cardiol 2008; 52: 997-1003 Boccanelli A, Mureddu GF, Cacciatore G, Clemenza F, Di Lenarda A, Gavazzi A, Porcu M, Latini R, Lucci D, Maggioni AP, Masson S, Vanasia M, De Simone G, AREA IN-CHF Investigators. Anti-remodelling effect of canrenone in patients with mild chronic heart failure (AREA IN-CHF study): final results. Eur J Heart Fail 2009; 11: 68-76 Masson S, Aleksova A, Favero C, Staszewsky L, Bernardinangeli M, Belvito C, Cioffi G, Sinagra G, Mazzone C , Bertocchi B, Vago T, Peri G, Cuccovillo I, Masuda N, Barlera S, Mantovani A, Maggioni AP, Franzosi MG, Disertori M,Latini R, on behalf of the GISSI-AF investigators. Predicting atrial fibrillation recurrence with circulating inflammatory markers in patients in sinus rhythm at high risk for atrial fibrillation: data from the GISSI atrial fibrillation trial. Heart 2010; 96: 1909-1914 Masson S, Latini L, Milani V, Moretti L, Rossi M G, Carbonieri E, Frisinghelli A, Minneci C, Valisi M, Maggioni A P, Marchioli R, Tognoni G, Tavazzi L, on behalf of the GISSI-HF Investigators. Prevalence and prognostic value of elevated urinary albumin excretion in patients with chronic HF. Data from the GISSI-Heart Failure (GISSI-HF) trial. Circ Heart Fail 2010; 3: 65-72 Masson S, Latini R, Carbonieri E, Moretti L, Rossi MG, Ciricugno S, Milani V, Marchioli R, Struck J, Bergmann A, Maggioni AP, Tognoni G, Tavazzi L, on behalf of the GISSI-HF Investigators. The predictive value of stable precursor fragments of vasoactive peptides in patients with chronic heart failure:data from the GISSI-heart failure (GISSI-HF) trial. Eur J Heart Fail 2010; 12 : 338-347

Enrico Bjrn Nicolis has attended the courses in Computer Science at the University of Milan. Education 1991-1999 Research fellow, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy Main fields of activity Data management and analysis of randomized clinical trials. Developing of database and tools for studies of population genetics, particularly for linkage analysis. Position from 2001 Head of the Bioinformatics Unit, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1999 Research fellow of the Laboratory of Clinical Drugs Evaluation from 1997 System administrator at the EDP center, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy from 1991 Research fellow at the Medical Informatics and Applied Statistics Unit, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy

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Selected publications Nobili A, Gebru F, Rossetti A, Schettino F, Zahn R W, Nicolis E, Macario G, Celani L, Acik V O, Farina ML, Naldi L. Doctorline: A private toll-free telephone medical information service. Five years of activity: Old problems and new perspectives. Ann Pharmacother 1998; 32: 120-125 Santoro E, Nicolis E, Franzosi MG.Telecommunication technology for the management of large scale clinical trials: The GISSI experience. Comput Methods Programs Biomed 1999; 60: 215-223 Tognoni G, Franzosi MG, Nicolis E, Barlera S, Specchia C, Chiodini B, Crociati L, Ferrario L, PROCARDIS Consortium. A trio family study showing association of the lymphotoxin-alfa N26 (804A) allele with coronary artery disease. Eur J Hum Genet 2004; 12: 770-774 Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: 221-227 Specchia C, Barlera S, Chiodini BD, Nicolis EB, Farrall M, Peden J, Collins R, Watkins H, Tognoni G, Franzosi MG, PROCARDIS Consortium. Quantitative trait genetic linkage analysis of body-mass index in familial coronary artery disease. Hum Hered 2008; 66: 19-24 Disertori M, Latini R, Maggioni AP, Barlera S, Di Pasquale G, Franzosi MG, Lucci D, Staszewsky L, Masson S, Baviera M, Nicolis E, Tognoni G, GISSI-AF Investigators. Valsartan for prevention of recurrent atrial fibrillation. N Engl J Med 2009; 360: 1606-1617

ACTIVITIES
The areas of interest of the Department of Cardiovascular Research include the experimental, clinical, genetic, epidemiological aspects of acute myocardial infarction, cardiac failure, cardiac arrhythmias, as well as the clinical and epidemiological investigation of cardiovascular prevention, hypertension and stroke. Following the successful experience of the GISSI-trials (Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto), the activation of large collaborative networks in the setting of the National Health Service hospitals and in general practice has become a key characteristics of the Department, which can now rely on the permanent collaboration of over 300 clinical groups and of several hundred general practitioners. Over the years, firm links have also been established with international leading research groups. The experimental research activity concerns the physiopathology, the pharmacological modulation and the prognostic role of the activation of the renin-angiotensin-aldosterone system, as well as other neurohormonal systems, in myocardial infarction and heart failure, the physiopathology, the pharmacological modulation and prognostic role of the activation of the inflammatory processes in myocardial infarction and heart failure; a more recent research topic is the cell therapy of experimental myocardial infarction. A model of cardiac arrest and cardiopulmonary resuscitation in the rat has been recently set up and is being used for assessing the role of inflammation in cardiac and brain injury after cardiac arrest. The activity in clinical research includes the clinical assessment of therapeutic strategies and of biomarkers of cardiovascular risk with large scale clinical trials in the field of acute coronary syndromes, congestive heart failure and atrial fibrillation. A recently developing area is the genetic epidemiology of myocardial infarction and heart failure. Several studies have been conducted in the area of clinical epidemiology and risk factors assessment of myocardial infarction. The collaboration with an european genetic network has allowed the participation to large GWAS (genome wide association studies) on coronary disease and myocardial infarction. The collaboration with a large network of General Practitioners in the area of cardiovascular prevention allowed to test new hypotheses through large scale clinical trials and to evaluate the actual transferability of evidence based interventions in the every day practice through epidemiological or outcome research studies. Pharmacoepidemiological studies through the analysis of a large sample of Local Health Units drug prescriptions were also performed. A research network of nurses has been developed with the main focus on the assessment of healthrelated quality of life of patients and on the epidemiology of nursing interventions and their implications for patients' well being and outcomes.

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MAIN FINDINGS
A subgroup analysis of patients enrolled in the GISSI-AF trial has shown that the risk of incident atrial fibrillation is predicted by circulating cardiac markers (natriuretic peptides and troponin T) and by left atrial function as assessed by echocardiography in patients in sinus rhythm. Predictors of atrial fibrillation could help in treating this arrhythmia which has a prevalence of 5-6% in the elderly and is associated with a 10-fold increase in risk of stroke. The PROCARDIS study has identified two single nucleotide polymorphisms (SNPs) at the LPA locus, strongly associated to coronary disease, with an odds ratio of 1.70 (95% CI 1.49-1.95) for rs10455872,and with an odds ratio of 1.92 (95% CI 1.48-2.49) for rs3798220. Both variants were strongly associated also with an increased level of lipoprotein(a), a reduced copy number in LPA, and a small lipoprotein(a) size. These common variants explain over a third of the variance in lipoprotein(a) levels in individuals of European descent. After adjustment for the plasma lipoprotein(a) level, the association between these genotypes and coronary disease was abolished, indicating a causal role of lipoprotein(a) in coronary disease. The GISSI- Prevenzione Genetic study confirmed the results of the PROCARDIS concerning the role of two SNPs located in the chromosome 9p21 region, that are independently associated with CAD and with type 2 diabetes (T2D) respectively.

NATIONAL COLLABORATIONS
AMD (Associazione Medici Diabetologi) - Lombardia ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri) AREU - Azienda Regionale Emergenza Urgenza - Lombardia Azienda Ospedaliera CTO/Maria Adelaide, Torino Centro Cardiologico Monzino IRCCS, Milano CINECA (Consorzio Interuniversitario per il Calcolo Automatico dell'Italia Nord-Orientale) CSeRMEG (Centro Studi e Ricerche in Medicina Generale) Dipartimento Cardio-Vascolare ed Endocrino-Metabolico, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo Dipartimento Cardiologico A. De Gasperis - Struttura Complessa di Cardiologia 2 Insufficienza Cardiaca e Trapianto, Azienda Ospedaliera Ospedale Niguarda Ca Granda, Milano Dipartimento di Cardiologia e UTIC, Istituto Clinico Humanitas IRCCS, Milano Dipartimento di Immunologia, Istituto Clinico Humanitas IRCCS, Milano Ematologia, Ospedale SantAnna, Torino Fondazione Don Gnocchi IRCCS, Milano Gruppi organizzati di MMG (FIMMG, CoS, Ass.Cu.M.I., AMISI) IFOM-FIRC, Milano IRC - Italian Resuscitation Council, Bologna Istituto di Anestesiologia e Rianimazione IRCCS, Ospedale Maggiore Policlinico, Mangiagalli, Regina Elena, Milano Istituto di Anestesia e Rianimazione, Ospedale San Gerardo, Monza Istituto di Ricerca in Cure palliative Lino Maestroni, Cremona Istituto Ortopedico Galeazzi, Milano Istituto Ortopedico Rizzoli, Bologna Laboratorio di Endocrinologia, Ospedale Luigi Sacco, Milano Regione Lombardia

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SIBioC (Societ Italiana di Biochimica Clinica e Biologia Molecolare) SIFO (Societ Italiana di Farmacia Ospedaliera) Stem Cell Research Institute, Ospedale San Raffaele, Milano Unit Operativa Semplice di Neuroanestesia e Neurorianimazione, Dipartimento di Medicina Perioperatoria e Terapie Intensive, Ospedale San Gerardo, Monza Universit degli Studi di Milano, Dipartimento di Medicina Interna Universit degli Studi di Milano Bicocca, Dipartimento di Biotecnologie e Bioscienze Universit degli Studi di Milano, Dipartimento di Scienze Farmacologiche Universit degli Studi di Torino, Dipartimento di Anatomia, Farmacologia e Medicina Forense Universit degli Studi di Torino, Dipartimento di Sanit Pubblica e Microbiologia Universit degli Studi di Verona, Dipartimento di Sanit Pubblica Universit degli Studi di Verona, Istituto di Anatomia Umana

INTERNATIONAL COLLABORATIONS
Cecomet (Centro de Epidemiologia comunitaria y Medicina tropical, Esmeraldas) Ecuador Cochrane Collaboration, Oxford, UK Clinical Trial Research Unit, Auckland University, Nuova Zelanda CNIC Centro Nacional de Investigaciones Cardiovasculares, Madrid , Spain CTSU (Clinical Trial Service Unit) /ISIS (International Studies on Infarct Survival), Oxford, UK Department of Cardiology, Italian Hospital of Buenos Aires, Argentina Department of Epidemiology, Harvard School of Public Health, Boston, USA Division of Genetics and Development, Guy's, King's and St Thomas' School of Medicine, King's College, London, UK DSAN SUPSI (Scuola Universitaria Professioni Sanitarie), Lugano CH ECLA (Estudios Cardiologicos de Latino-America) Karolinska Institutet, Stockholm, Svezia Laerdal Foundation for Acute Medicine, Stavanger, Norway PHRI (Population Health Research Institute), McMaster University, Hamilton, Ontario, Canada SIOP Europe (European Society for Paediatric Oncology) University of Cambridge, UK University of Minnesota, Minneapolis, USA University of Oslo, Norvegia University Medical Center, Groningen, Olanda Wellcome Trust Centre for Human Genetics, University of Oxford, UK WONCA (World Organization of Family Doctors)

EDITORIAL BOARD MEMBERSHIP

Assistenza Infermieristica e Ricerca, European Journal of Cancer Care, European Journal of Oncology Nursing (Paola Di Giulio) European Heart Journal, International Journal of Health Services, Italian Journal of Cardiology(Gianni Tognoni) Italian Resuscitation Council Edizioni (Giuseppe Ristagno) Journal of Cardiac Failure, Journal of Cardiovascular Medicine (Roberto Latini)

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PEER REVIEW ACTIVITIES

American Heart Journal, American Journal of Cardiology, American Journal of Medicine, Archives of Medical Research, Atherosclerosis Thrombosis and Vascular Biology, Biomarkers in Medicine, Canadian Medical Association Journal, Cardiovascular Drugs and Therapy, Cardiovascular Research, Circulation, Circulation Research, Clinical Pharmacology and Therapeutics, Critical Care Medicine, European Heart Journal, European Journal of Cancer Care, European Journal of Cardiovascular Nursing, European Journal of Oncology Nursing, Free Radical Biology & Medicine, Health and Quality of Life, Heart, Heart Vessels, International Journal of Cardiology, International Journal Diabetes in Developing Countries, ISRN Nursing (International Scholarly Research Network), International Journal of Obesity, Intensive Care Medicine, Italian Journal of Cardiology, Journal of American College of Cardiology, Journal of Cardiac Failure, Journal of Clinical Laboratory Analysis, Journal of Internal Medicine, Journal of Cardiovascular Medicine, The Lancet, Life Sciences, Metabolism, PLoS Medicine, PharmacoEconomics, Pharmacological Research, Postgraduate Medical Journal, Redox Report, Value in Health.

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Comitato Etico ASL di Milano Comitato Etico della Regione Lombardia Comitato Etico della Provincia di Trento Comitato Etico della Provincia di Verona Comitato Etico Scientifico dellA.O. Fatebenefratelli e Oftalmico di Milano International Liaison Committee on Resuscitation, Task Force of the 2010 American Heart Association Guidelines for Cardiopulmonary Resuscitation

EVENT ORGANIZATION
Investigator's Meeting Riunione di avvio dello studio REGIA - Rischio Emorragico GInocchio e Anca Studio osservazionale prospettico di coorte sullincidenza degli eventi emorragici nei pazienti sottoposti ad interventi di sostituzione protesica di ginocchio ed anca 28/01/10, Istituto di Ricerche Farmacologiche Mario Negri, Milano Investigator's Meeting Riunione per la presentazione dei risultati dello studio GLICINE SPIDER Gruppo Lombardo per lo studio delliperglicemia nelle sindromi coronariche acute Studio osservazionale prospettico sulla gestione delliperglicemia in corso di sindrome coronarica acuta 08/09/10, Istituto di Ricerche Farmacologiche Mario Negri, Milano Investigator's Meeting Riunione di avvio dello studio BeTACTIC - Best Therapy After Cardiac Transplantation, the Italian Challenge 28/09/10, Centro Congressi Stella Polare Porta Sud, Nuovo Polo della Fiera di Milano, PeroRho Master di I Livello in Ricerca Clinica dellUniversit degli Studi di Milano, Facolt di Medicina e Chirurgia, Dipartimento di Medicina Interna (Anno Accademico 2010-2011). Istituto di Ricerche Farmacologiche Mario Negri, Milano

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02/11/10 03/11/10 04/11/10 09/11/10 10/11/10 11/11/10 15/11/10 16/11/10 17/11/10 22/11/10 29/11/10 30/11/10 01/12/10 02/12/10 09/12/10

Introduzione al corso La ricerca clinica oggi: profit e no-profit Corso di introduzione alla statistica medica La variabilit dei fenomeni biologici Elementi di statistica descrittiva Il disegno dello studio in epidemiologia Farmacovigilanza Misure di rischio in epidemiologia Monitoraggio degli studi clinici no-profit Il disegno degli studi clinici Report delle reazioni avverse negli studi clinici no profit Le interazioni tra farmaci Analisi della sopravvivenza Inferenza statistica Legislazione sulla sperimentazione clinica e ruolo dei Comitati Etici Esercitazione di inferenza statistica Test diagnostici Esercitazioni di statistica descrittiva Ricerca clinica nel capo dell'epilessia. Ricerca clinica nell'ictus Studi di fase 2: obiettivi, disegno e stima del campione in oncologia Ricerca traslazionale. Outcome research Dalla preclinica alla clinica: sviluppo di nuovi farmaci cardiovascolari I bias negli studi clinici controllati Revisioni sistematiche e metanalisi La ricerca bibliografica oggi Regolamentazione dei farmaci in Europa Ricerca in medicina generale Monitoraggio degli studi clinici profit Report delle reazioni avverse Internet e le nuove tecnologie per l'aggiornamento medico-scientifico

CONFERENCE AND WORKSHOP CONTRIBUTIONS

Associazione Internazionale dei Lions Clubs Distretto 108 Ta 1 Italy. Il presente e il futuro delle cellule staminali. Oltre lorizzonte, 23/01/10, Verona, Italy - Cellule per riparare un cuore malato: a che punto siamo? American Heart Association International Liaison Committee on Resuscitation. 2010 International Consensus on CPR & ECC Science with treatment and recommendations. 31/0106/02/10, Addison, Texas USA - Automated vs manual defibrillation European Society of Anaestesiology ESA. 6th EuroNeuro Congress, 04-06/02/10, Porto, Portugal - Evaluation of anesthesiological strategies in elective craniotomy: the Neuromorfeo trial Amici dellIstituto Mario Negri (Delegazione di Genova e Riviera di Levante) Comune di Lavagna. Le malattie cardio-cerebrovascolari: riconoscerle e prevenirle. 16/02/10, Auditorium G.B. Campodonico Lavagna, Italy - Il contributo della genetica alla prevenzione dellinfarto miocardico

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INTERcardio Onlus UO Cardiologia Interventistica, Azienda Ospedaliera S. Camillo Forlanini. INTERcardio 2010. Lespansione delle tecnologie, il ruolo della clinica, oggi. 2123/04/2010, SGM Conference Center Roma, Italy - Atrial fibrillation: new approaches. New anticoagulant drugs Regione Lombardia. Il profilo epidemiologico e il carico assistenziale del diabete nelle ASL della Regione Lombardia. 17/05/10, I.ReF. Istituto Regionale lombardo di Formazione per lamministrazione pubblica, Milano, Italy - Il profilo epidemiologico della popolazione con diabete in Regione Lombardia: analisi dei database amministrativi - Il carico assistenziale della popolazione con diabete in Regione Lombardia: analisi dei ricoveri ospedalieri - Profilo farmaco-epidemiologico della popolazione diabetica: il trattamento diabetico e i farmaci cardiovascolari ANMDO Associazione Nazionale dei Medici delle Direzioni Ospedaliere. 36 congresso Nazionale ANMDO Progettare e Costruire il Futuro. 19-22/05/10, Royal Continental Hotel, Napoli, Italy - Terapia anticoagulante: novit organizzative in vista? Associazione Nazionale Medici Cardiologi Ospedalieri. 41 Congresso Nazionale di Cardiologia, 19-22/05/10, Fortezza da Basso, Firenze, Italy - Infiammazione e scompenso cardiaco GISSI-HF & CORONA - Qualit di vita Depressione Funzioni cognitive QDF - Rete Infermieri GISSI-HF SMART-Organizing and Scientific Committee. 21 SMART Simposio Mostra Anestesia, Rianimazione e Terapia Intensiva, 26-28/05/10, MIC Convention Centre, Milano, Italy - Research in emergency: where is it going? - Whats new in defibrillation - The adrenergic discharge in anesthesia: biohumoral markers Italian Resuscitation Council. IRC 2010 Cardiac arrest, trauma and paediatric emergencies: a multiprofessional health care approach. 04-05/06/10, Sheraton Hotel & Conference Center, Catania, Italy - Is epinephrine the best drug to protect cerebral perfusion during CPR? Committee for European Education in Anaesthesiology - Universit degli Studi di Catania. Anestesia, medicina perioperatoria e terapia intensiva. Corso 2 Ciclo 1, 18-20/06/10 Aula G. Pero - Policlinico G. Rodolico, Catania, Italy - Amplitude Spectrum Area (AMSA) quale indicatore del successo della defibrillazione - Biomarkers del danno cardiaco - Ruolo della capnometria negli stati di shock European Society of Cardiology. Frontiers in Cardiovascular Biology, 16-18/07/10, Berlino, Germany - Effects of dipeptildyl peptidase-4 (DDP-4) inibition on angiogenesis and hypoxy injury in type 2 diabetes - Ex-vivo expanded bone marrow human CD34+ hematopoietic stem cells for repairing myocardial ischemic injury in immunodeficient mice European Society of Cardiology. ESC Congress 2010, 29/08-01/09/10, Stoccolma, Sweden - Tubular damage and outcome in chronic heart failure

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- Prognostic value of osteoprotegerin in chronic heart failure: the GISSI-HF trial - The long-chain pentraxin-3 (PTX3) predicts short-term functional recovery of rehabilitation in patients with cardiac surgery - The added value of depression to NYHA class: the results of the QDF (Quality of life, Depression and Cognitive Function) Study School of Medicine, University Hospital Policlinico di Catania. International symposium of mediterranean school of intensive and critical care medicine, 13/09/10, Catania, Italy - Clinical trials in cardiovascular care - Survival and neurological outcome after nasopharyngeal cooling in experimental model ISMETT Centre for transplantation. International symposium of mediterranean school of intensive and critical care medicine, 14/09/10, Palermo, Italy - Stem cell therapy in cardiac diseases - Hypotherma improves ventricular myocyte contractility European Society for Perioperative Care of the Obese Patients (ESPCOP). Multidisciplinary approach to the obese patient. 2nd Annual Meeting, 18/09/10, Pordenone, Italy - Cardiac arrest in an experimental model of obesity European Society of Cardiology. Biomarkers and cardiovascular personalized medicine. Fifth Annual Meeting: Transatlantic Heart Failure Biomarker Working Group, 16-17/10/10, Cannes, France - Update on novel CV biomarkers - Findingh the high risk CVD patient novel markers Centro di Ricerche del Parco Tecnologico Padano. Genomics for research and molecular diagnostics. European workshop - 5th edition, 21-22/10/10 Lodi (MI), Italy - Common genetics variants on chromosome 9p21 are associated with myocardial infarction and type 2 diabetes in an Italian population European and Mediterranean School of Intensive and Critical Care. Anaesthesia Pharmacology Intensive Care and Emergency. 23rd Annual Meeting, 05-07/11/10 Catania, Italy - Nasopharyngeal cooling for neurological and myocardial outcome in an experimental model - Amplitude spectrum area (AMSA) as predictor of successful defibrillation - Advances in thoracic compression devices - Whats new in defibrillation? EATRIS European Advanced Translational Research Infrastructure in Medicine. Introductory PhD Couse in Translational Medicine, 08-12/11/10 Milano, Italy - Clinical trial in cardiology - Mechanisms by which selective head cooling during cardio-pulmonary resuscitation (CPR) decreases cerebral and myocardial ischemic injury American Heart Association. AHA Scientific Session 2010, 13-17/11/10, Chicago, Illinois, USA - Enhanced cardioprotection by histone deacetylase inhibitor valproic acid-preconditioned human cord blood (UCB)-derived CD34+ cells - Evaluation of the efficacy of aminaftone in a rat model of monocrotaline-induced pulmonary hypertension - High dose of epinephrine administered during cardiopulmonary resuscitation leads to greater oxidative stress following resuscitation from cardiac arrest

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- High sensitivity cardiac troponin T in patients with chronic heart failure: prognostic value of changes over time and effects of randomized therapy - How closely left atrial volume and circulating natriuretic peptides levels relate to the duration of atrial fibrillation recurrences. The results of a GISSI-AF Trial substudy - Worse hemodynamic and greater oxidative stress response in obese rats compared to normal rats following resuscitation from cardiac arrest - AMSA-based shock decision: a human retrospective analyses during pre-hospital CRP intervention - Cold saline infusion during CRP and continued following return of spontaneous circulation does not impair resuscitation nor induce heart failure by volume overload after cardiac arrest - Comparison of defibrillation efficacy between two pads placements in a pediatric pig model of cardiac arrest - Preserved heart rate variability during rapid head cooling correlated to better survival and neurological outcomes in a pig model of cardiac arrest - Selective head cooling initiated during CPR and continued following return of spontaneous circulation improves coronary perfusion pressure and myocardial tissue perfusion after cardiac arrest - Association of plasma n-3polyunsaturated acids levels with metabolic and infiammatory biomarkers in patients with chronic heart failure. Data from the GISSI-HF trial European Resuscitation Council. Resuscitation 10th Scientific Congress. The Guidelines Congress, 02-04/12/10, Porto, Portugal - AMSA for monitoring depth of chest compression during out-of-hospital cardiopulmonary resuscitation

GRANTS AND CONTRACTS

AIFA (Agenzia Italiana del Farmaco), Azienda Ospedaliera Ospedale Niguarda Ca Granda Milano, Azienda Ospedaliera San Gerardo Monza, Chiesi Farmaceutici, Centro Cardiologico Monzino IRCCS Milano, Comunit Europea, CONGENIA, Consorzio Mario Negri Sud Santa Maria Imbaro, Fondazione CARIPLO, Fondazione Don Gnocchi Milano, Fondazione San Raffaele Milano, Heart Care Foundation Firenze, Fondazione Humanitas per la Ricerca Rozzano, Centro Nacional de Investigaciones Cardiovasculares (CNIC) Madrid, International Biomedical System SpA Trieste, IRC-Italian Resuscitation Council Bologna, Istituto Dermopatico dellImmacolata Roma, LACHIFARMA, MEDESTEA Research & Production SpA, Laerdal Foundation for Acute Medicine Stavanger, Ministero della Salute, Novartis Pharma, Ospedale Casa Sollievo della Sofferenza IRCCS San Giovanni Rotondo, Oxford University, Population Health Research Institute-Mc Master University, Pfizer Italia, Regione Lombardia, ROCHE Diagnostics, Sanofi-Aventis, Sigma Tau, SPA Societ Prodotti Antibiotici SpA, Takeda Italia SpA, Universit degli Studi Milano Bicocca, Universit degli Studi Torino, University Medical Center di Groningen.

SCIENTIFIC PUBLICATIONS (2010)

Balducci C, Tonini R, Zianni E, Nazzaro C, Fiordaliso F, Salio M, Vismara L, Gardoni F, Di Luca M, Carli M, Forloni G. Cognitive Deficits Associated with Alteration of Synaptic Metaplasticity Precede Plaque Deposition in AbetaPP23 Transgenic Mice. J Alzheimers Dis 2010; 21: 1367-1381

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Burgess S, Thompson SG, CRP CHD Genetics Collaboration Bayesian methods for meta-analysis of causal relationships estimated using genetic instrumental variables Statist Med 2010; 29: 1298-1311 Chiodini BD, Specchia C, Gori F, Barlera S, DOrazio A, Pietri S, Crociati L, Nicolucci A, Franciosi M, Signorini S, Brambilla P, Franzosi MG, on behalf of GISSI-Prevenzione Investigators and SiBioC-GISSI Prevenzione Group Adiponectin gene polymorphisms and their effect on the risk of myocardial infarction and type 2 diabetes: an association study in an Italian population Ther Adv Cardiovasc Dis 2010; 4: 223-230

Deban L, Castro Russo R, Sironi M, Moalli F, Scanziani M, Zambelli V, Cuccovillo I, Bastone A, Gobbi M, Valentino S, Doni A, Garlanda C, Danese S, Salvatori G, Sassano M, Evangelista V, Rossi B, Zenaro E, Constantin G, Laudanna C, Bottazzi B, Mantovani A Regulation of leukocyte recruitment by the long pentraxin PTX3 Nat Immun 2010; 11 : 328-334 Diomede L, Cassata G, Fiordaliso F, Salio M, Ami D, Natalello A, Doglia SM, De Luigi A, Salmona M Tetracycline and its analogues protect Caenorhabditis elegans from amyloid-induced toxicity by targeting oligomers Neurobiol Dis 2010; 40: 424-431 Disertori M, Lombardi F, Barlera S, Latini R, Maggioni AP, Zeni P, Di Pasquale G, Cosmi F, Franzosi MG, on behalf of the GISSI-AF Investigators Clinical predictors of atrial fibrillation recurrence in the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto MiocardicoAtrial Fibrillation (GISSI-AF) trial Am Heart J 2010; 159: 857-863 Dupuis J, Langenberg C, Prokopenko I, et al, on behalf of the MAGIC Investigators New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk Nat Genet 2010; 42: 105-116 Fosgerau K, Ristagno G, Jayatissa M, Axelsen M, Gotfredsen JW, Weber UJ, Kober L, Torp-Pedersen C, Videbaek C Increased susceptibility to cardiovascular effects of dihydrocapcaicin in resuscitated rats. Cardiovascular effects of dihydrocapsaicin BMC Cardiovasc Disord 2010; 10: 39 Franzosi MG, Latini R Beta-adrenoceptor antagonists and antianginal drugs In: Side effects of drugs. Annual 32. Elsevier, Amsterdam, 2010; 363-369 Ghezzi P, Bernaudin M, Bianchi R, Blomgren K, Brines M, Campana W, Cavaletti G, Cerami A, Chopp M, Coleman T, Digicaylioglu M, Ehrenreich H, Erbayraktar S, Erbayraktar Z, Gassmann M, Genc S, Gokmen N, Grasso G, Juul S, Lipton SA, Hand CC, Latini R, Lauria G, Leist M, Newton SS, Petit E, Probert L, Sfacteria A, Siren AL, Talan M, Thiemermann C, Westenbrink D, Yaqoob M, Zhu C. Erythropoietin: not just about erythropoiesis Lancet 2010; 375: 2142 Ghio S, Scelsi L, Latini R, Masson S, Eleuteri E, Palvarini M, Vriz O, Pasotti M, Gorini M, Marchioli R, Maggioni AP, Tavazzi L, for the GISSI-HF Investigators Effects of n-3 polyunsaturated fatty acids and of rosuvastatin on left ventricular function in chronic heart failure: a substudy of GISSI-HF trial Eur J Heart Fail 2010; 12: 1345-1353 Gori F, Specchia C, Pietri S, Crociati L, Barlera S, Franciosi M, Nicolucci A, Signorini S, Brambilla P, Franzosi MG, for GISSI Prevenzione Investigators and SIBioC-GISSI Prevenzione Group Common genetic variants chromosome 9p21are associated with myocardial infarction and type 2 diabetes in an Italian population BMC Med Genet 2010; 11: 60

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Masson S, Aleksova A, Favero C, Staszewsky L, Bernardinangeli M, Belvito C, Cioffi G, Sinagra G, Mazzone C, Bertocchi F, Vago T, Peri G, Cuccovillo I, Masuda N, Barlera S, Mantovani A, Maggioni AP, Franzosi MG, Disertori M, Latini R, on behalf of the GISSI-AF Investigators Predicting atrial fibrillation recurrence with circulating inflammatory markers in patients in sinus rhythm at high risk for atrial fibrillation: data from the GISSI atrial fibrillation trial Heart 2010; 96: 1909-1914 Masson S, Latini L, Milani V, Moretti L, Rossi MG, Carbonieri E, Frisinghelli A, Minneci C, Valisi M, Maggioni AP, Marchioli R, Tognoni G, Tavazzi L, on behalf of the GISSI-HF Investigators Prevalence and prognostic value of elevated urinary albumin excretion in patients with chronic HF. Data from the GISSI-Heart Failure (GISSI-HF) trial Circ Heart Fail 2010; 3: 65-72 Masson S, Latini R, Anand IS An update on cardiac troponins as circulating biomarkers in heart failure Curr Heart Fail Rep 2010; 7: 15-21 Masson S, Latini R, Carbonieri E, Moretti L, Rossi MG, Ciricugno S, Milani V, Marchioli R, Struck J, Bergmann A, Maggioni AP, Tognoni G, Tavazzi L, on behalf of the GISSI-HF Investigators The predictive value of stable precursor fragments of vasoactive peptides in patients with chronic heart failure: data from the GISSI-heart failure (GISSI-HF) trial Eur J Heart Fail 2010; 12: 338-347 Masson S, Solomon S, Angelici L, Latini R, Anand IS, Prescott M, Maggioni AP, Tognoni G, Cohn JN, on behalf of the Val-HeFT Investigators Elevated plasma renin activity predicts adverse outcome in chronic heart failure, independently of pharmacologic therapy: Data from the Valsartan Heart Failure trial (Val-HeFT) J Card Fail 2010; 16: 964-970 Mauri T, Masson S, Pradella A, Bellani G, Coppadoro A, Bombino M, Valentino S, Patroniti N, Mantovani A, Pesenti A, Latini R Elevated plasma and alveolar levels of soluble receptor for advanced glycation endproducts are associated with severity of lung dysfunction in ARDS patients Tohoku J Exp Med 2010; 222: 105-112 Mehta SR, Tanguay JF, Eikelboom JW, Jolly SS, Joyner CD, Granger CB, Faxon DP, Rupprecht HJ, Budaj A, Avezum A, Widimsky P, Steg PG, Bassand JP, Montalescot G, Macaya C, Di Pasquale G, Niemela K, Ajani AE, White HD, Chrolavicius S, Gao P, Fox KA, Yusuf S, on behalf of the CURRENT-OASIS 7 trial investigators Double-dose versus standard-dose clopidogrel and high-dose versus low-dose aspirin in individuals undergoing percutaneous coronary intervention for acute coronary syndromes (CURRENT-OASIS 7): a randomised factorial trial Lancet 2010; 376:1233-1243 Rischio and Prevenzione Investigators Efficacy of n-3 polyunsaturated fatty acids and feasibility of optimizing preventive strategies in patients at high cardiovascular risk: rationale, design and baseline characteristics of the Rischio and Prevenzione study, a large randomised trial in general practice Trials 2010; 11: 68 Rsj H, Masson S, Latini R, Flyvbjerg A, Milani V, La Rovere MT, Revera M, Mezzani A, Tognoni G, Tavazzi L, Omland T, on behalf of the GISSI-HF Investigators Prognostic value of chromogranin A in chronic heart failure: data from the GISSI-Heart Failure trial Eur J Heart Fail 2010; 12: 549-556 Rysland R, Masson S, Omland T, Milani V, Bjerre M, Flyvbjerg A, Di Tano G, Misuraca G, Maggioni AP, Tognoni G, Tavazzi L, Latini R, on behalf of the GISSI-HF Investigators Prognostic value of osteoprotegerin in chronic heart failure: The GISSI-HF trial Am Heart J 2010; 160: 286-293 Soranzo N, Sanna S, Wheeler E, et al Common variants at 10 genomic loci influence hemoglobin A1C levels via glycemic and nonglycemic pathways Diabetes 2010; 59: 3229-3239

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Shuster M, Lim SH, Deakin CD, Kleinman ME, Koster RW, Morrison LJ, Nolan JP, Sayre MR, on behalf of the CPR Techniques and Devices Collaborators Part 7: CPR techniques and devices: 2010 International Consensus on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science With Treatment Recommendations Circulation 2010; 122 Suppl 2: S338-S44 Sud S, Friedrich J O, Taccone P, Polli F, Adhikari NKJ, Latini R, Pesenti A, Gurin C, Mancebo J, Curley MAQ, Fernandez R, Chang MC, Beuret P, Voggenreiter G, Sud M, Tognoni G, Gattinoni L Prone ventilation reduces mortality in patients with acute respiratory failure and severe hypoxemia: systematic review and meta-analysis Intensive Care Med 2010; 36: 585-599 The CURRENT-OASIS 7 Investigators Dose comparisons of clopidogrel and aspirin in acute coronary syndromes N Engl J Med 2010; 363: 930-942 The FUTURA/OASIS-8 Trial Group Low-Dose vs standard-dose unfractionated heparin for percutaneous coronary intervention in acute coronary syndromes treated with fondaparinux: The FUTURA/OASIS-8 Randomized Trial. JAMA 2010; 304: 1339-1349 Tonutti L, Manzi L, Tacconi MT, Bazzoni G Eicosapentaenoic acid inhibits endothelial cell migration in vitro J Angiogenes Res 2010; 2: 12 Visigalli I, Delai S, Politi LS, Di Domenico C, Cerri F, Mrak E, D'Isa R, Ungaro D, Stok M, Sanvito F, Mariani E, Staszewsky L, Godi C, Russo I, Cecere F, Del Carro U, Rubinacci A, Brambilla R, Quattrini A, Di Natale P, Ponder K, Naldini L, Biffi A Gene therapy augments the efficacy of hematopoietic cell transplantation and fully corrects mucopolysaccharidosis type I phenotype in the mouse model Blood 2010; 116: 5130-5139 Wallentin L, Yusuf S, Ezekowitz MD, Alings M, Flather M, Franzosi MG, Pais P, Dans A, Eikelboom J, Oldgren J, Pogue J, Reilly PA, Yang S, Connolly SJ, on behalf of the RE-LY investigators Efficacy and safety of dabigatran compared with warfarin at different levels of international normalised ratio control for stroke prevention in atrial fibrillation: an analysis of the RE-LY trial Lancet 2010; 376: 975-983

LAY PRESS SELECTION (2010)

Aspromonte N, Di Tano G, Latini R, Masson S, Valle R, Emdin M Ruolo dei biomarcatori per la stratificazione prognostica e la personalizzazione del follow-up nel paziente con scompenso cardiaco G Ital Cardiol 2010; 11: 17s-23s

De Berardis G, Sacco M, Evangelista V, Filippi A, Giorda C B, Valentini U, Tognoni G, Nicolucci A Studio clinico randomizzato sull'efficacia dell'aspirina a basse dosi per la prevenzione degli eventi cardiovascolari nei soggetti con diabete mellito trattati con statine (ACCEPT-D: studio di combinazione di aspririna e simvastatina per la prevenzione di eventi cardiovascolari nel diabete) Ricerca & Pratica 2010; n. 152: 63 Gori F Prevenire l'infarto Elisir di Salute 2010; n. 4 : 30-31 Maione A, Nicolucci A, Craig J C, Tognoni G, Palasciano G, Pugliese G, Procaccini D A, Gesualdo L, Pellegrini F, Strippoli GFM, et al

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Effetti cardio-renali dell'inibizione del sistema renina-angiotensina in soggetti a rischio cardio-renale: studio di confronto randomizzato tra ACE-inibitori, antagonisti recettoriali e terapia combinata con le due classi di farmaci in soggetti con uno o pi fattori di rischio cardiovascolare, positivi allo screening per albuminuria, diabetici e non diabetici Ricerca & Pratica 2010; n. 152: 64 Marchioli R, Filippi A, Maggioni AP, Cricelli C, Modena MG, Roncaglioni MC, Romero M, Tombesi M, Monte S, Tognoni G, et al Epidemiologia del rischio cardiovascolare in Italia: risultati preliminari dello Studio Rischio Cardiovascolare Assoluto-Epidemiologia (RIACE) Ricerca & Pratica 2010; n. 152: 65 Staszewsky L Terapia farmacologica della fibrillazione atriale. Un crescente problema di salute pubblica Informazioni sui Farmaci 2010; 34: 78-84

RESEARCH ACTIVITIES Laboratory of Cardiovascular Clinical Pharmacology The effects of mesoangioblasts and of different progenitor cells on injury after experimental myocardial infarction in the mouse
Many studies have demonstrated that autologous and homologous cells of various origins can repair myocardium damaged due to an acute ischemic insult. Mesangioblasts are potentially interesting when compared with bone marrow precursors because (a) they are easily expanded and (b) obtainable by a biopsy of skeletal muscle in man. Mesoangioblasts isolated from human heart biopsies migrate and home in the heart of immunodeficient mice after coronary ligation, and they survive for at least 4 weeks; however, there are no evidences of myocardial regeneration, and improvement in cardiac function was modest. Experiments are being conducted with mesoangioblasts transfected with lentivirus carrying genes for PlGF and/or MMP-9. The same model of coronary ligation in immunodeficient mice is being used for testing in vivo the angiogenic potential of other human progenitor cells such as CD34+ (collaboration with F. Fagioli, Ospedale S. Anna, Torino), CD133+ (collaboration with M. Pesce and G. Pompilio, Centro Cardiologico Monzino, Milano). Studies are ongoing using genetically modified mesoangioblasts (transfected with lentivirus codifying for PlGF or MMP9) derived from mice in the treatment of experimental myocardial infarction in immunocompentent mice. These studies have been concluded end 2010.

Pulmonary injury by hydrochloric acid in the mouse: a model of Aspiration pneumonitis to test protective interventions
Aspiration pneumonitis (AP) occurs when the acid content of the stomach makes his way through the larynx in the lower respiratory tract. Patients with consciousness disturbance are at risk for this event. Specifically, it has been shown that Pulmonary Aspiration can complicate between 0.47-1.41% general anesthesia procedures. The course of AP can be extremely variable, ranging from the silent aspiration characterized by a modest desaturation to the dramatic sequelae of Acute Lung Injury (ALI) and Acute Respiratory Distress Syndrome (ARDS), requiring prolonged mechanical ventilation and potentially leading to death. In a murine model of monolateral acid instillation established in our laboratory, we have shown the protective effect of exogenous pulmonary surfactant instillation. We are currently working on a model of ventilation-induced lung injury (VILI) to assess the

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effect of exogenous surfactant. A study completed recently has shown that PTX3 modulates inflammation in a murine model of ALI by interacting with P-selectin. The effects of a synthetic surfactant have been compared with those of surfactanct of animal origin in the model of lung acid injury: the two agents have been shown to be equivalent.

Roles of macrophages in cardiac ischemia/reperfusion injury and in cardiac repair

Macrophages either resident or from blood-borne monocytes play several key roles in the response of the heart to ischemic injury. They may be useful in particular during cardiac repair, when collagen is deposited in the scar and neoangiogenesis occurs, stimulated by growth factors produced by macrophages. The aim of the project is to assess the relevance of macrophages in myocardial repair and scar formation after myocardial infarction, in the attempt to dissect the role of inflammatory cells in myocardial injury vs repair. Experiments of experimental infarction in transgenic mice expressing human Dyphtheria toxin receptor (hDTR) in (CD11b-DTR) have shown (a) the halving of circulating monocytes, and hence of cardiac macrophages, but at the same time (b) serious wasting of mice receiving Dyphtheria toxin right after cardiac ischemia. The high mortality did not allow to continue the experiments, and another model based on clodronate liposomes is now being tested. We are now running experiments to evaluate the structural and functional consequences of the reduction in circulating monocytes and in cardiac macrophages induced by clodronate. The animal treated with clodronate showed an elevated mortality after myocardial infarction, due to higher vulnerability. We also observed thrombi in the ventricles of mice treated with clodronate and subjected to coronary artery ligation. We currently running adoptive transfer experiments with mice lacking p50 protein, in collaboration with A. Sica (Istituto Humanitas, Rozzano, MI).

Effects of dipeptidyl peptidase-4 (DPP-4) inhibition on endothelial progenitor cells and hypoxic injury in type 2 diabetes
The present study aimed at evaluating whether the administration to diabetic ob/ob mice of a dipeptidyl peptidase-4 (DPP-4) inhibitor, an antihyperglycemic drug, with the ability to preserve the active form of the SDF-1 (Stromal cell-Derived Factor-1), mediate progenitor cell mobilization, increases EPC (Endothelial Progenitors Cell) circulating levels, which, in turn, may contribute to the regeneration of blood vessels and reducing myocardial ischemic stress induced by strenuous exercise. DPP-4 inhibitor improved glucose tolerance, increased level of circulating EPCs and the production of new capillaries in ob/ob mice. DPP-4 inhibitor treatment in ob/ob mice did not influence the level SDF-1 assessed in the blood by enzyme immunoassay or in myocardium by western blot. In conclusion, the inhibitor of DPP-IV in ob/ob mice improved glucose tolerance in response to an oral glucose challenge and re-established an adequate capillary network in the myocardium of diabetic ob/ob mice by the mobilization/homing of endogenous EPCs reducing the susceptibility to myocardial ischemic injury induced by forced swimming in diabetic ob/ob mice. Nowadays we are trying to identify other physiological substrates of DPP-IV, than SDF-1, involved in the phenomena previously described.

Effects of propionyl-L-carnitine on vascular damage and cardiovascular oxidative stress in type 2 diabetic mouse
The present study aimed at evaluating whether propionyl-L-carnitine treatments (PLC, 200 mg/kg and 1 g/kg) for 8 weeks in obese/diabetic ob/ob mice improved mobilization of endothelial progenitor cells (EPCs) and diminished the vascular damage in myocardium and aorta by reducing superoxide anion production at cellular levels in a model of swimming exercise-induce oxidative stress. PLC did not improve the glucose tolerance in ob/ob mice and did not attenuate the ischemic damage induced by forced swim but significantly reduced the

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production of superoxide anion and increased the number of capillaries in the myocardium of ob/ob mice. Similarly, PLC diminished the reactive oxygen species and the expression of a marker of endothelial damage (ICAM-1, intercellular adhesion molecule-1) in the thoracic aorta of ob/ob mice. In conclusion, two different treatment regimens with PLC reduced oxidative stress in myocardium and thoracic aorta of ob/ob diabetic mice, exerting an angiogenic activity in the diabetic myocardium. Furthermore PLC significantly reduced the expression on endothelial cell membrane of ICAM-1, an adhesion molecule involved in the migration of inflammatory cells and in the detrimental process of atherosclerotic plaque formation.

Preclinical and clinical studies in cardiac arrest and cardiopulmonary resuscitation

Cardiovascular disease remains the leading cause of death in the Western world with 350,000 Americans and 700,000 Europeans sustaining cardiac arrest each year. Instead of the initial success of cardiopulmonary resuscitation, the majority victims die within 72 hours because of severe heart contractile failure due to post-resuscitation myocardial dysfunction. Furthermore, cardiac arrest and cardiopulmonary resuscitation represent a condition of systemic ischemia-reperfusion injury causing multi-organ damage. For this purpose we are currently studying a preclinical model of cardiac arrest and cardiopulmonary resuscitation (CPR) in intact rats or in rats with metabolic syndrome (i.e. obesity, diabetes) aiming to: (a) evaluate inflammatory response and organ dysfunction after return of spontaneous circulation; (b) evaluate success of cardiopulmonary resuscitation manoeuvres and survival after new pharmacological approaches ( i.e. n-3 PUFA, pentraxin PTX3, carbon monoxide). Moreover, the severity of post-resuscitation myocardial dysfunction has been recognized to be related, partially, to the magnitude of the total electrical energy delivered with debrillation. Consequently, the development of a non-invasive and real-time monitoring that allow prediction of outcome of the defibrillation attempt is therefore of great importance in decreasing the debrillation energy. At present, we are evaluating a clinically applicable method based on electrocardiographic analysis of ventricular fibrillation waveform aiming to asses a non-invasive approach in order to guide the priority of interventions, namely chest compression or debrillation (collaborating institutions: Emergency Department, San Gerardo Hospital, Monza and Azienda Regionale Emergenza Urgenza - Lombardia).

GISSI-HF: biohumoral substudy and urinary markers of renal injury

The GISSI-HF trial was designed to assess whether two treatments (a statin and n-3 polyunsaturated fatty acids or PUFA) can improve the prognosis of patients with heart failure of any etiology, with preserved or compromised left ventricular ejection fraction. Main results have been published (Lancet 2008; 372: 1223-1230 and Lancet 2008; 372: 1231-1239). Microalbuminuria (defined as the ratio between urinary concentrations of albumin and creatinine) is being measured in more than 2000 patients enrolled in the GISSI-HF trial as an indicator of renal endothelial dysfunction. Microalbuminuria (albumin/creatinine = 30-299 mg/g) is present in 19% of the patients and is a robust marker of bad outcome. New and early markers of renal tubular injury (KIM-1, N-GAL e NAG) have been assayed in the urine samples. Preliminary analyses indicate that these markers are strongly associated with death, independently of microalbuminuria or renal glomerular filtration rate. Results are being submitted for publication.

PTX-3, a novel long pentraxin is a marker of severity of disease and of outcome in cardiovascular diseases, independent of C-reactive protein
PTX-3 is a novel long pentraxin whose expression is induced by cytokines in endothelial and

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mononuclear cells, mostly in striated muscle and heart, while C-reactive protein (CRP) is mainly synthesized in the liver. PTX3 was shown to peak in plasma around 7 h after onset of symptoms of MI and to be an independent predictor of 3-month mortality. PTX3 has been assayed with a more accurate method in 1200 patients with symptomatic heart failure (GISSIHF) and in 380 patients with atrial fibrillation (GISSI-AF) to explore its role in other cardiovascular diseases. Inflammatory markers do not seem to predict recurrence of AF (Heart 2010; 96: 1909-14). First results indicate that PTX3 is associated with different clinical characteristics in patients with heart failure, including advanced age, ventricular dysfunction, functional class (NYHA class), and comorbidities such as atrial fibrillation and diabetes. PTX3 independently predicts mortality and morbidity in the patients with chronic heart failure. Plasma concentrations of PTX3 have been measured, in collaboration with the laboratory headed by A. Mantovani (Istituto Clinico Humanitas, Rozzano) in samples collected in the GISSI-HF trial, but also in samples collected in another large-scale clinical study (CORONA), that enrolled elderly patients with heart failure of ischemic origin, randomized to rosuvastatin or placebo. Results from both trials will be examined together to evaluate the effect of a statin on PTX3 circulating levels and the prognostic value of this marker in a population of 2690 patients (manuscript in preparation).

Echocardiographic and biohumoral substudy GISSI-AF trial

The GISSI Atrial Fibrillation trial (GISSI-AF) tested the efficacy of valsartan, an angiotensin II AT1-receptor blocker, in the prevention of atrial fibrillation recurrence in 1400 patients. A substudy of the GISSI-AF has recently been concluded; it evaluated the potential role of biohumoral factors and cardiac structural remodelling in the reoccurrence and severity of atrial fibrillation. In approx. 400 patients three serial echocardiographic exams (at randomization, 6 months and 1 year) and contemporaneous blood collection were performed. Left ventricular and atrial dimensions were determined by echocardiography, whereas plasma levels of natriuretic peptides (BNP, NT-proBNP and MR-proANP), troponin T (high sensitive method), stable vasopactive peptides (endothelin-1, Adrenomedullin and vasopressin), and inflammatory markers (C-reactive protein, interleukin-6 and pentraxin-3) have been measured. Besides giving clues on the pathophysiology of atrial fibrillation, the most common arrhythmia in elderly, this substudy aims at providing mechanical insights of the potential benefits of the study drug. The study was completed on January 31, 2008 and results being analyzed. Two papers, one on cardiac markers and the other on inflammatory markers, are published, the first echocardiographic paper is being submitted.

CandHeart: effects of candesartan on BNP and left ventricular function in patients with symptomatic heart failure
Candesartan, an antagonist of angiotensin II type 1 receptors, significantly reduces mortality and morbidity in heart failure, as shown by the CHARM trials. The principal objective of the CandHeart trial is to assess the effects of Candesartan on circulating levels of brain natriuretic peptide (BNP) in patients suffering from CHF with depressed or preserved left ventricular (LV) systolic function. The study enrolled 514 patients in 70 clinical centers with a follow-up of 1year. Serial circulating blood samples and echocardiographic examinations have been performed at baseline and after 3 and 12 months (end of study). Besides BNP, other prognostic biomarkers such as aldosterone and microalbuminuria have been assayed. A paper is being submitted showing a beneficial effect of candesartan on NYHA class, echocardiographic variables and circulating aldosterone.

DyDa: left ventricular dysfunction in diabetes. Prevalence and incidence of left ventricular dysfunction in diabetics patients without clinical cardiac disease

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This is a prospective, multicentric, national and epidemiological trial aimed at evaluating the prevalence of left ventricular dysfunction (systolic or diastolic) in 1000 patients with type 2 diabetes mellitus but no clinical cardiovascular disease at enrolment. The incidence of left ventricular dysfunction is monitored during a 2-year follow-up using ECG and echocardiography. The Biomarker Core Laboratory evaluated the biohumoral profile of these patients at study entry, measuring the circulating levels of brain natriuretic peptide, C-reactive protein, microalbuminuria and glycated hemoglobin. Enrolment started in July 2006 and ended in March 2008 with 970 patients recruited. The assays of the biomarkers are concluded and first data on the association between theses markers and baseline clinical characteristics are under analysis. The follow-up phase of the study should be concluded in 2010 to get data on incident left ventricular dysfunction. Two manuscripts on baseline data have been published.

Albumin Italian Outcome Sepsis Study. The ALBIOS Study (AIFA)

ALBIOS is a multicenter, controlled, randomized clinical trial that compares the efficacy of human albumin and a crystalloid solution for volume replacement in patients with severe sepsis or septic shock. The primary endpoint is survival at 28 and 90 days after enrolment. Secondary endpoints include the number of organ dysfunctions, severity of organ dysfunction (SOFA scale), and lengths of stay in (intensive care unit) ICU and in hospital. More than 150 ICU in Italy are expected to participate to this large study, coordinated by the Ospedale Maggiore Policlinico in Milan and the Consorzio Mario Negri Sud. A group of 48 ICUs participates to a biomarkers substudy, coordinated by the laboratory of Clinical Cardiovascular Pharmacology, with the aims of enrolling 800 patients. Serial blood samples are collected to measure the possible effects of albumin on markers of inflammation, infection, cardiac function and coagulation. 1350 patients have been randomized by the end of 2010, and blood samples collected in at least 600 of them.

Evaluation of different anesthesiological strategies for supratentorial neurosurgery. The NeuroMorfeo Study (AIFA)
The aim of the study was to evaluate whether an anesthesia with volatile anesthetics is equivalent to endovenous anesthetics for elective supratentorial surgery. This was a multicenter, randomized, controlled and opened study, based on a design of equivalence for comparison of different anesthesiological strategies (see Laboratory of Clinical Drug Evaluation for details). The biohumoral response to surgical stress was measured as an indicator of homeostasis and neurovegetative status. The urinary excretion of catecholamines and cortisol and the plasma concentration of cortisol were be measured in a central laboratory in collaboration with the Laboratory of Pharmacokinetics and Clinical Chemistry from the Istituto Mario Negri at Ranica. A manuscript is being submitted for publication.

Prevalence of asymptomatic cardiac dysfunction and heart failure in a population of elderly subjects from Lazio. The PREDICTOR Study
This observational study aims at evaluating the prevalence of asymptomatic cardiac dysfunction and heart failure in a random sample of elderly subjects from the Lazio area. The secondary objective is to identify clinical, biohumoral (natriuretic peptides) and non-invasive instrumental (echocardiography and ECG) markers of asymptomatic cardiac dysfunction and heart failure. The population under observation is a randomly selected sample of elderly subjects (age ranging from 65 and 84 years) resident in the area of 10 hospital cardiology centers. In a first phase, 1000 subjects have been recruited; a second phase has recently started with the objective of enrolling another 2000 subjects. Blood samples are collected for each subject and stored in the biobank in the Laboratory of Clinical Cardiovascular Pharmacology. Preliminary data obtained in the first 1000 subjects show a good pathophysiological agreement between the circulating levels of NT-proBNP and indexes of left or right asymptomatic cardiac dysfunction or the

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severity of heart failure. By the end of 2010, more than 2000 subjects have been enrolled, a figure close to the final objective of this study. A paper is in preparation. Further assays are being performed on new markers of cardiac dysfunction and of cardiovascular risk.

OPERA: Omega-3 Fatty Acids for Prevention of Post-Operative Atrial Fibrillation

Peri-operative administration of n-3 polyunsaturated fatty acids (PUFA) may significantly reduce the incidence of post-operative atrial fibrillation (AF) in patients undergoing cardiac surgery (CAS). The aim is to determine, using a randomized, double-blind, placebo-controlled, clinical trial, whether peri-operative administration of n-3 PUFA (8 g total pre-op and then 2 g/d for 14 days or until hospital discharge) reduces the incidence of AF in 1,516 patients undergoing CAS. It is a multinational, multicenter, double-blind, parallel design clinical trial enrolling 1516 patients undergoing CAS in 40-50 major medical centers in the U.S., Argentina and Italy. A core laboratory for Italy and Argentina is at Mario Negri. By the end of 2010, 137 pts have been enrolled, and their biologic samples have been collected.

GISSI-outliers: the CAPIRE study

Aim of the CAPIRE study is to analyze in patients undergoing coronary multislice tomography the extreme populations: patients without risk factors for coronary disease, but with severe coronary lesions, and patients with risk factors, but no major coronary lesions. A panel of biomarkers and genetic polymorphisms will be assayed in the attempt to identify protective factors/mechanisms.

Laboratory of Clinical Drug Evaluation PROCARDIS: A genome-wide strategy to identify susceptibility loci in precocious coronary artery disease
The PROCARDIS research programme, a genome-wide strategy to identify susceptibility loci in precocious coronary artery disease (CAD) supported by the 5th Framework Programme of the EC, was initiated as a collaboration between the Universities of Oxford and Munster, the Karolinska Institute, and the Mario Negri Institute with the support of the GISSI group. The objectives of the first stage of this programme were to collect a minimum of 2000 affected sibling pairs (ASPs) and families with precocious CAD and to apply genome-wide linkage mapping techniques, to identify chromosomal regions linked to the susceptibility to early-onset CAD. The PROCARDIS collected 2036 CAD families from four European countries, in order to maximise the power of detecting genes that confer modest risks. A genome-wide linkage scan identified three promising regions for intensive study. Extensive clinical and biochemical intermediate phenotype data have also been collected and assessed. The second stage of PROCARDIS, supported by the EC 6th Framework Programme, is conducting a large GWAS (genome wide association study), where the patients with myocardial infarction enrolled in the first stage are compared with control subjects to identify novel candidate genes. The results are replicated in different populations. The PROCARDIS study identified risk loci for coronary disease by using a novel gene chip consisting of 48,742 SNPs for 2100 candidate genes that were selected for their potential relevance to coronary artery disease. With this gene chip, PROCARDIS confirmed the previous identification of three chromosomal regions that were correlated with the risk of coronary disease: 6q2627, 9p21, and 1p13. In the chromosome 9p21 region PROCARDIS has identified two SNPs, that are independently associated with CAD and with type 2 diabetes (T2D) respectively. Recently, the PROCARDIS study has identified two single nucleotide polymorphisms (SNPs) at the LPA locus, strongly associated to coronary disease. Both variants were associated also with an increased level of lipoprotein(a), a

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reduced copy number in LPA, and a small lipoprotein(a) size. These common variants explain over a third of the variance in lipoprotein(a) levels in individuals of European descent. After adjustment for the plasma lipoprotein(a) level, the association between these genotypes and coronary disease was abolished, indicating a causal role of lipoprotein(a) in coronary disease.

GISSI-HF Genetic Substudy

The GISSI (Gruppo Italiano per lo Studio della Sopravvivenza nell'Insufficienza cardiaca) is a collaborative group endorsed by ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri) and by the Istituto Mario Negri, active from 20 years in the cardiovascular research field. The GISSI-HF was the fifth large scale clinical trial conducted by the Group and was a prospective, multicenter, randomized, double blind, placebo controlled study, with randomized allocation of patients with a clinical diagnosis of heart failure to n-3 PUFA and/or to rosuvastatin to assess the effects of long-term administration of n-3 PUFA and/or rosuvastatin on all-cause mortality and cardiovascular hospitalizations. The study randomized more than 7000 patients with the participation of 357 departments of cardiology; in a follow-up time of four years, 27% of patients in the PUFA group died, compared with 29% in the placebo group, meaning a relative risk reduction of 9% in the PUFA group. A higher proportion of patients in the placebo group died or were admitted to hospital for cardiovascular reasons than in the PUFA group. Statin treatment with rosuvastatin did not affect clinical outcomes in patients with chronic heart failure. Several substudies focus on possible mechanistic effects of the study treatments. Among them a genetic substudy conducted by nearly 100 Centres that have included 2500 patients, gives the opportunity to improve knowledge on the role of genetic factors involved in heart failure, through a collection of blood samples of a large population of patients, involving cases of heart failure of different etiologies, i.e. non-ischaemic and ischaemic heart disease. The role of genetic factors in causes, evolution, prognosis and treatment of heart failure is largely unexplored, with the exception of heart failure originated by specific cardiomyopathies (such as dilated, hypertrophic, arrhythmogenic right ventricular cardiomyopathies), for which the role of heritable gene mutations is increasingly well understood. Heart failure (HF) is a syndrome with different etiologies, and more than one half is caused by coronary heart disease (CHD). The objective of the genetic substudy is 1) to assess the relationships between the polymorphysms of various candidate genes and the clinical outcome in patients enrolled in GISSI-HF study; 2) to assess whether these relationships are modified by the experimental treatments.

GISSI-Prevenzione-Genetic Study
Myocardial infarction is a multifactorial disease. While the role of known risk factors on coronary heart disease susceptibility is well defined, the impact of the genetic components and its interaction with environmental factors need investigation. The GISSI-Prevenzione trial investigated the effects of pharmacological treatments with n-3 PUFA and pravastatin on morbidity and mortality after myocardial infarction. During the study more than 8000 samples of a large population of patients affected by this disease have been collected and stored with the collaboration of SIBioC (Societ Italiana di Biochimica Clinica e Biologia Molecolare). The GISSI-Prevenzione-Genetic Study investigates the role of genetic factors in ischaemic heart disease. The objectives of the project are 1) to assess the relationships between the polymorphysms of various candidate genes and the clinical outcome in patients enrolled in the large clinical trial GISSI-Prevenzione study; 2) to assess whether these relationships are modified by the pharmacological treatments. According to these objectives, we investigated the relationship between APOE, mortality and the response to treatment in 3300 myocardial infarction survivors randomized to pravastatin or no treatment. We found that epsilon 4 allele is a determinant of pravastatin response in terms of survival (Eur Heart J 2007; 28:1977-1983).

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Association studies in the same population on the adiponectin gene variants, the CRP (Creactive protein) gene variants, some genetic variants on Chromosome 9p21 have been conducted. A genetic assessment of PTX3 protein, a novel long pentraxin whose expression is induced by cytokines in endothelial and mononuclear cells, and involved in the atherogenesis process, is ongoing in collaboration with the Istituto Clinico Humanitas and the IRCCS San Giovanni Rotondo.

Evaluation of different anesthesiological strategies for supratentorial neurosurgery. The NeuroMorfeo Study (AIFA)
NeuroMorfeo is a multicenter, randomized, controlled and open study, based on an equivalence design, with the aim to assess whether an anesthesia with volatile anesthetics is equivalent to endovenous anesthetics for elective supratentorial surgery. The study is financed by the AIFA and is coordinated by the San Gerardo Hospital in Monza, with the support of the Department of Cardiovascular Research of the Mario Negri Institute. The study recruitment has been completed by 14 neurosurgical centers in Italy that have randomized 411 patients admitted for elective intracranial surgery with supratentorial lesions without signs of endocranial hypertension (range of age 18-75 years). Statistical analyses and drafting of manuscripts are ongoing.

BeTACTIC Study: Best Therapy After Cardiac Transplantation, the Italian Challenge
BeTactic is a multicenter, randomized, no-profit trial funded by the National Health Service. The study compares efficacy and safety of Everolimus (Ev) and Mycophenolate (MMF) in association with Cyclosporine (CyA) in pts with acute multiple/late rejection, cardiac allograft vasculopathy (CAV), renal dysfunction after cardiac transplantation (HTx). Survival after HTx has improved, while the attrition rate beyond the 1st year did not change substantially. CAV and cancer are the leading causes of death late after HTx. Many factors as acute rejections and citomegalovirus infections are involved in CAV pathogenesis. Cancer shows higher incidence in immunosuppressed pts. Significant morbidity/mortality derive from renal insufficiency and vascular complications. Ev and MMF were adopted due to better efficacy vs Azathioprine in de novo HTx. However, Ev and MMF have not been tested in a head to head comparison late after HTx. The planned length of the BeTACTIC study is 5 yrs. Patients will be enrolled at least 1yr after HTx. A total of 400 pts will be randomized in 11 Transplant Centers in Italy. BeTACTIC is coordinated by the Cardiology Depatment, Trapianti e Insufficienza Cardiaca, Ospedale Niguarda Ca' Granda di Milano, Milano, in collaboration with the Laboratory of Clinical Drug Evaluation of the Istituto Mario Negri.

REGIA - Rischio Emorragico GInocchio e Anca Assessment of the hemorrhagic risk of treatment with low molecular weight heparins, oral anticoagulants, antiplatelet drugs in patients undergoing total hip or knee replacement surgery.
Major orthopedic surgery is as a high-risk event for venous thromboembolism (VTE). The anticoagulant prophylaxis reduces the risk of postoperative VTE by 50 to 70%. Major bleeding is a possible complication of thromboprophylaxis with an estimated frequency of 1% to 3% in randomized clinical trials (RCT). However, in clinical practice, the estimates may be argued since: 1) bleeding rates are probably underestimated in RCT, due to frequent exclusion of the high risk patients; 2) definition of bleeding is non-standardized and can vary from study to study; 3) the type of intervention, of anesthesia, of prophylactic agent and the timing of administration in relation to surgery may influence bleeding rate. There is scarce information on the frequency of

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bleeding after hip or knee replacement in routine practice in Italy. The objectives of the study are the incidence of major and minor bleedings in the first three months after surgery. The REGIA study is funded by the National Health Service and will collect data on bleedings in near 4000 patients admitted for hip or knee replacement in one year in the three participating hospitals (Istituto Ortopedico Galeazzi, Milano; Istituto Rizzoli, Bologna; CTO Maria Adelaide, Torino).

Collaboration with the Population Health Research Institute (PHRI)

The Population Health Research Institute (PHRI), McMaster University, Hamilton, Ontario, is the coordinating center of a multinational network of cardiology clinics that collaborate to multicenter large scale clinical trials (nearly 40 Countries and more than 600 cardiology clinics). The Laboratory of Clinical Drug Evaluation is responsible for the scientific coordination in Italy of some of these trials (INTER-HEART, CURE, ACTIVE, RE-LY, CURRENT, OASIS-8 FUTURA, AVERROES, RE-LY Registry, RIVAL). The state of advancement of the studies is as follows:

RE-LY Study (Randomized Evaluation of Long term anticoagulant therapY)

Non-valvular atrial fibrillation is implicated in nearly 15% of strokes. Dose-adjusted warfarin decreases the risk of stroke by 62%. However, in practice, the risk of bleeding, the variability of anticoagulation intensity and the need of frequent monitoring and dose adjustments limit the treatment with warfarin, leaving patients outside the therapeutic range almost half the time. Underuse of warfarin in patients with atrial fibrillation at high risk of bleeding calls for safer, more reliable alternatives. For these reasons an international multicentre, randomized, active controlled, parallel group, non-inferiority, clinical trial (RE-LY study) was designed to evaluate the efficacy and safety of dabigatran etexilate, a direct thrombin inhibitor, compared with open label adjusted warfarin for the prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation. The study recruited more than18000 patients. The median duration of the follow-up period was 2.0 years. The results, published on New Engl J Med in 2009, showed that dabigatran given at a dose of 110 mg was associated with rates of stroke and systemic embolism similar to those associated with warfarin, as well as lower rates of major hemorrhage. Dabigatran administered at a dose of 150 mg, as compared with warfarin, was associated with lower rates of stroke and systemic embolism but similar rates of major hemorrhage. The direct thrombin inhibitor dabigatran may offer fixed oral dosing without need for coagulation monitoring, rapid onset and offset of action, stable pharmacokinetics with little potential for drug interactions, and no known food interactions. Several secondary and subgroup analyses of the RE-LY study are ongoing.

AVERROES Study (Apixaban VErsus ASA to Reduce the Rate Of Embolic Stroke)
Although vitamin K antagonists (VKA) are effective for preventing stroke or systemic embolism in patients with atrial fibrillation (AF), complexity of use and bleeding risk limit their potential benefit. Many patients not treated with VKA receive aspirin. There are two main groups of patients with AF who could benefit from a better antithrombotic than ASA: (1) Those not expected to do well on VKA; and (2) Those with only a moderate risk for stroke. Aspirin is presently the only alternative to VKA to prevent stroke in patients with AF but is relatively ineffective, reducing the risk of stroke or systemic embolism by about one-fifth compared with a two-thirds reduction by VKA. New treatments that are more effective than ASA but do not share the many limitations of VKA are required. AVERROES is the only study

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of a new anticoagulant that directly addresses this important unmet need. 5600 patients with AF not suitable for VKA, have been randomized to receive a new oral anticoagulant, Apixaban, or aspirin. Apixaban, an oral inhibitor of the Xa factor, is simple to use, convenient, does not require laboratory monitoring, does not interact with foods, and has very few drug interactions. AVERROES trial has been completed, showing that Apixaban is an effective alternative to ASA for those unsuitable for oral anticoagulation and also for those at moderate risk.

CURRENT OASIS-7 Study

OASIS 7 is a randomized, multinational, 2X2 factorial design, parallel-group, double-blind study, comparing a high loading dose regimen of clopidogrel versus standard dose and high dose regimen of aspirin versus standard dose, in patients with acute coronary syndrome (ACS) managed with an early invasive strategy. The study published on the New Engl J Med in 2010, recruited more than 25000 patients in 800 clinical centers worldwide, 17232 of them undergone planned angioplasty (PCI). The primary objective of the study was to determine whether a high dose regimen of clopidogrel is superior to a standard dose of clopidogrel in preventing CV death, myocardial infarction or stroke and to determine if high dose of aspirin is as safe as low dose in terms of TIMI major bleeding rate. In terms of efficacy, there was no significant difference in the primary outcome or its components, although there was a numerical reduction with the higher dose of aspirin. However, doubling the loading and maintenance doses of clopidogrel in ACS patients undergoing planned PCI significantly reduced stent thrombosis and cardiovascular events, largely driven by reductions in MI, without a significant increase in major bleeding.

FUTURA OASIS-8 Study

The FUTURA-OASIS-8 (FondaparinUx Trial with Unfractionated heparin (UHF) during Revascularization in Acute coronary syndromes) study will expanded the safety experience in UA/NSTEMI patients initially treated with fondaparinux and undergone PCI with adjunctive UFH, while also addressing the question of the optimal dosing strategy with adjunctive UFH during the procedure. The study design consisted of: An international, prospective cohort study of high risk patients presenting to hospital with UA/NSTEMI who are treated with s.c. fondaparinux as initial medical therapy and referred for early coronary angiography and potentially PCI. A double-blind, international, randomized, parallel-group study evaluating standard versus low dose adjunctive i.v. UFH in those patients where a PCI procedure is indicated. The main outcome measure was a composite of major bleeding, minor bleeding, or major vascular access-site complications up to 48 hours after PCI. The results, published on JAMA in September 2010, showed that low-dose compared with standard-dose unfractionated heparin did not reduce major peri-PCI bleeding and vascular access-site complications. Therefore, patients with acute coronary syndromes treated with fondaparinux and undergoing PCI, should receive the guideline-recommended standard dose of unfractionated heparin.

RE-LY AF Registry: Risk Factors, Treatments and Outcomes for Emergency Department Patients with Atrial Fibrillation in Multiple Regions of the World
Due to variations in medical practice and access to care, there will be geographical variation in presentation and management of patients with atrial fibrillation. They will have different predisposing conditions, presenting symptoms, rates of adverse outcomes and will be managed differently. The RE-LY AF Registry has the following objectives: 1. To determine variations in the predisposing conditions for atrial fibrillation and atrial flutter (AF/flutter) between different regions of the world and practice settings. 2. To document regional variations in the management of AF/flutter and associated cardiovascular disease, including the frequency of

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anti-thrombotic and anti-hypertensive therapy and the degree of INR control. 3. To document differences in the adverse cardiovascular outcomes of AF/flutter. The study will recruit 15000 patients with documented atrial fibrillation or atrial flutter at the time of an emergency department visit for any reason, in approximately 300 participating clinical centres in multiple regions of the world. Patients will be followed-up at one year after the enrolment in the study.

RIVAL: Radial vs. Femoral PCI Access Site Substudy

Major bleeding is the major common complication in patients with acute coronary syndrome (ACS). Major bleeding is independently associated with an increased risk of ischemic events (MI and stroke) and mortality. A significant proportion of major bleeding events can be related to the femoral arterial puncture site for invasive procedures. Femoral access has been the dominant method of access for coronary procedures for many years. Radial artery vascular access has been developed as an alternative approach with potentially less major bleeding. The RadIal Vs. femorAL (RIVAL) access site study was started as a randomized sub-study of the OASIS 7-CURRENT and extended beyond its conclusion in order to reach the recruitment goal of 6,000 patients randomized to to either radial or femoral approach for vascular access for coronary angiography and PCI. The primary objective of RIVAL is to determine if radial instead of femoral access for coronary angiography or intervention in patients with acute coronary syndromes reduces the rate of death, MI, stroke or non-CABG related major bleeding. The study has been completed with 7021 patients randomized and data analysis is ongoing.

Laboratory of General Practice Research Risk and Prevention Study (R&P)

R&P is a study on the optimization of cardiovascular prevention of subjects at high risk performed at national level by General Practitioners. Study objective and design - Controlled clinical trial, double-blind and randomised, of the efficacy of a n-3 PUFA treatment in reducing the incidence of cardiovascular events, both fatal and non-fatal, in a population defined as at high risk by participating GPs. - Practicability and overall yield of the preventive interventions adopted (outcome study) The epidemiological and care history of this population shall form the object of a specific evaluation according to a plan of formal predefined analyses. Study population Inclusion criteria Among the subjects deemed by GPs to be at high cardiovascular risk, patients are selected if presenting: - multiple risk factors (e.g. hypertension, hypercholesterolemia, diabetes, smoking, family history of myocardial infarction, obesity, sex and old age) - previous cardio-cerebrovascular events or clinical manifestations of atherosclerotic disease (stroke, TIA, peripheral arteriopathy, previous arterial revascularisation procedures, angina pectoris). Exclusion criteria - serious co morbidity with an unfavourable prognosis over the short term (e.g. cancer) - expected non-compliance over a long period of time; contraindications (known allergies to n3PUFA) - indications (previous MI) for treatment with n-3 PUFA. Efficacy measures The primary objective is to evaluate if a long-term administration of n-3 PUFA is more effective than the corresponding placebo in reducing cardiovascular mortality and hospitalization for cardiovascular causes (primary end-point).

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Randomisation is central, stratified by GP. The experimental treatment consists of one capsule containing 1g of n-3 PUFA, or the corresponding placebo, to be taken daily. The duration of follow-up is 5 years. In order to document with sufficient statistical reliability that the experimental treatment with n3 PUFA reduces of 15% the incidence of the events considered in the primary end-point, a total of 12,000 patients is required. Up-date of the study: From February 2004 to March 2007 12,521 patients have been enrolled by a network of 860 GPs. The Local Health Authorities involved are 57 and in each one investigators meeting has been organized. The characteristics of the population so far enrolled are the following: mean age 65 years, males 62%, hypertension 79%, hypercholesterolaemia 62%, diabetes 56%, smokers 16%, obesity 35%, family history of premature myocardial infarction 20%. Twenty five% of patients have a clinical manifestation of atherosclerotic diseases, 50% have diabetes in association with another risk factor and 23% have multiple risk factors. The trial will continue until the minimum expected number of 1,383 events will occurred. More information available on the website www.rischioeprevenzione.it.

Epidemiological and clinical profile of diabetic patients in Lombardy Region using administrative databases.
The study is part of an ongoing pharmacoepidemiological project in collaboration with the Health Department of the Lombardy Region. Its main objective is the definition of a model to assess and control the use of health resources of diabetic patients by means of integrated administrative database. Specific aims of the study are: To describe prevalence, incidence, hospitalization and mortality of the diabetic population each year, from 2000 to 2009. To assess the prescriptions of both anti-diabetic and cardiovascular drugs Diabetic patients have been identified each year if they met one of the three following criteria: a prescription of an A10 drug: insulin and/or oral glucose lowering agent ; - the occurrence of at least one hospitalization with Disease Related Group (DRG)=294 or DRG=295; presence of the exemption code number 013.250 indicating diabetes. Data from prescription database, hospital admission and outpatient clinic visits and examinations were also included in the analysis via linkage to the personal identification number (national identifiers). Results are in the processing phase.

GLICINE-SPIDER Study
Glicine-Spider is an observational study, carried out in the Coronary Care Unit (CCU) of Lombardy. The protocol is a collaboration between the ANMCO (Italian Association of Hospital Cardiologists) Lombardia , AMD (Association of Medical Diabetologists) Lombardia and the Mario Negri Institute. The study is coordinated by the General Practice Research Laboratory and the Clinical Drug Evaluation Laboratory. Hyperglycemia at the onset of an acute coronary syndrome (ACS) constitutes a negative prognostic factor in diabetic and non-diabetic patients and a poor control of blood glucose in the early hours after hospital admission for ACS is an additional unfavourable prognostic factor. Recent guidelines, although recognizing the importance of controlling blood glucose in ACS, do not clearly define therapeutic strategies to apply and target range. Patients with and without diabetes hospitalized in CCU for a confirmed ACS. The aim of the study is to describe in a large sample of patients hospitalized in CCU for a ASC: the prevalence of diabetes and hyperglycemia the type of treatment and blood glucose control during the acute phase

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the incidence of mortality and cardiovascular complications occurred during the hospitalization according to diagnosis and blood glucose level From May 2009 to April 2010, 1,282 patients have been included from 31 CCUs. The data analysis is in progress.

FOCUS Study (Fixed Dose Combination Drug for Secondary Cardiovascular Prevention. Improving Equitable Access and Adherence to Secondary Prevention Therapy with a Fixed-Dose Combination Drug)
The study is supported by the 7th Framework Programme of the EC. Several randomized controlled trials and metaanalyses have demonstrated that the long term administration of aspirin, statins beta-blockers, and agiontensin converting enzyme inhibitors (ACE inhibitor) improve prognosis in high risk patients, particularly those recovering from an acute coronary event. However, wide variability in the pattern of prescription among physicians, limited access to expensive drugs in emerging countries, and poor adherence to medications limit the use of these drugs and the efficacy of cardiovascular prevention. A Fixed Dose Combination (FDC) pill for cardiovascular prevention was first proposed by Wald and Law in 2000 and supported by the WHO. During the last few years this concept, particularly in the field of primary prevention has been questioned by some experts while the potential role of a polypill for secondary cardiovascular prevention is receiving increasing attention. However, a direct proof of the polypill effect on patients adherence is still lacking. The global objective of the FOCUS consortium is to make FDC drugs for secondary cardiovascular prevention available throughout the world at a low price, in order to improve access to treatment in developing countries improving adherence to medication. The Study is international, multicenter in two phases: Phase 1 is a descriptive, non-interventional study. Its aim is to provide a comprehensive analysis of factors precluding adequate secondary prevention, including health system characteristics, drugs affordability and availability, as well as patients characteristics. Phase 2 is an interventional, randomized, two-arm study. Patients will be randomized to receive a FDC of ramipril, simvastatin and acetilsalycilic acid or the three medications separately. The primary objectives is to compare the adherence to treatment in post myocardial infarction patients receiving a FDC vs those with conventional treatment (3 drugs separately). Secondary objectives are to evaluate the effect of a FDC on blood pressure control and lipid profile and the safety and tolerability of FDC treatment. Two countries in Europe (Spain and Italy) and three in South America (Argentina, Brazil e Paraguay) are involved in the Study. A total number of 4,000 subjects will be included in phase 1 and 1,340 in the phase 2. The study will start on March 2011.

The stratification of global cardiovascular risk in hypertensive patients of the district of Borbon Ecuador
The Laboratory is involved in a collaborative project with the Cecomet (Centro de Epidemiologia comunitaria y Medicina tropical) in Esmeralda, Ecuador, on the prevalence and treatment of hypertension in the district of Borbon, a rural zone of Ecuador in the northern part of the country. In this area, 36% of the adult population is affected by hypertension and more than half of hypertensive patients present blood pressure levels > 160/110 mmHg. From 2001, in the District is ongoing an intensive follow-up of the hypertensive population with the following aims: to evaluate the global cardiovascular risk of the population, to better control blood pressure levels increasing the number of subjects treated with hypertensive therapy (in particular those at high cardiovascular risk) and monitoring of the clinical complications. Preliminary data show that: Patients treated with hypertensive therapy are increased from 39% to 59%

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Antihypertensive drugs are mainly prescribed to subjects with high blood pressure levels (80% of those with systolic blood pressure >180mmHg are actually under treatment) or at high cardiovascular risk (82%) Blood pressure control is improved (patients with systolic blood pressure levels > 180mmHg decreased from 33% to 24% and those with levels <160-179 increased from 26% to 34%) The fraction of patients at high or very high cardiovascular risk is decreased from 40% to 33% However, the compliance to antihypertensive treatment is still unsatisfactory since only half of the subjects are compliant with the prescribed therapy.

Laboratory of Medical Statistics

The Laboratory of Medical Statistics develops applied research in three main fields: controlled clinical trials, observational studies and genetic epidemiology.

Controlled clinical trials

The laboratory deals with planning, management and statistical analysis of controlled clinical trials, carried out in the different laboratories of the Cardiovascular Research Department, by means of the GISSI trials sound experience. At present, GISSI trials focus on GISSI-HF and GISSI-AF clinical trials, concerning heart failure and atrial fibrillation and their respective subprojects aiming to assess the role of biomarkers, of echocardiographic and electrocardiographic parameters on the patients prognosis. Recently, the superiority trial BeTACTIC that will randomize about 400 patients undergone heart transplantation has been activated. A large trial concerning cardiovascular prevention, Risk & Prevention study (Rischio & Prevenzione) which has included about 12000 patients is still ongoing. Statistical methodology applied to clinical studies has a leading and developing role as far as methods are concerned (e.g.: missing data management; development of prognostic risk scores, development of forecasting models for biomarkers based on Reclassification techniques and on Discriminations Indices etc.). Moreover, clinical trial management implies the setup of data planning and screening methods, the ad interim analysis and the choice of the best study design (superiority, non-inferiority and equivalence studies).

Observational studies
The activation of observational studies allows to characterize the epidemiological profile of categories of patients followed in their natural clinical course. The prospective observational study GLICINE-SPIDER has evaluated the risk profile of 1300 patients with hyperglycemia at the onset of an acute coronary syndrome (ACS) in the hospitals of the Lombardia region. The aim of the cohort study REGIA, presently ongoing, is the evaluation of the incidence of major and minor hemorrhages and the characterization of the risk profile of about 4000 patients undergoing hip and knee replacement surgery. From a strictly epidemiologic point of view, the epidemiologic and health-care history of diabetes mellitus in Regione Lombardia has been investigated by means of administrative databases.

Genetic Epidemiology
The laboratory has recently developed specific skills on genetic epidemiology analysis. These studies are carried out together with the laboratory of Clinical Drugs Evaluation. Statistical

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analysis techniques concerning cardiovascular genetics have been developed in the last five years. The study of the genetic component of multifactorial diseases, such as the cardiovascular disease, has been dealt with in the PROCARDIS study, by means of the genome-wide screening. This technique aims at identifying genes that can cause coronary disease. PROCARDIS database gave the opportunity of studying some quantitative traits such as the level of lipids or body max indexes. During the second step of the PROCARDIS project, supported by the 6th Framework Program of EEC, a screening on the whole genome has been carried out by means of the genome-wide association technique. For this project about 1 million of polymorphisms (SNPs) have been analyzed in order to identify a possible relationship with coronary disease. Concerning the GISSI-Genetic Prevention study, the laboratory has developed statistics genetics techniques to analyze case control studies in order to assess the association of genetic variants linked to adiponectin, HsCRP, PTX3 with coronary disease. Besides, the association between some polymorphisms of chromosome 9p area and coronary disease has been evaluated in diabetic patients. With regard to the GISSI-HF genetic substudy that has included about 2500 patients to evaluate the role of genetic variants involved in heart failure, the association of four polimorphisms of the adiponectin gene has been investigated by a case-control design.

Laboratory of Clinical Pharmacology Quality of Life, Depression and Cognitive problems in heart failure patients (QDF-GISSI-HF)
The QDF project is a sub-project of the GISSI-HF study. The aims of the study are 1) to describe the evolution of depression, cognitive problems and the quality of life in a sample of 1500 heart failure patients; 2) to assess the use of common instruments that measure QDF variables; 3) to compare the assessment of the instrument (Geriatric Depression scale, Mini Mental State Examination, Kansas City Cardiomiopathy Questionnaire) with the clinical perception of the nurses; 4) to describe if assessed or perceived patients' problems (low quality of life, high depression or compromised cognitive function) lead to any caring intervention. The baseline clinical characteristics of the 1564 patients included in the QDF study are closely comparable with those of the GISSI-HF population. The study instruments could be validly administered to the greatest majority of patients (KCQQ 97.2%, GDS 94.9%, MMSE 80.6% of patients >70 years). The nurses network nested in a major clinical trial, has produced one of the largest prospective cohort of HF patients who are comprehensively assessed and prospectively monitored, to allow an integrated evaluation of the relevance and implications of QDF measurements also on the clinical outcomes of this population. Manuscripts with the study results are in preparation.

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DEPARTMENT OF MOLECULAR BIOCHEMISTRY AND PHARMACOLOGY

STAFF
Head Mario SALMONA, Food Technology D, Ph.D.

Laboratory of Biochemistry and Protein Chemistry Head Ph.D. Laboratory of Molecular Biology Head Pharmacogenomics Unit Head Maddalena FRATELLI, Biol.Sci.D. Enrico GARATTINI, M.D. Mario SALMONA, Food Technology D,

Gene Structure and Regulation Unit Head Mineko TERAO, Bioch.D., Ph.D.

Laboratory of Pharmacodynamics and Pharmacokinetics Head Marco GOBBI, Pharm.D.

Laboratory of Molecular Pathology Head Lavinia CANTONI, Biol.Sci.D.

Laboratory of Translational Proteomics Head Laboratory of Systems Biology Head Gianfranco BAZZONI, M.D. Valentina BONETTO, Chem.Pharm.D.

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CURRICULA
Mario Salmona obtained his doctorate degree in Biochemistry and Food Technology at the University of Milan in 1971. His background is in biochemistry, biophysics and pharmacology. His scientific interests relate to problems of human and animal diseases originating from the aberrant folding of proteins. In this context, a major portion of his studies was devoted to the etiopathogenesis and therapy of prion diseases. He has published over 200 articles on peer reviewed scientific journals. 1971-1975 Ph.D in Pharmacology, Mario Negri Institute 1975 Visiting Fellow in the Department of Biology of the Weizmann Institute of Science, Rehovot, Israel 1976-1997 Head, Laboratory of Enzyme Research, Mario Negri Institute 1995 to date Dean of the School of Advanced Pharmacology, Mario Negri Institute 1997 to date Head, Department of Biochemistry and Molecular Pharmacology, Mario Negri Institute 2003 to date Member of the American Society of Biochemistry and Molecular Biology Referee for international scientific journals.
Selected publications Airoldi C, Colombo L, Manzoni C, Sironi E, Natalello A, Doglia S M, Forloni G, Tagliavini F, Del Favero E, Cantu' L, Nicotra F, Salmona M Tetracycline prevents A oligomer toxicity through an atypical supramolecular interaction Org Biomol Chem, 2010 9: 463-72 Urru S A M, Veglianese P, De Luigi A, Fumagalli E, Erba E, Gonella Diaza R, Carr A, Davoli E, Borsello T, Forloni G, Pengo N, Monzani E, Cascio P, Cenci S, Sitia R, Salmona M A new fluorogenic peptide determines proteasome activity in single cells J Med Chem 2010 53 : 7452-7460 Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, Tapella L, Colombo L, Manzoni C, Borsello T, Chiesa R, Gobbi M, Salmona M, Forloni G Synthetic amyloid-beta oligomers impair long-term memory independently of cellular prion protein Proc Natl Acad Sci U S A. 2010 107 : 2295-2300 Di Fede G, Catania M, Morbin M, Rossi G, Suardi S, Mazzoleni G, Merlin M, Giovagnoli A R, Prioni S, Erbetta A, Falcone C, Gobbi M, Colombo L, Bastone A, Beeg M, Manzoni Claudia, Francescucci B, Spagnoli A, Cantu' L, Del Favero A, Levy E, Salmona M, Tagliavini F A recessive mutation in the APP gene with dominant-negative effect on amyloidogenesis Science 2009 323 : 1473-1477 Saracino GA, Villa A, Moro G, Cosentino U, Salmona M. Spontaneous beta-helical fold in prion protein: The case of PrP(82-146). Proteins. 2009 Jun;75(4):964-76 De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Limido L, Forloni G, Tagliavini F, Salmona M. The efficacy of tetracyclines in peripheral and intracerebral prion infection. PLoS ONE. 2008;3(3):e1888

Gianfranco Bazzoni got his Medicine and Surgery degree in 1988 (at the University of Milan) and the specialisation in Pharmacological Research in 1992 (at the Mario Negri Institute, Milan). His area of expertise is cell biology, with focus on the processes of cell adhesion and migration. 1988-2000 Research Fellow, Mario Negri Institute 1993-1997 Post-doctoral Fellow, Dana Farber Cancer Institute and Harvard Medical School, Boston, MA 2000-2002 Research Scientist, Mario Negri Institute 2003 Head, Unit of Cell Adhesion, Mario Negri Institute 2004 to date Head, Laboratory of Systems Biology, Mario Negri Institute 2004 Regular Member of The American Physiological Society, Bethesda, MD Referee for international scientific journals
Selected publications Paris L, Bazzoni G The protein interaction network of the epithelial junctional complex: a system-level analysis Mol Biol Cell 19: 54095421, 2008 Paris L, Tonutti L, Vannini C, Bazzoni G Structural organization of the tight junction. Biochim Biophys Acta 1778: 646-659, 2008 Huang H, Cruz F, Bazzoni G Junctional adhesion molecule-A regulates cell migration and resistance to shear stress. J. Cell Physiol 209; 122-130, 2006

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Martinez-Estrada OM, Manzi L, Tonetti P, Dejana E, Bazzoni G Opposite effects of Tumor Necrosis Factor and soluble fibronectin on Junctional Adhesion Molecule-A in endothelial cells Am J Physiol (Lung Cell Mol Physiol) 288: L1081-L1088, 2005 Bazzoni G, Tonetti P, Manzi L, Cera MR, Balconi G, Dejana E Expression of Junction Adhesion Molecule-A prevents spontaneous and random motility. J Cell Sci 118: 623-632, 2005 Bazzoni G, Dejana E Endothelial cell-to-cell junctions: molecular organization and role in vascular homeostasis. Physiol Rev 84: 869-901, 2004

Valentina Bonetto has got the degree in Pharmaceutical Chemistry and Technology at the University of Padua, Italy in 1993. She has got the Ph.D in Medical Biochemistry and Biophysics at Karolinska Institutet, Stockholm, Sweden. Her principal lines of research are: 1) Study of the pathogenetic mechanisms at the basis of amyotrophic lateral sclerosis (ALS); 2) Identification of biomarkers of ALS; 3) Role of the oxidative modification in neurological disorders. These issues are investigated by different experimental approaches, including proteomics and mass spectrometry. 2000-2009 Research Scientist, Laboratory of Biochemistry and Protein Chemistry, Mario Negri Institute 2002-2009 also Assistant Telethon Scientist at Dulbecco Telethon Institute 2007-2009 Head, Unit of Medical Biochemistry, Laboratory of Biochemistry and Protein Chemistry, Mario Negri Institute From 2009 to date, Head Laboratory of Translational Proteomics and Associate Telethon Scientist. She is author of 33 publications from 1994 to 2010, in peer-reviewed journals. She is reviewer for scientific journals in the field of Proteomics and Neuroscience.
Selected publications Massignan T, Biasini E, Lauranzano E, Veglianese P, Pignataro M, Fioriti L, Harris DA, Salmona M, Chiesa R, Bonetto V Mutant prion protein expression is associated with an alteration of the Rab GDP dissociation inhibitor alpha (GDI)/Rab11 pathway. Mol Cell Proteomics, 9:611-622, 2010 Basso M, Samengo G, Nardo G, Massignan T, DAlessandro G, Tartari S, Cantoni L, Marino M, Cheroni C, De Biasi S, Giordana MT, Strong MJ, Estevez AG, Salmona M, Bendotti C, Bonetto V Characterization of detergent-insoluble proteins in ALS indicates a causal link between nitrative stress and aggregation in pathogenesis PLoS ONE 4:e8130, 2009 Nardo G, Pozzi S, Mantovani S, Garbelli S, Marinou K, Basso M, Mora G, Bendotti C, Bonetto V Nitroproteomics of peripheral blood mononuclear cells from patients and a rat model of ALS Antioxid. Redox Signal 11: 1559-1567, 2009 Massignan T, Casoni F, Basso M, Stefanazzi P, Biasini E, Tortarolo M, Salmona M, Gianazza E, Bendotti C, Bonetto V Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse Biochem. Biophys. Res. Commun. 353: 719-25, 2007 Basso M, Massignan T, Samengo G, Cheroni C, De Biasi S, Salmona M, Bendotti C, Bonetto V Insoluble mutant SOD1 is partly oligoubiquitinated in amyotrophic lateral sclerosis mice J. Biol. Chem. 281:33325-33335, 2006 Casoni F, Basso M, Massignan T, Gianazza E, Cheroni C, Salmona M, Bendotti C, Bonetto V Protein nitration in a mouse model of familial amyotrophic lateral sclerosis: Possible multifunctional role in the pathogenesis. J. Biol. Chem., 280: 16295-16304, 2005

Lavinia Cantoni obtained her degree in Biological Sciences in 1973 at the University of Milan. Then she specialized in pharmacological research at the Mario Negri Institute (1974-1977). Research areas 1) biochemical-molecular mechanisms activated by oxidative stress 2) drug metabolism 3) porphyrias. 1977-1978 Post-doctoral Fellow, Medical Research Council, Toxicology Unit, Carshalton, UK (Winner of a Welcome Trust Research Fellowship) 1979-1982 Research Scientist, Mario Negri Institute 1980-1990 Visiting Scientist, Toxicology Unit, Carshalton, UK, and Cornell Medical Center, New York, NY (short periods) 1983-1997 Head, Unit of Heme and Hemoprotein Metabolism, Mario Negri Institute 1998 to date, Head, Laboratory of Molecular Pathology, Mario Negri Institute 1975 to date Member of the National Roll of Biologists 1983 to date Member of the Italian Toxicology Society Referee for international scientific journals.

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Selected publications Tartari S, DAlessandro G, Babetto E, Rizzardini M, Conforti L, Cantoni L. Adaptation to G93Asuperoxide dismutase 1 in a motor neuron cell line model of amyotrophic lateral sclerosis. The role of glutathione FEBS J. 276: 2861-2874, 2009 Raimondi A, Mangolini A, Rizzardini M, Tartari S, Massari S, Bendotti C, Francolini M, Borghese N, Cantoni L, Pietrini G. Cell culture models to investigate the selective vulnerability of motoneuronal mitochondria to familial ALS-linked G93ASOD1 Eur. J. Neurosci. 24: 387-399, 2006 Babetto E, Mangolini A, Rizzardini M, Lupi M, Conforti L, Poletti A, Rusmini P, Cantoni L. Tetracycline-regulated gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases Mol. Brain Res. 140: 63-72, 2005 Cantoni L,Valaperta R, Ponsoda X, Castell JV, Barelli D, Rizzardini M, Mangolini A, Hauri L, Villa P. Induction of hepatic heme oxygenase-1 by diclofenac in rodents: role of oxidative stress and cytochrome P-450 activity J. Hepatology 38: 776-783, 2003 Cantoni L, Rozio M, Mangolini A, Hauri L, Caccia S. Hyperforin contributes to the hepatic CYP3A-inducing effect of Hypericum perforatum extract in the mouse. Toxicol.Sci. 75:25-30, 2003 Rizzardini M, Zappone M, Villa P, Gnocchi P, Sironi M, Diomede L, Meazza C, Monshouwer M, Cantoni L. Kupffer cell depletion partially prevents hepatic heme oxygenase 1 messenger RNA accumulation in systemic inflammation in mice: role of interleukin 1 beta Hepatology 27: 703-710, 1998

Enrico Garattini obtained his degree in Medicine and Surgery with full marks (110/110) in 1982 at the University of Milan. His scientific interests relate to problems of Cellular Biology and Molecular Biology. 1982-1990 Research Fellow of the National Research Council, Mario Negri Institute 1983-1987 Postdoctoral Researcher at the Roche Institute of Molecular Biology, Department of Neurosciences Nutley, New Jersey, US 1991-1997 Senior Researcher Regione Lombardia and Head of the Molecular Biology Unit, Mario Negri Institute 1997 to date Head, Laboratory of Molecular Biology, Mario Negri Institute From 2005 Dean, Advanced School of Pharmacology (Philosophy Doctor), Mario Negri Institute Member of the Editorial Board of the European Journal of Cancer and of Current Cancer Therapy Reviews Member of the American Society of Biochemistry and Molecular Biology (ASBMB)
Selected publications Gianni M, Boldetti A, Guarnaccia V, Rambaldi A, Parrella E, Raska I Jr, Rochette-Egly C, Del Sal G, Rustighi A, Terao M, Garattini E Inhibition of the peptidyl-propyl-isomerase Pin1 enhances the responses of acute myeloid leukemia cells to retinoic acid via stabilization of RAR and PML-RAR. Cancer Res 69 : 1016-1026, 2009 Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C, Garattini E Role of the molybdo-flavoenzyme, aldehyde oxidase homolog 2, in the biosynthesis of retinoic acid: generation and characterization of a knock-out mouse Mol Cell Biol 29: 357-77, 2009 Gianni M, Parrella E, Raska I Jr, Gaillard E, Nigro EA, Gaudon C, Garattini E, Rochette-Egly C. P38MAPK-dependent phosphorylation and degradation of SRC-3/AIB1 and RARalpha-mediated transcription EMBO J. 25:739-51, 2006 Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I, Carminati P, Terao M, Pisano C ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in myeloid leukemia cells: Modulation of intracellular calcium homeostasis Blood 103: 194-207, 2004 Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with selective expression in the olfactory mucosa J Biol Chem 279: 50482-50498, 2004 Pisano C, Kollar P, Gianni M, Kalac Y, Giordano V, Ferrara F F, Tancredi R, Devoto A, Rinaldi A, Rambaldi A, Penco S, Marzi M, Moretti G, Vesci L, Tinti O, Carminati P, Terao M, Garattini E Bis-indols a novel class of molecules enhancing the cytodifferentianting properties of retinoids in myeloid leukemia cells

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Blood 100: 3719-3730, 2002

Marco Gobbi got his degree in Pharmacy at the University of Milan, Italy, in 1989. His main fields of interest are: i) neurodegenerative diseases associated to misfolding and aggregation of peptides/proteins, such as beta-amyloid and prions; ii) alterations of synaptic transmission in the CNS, either due to diseases or to the effects of drugs on receptors and transporters; iii) nanoparticles for diagnostic and therapeutic purposes. These research fields are investigated by a close integration of pharmacodynamics (e.g. biomolecular interactions, mainly using surface plasmon resonance) and pharmacokinetics studies. 1981-1995 Researcher, Laboratory of Neuropharmacology and, from 1988, in the Laboratory of Receptor Pharmacology, Mario Negri Institute 1995-2010 Head, Unit of Synaptic Transmission, Mario Negri Institute From 2010, Head, Laboratory of Pharmacodynamics and Pharmacokinetics Co-author in more than 100 scientific publications on peer-reviewed international journals. First or last author in more than 50 of them. Reviewer for international scientific journals operating in the Neuroscience/Neuropharmacology/Biochemistry fields.
Selected publications Gobbi M, Re F, Canovi M, Beeg M, Gregori M, Sesana S, Sonnino S, Brogioli D, Musicanti C, Gasco P, Salmona M and Masserini ME. Lipid-based nanoparticles with high binding affinity for amyloid-beta1-42 peptide. Biomaterials 31:65196529 (2010). Caccia S and Gobbi M St. John's Wort components and the brain: Uptake, concentrations reached and the mechanisms underlying pharmacological effects. Curr Drug Metab 10(9):1055-1065 (2009). Gesuete R, Storini C, Fantin A, Stravalaci M, Zanier ER, Orsini F, Vietsch H, Mannesse MLM, Ziere B, Gobbi M and De Simoni MG Recombinant C1 inhibitor in brain ischemic injury. Ann Neurol 66(3):332-342 (2009). Colleoni S, Jensen AA, Landucci E, Fumagalli E, Conti P, Pinto A, De Amici M, Pellegrini-Giampietro DE, De Micheli C, Mennini T and Gobbi M. Neuroprotective effects of the novel glutamate transporter inhibitor (-)-3-hydroxy-4,5,6,6atetrahydro-3aH-pyrrolo[3,4-d]-isoxazole-4-carboxylic acid, which preferentially inhibits reverse transport (glutamate release) compared with glutamate reuptake. J Pharmacol Exp Therap 326:646-656 (2008). Gobbi M, Colombo L, Morbin M, Mazzoleni G, Accardo E, Vanoni M, Del Favero E, Cant L, Kirschner DA, Manzoni C, Beeg M, Ceci P, Ubezio P, Forloni G, Tagliavini F and Salmona M. Gerstmann-Strussler-Scheinker disease amyloid protein polymerizes according to the "dock-and-lock" model. J Biol Chem 281:843-849 (2006). Crespi D, Mennini T, Gobbi M. Carrier-dependent and Ca(2+)-dependent 5-HT and dopamine release induced by (+)amphetamine, 3,4-methylendioxymethamphetamine, p-chloroamphetamine and (+)-fenfluramine. Br J Pharmacol 121:1735-1743 (1997).

Maddalena Fratelli got her degree in Biological Sciences at the University of Pisa and at the Scuola Normale Superiore di Pisa in 1983. Then the specialization in Pharmacological Research at the Mario Negri Institute in 1986. Her main fields of interest are: 1. High throughput genomic systems for the study of drug action and pharmacoresistance. 2. Redox regulation of protein function and gene expression: glutathionylation and gene expression profiling of glutathione dependent responses to oxidant challenge. 1988-1989 Postdoctoral Research Fellow in the Medical Research Council, Neurobiology Unit, Cambridge, UK. Since 1995, Head, Unit of Mediators of inflammation, Laboratory of Neuroimmunology, Mario Negri Institute Since 2005, Head, Unit of Pharmacogenomics, Laboratory of Molecular Biology, Mario Negri Institute
Selected publications Garattini E, Fratelli M, Terao M. The mammalian aldehyde oxidase gene family Hum Genomics. 4: 119-30, 2009 Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi P. Gene expression profiling reveals a signaling role of glutathione in redox regulation. Proc Natl Acad Sci U S A 102:13998-4003, 2005 Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, Latini R, Xie QW, Smart J, Su-Rick CJ, Pobre E, Diaz D, Gomez D, Hand C, Coleman T, Cerami A. Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor Proc Natl Acad Sci U S A 101:14907-12, 2004

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Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P, Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S, Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M. Derivatives of erythropoietin that are tissue protective but not erythropoietic Science 305:239-42, 2004 Fratelli M, Minto M, Crespi A, Erba E, Vandenabeele P, Del Soldato P, Ghezzi P. Inhibition of nuclear factor-kappaB by a nitro-derivative of flurbiprofen: a possible mechanism for antiinflammatory and antiproliferative effect Antioxid Redox Signal. 5:229-35, 2003 Fratelli M, Demol H, Puype M, Casagrande S, Eberini I, Salmona M, Bonetto V, Mengozzi M, Duffieux F, Miclet E, Bachi A, Vandekerckhove J, Gianazza E, Ghezzi P. Identification by redox proteomics of glutathionylated proteins in oxidatively stressed human T lymphocytes Proc Natl Acad Sci U S A 99:3505-10, 2002

Mineko Terao obtained her doctorate degree in Pharmaceutical Science from the Kobe Womens College of Pharmacy, Japan in 1978. Her scientific interests relate to problems of Cellular Biology and Molecular Biology. 1983 Ph.D in Molecular Biology, Kyoto University, Japan 1982-1983 Research Fellow, Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan 1983-1987 Postdoctoral Associate of the Institute for Cancer Research, Philadelphia, US From 1987 Visiting Scientist of Mario Negri Institute From 1998 Head of the Unit of Gene Structure and Regulation, Mario Negri Institute
Selected publications Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C, Garattini E Role of the molybdoflavoenzyme aldehyde oxidase homolog 2 in the biosynthesis of retinoic acid: generation and characterization of a knockout mouse Mol Cell Biol 29 : 357-377, 2009 Terao M, Kurosaki M, Barzago MM, Varasano E, Boldetti A, Bastone A, Fratelli M, Garattini E. Avian and canine aldehyde oxidases. Novel insights into the biology and evolution of molybdo-flavoenzymes. J Biol Chem. 281: 19748-61, 2006 Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I, Carminati P,Terao M, Pisano C. ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in myeloid leukemia cells: Modulation of intracellular calcium homeostasis. Blood 103: 194-207, 2004 Vila R, Kurosaki M, Barzago M M, Kolek M, Bastone A, Colombo L, Salmona M, Terao M, Garattini E. Regulation and biochemistry of mouse molybdo-flavoenzymes. The DBA/2 mouse is selectively deficient in the expression of aldehyde oxidase homologues 1 and 2 and represents a unique source for the purification and characterization of aldehyde oxidase. J Biol Chem 279: 8668-8683, 2004 Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with selective expression in the olfactory mucosa J Biol Chem 279: 50482-50498, 2004 Parrella E, Gianni M, Cecconi V, Nigro E, Barzago MM, Rambaldi A, Rochette-Egly C, Terao M and Garattini E. Phosphodiesterase 4 inhibition by piclamilast potentiates the cyto-differentiating action of retinoids in myeloid leukemia cells. J Biol Chem 279: 42026-42040, 2004

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ACTIVITIES The Department comprises six laboratories. Research is heterogeneous in terms of scientific interests and aims, but it is unified by the structural and functional study of specific, pharmacologically important gene products, using a common body of techniques. Classical biochemistry and molecular biology methods are used to define proteins that might be targets for the pharmacological activity of drugs. Potential direct interactions between drugs and proteins are studied at the molecular level by a variety of approaches ranging from animal studies to computer simulation. MAIN FINDINGS Identification of the molecular mechanisms responsible of oligomer formation of amyloidogenic proteins. Identification of tetracyclines as potential therapeutic agents for prion diseases. Synthesis, biological and chemico-physical characterization of peptides deduced from prion protein sequence. Identification of a correlation between cholesterol synthesis and prion protein production. Protein identifications by mass spectrometry and data base searching using a combination of techniques. Characterization of the Pentraxin 3 role in the organization of the cumulus oophorus extracellular matrix and in female fertility. System-level analysis of protein interactions in the epithelial junctional complex. Proteomic analysis of the aggregates isolated from spinal cord of a mouse model of amyotrophic lateral sclerosis (ALS) Identification of nitroprotein biomarkers in peripheral blood mononuclear cells of ALS patients and a rat model of ALS Proteomic analysis of a cellular model of fatal familial insomnia Development of constitutive and conditional motor neuronal cell models to unravel the toxicity of mutant G93A superoxide dismutase 1 responsible for some forms of familial amyotrophic lateral sclerosis. Drugs or exogenous compounds impairing the electron transport chain are a risk factor to motor neurons of individuals carrying mutant forms of superoxide dismutase 1. Mitochondrial damage due to mutant G93A superoxide dismutase 1 occurs selectively in motor neurons. Synthesis of glutathione, the main cellular antioxidant, is altered in motor neuronal cells by human G93A mutant superoxide dismutase 1. Identification and characterization of a novel class of retinoids endowed with strong and selective apoptogenic activity on the neoplastic cell. Pre-clinical development of these agents for the treatment of acute leukemia. Identification and characterization of novel retinoid-based pharmacological combinations for the treatment of acute myelogenous leukemia.

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Molecular cloning and characterization of the cDNAs and genes of four novel members of the mammalian molybdo-flavoprotein family. Definition of a novel gene cluster on human chromosome 2 and mouse chromosome 1. Development of knok-out animals for molybdo-flavoproteins: AOX1, AOH1, AOH2, AOH3. Creation of integrated instruments for the rationalization of Microarray analysis processes. The treatment with a non haematopoietic derivate of Erythropoietin (CEPO) reduces motor neuron loss and clinical progression in a mouse model of ALS related to alterations in vesicle trafficking, the wobbler mouse. Treatment with a soluble TNF receptor in the wobbler mouse, reduces motor neuron degeneration and the phosphorylation of the two main stress kinases (p38 e JNK) activated by TNF receptors. Riluzole treatment reduces motor neuron loss and clinical progression of wobbler mouse by increasing the endogenous BDNF expression . Oxidative stress, glial activation and inflammation occur in the retinopathy as well as in cerebral and spinal cord dysfunction in the mnd mouse, a model of progressive epilepsy with mental retardation related to mutation in the CLN8 gene. These findings provide further evidence for the implication of TNF death receptor signalling in the pathology of Neuronal Ceroid Lipofuscinosis Recombinant C1-inhibitor binds with high affinity with Mannose Binding Lectins, an interaction possibly underlying its superior anti-ischemic properties in animal models. Confirmation and characterization of the binding of -amyloid oligomers to prion protein. Characterization of the binding properties of a new peptide inhibitor of -amyloid aggregation. Development of new protocols to evaluate, by surface plasmon resonance, the interaction between nanoparticles and their putative targets: application to nanoparticles functionalized to bind -amyloid. Confirmation and characterization of the binding of pentraxin-3 to P-selectin, a new mechanism involved in the leukocyte recruitment at sites of inflammation. Determination of the binding properties of new FGF-2 ligands with antiangiogenic activities. NATIONAL COLLABORATIONS Advanced Biology Center, Genoa Centro Clinico Nemo, Ospedale Niguarda, Milan Center of Excellence on Neurodegenerative Diseases, University of Milan, Segrate, Milan Dip. Anatomia, Farmacologia, Medicina Legale, University of Turin Dip. Biochimica, University of Pavia Dip. Biotecnologie, Universit degli Studi, Milan Dip. Chimica Biochimica e Biotecnologie per la Medicina, Universit degli Studi, Milan Dip. Chimica Farmaceutica e Tossicologica, Universit degli Studi, Milan

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Dip. Chimica Organica e Industriale Universit degli Studi, Milan Dip. Farmaco-Chimico, Universit degli Studi, Messina Dip. Farmaco-Chimico-Tecnologico, University of Siena Dip. Farmacologia Medica, Universit degli Studi, Milan Dip. Medicina Sperimentale, Universit Bicocca, Monza Dip. Scienze Biochimiche, University of Florence Dip. Scienze Farmaceutiche, University of Catania Dip. Scienze Farmaceutiche, University of Genoa Dip. Scienze Farmacologiche, Universit degli Studi, Milan Dip. Scienze Fisiologiche e Farmacologiche, University of Pavia Dip. Scienze Molecolari, University of Milan Dip. Studi pre-clinici, University of Milan Facolt di Biologia, Universit degli Studi, Milan Facolt di Chimica, Universit degli Studi, Milan Facolt di Chimica, University of Ferrara Fondazione S. Maugeri, Milan Fondo Edo Tempia, Biella IFOM Fondazione Istituto FIRC di Oncologia Molecolare, Milan IRCCS Fondazione "Istituto C. Mondino", Laboratorio di Neurobiologia Sperimentale, Pavia Istituto di Biologia Molecolare Buzzati Traverso, Naples Istituto di Biomedicina e Immunologia Molecolare CNR, Palermo Istituto di Chimica del Riconoscimento Molecolare CNR, Milan Istituto di Endocrinologia, Centro di Eccellenza per le Malattie Neurodegenerative, Universit degli Studi, Milan Istituto di Clinica Neurologica, Ospedale Maggiore Policlinico, Milan Istituto Clinico Humanitas, Milan Istituto di Neuroscienze C.N.R., Pisa Istituto Nazionale dei Tumori, Milan Istituto Nazionale dei Tumori, Naples Istituto Nazionale Neurologico "C. Besta", Milan Istituto Oncologico Europeo, Milan Istituto Regina Elena, Rome Istituto Toscano Tumori, Florence Nanovector srl, Turin Newron Pharmaceuticals, Milan Ospedale Maggiore Policlinico, Milan Ospedale Pediatrico Bambino Gesu', Rome Ospedale Pediatrico "Gaslini", Genoa Ospedale S. Gerardo, Monza, Milan Ospedale San Matteo, Pavia Sigma-Tau, Pomezia, Rome Zambon, Milan

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INTERNATIONAL COLLABORATIONS The Babraham Institute, Cambridge, UK Boston College, Boston, MA, USA Burke Medical Research Institute, White Plains, new York, USA Case Western Research University, Cleveland, OH, USA Dept. de Quimica-Fisica de Macromoleculas Biologicas, CSIC, Madrid, Spain Division of Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster LA1 4YQ, UK Faculdad de Ciencias Medicas, Universidad de Santiago de Chile, Chile ETH, Zurig, Switzerland FMP, Berlin, Germany Giessen Polyclinic University, Giessen, Germany Houston University, TX, USA Keio University, Tokyo, Japan IBSN CNRS, Marseille, France Indiana University, Indianapolis, IN, USA Institut de Genetique et Biologie Moleculaire et Cellulaire, Strasbourg, France Institut Pasteur, Paris, France John Innes Centre, Norwich, UK Laboratoire de Physico-Chimie, Pharmacotechnie et Biopharmacie, , Univ Paris-Sud 11, Chatenay-Malabry, France Lundbeck, USA Mayo Clinic College of Medicine, Jacksonville, FL, USA National Institute of Health, Bethesda, MD, USA Nippon University, Tokyo, Japan Pepscan System BV, Lelystad, Holland Polichem S.A., Lugano, Switzerland Politecnico di Zurigo (ETH), Switzerland Technical University Braunschweig, Germany The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel Trinity College, Dublin, Ireland Universidad de La Laguna, Tenerife, Spain Universidad Nova, Lisbon, Portugal Universitat des Saarlandes, Hamburg, Germany Universitat Freiburg, Germany Universit Paris, France Universit Victor Segalen Bordeaux 2, Bordeaux, France University of Aberdeen, UK University of Amsterdam, The Netherlands University of Birmingham, UK University of Cardiff, UK University of Glasgow, UK University of Gottingen, Germany University of Muenster, Germany

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University of Patrasso, Grece University of Southampton, UK University of Sussex, UK University of Vienna, Austria Waring-Webb Institute, University of Colorado, Denver CO, USA Weizmann Institut, Rehovot, Israel Westfaelische Wilhelms-Universitaet Muenster, Germany EDITORIAL BOARD MEMBERSHIP Neurobiology of Lipids (L. Diomede) European Journal of Cancer (E. Garattini) PEER REVIEW ACTIVITIES American Journal Physiology, Antioxidants and Redox Signaling, BBA-Proteomics, Biochemical Journal, Biochemical Pharmacology, Biochimica Biophysica Acta, BMCBiochemistry, Brain Research, Cancer Research, Cell Death and Differentiation, Cell Research, Cellular and Molecular Life Sciences, Circulation, Drug Investigation, European Journal of Cancer, European Journal of Immunology, European Journal of Neuroscience, International Journal of Cancer, Journal of Cell Biology, Journal of Hepatology, Journal of Immunology, Journal of Investigative Dermatology, Journal of Lipid Mediators, Journal of Neurochemistry, Journal of Neuroimmunology, Journal of Translational Medicine, Neuroscience, Neuroscience Letters, Pharmacological Research, Physiological Genomics, PLoS ONE, Proceedings of the National Academy of Sciences, Life Sciences, Proteomics, Proteome Science. CONFERENCE AND WORKSHOP CONTRIBUTIONS Meeting: 7th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology, Nanoparticles against ALZHEIMER'S disease: PEG-PACA nanoparticles are able to link the A-peptide and influence its aggregation kinetic, 811 March, Malta, Valetta Congress: Microscale separation, Interaction between PEG-PACA nanoparticles and amyloid peptide highlighted by Capillary Electrophoresis. 21-25 May, Prague, Rpublique Tchque

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Workshop: Italian-Spanish Joint Workshop, NMR characterization of the interaction between A peptide and small ligands for the development of the new antiAlzheimers drugs, 22 April, Milan, Italy Conference: 3rd European Conference for Clinical Nanomedicine, Nanoparticles against ALZHEIMER'S disease: PEG-PACA nanoparticles are able to link the Apeptide and influence its aggregation kinetic, 10-12 May, Basel, Switzerland Congress: 7th FENS Forum of European Neuroscience, Biophysical and biological features of amyloid-beta 1-42 in its initial, oligomeric and fibrillar state, prepared by a novel synthetic technique, In vitro characterization of nanoparticles functionalized for targeting Amyloid-beta 1-42 peptide. 3-7 July, Amsterdam, The Netherlands Symposium: EFMC-ISMC 2010 - XXIst International Symposium on Medicinal Chemistry, Novel [1[benzothieno[3,2-d]pyrimidin-4(3H)-ones. Ligands for 5-HT7 receptor, 5 September, Brussels, Belgium Course: 2nd Training course Pharmacokinetics of Nanoparticles and their passage through the blood-brain barrier, Nanoparticles in biomedicine, NPs and stem cells tracking, NPs and interaction to biological targets, 22-24 September, Milan, Italy Congress: Nanotechitaly 2010, Synthesis and functionalization of polymeric nanoparticles for clinical imaging application, Stability of novel polymer-based nanoparticles designed for drug delivery and diagnostic purposes Nanotech Italy 2010, 20-22 October, Venice, Italy Congress: Neuroscience, [11C]PIB Shows Increased Binding in the Brain of the Transgenic APP23 Mice Even With Modest Specific Radioactivity, 13-15 November, San Diego, USA GRANTS AND CONTRACTS Agenzia Italiana del Farmaco, Rome, Italy Associazione Italiana Ricerca sul Cancro (AIRC), Milan, Italy Biotecnologie BT - Perugia, Italy Comunit Europea (EU), Bruxelles, Belgium Consiglio Nazionale delle Ricerche (CNR), Palermo, Italy Fondazione Don Gnocchi, Milan, Italy Fondazione Cariplo, Milan, Italy Fondazione Mariani, Milan, Italy Fondazione Monzino, Milan, Italy Fondazione Weizmann-Pasteur-Negri, Paris, France Istituto Auxologico Italiano, Milan, Italy Istituto Nazionale Neurologico "C. Besta", Milan, Italy Lundbeck A/S, Copenhagen, Denmark Ministero della Salute, Roma, Italy

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Ministero dell'Istruzione, Universit e Ricerca Scientifica (MIUR), Rome, Italy North Shore University Hospital, NY, USA Perfetti-Van Melle, Lainate, Milan, Italy Sigma Tau, Pomezia, Rome, Italy Telethon, Milan, Italy Universit di Firenze, Italy Universit di Milano-Bicocca, Italy Universit di Siena, Italy Zambon Group, Bresso (Mi), Italy SCIENTIFIC PUBLICATIONS (2010)
Airoldi C, Colombo L, Manzoni C, Sironi E, Natalello A, Doglia S M, Forloni G, Tagliavini F, Del Favero E, Cantu' L, Nicotra F, Salmona M Tetracycline prevents A oligomer toxicity through an atypical supramolecular interaction Org Biomol Chem, 2010 9: 463-72 Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, Tapella L, Colombo L, Manzoni C, Borsello T, Chiesa R, Gobbi M, Salmona M, Forloni G Synthetic amyloid- oligomers impair long-term memory independently of cellular prion protein PNAS 2010 107 : 2295-2300 Bate C, Tayebi M, Salmona M, Diomede L, Williams A Polyunsaturated fatty acids protect against prion-mediated synapse damage in vitro Neurotox Res 2010 17 : 203-214 Bazzoni G Pathobiology of Junctional Adhesion Molecules Antioxid Redox Signal, 2010 [Epub ahead of print] Beeg M, Stravalaci M, Bastone A, Salmona M, Gobbi M A modified protocol to prepare seed-free starting solutions of A1-40/1-42 from the corresponding depsi-peptides Anal Biochem, 2010 [Epub ahead of print] Bigini P, Steffensen K R, Ferrario A, Diomede L, Ferrara G, Barbera S, Salzano S, Fumagalli E, Ghezzi P, Mennini T, Gustafsson J - A Neuropathologic and biochemical changes during disease progression in liver X receptor -/- mice, a model of adult neuron disease J Neuropathol Exp Neurol 2010 69 : 593-605 Brambilla D, Verpillot R, Taverna M, De Kimpe L, Le Droumaguet B, Nicolas J, Canovi M, Gobbi M, Mantegazza F, Salmona M, Nicolas V, Scheper W, Couvreur P, Andrieux K New method based on capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) to monitor interaction between nanoparticles and the amyloid- peptide Anal Chem 2010 82 : 10083-10089 Colombo G, Margosio B, Ragona L, Neves M, Bonifacio S, Annis D S, Stravalaci M, Tomaselli S, Giavazzi R, Rusnati M, Presta M, Zetta L, Mosher D F, Ribatti D, Gobbi M, Taraboletti G Non-peptidic thrombospondin-1 mimics as fibroblast growth factor-2 inhibitors: an integrated strategy for the development of new antiangiogenic compounds J Biol Chem 2010 285 : 8733-8742 Cooper I, Malina K C, Cagnotto A, Bazzoni G, Salmona M, Teichberg V I Interactions of the prion peptide (PrP 106-126) with brain capillary endothelial cells: coordinated cell killing and remodeling of intercellular junctions J Neurochem 2010 [Epub ahead of print] Corrado L, Gagliardi S, Carlomagno Y, Mennini T, Ticozzi N, Mazzini L, Silani V

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VPS54 genetic analysis in ALS Italian cohort Eur J Neurol 2010 [Epub ahead of print] Deban L, Castro Russo R, Sironi M, Moalli F, Scanziani M, Zambelli V, Cuccovillo I, Bastone A, Gobbi M, Valentino S, Doni A, Garlanda C, Danese S, Salvatori G, Sassano M, Evangelista V, Rossi B, Zenaro E, Constantin G, Laudanna C, Bottazzi B, Mantovani A Regulation of leukocyte recruitment by the long pentraxin PTX3 Nat Immun 2010 11 : 328-334 De Paola M, Visconti L, Vianello E, Mattana F, Banfi G, Corsi M M, Beghi E, Mennini T Circulating cytokines and growth factors in professional soccer players: correlation with in vitro-induced motor neuron death Eur J Neurol, 2010 [Epub ahead of print] Diana V, Botti F, Fumagalli E, Calcagno E, De Paola M, Cagnotto A, Invernici G, Parati E, Curti D, Mennini T Neural precursor-derived astrocytes of wobbler mice induce apoptotic death of motor neurons through reduced glutamate uptake Exp Neurol 2010 225 : 163-172 Diomede L, Cassata G, Fiordaliso F, Salio M, Ami D, Natalello A, Doglia S M, De Luigi A, Salmona M Tetracycline and its analogues protect Caenorhabditis elegans from amyloid-induced toxicity by targeting oligomers Neurobiol Dis 2010 40 : 424-431 Galizzi G, Russo Domenica, Deidda A, Cascio C, Passantino R, Guarneri R, Bigini P, Mennini T, Drago G, Guarneri P Different early ER-stress responses in the CLN8mnd mouse model of neuronal ceroid lipofuscinosis Neurosci Lett 2010 [Epub ahead of print] Giaccone G, Morbin M, Moda F, Botta M, Mazzoleni G, Uggetti A, Catania M, Moro M L, Redaelli V, Spagnoli A, Rossi R S, Salmona M, Di Fede G, Tagliavini F Neuropathology of the recessive A673V APP mutation: Alzheimer disease with distinctive features Acta Neuropathol 2010 120 : 803-812 Giorgetti S, Raimondi S, Pagano K, Relini A, Bucciantini M, Corazza A, Fogolari F, Codutti L, Salmona M, Mangione P, Colombo Lino, De Luigi A, Porcari R, Gliozzi A, Stefani M, Esposito G, Bellotti V, Stoppini M Effect of tetracyclines on the dynamics of formation and destructuration of 2-microglobulin amyloid fibrils J Biol Chem 2010 [Epub ahead of print] Gobbi M, Re F, Canovi M, Beeg M, Gregori M, Sesana S, Sonnino S, Brogioli D, Musicanti C, Gasco P, Salmona M, Masserini M Lipid-based nanoparticles with high binding affinity for amyloid-1-42 peptide Biomaterials 2010 31 : 6519-6529 Lacivita E, De Giorgio P, Lee I T, Rodeheaver S I, Weiss B, Fracasso C, Caccia S, Berardi F, Perrone R, Zhang M R, Maeda Y, Higuchi M, Suhara T, Schetz J A, Leopoldo M Design, synthesis, radiolabeling and in vivo evaluation of carbon-11 labeled N-[2-[4-(3-cyanopyridin-2-yl)piperazin1-yl]ethyl]-3-methoxybenzamide, a potential Positron Emission Tomography tracer for the dopamine D4 receptors J Med Chem 2010 53 : 7344-7355 Martinez de la Torre Y, Fabbri M, Jaillon S, Bastone A, Nebuloni M, Vecchi A, Mantovani A, Garlanda C Evolution of the pentraxin family: the new entry PTX4 J Immunol 2010 184 : 5055-5064 Massignan T, Biasini E, Lauranzano E, Veglianese P, Pignataro M, Fioriti L, Harris D A, Salmona M, Chiesa R, Bonetto V Mutant prion protein expression is associated with an alteration of the Rab GDP dissociation inhibitor alpha (GDI)/Rab11 pathway Mol Cell Proteomics 2010 9 : 611-622 Massignan T, Stewart R S, Biasini E, Solomon I H, Bonetto V, Chiesa R, Harris D A A novel, drug-based, cellular assay for the activity of neurotoxic mutants of the prion protein J Biol Chem 2010 285 : 7752-7765

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Melo T, Bigini P, Sonnewald U, Balosso S, Cagnotto A, Barbera S, Uboldi S, Vezzani A, Mennini T Neuronal hyperexcitability and seizures are associated with changes in glial-neuronal interactions in the hippocampus of a mouse model of epilepsy with mental retardation J Neurochem 2010, 115: 1445-1454 Mennini T, Testa R Are descending control pathways of the lower urinary tract and pain overlapping systems? Cent Nerv Syst Agents Med Chem 2010 10 : 113-147 Mourtas S, Canovi M, Zona C, Aurilia D, Niarakis A, La Ferla B, Salmona M, Nicotra F, Gobbi M, Antimisiaris S G Curcumin-decorated nanoliposomes with very high affinity for amyloid-1-42 peptide Biomaterials 2010 [Epub ahead of print] Oliva A, Sanchez Ashen D, Salmona M, Farina J B, Llabres M Solid-state stability studies of cholecystokinin (CCK-4) peptide under nonisothermal conditions using thermal analysis, chromatography and mass spectrometry Eur J Pharm Sci 2010 39 : 263-271 Provenza G, Provenzano M, Visai L, Burke F M, Geoghegan J A, Stravalaci M, Gobbi M, Mazzini G, Arciola C R, Foster T J, Speziale P Functional analysis of a murine monoclonal antibody against the repetitive region of the fibronectin-binding adhesins fibronectin-binding protein A and fibronectin-binding protein B from Staphylococcus aureus FEBS J 2010 277 : 4490-4505 Stravalaci M, Beeg M, Salmona M, Gobbi M Use of surface plasmon resonance to study the elongation kinetics and the binding properties of the highly amyloidogenic A(1-42) peptide, synthesized by depsi-peptide technique Biosens Bioelectron 2010 [Epub ahead of print] Taylor M, Moore S, Mayes J, Parkin E, Beeg M, Canovi M, Gobbi M, Mann D M A, Allsop D Development of a proteolytically stable retro-inverso peptide inhibitor of beta-amyloid oligomerization as a potential novel treatment for Alzheimer's disease Biochemistry 2010 49 : 3261-3272 Terao M, Fratelli M, Kurosaki M, Zanetti A, Guarnaccia V, Paroni G, Tsykin A, Lupi M, Gianni' M, Goodall GJ, Garattini E. Induction of MIR-21 by retinoic acid in estrogen-receptor-positive breast carcinoma cells: biological correlates and molecular targets. J Biol Chem. 2010 [Epub ahead of print] Tonutti L, Bononi E, Paris L, Breillout F, Corvaia N, Bailly C, Bazzoni G Effect of microtubule-targeting drugs on cell-cell and cell-matrix junctions in tumor epithelial cells Anticancer Drugs, in press 2010 Tonutti L, Manzi L, Tacconi M T, Bazzoni G Eicosapentaenoic acid inhibits endothelial cell migration in vitro J Angiogenes Res 2010 2 : 12-19 Urru S A M, Veglianese P, De Luigi A, Fumagalli E, Erba E, Gonella Diaza R, Carr A, Davoli E, Borsello T, Forloni G, Pengo N, Monzani E, Cascio P, Cenci S, Sitia R, Salmona M A new fluorogenic peptide determines proteasome activity in single cells J Med Chem 2010 53 : 7452-7460

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RESEARCH ACTIVITIES Laboratory of Biochemistry and Protein Chemistry Development of new therapeutic strategies for the treatment of central and peripheral amyloidosis The development of an effective strategy for the prevention and cure of Alzheimer disease and systemic amyloidosis is of great importance due to the absence of an effective therapy. Their severity affects seriously the life of patients and their relatives. The formation of amyloid fibrils and their deposition in specific tissues were for longtime considered the cause of the disease, however recent studies showed that soluble oligomeric species are the actual culprits of the toxicity. The kinetics of protein aggregation due to conformational modifications and the comprehension of genetic, biochemical and structural determinants at the basis of this transformation are very important for unveiling the pathogenic process and the development of therapeutic strategies. Aiming at developing simple models that enable monitoring the conformational changes that preceds fibril deposition, we have designed and developed a variety of synthetic peptides as deduced from the primary sequence of human amyloidogenic proteins in their wild-type or mutated forms. In collaboration with the Istituto Neurologico Carlo Besta of Milan we have identified a mutated form of beta-amyloid (A673V) that displays amazing biological features since it binds to wild-type beta-amyloid and inhibits amyloid formation and the onset of the disease. This observation opens new therapeutic perspectives both for genetic and sporadic forms of Alzheimer disease based upon the use of protein fragments containing this mutation or peptide-mimetic compounds. Moreover, we have synthesized several Abeta peptides containing the same mutation and we have evaluated its importance in the aggregation and amyloidogenic properties. Similar studies have been carried out with prion protein and some amyloidogenic proteins responsible of peripheral amyloidosis. The first approach for the development of candidate drugs contemplates the development of molecules capable to interfere with oligomeric species following direct interaction with protein molecules disrupting its beta-sheet conformation or the fibrillary aggregates. This activity requires in vitro studies with cell free models to determine the conformational features of amyloidogenic peptides, their secondary structure, the hydrogen-deuterium exchange, the resistance to digestion by proteases, the aggregation propensity and amyloidogenic characteristcs. To understand the molecular and biochemical mechanisms of action underlying the cause of the cytotoxic action, peptides are used for in vitro studies in variety of cellular models trying to correlate their physical features and the biological effect. Moreover, the subcellular distribution of peptides and their molecular targets are also investigated. We have reported that tetracyclines are new candidates as anti-amyloidogenesis drugs, in particular they disrupt amyloid tangles and increase the sensitivity of PrP to proteinase K digestion. Tetracycline are able to inhibit neuronal cell death and astroglial prolipheration induce by PrP peptides and, in animal model of disease, they prolong the survival of animals inoculated with PrP.

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The nematode Caenorhabditis elegans as experimental model to investigate in vivo the molecular mechanisms underlying the aggregation of amyloidogenic proteins The description of the molecular events underlying the in vivo amyloidogenesis is crucial for the design of effective therapeutic strategies. To this end, in our laboratory we use Caenorhabditis elegans as experimental model since it offers the unique opportunity to analyze the genetic and molecular functions of human disease-related genes in vivo. This nematode offers also the major advantages of the easy generation of transgenic strains expressing human genes, the production of distinct phenotypes offers insight into the biology of the disease and help to elucidate fundamental cellular processes related to it. In particular, it is possible to correlate the phenotype of the transgene with the disease insurgence, the degeneration, the protein expression and its aggregation into the oligomeric or fibrillar forms. Different transgenic strains expressing various fragments of human amyloid in neurons or in muscles are available in our laboratory. We also developed new transgenic strains expressing A-V or A-T mutated peptides in position 2 under a neuronal promoter, to evaluate their in vivo effects. The expression of these peptides results in the cytoplasmic amyloid inclusion and in the appearance of progressive phenotypes related to the disease. In these C. elegans strains, amyloid aggregates were observed and they are similar to those observed in the brain of patients with AD or in muscles of patients with sporadic forms of Inclusion Body Myositis, the most common myopathy. These models were already used to study the relationship between A sequence, amyloid formation and toxicity. A transgenic C. elegans strain producing only the oligomeric form of the amyloid protein was also available representing a good predictive model for the investigation of drugs specifically interfering with oligomers. C. elegans is also applied to investigate the molecular mechanisms underlying some systemic amyloidosis, like those caused by tissue deposition of immunoglobulin light chain or 2 microglobuline. Using this multidisciplinary genomic and molecular integrated approach, we will obtain important informations for the development and validation of innovative therapeutic strategies and for the comprehension of the in vivo molecular functions of genes related to human amyloidosis. Nanoparticles in pharmacology: new diagnosis and therapy systems The clinical development of molecules with promising therapeutic activity for the treatment of diseases with unfavorable prognosis, is sometimes limited by the molecules scarse bioavailability, by a rapid clearance, or the difficulty to cross certain biological barriers, and last but not least, by the onset of severe side effects. To overcome those hurdles the usage of biocompatible and biodegradable nanoparticles (NPs) has been suggested. These NPs protect the active compounds loaded in the NPs and act as controlled release devices. Various types of NPs, both lipidic and polymeric, have been used in our laboratories to enhance the release kinetics of the loaded molecules. Modifying the NPs surfaces with particular peptides, antibodies and ligands, it has been possible to change their biodistribution with respect to healthy and tumoral tissues Our laboratory has evaluated, within the European project Nanoparticles for therapy and diagnosis of Alzheimer Disease (NAD), the ability of different NPs of lipidic and

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polymeric nature to cross the blood-brain-barrier in vivo, to deliver anti-amiloidogenic drugs to the brain. Furthermore, in collaboration with Politecnico di Milano and the Swiss Federal Institute of Technology (ETH), new polymeric NPs have been synthesized and their stability has been evaluated in biological fluids. Utilizing non degradable NPs loaded with fluorescent dyes and/or paramagnetic molecules, biodistribution studies have been performed in vivo employing Optical Imaging techniques, Magnetic Resonance Imaging, optical and fluorescence microscopy. These results will represent the basis for the design of NPs for early diagnosis of specific diseases and for monitoring the therapys efficacy. A deeper analysis of the parameters influencing the in vitro and in vivo drug release from NPs are currently in progress. All obtained data will be used for the development of mathematical models able to describe the pharmacokinetics of the NPs and of the released compounds. Preclinical imaging to improve the translation of results from mice to patients One of the main goal of the modern pharmacological research is to translate the results obtained form preclinical models (cells and animals) to the clinical practice. The use of non invasive instruments of screening has been more and more taking place either for the diagnosis or to follow the efficacy of therapy in different clinical fields. The recent development of in vivo imaging instruments dedicated to small rodents may therefore allow to perform the same strategy of investigation already at preclinical level. The Department of Biochemistry and Molecular Pharmacology has been developing a series of experimental procedures aimed at coupling the results obtained by different in vivo analyses (Magnetic Resonance Imaging, micro Computerized Tomography, Fluorescent Molecular Tomography, Ocular Coherence Tomography) to the data obtained by ex vivo studies (histology and/or immunohistochemistry). The integration of these two areas can be identified as preclinical imaging. This approach has been recently exploited to better investigate the clinical progression of an interesting of model of neuronal ceroid lipofuscinosis, the mnd mouse. Analyses carried out by fluoro-angiography and ocular coherence tomography allowed us to characterize the progressive ocular inflammation and retinal degeneration affecting mnd mice by simply following the same group animals during the time. Histological characterization, performed by sacrificing animals at different time points, confirmed these data and highlighted that lipofuscin accumulation, apoptosis of retinal cells and reactive gliosis, are the cellular bases for the alteration revealed by in vitro imaging analyses. A series of experiments will be carried out, by three different degree of resolution, to better characterize this peculiar accumulation of autofluorescent ceroid and lipofuscin-like material in brain and in eyes of mnd mice. In collaboration with the Department of Oncology, we investigated about the anatomical localization of autofluorescent material by a non invasive approach (fluorescent molecular tomography). Such strategy allowed us to follow the progressive deposition of autofluorescent material in the same group of mice at different ages. A marked fluorescence was first observed in the posterior area of forebrain and in the cerebellar region. In older animals the fluorescent signal spread in the whole brain parenchyma and in other peripheral organs.

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Histological analysis (by the observation of the autofluorescence in 20 m thick sections) confirmed the reliability of in vivo imaging and evidenced a selective deposition of autofluorescent material in neurons. Finally, electron microscopy studies (in collaboration with the Department of Cardiovascular Diseases) showed that lipofuscin-like bodies were mainly segregated in swollen lysosomes and distributed in the whole cytoplasm of neurons. Contrast Enhanced (MnCl2) MRI experiments have demonstrated that in the hippocampus of mnd micea marked process of hyperexcitability occurs before motor symptom onset. Hippocampal hyperexcitabilty was further confirmed by EEG analysis (in collaboration with the Laboratory of Experimental Neurology) and by c-fos immunohistochemistry. The body of knowledge emerging from all these experiments allow us to propose the mnd mouse as a reliable model of Epilepsy with mental retardation, one of the most common form of neuronal ceroid lipofuscinosis and associated with mutation(s) of the same gene (cln8) responsible for the phenotypic changes found in mnd mice. In collaboration with the Unit of Cancer Clinical Pharmacology we evaluated, by Gadolinium enhanced MRI, the growth of an orthotopically implanted human glioblastoma in the brain parenchyma of nude mice. In addition, the internalization of paramagnetic and fluorescent probes in human foetal stem cells has allowed to track the fate of these cells once transplanted in cerebral ventricles of healthy and diseased mice (more specifically in a model of amyotrophic lateral sclerosis: the wobbler mouse). Other studies with dual paramagneticfluorescent are now in progress to follow the route of human fetal stem cells by MRI and fluorescent molecular tomography in the same group of animals at different times after transplantation. Laboratory of Molecular Biology The family of molybdo-enzymes Molybdo-enzymes are proteins requiring a molybdo-pterin cofactor (molybdenumcofactor, MoCo) for their catalytic activity. Until a few years ago, it was believed that the family of molybdo-enzymes consisted only of three members: sulfite oxidase, aldehyde oxidase and xanthine oxidoreductase. In the last few years of research, our laboratory has determined the structure of the genes coding for different molybdoenzymes in rodents and humans. In particular, we demonstrated that rodents are endowed with four different aldehyde oxidase (AOX1, AOX3, AOX4 and AOX3L1) characterized by remarkable structural and functional similarity. The physiological substrate(s) and the physiological function(s) of this group of protein have not yet been identified, although it is known that aldehyde oxidases can oxidize aliphatic and aromatic aldehydes into the corresponding carboxylic acids and to hydroxylate different types of n-heterocyclic aromatic rings. The four different aldehyde oxidases of rats and mice are the product of an equivalent number of genes located at the short distance one from the other on the same chromosome. These genes originated through a number of a synchronous gene duplication events. Our studies aimed at the determination of the evolutionary processes underlying the development of the genes coding for aldehyde oxidases allowed us to establish that the natural history of

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this gene family is made of duplication and suppression events. These evolutionary processes resulted in the presence of variable number of aldehyde oxidases in different genomes. Man is characterized by the presence of a single active gene (AOX1) and two inactive pseudo genes clustered on chromosome 2. In the last years we have focused on the functional definition of the different mouse aldehyde oxidases and our long term aim is to establish the reasons underlying the disparity in the number of these enzymes between humans and rodents. To this purpose, we generated two knockout animals for the AOX4 and AOX3L1 genes. The AOX4 knockout mouse was characterized phenotypically demonstrating minimal alterations of the epidermis. Indeed, the AOX4 knockout animal shows epidermal hypertrophy, which is associated with a peculiar fragility of the corneal layer. At the biochemical level, we observed a deficiency in the synthesis of retinoic acid in the two organs where AOX4 is present in significant amounts (skin and Harderian glands). This observation is in line with the idea that AOX4 may have a role in the metabolism of retinaldehyde to retinoic acid, the active metabolite of vitamine A. Recently we gathered novel data indicating a role for AOX4 in the control of the adipose tissue homeostasis. The observation is of particular importance also in man as human AOX1 seems to exert a similar effect in the synthesis and deposition of lipids. Currently we are performing similar studies in a knockout mouse for AOX3L1. Retinoids in the treatment and chemoprevention of myeloid leukemia and mammary carcinoma Our laboratory has a long standing interest in defining the therapeutic potential of natural and synthetic derivatives of retinoic acid, the active metabolite of vitamin A. These compounds, commonly defined as retinoids, are characterized by cytodifferentiating, anti-proliferative and apoptotic effects which are at the bases of their therapeutic activity in the context of myeloid leukemia and mammary carcinoma. Retinoids are very active therapeutic agents, although they are endowed with dose limiting side effects, particularly chronic administration. A rational clinical use of retinoids calls for a better knowledge of the mechanisms of action underlying the antineoplastic action exerted by these compounds. In-depth knowledge is of fundamental value for the design of novel retinoid-based treatment strategies characterized by increased therapeutic index. We have a long-standing interest in the definition of the molecular mechanisms regulating the activity of retinoic acid nuclear receptors, as they may lead to the identification of pharmacological targets to be modulated in a specific manner. Indeed, we believe that knowledge in this field may lead to the development of rational combinations between retinoids and other pharmacologically active agents to be used in the treatment of different tumor types. Such an approach has led us to the recent identification of the prolyl-isomerase, PIN1 as a negative regulator of the retinoic acid receptor, RAR. Pharmacological inhibitors PIN1 proved to be particularly effective in sensitizing the leukemic cell to the anti-neoplastic activity of retinoids. These results open up the possibility to develop combinations based on PIN1 inhibitors and retinoids for the treatment of acute myeloid leukemia. Following the same type of logic, we have recently demonstrated that the inhibition of the microRNA, miR21 in mammary carcinomas positive for estrogen receptor is of the utmost importance in potentiating the anti-proliferative activity of retinoids in this particular type of tumor. Finally, we observed that the peculiar subgroup of mammary cancer positive for HER2 may benefit

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from retinoid-based treatment or associations between retinoids and inhibitors of HER2 receptor tyrosine kinase activity. Laboratory of of Pharmacodynamics and Pharmacokinetics Misfolding proteins and related diseases One of the laboratorys main research fields regards the diseases associated with protein misfolding, i.e. the formation of aberrant tertiary conformations of proteins or peptides, as a consequence of mutations, stress or aging. Besides the loss of the proteins physiological properties, the misfolding often results in new biochemical properties, particularly the propensity to aggregate and form amyloid-like deposits. We are particularly interested in Alzheimers disease (AD), in which there is aggregation of amyloid- (A) peptides (A1-40 and A1-42, detectable in the amyloid plaques typical of AD brain), and in spongiform encephalopathies, due to misfolding and aggregation of the prion protein (PrP). Recent studies suggest that misfolding and the consequent propensity to form toxic aggregates is common to different proteins and results in different diseases (e.g. alpha-synuclein for Parkinson disease, poly-Q expansions for Huntington disease, superoxide dismutase in amiotrophic lateral sclerosis, transthyretin in systemic amyloidosis). Better knowledge of the molecular and cellular mechanisms involved in these events is needed for the development of useful therapeutic strategies. We have investigated the kinetics and the mechanisms underlying protein aggregation in different experimental conditions, using different biochemical and chemical-physical techniques. In particular, we applied surface Plasmon resonance (SPR) to analyze the polymerization kinetics of A1-42 fibrils (Stravalaci et al., Biosens Bioelectron, 2011), to investigate the effects of A mutations (Di Fede et al., Science, 2009), and to test molecules with potential anti-amyloidogenic activities (Di Fede et al., Science, 2009, Taylor et al., Biochemistry, 2010). We have also optimized experimental protocols to study the different aggregation states of amyloidogenic peptides, including the soluble oligomers thought to be the most toxic species. This enabled us, for example, to describe the interaction between PrP and A1-42 oligomers (Balducci et al., PNAS, 2010). Our laboratory is a partner in a European network (FP7), which plans to develop nanoparticles for therapy and diagnosis of Alzheimer disease (NAD), focusing on A as the target. We have evaluated the affinities of functionalized nanoparticles for A1-42 fibrils, immobilized on the SPR sensor surface (Gobbi et al., Biomaterials, 2010; Mourtas et al., Biomaterials, 2011). We are also involved in examining the pharmacokinetic properties of these nanoparticles, especially their blood-brain barrier passage. By studying both pharmacodynamic and pharmacokinetic properties of nanoparticles our laboratory adds value in this expanding field of research . Q10 coenzyme in amyotrophic lateral sclerosis Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disease involving brainstem and spinal cord motoneurons, leading to complete paralysis of skeletal muscles and early death, usually from respiratory failure. Mutations in the copper-zinc superoxide dismutase (SOD1) gene are responsible of some forms of familial ALS, and on this basis transgenic rodents have been generated, contributing significantly to our

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understanding of the pathogenic mechanisms of the disease. For example, transgenic mice carrying the mutant SOD1, and showing motoneuron degeneration, have increased oxidative stress associated with mitochondrial damage. The Q10 coenzyme (CoQ10, or ubiquinone) is a major component of the mitochondrial respiratory chain, and its reduced form ubiquinol has antioxidant proprieties. We are therefore investigating whether there is a correlation between CoQ10 levels (measured in plasma and CNS) and the disease progression and survival in SOD1 transgenic mice. We shall also investigate whether Q10 supplementation has positive effects on the disease progression. These studies are carried out in collaboration with the Laboratory of Molecular Neurobiology (headed by Dr. C. Bendotti), which is leading most of the research on ALS carried out at the Mario Negri Institute.

Synaptopathies A number of observations suggest that the neuronal degeneration in misfolding diseases is preceded by more subtle cellular dysfunctions of synaptic transmission (synaptopathies), in many cases thought to be caused by the toxic aggregates described above. In collaboration with other laboratories of the Mario Negri Institute (e.g. the Neurobiology of Prions laboratory , headed by Dr. R. Chiesa), we are characterizing glutamatergic and GABAergic transmission in brains of transgenic animal models of these diseases, using the biochemical methods available in our laboratory.

Glutamatergic and serotoninergic neuropharmacology Our laboratory has long experience in neuropharmacological studies, particularly on the glutamatergic and serotoninergic neurotransmission systems. Specific biochemical techniques to analyze the main mechanisms of synaptic transmission (neurotransmitter release, binding to receptors and reuptake by specific transporters) are used in collaboration with chemico-pharmaceutical departments, for the characterization of new molecules acting on these mechanisms.

Molecular interactions SPR, an advanced technique specifically developed for the study of molecular interactions, enables us to make useful contributions to different projects in collaboration with other laboratories of the Institute (Gesuete et al., Annals of Neurology, 2009; Colombo et al., J Biol Chem, 2010; Deban et al., Nature Immunology, 2010; Provenza et al., FEBS J, 2010). In particular, one of these projects, carried out in collaboration with the Inflammation and Nervous System Disease Laboratory headed by Dr. M.G. De Simoni, is investigating the idea that MBL may play a role in ischemia-induced damage (Gesuete et al., Annals of Neurology, 2009), where MBL inhibitors might have significant antiischemic effects. Studies include the synthesis of new potential MBL ligands (Department of Organic and Industrial Chemistry, University of Milan, headed by Dr. A. Bernardi), evaluation of their ability to interact with MBL in vitro (through SPR studies in our laboratory) and their anti-ischemic effects in vivo (De Simonis laboratory). The laboratory is currently collaborating with the Mario Negri Institute in Bergamo (laboratories of Dr. M. Morigi and Dr. M. Noris) in research into new molecular mechanisms involved in the haemolytic uremic syndrome (HUS) and thrombotic

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thrombocytopenic purpura (TTP). We are using SPR to investigate whether new protein-protein interactions contribute to the link between thrombosis and complement cascade activation. Our laboratory is a partner in a multicentre triennial project sponsored by the Regione Lombardia (Metadistretti) entitled Miniaturized System for Molecular Diagnostic and Proteomic of Sepsis Based on Integration of Surface Plasmon Resonance, aiming at identifying new biomarkers of sepsis and exploiting new SPR systems as low cost, rapid diagnostic tools. Our laboratory will be responsible for biomarker identification and validation, measuring them in plasma with our conventional SPR system.

Laboratory of Molecular Pathology In vitro models for investigating motor neuron pathologies Mutant forms of specific proteins play a key role in many neurodegenerative diseases. Experimental models in vivo and in vitro are sorely needed to study the effects of these toxic proteins. The motor neuronal cell line NSC-34, a widely used model to study motor neuron degeneration, is available in the laboratory. We have applied the pTet-Off system to control gene expression through the level of tetracyclines to the NSC-34 cell line establishing a new cell line (NSC-34 tTA40) that stably expresses the transactivating protein tTA. This cell line is suitable to study the pathogenic mechanisms of motor neuron diseases after transient/stable transfection with genes of interest for these pathologies. The NSC-34 and the NSC-34 tTA40 cell lines were used to obtain in vitro models to study the pathogenic mechanisms of amyotrophic lateral sclerosis (ALS). Mutant forms of superoxide dismutase 1 are responsible for some of the familial forms of ALS. We developed NSC-34-based cell lines expressing constitutively or conditionally human G93A mutant superoxide dismutase 1 (G93ASOD1). Novel intracellular targets in the selective degeneration of motor neurons in amyotrophic lateral sclerosis Amyotrophic lateral sclerosis (ALS) is a rapidly fatal neurodegenerative disease characterized by loss of motor neurons. The management of this disease remains essentially supportive and symptomatic. Understanding the mechanisms underlying the disease is a way to favor more efficient therapeutic strategies. We utilized our cell models to investigate the biochemical-molecular mechanisms underlying the alterations of mitochondrial morphology observed in the early stages of the disease in the motor nerve terminals of ALS patients and in the murine models of the disease. We showed that motor neurons are selectively susceptible to mitochondrial damage induced by a mutant form of human superoxide dismutase 1 (G93ASOD1) and that this damage was modulated by the extent of expression of the mutant protein. Furthermore the expression of G93ASOD1 protein increased the susceptibility of motor neurons to inhibitors of the electron transport chain (ETC) and to oxidants. Exposure to drugs or exogenous compounds impairing the ETC could thus be a risk factor to motor neurons of individuals carrying mutant superoxide dismutase 1. We have shown that in motor neuronal cells the activity of glutamate cysteine ligase, the rate limiting enzyme for glutathione biosynthesis, was modulated by the level of

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G93ASOD1. As a consequence, the level of glutathione, the main cell antioxidant, increased or decreased. A variation in the level of glutathione may influence the formation of nitrated proteins, a pathogenic mechanism in ALS, which was investigated in collaboration with the laboratory of Translational Proteomics. Cytochrome P-450 superfamily Cytochrome(s) P-450 have evolved into a large superfamily which plays a major role in the metabolism of drugs and other chemicals. The majority of existing drugs depends on the P-450 system for terminating their biological effects or for side effects or adverse reaction. The laboratory has a long-standing interest in the induction/degradation mechanisms of specific cytochrome P-450 families due to drug administration or to disease states. Activation of enzymes of the heme metabolic pathway (heme oxygenase system, biliverdin reductase) as a protective response to stress The enzymatic system of heme oxygenase (HO) is devoted to cellular degradation of heme containing molecules, like cytochromes and hemoglobin, and to recycling of iron. Products formed by the catalytic activity of HO - carbon monoxide and bile pigments are important regulating factors in the cell. An increase of HO activity (which is usually sustained by activation of the inducible form HO-1) is now considered a protective mechanism against untoward stimuli particularly when oxidative stress is involved. In the past, the laboratory of Molecular Pathology identified cytokines as inducers of HO activity and as transcriptional activators of the HO-1 gene. We are currently investigating the functional significance of HO-1 activation in neurodegeneration.
Laboratory of Translational Proteomics

Nitrative stress and protein aggregation in amyotrophic lateral sclerosis: a proteomic approach The molecular mechanisms at the basis of neurodegenerative diseases including the genetically linked ones, such as amyotrophic lateral sclerosis (ALS), are still unknown. However, there is evidence that nitrative stress and protein aggregation play central roles in the pathogenesis of such diseases. In collaboration with the Laboratory of Molecular Neurobiology at the Mario Negri Institute in Milano we conducted proteome analysis of an animal model of familial ALS (fALS). We focused the attention on the analysis of protein expression changes and protein modifications, such as tyrosine nitration (marker of nitrative stress), in a transgenic mouse, which over-express human mutated (G93A) superoxide dismutase (SOD1). We analyzed, by proteomic tools, spinal cord of presymptomatic G93A SOD1 mice, we identified nitrated proteins and quantified the level of nitration for each protein in comparison with healthy controls. We revealed that there was a substantial increase of the nitration level in at least five proteins: actin, alpha and gamma enolase, ATP synthase and a chaperone protein, HSC71. The alteration of the function of these proteins, due to the oxidative modification, may have important consequences on the cellular metabolism/catabolism, signaling pathways and phosphorylation cascades, therefore may be at the basis of the molecular mechanisms leading to neurodegeneration. Regarding the aggregation studies, we carried out a proteomic analysis of the protein composition of the insoluble

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fraction, as a model of protein aggregates, from the fALS mouse model at different disease stages. We identified several proteins enriched in the detergent-insoluble fraction already at a preclinical stage, including intermediate filaments, chaperones and mitochondrial proteins. Aconitase, HSC70 and cyclophilin A were also significantly enriched in the insoluble fraction of spinal cords of ALS patients. Moreover, we found that the majority of proteins in mice and HSP90 in patients were tyrosine-nitrated. In collaboration with the Laboratory of Molecular Pathology at the Mario Negri Institute in Milan we therefore investigated the role of nitrative stress in aggregate formation in fALS-like murine motor neuron-neuroblastoma (NSC-34) cell lines. By inhibiting nitric oxide synthesis the amount of insoluble proteins, particularly aconitase, HSC70, cyclophilin A and SOD1 can be substantially reduced. In conclusions, analysis of the insoluble fractions from cellular/mouse models and human tissues revealed novel aggregation-prone proteins and suggests that nitrative stress contribute to protein aggregate formation in ALS. Identification of biomarkers of ALS In collaboration with the Laboratory of Molecular Neurobiology at the Mario Negri Institute, Fondazione Salvatore Maugeri, IRCCS, Milano, and NEuroMuscular Omnicentre (NEMO), Niguarda Ca Granda Hospital, Milano we are conducting a series of studies with the aim to identify biomarkers of ALS useful in diagnosis and prognosis and to unravel pathogenetic mechanisms, still unknown. Increased levels of nitrotyrosine in the central nervous system have been found in patients and mouse models of fALS, suggesting a possible use of nitrated proteins as biomarkers. We analyzed peripheral blood mononuclear cells (PBMCs), easily accessible samples, from sporadic ALS (sALS) patients and a rat model of fALS (a) to establish whether an increased level of nitrated proteins was present in PBMCs, too, and (b) to identify possible candidate biomarkers. With a proteomic approach, we identified for the first time the major over-nitrated proteins in PBMCs from patients and rats at different disease stages. In the rats, their increased levels were measured already at a presymptomatic stage. Among them, actin, ATP synthase, and vinculin overlap between sALS patients and the rat model. Both in patients and in rats the nitration level is at least twice than the one in the controls. Interestingly, in a previous study, actin and ATPase have been found over nitrated in the spinal cord of a mouse model of fALS before disease onset, suggesting their possible involvement in motor neuron degeneration. In conclusion, we observed that an increased level of nitrated proteins was not restricted to the spinal cord but also was present in peripheral cells of patients and an animal model, and that nitrated proteins are promising candidate biomarkers for early diagnosis of ALS. We are now carrying out a global proteomic analysis of PBMC of sALS patients to identify other protein alterations to be used as disease biomarkers. Proteomic analysis of a cellular model of fatal familial insomnia The prion protein (PrP) is a glycosyl-phosphatidyl-inositol (GPI)-anchored membrane glycoprotein that plays a vital role in prion diseases, a class of fatal neurodegenerative disorders of humans and animals. Approximately 20% of human prion diseases display autosomal dominant inheritance and are linked to mutations in the PrP gene on chromosome 20. PrP mutations are thought to favor the conformational conversion of PrP into a misfolded isoform that causes disease by an unknown mechanism. The PrP

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mutation D178N/M129 is linked to fatal familial insomnia, which causes severe sleep abnormalities and autonomic dysfunction. In collaboration with the Laboratory of Neurobiology of Prions at the Mario Negri Institute in Milan we found that mutant PrP accumulates abnormally in the endoplasmic reticulum and Golgi of transfected neuroblastoma N2a cells. We then investigated the impact of intracellular PrP accumulation on cellular homeostasis, we did a 2D gel-based differential proteomic analysis. We used wide-range immobilized pH gradient strips, pH 4-7 and 6-11, to analyze a large number of proteins. We found changes in proteins involved in energy metabolism, redox regulation and vesicular transport. Rab GDP dissociation inhibitor alpha (GDI) was one of the proteins that changed most. GDI regulates vesicular protein trafficking by acting on the activity of several Rab proteins. We found a specific reduction in the level of functional Rab11 in mutant PrP expressing cells, associated with impaired post-Golgi trafficking. Our data are consistent with a model by which mutant PrP induces over-expression of GDI activating a cytotoxic feedback loop which leads to protein accumulation in the secretory pathway. Laboratory for the Study of Biological Systems System-level analysis of protein interactions in the epithelial junctional complex Inter-cellular junctions form the apical junctional complex and mediate adhesion between adjacent cells, thus representing the cellular basis for tissue cohesion (for instance, the epithelial lining of the intestine). In order to acquire system-level understanding of the apical junctional complex, we have studied (using a methodological approach of network analysis) all the protein interactions that have been described at the junctions in epithelial cells of human origin. We also found that proper hubs (i.e., very rare proteins with an exceedingly high number of interactions with other proteins) were absent from the junctional network. Nevertheless, we observed that the most connected (albeit non-hub) proteins were also essential proteins. In addition, we have detected modules within the junctional networks (i.e., densely inter-connected groups of proteins). Analysis of the modules has highlighted general organizing principles of the junctional complex, thus confirming the usefulness of network analysis for studying the components and the interactions of the cell.

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DEPARTMENT OF EPIDEMIOLOGY
STAFF

Head Laboratory of General Epidemiology

Head

Carlo LA VECCHIA, M.D.

Carlo LA VECCHIA, M.D.

Cancer Epidemiology Unit

Head Cristina BOSETTI, Mat.Sci.D.

Lifestyle Habits and Prevention Unit

Head Liliane CHATENOUD, Biol.Sci.D.

Epidemiology for Clinical Research Unit

Head Silvano GALLUS, Comp.Sci.D.

Laboratory of Epidemiological Methods

Head Eva NEGRI, Mat.Sci.D.

Laboratory of Epidemiology of Chronic Diseases

Head Alessandra TAVANI, Biol.Sci.D.

Laboratory of Medical Informatics

Head Eugenio SANTORO, Comp.Sci.D.

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CURRICULA
Carlo La Vecchia received his medical degree from the University of Milan and a master of science degree in clinical epidemiology from Oxford University. He is recognized worldwide as a leading authority in cancer etiology and epidemiology. Work experiences: Presently, he is Chief of Epidemiology at the Mario Negri Institute in Milan, Italy and on the faculty of Medicine at the University of Milan. Dr. La Vecchia serves as an editor for numerous clinical and epidemiologic journals. He is among the most renowned and productive epidemiologists in the field with over 1,560 peer-reviewed papers in the literature and is among the most highly cited medical researchers in the world, according to ISIHighlyCited.comsm, the developer and publisher of the Science Citation Index. Dr. La Vecchia is an Adjunct Professor of Medicine at Vanderbilt Medical Center and the Vanderbilt-Ingram Cancer Center and of Epidemiology at the University of Lausanne, CH. Dr. La Vecchia is a temporary advisor at the International Agency for Research on Cancer IARC/WHO and at the World Health Organization in Geneva, and a registered journalist in Milan. He was Adjunct Associate Professor of Epidemiology at Harvard School of Public Health between 1996 and 2001. Areas of interest: Dr. La Vecchias main fields of interest include cancer epidemiology and the risk related to diet, tobacco, oral contraceptive use and occupational or environmental exposure to toxic substances; and analysis of temporal trends and geographical distribution of mortality from cancer, cardiovascular diseases, perinatal and other selected conditions. Eva Negri got a degree in Mathematics in 1985 at the University of Milan, School of Mathematics. Work experiences: Since 2007: Laboratory Chief, Unit of Epidemiologic Methods, Department of Epidemiology; 1992-2006: Unit Chief, Unit of Epidemiologic Methods, Laboratory Epidemiology; since 1990-1992: Researcher at the Laboratory of Epidemiology; 1984-1990: Collaborator of the Laboratory of Epidemiology. Areas of interest: Design, conduction and analysis of epidemiologic studies on chronic diseases (e.g. cancer and myocardial infarction) and injuries, analysis of mortality of cohorts of workers, analysis of temporal trends and geographic distribution of mortality from cancer, cardiovascular disease, injuries and other selected conditions, analysis of national health surveys, application of linear modeling techniques to the analysis of epidemiological data, collaborative re-analyses and meta-analyses of epidemiological studies. Awards: EEC scholarship for postgraduate training in Epidemiology (1988).
Selected publications Negri E, La Vecchia C, Pelucchi C, Tavani A The risk of acute myocardial infarction after stopping drinking Prev Med 2005; 40: 725-728 Negri E, Pelucchi C, Talamini R, Montella M, Gallus S, Bosetti C, Franceschi S, La Vecchia C Family history of cancer and the risk of prostate cancer and benign prostatic hyperplasia Int J Cancer 2005; 114: 648-652 Negri E, Little D, Boiocchi M, La Vecchia C, Franceschi S. B-cell non-Hodgkins lymphoma and hepatitis C virus infection: A systematic review Int J Cancer 2004; 111: 1-8 Negri E, Ron E, Franceschi S, La Vecchia C, Preston-Martin S, Kolonel L, et al. Risk factors for medullary thyroid carcinoma: A pooled analysis Cancer Causes Control 2002; 13: 365-372 Levi F, La Vecchia C, Boyle P, Lucchini F, Negri E Western and eastern European trends in testicular cancer mortality Lancet 2001; 357: 1853-1854

Alessandra Tavani - degree in Biological Sciences, University of Milan, Italy (July 1977); Pharmacological Research Specialist, Mario Negri Institute for Pharmacological Research, Milan, Italy (July 1979). Work experiences: 1979-81: Researcher at the laboratory of Drug Metabolism, Mario Negri Institute for Pharmacological Research. 1981: Researcher at the Unit for Research on Addictive Drugs (director prof. H.W. Kosterlitz), University of Aberdeen, Scotland, U.K. 1982-1990: Head of the Unit of Opioid Neuropharmacology, Mario Negri Institute for Pharmacological Research. 1990: Researcher at the Unit of Clinical Perinatal Pharmacology, Mario Negri Institute for Pharmacological Research. From 1991-2006: Head of the Unit of Epidemiology of Chronic Diseases of the Laboratory of Epidemiology, Mario Negri Institute for Pharmacological Research. 2007-: Head of the Laboratory of Epidemiology

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of Chronic Diseases of the Department of Epidemiology, Mario Negri Institute for Pharmacological Research. Areas of interest: Epidemiology of cancer and coronary heart disease. Organization of case-control studies and cohort studies on cancer and coronary heart disease, including biological sample collection. Analyses of risk factors related to genetic factors and lifestyles, particularly coffee, diet, physical activity. Awards: "Rafaelsen Scholar Award" from the Collegium Internationale Neuro-Psychopharmacologicum (CINP), 16th Meeting, Munich (F.R.G.), 1988.
Selected publications Bravi F, Scotti L, Bosetti C, Gallus S, Negri E, La Vecchia C, Tavani A. Coffee drinking and endometrial cancer risk: a metaanalysis of observational studies. Am J Obstet Gynecol 2009; 200: 130-135 Herrero R, Castellsague X, Pawlita M, Lissowska J, Kee F, Balaram P, Rajkumar T, Sridhar H, Rose B, Pintos J, Fernandez L, Idris A, Sanchez M J, Nieto A, Talamini R, Tavani A, et al. Human papillomavirus and oral cancer: The International Agency for Research on Cancer Multicenter Study. J Natl Cancer Inst 2003; 95: 1772-1783 Galeone C, Tavani A, Pelucchi C, Turati F, Winn D M, Levi F, Yu G - P, Morgenstern H, Kelsey K, Dal Maso L, Purdue M, McClean M, Talamini R, Hayes R B, Franceschi S, Schantz S, Zhang Z F, Ferro G, Chuang S - C, Boffetta P, La Vecchia C, Hashibe M. Coffee and tea intake and risk of head and neck cancer: pooled analysis in the international head and neck cancer epidemiology consortium. Cancer Epidemiol Biomarkers Prev 2010 ; 19: 1723-1736 Galeone C, Turati F, La Vecchia C, Tavani A. Coffee consumption and risk of colorectal cancer: a meta-analysis of casecontrol studies. Cancer Causes Control 2010 ; 21: 1949-1959 Galeone C, Turati F, La Vecchia C, Tavani A. Coffee consumption and risk of colorectal cancer: a meta-analysis of casecontrol studies. Cancer Causes Control 2010 ; 21: 1949-1959

Eugenio Santoro got his degree in Computer Science in 1990 at the Milan University. He started to work at the Mario Negri Institute in 1985 as a research fellow. He was Head of the Applied Statistics and Informatics Unit and of the Applied Statistics and Informatics laboratory, which was part of the Department of Cardiovascular Research. Since 2001 he is Head of the Laboratory of Medical Informatics that is currently part of the Department of Epidemiology. His main areas of interest have been biostatistics and clinical informatics with the development of software for data management and data analyses of large scale clinical trials in cardiology, such as the GISSI studies (Gruppo Italiano per lo Studio della Sopravvivenza nellInfarto miocardico). His main current area of interest is the Internet, and more recently the web 2.0, the social media, and their application in the medical field, in clinical research, and in medical education through the development of health related websites. He is author or co-author of more than 200 scientific papers published in peer reviewed journals, and of more than 70 scientific abstracts submitted to the main international meetings in the cardiology and in the computer science fields. He is also author of three books (available in Italian) about the use of the Internet in medicine (Web 2.0 and medicine, Guida alla medicina in rete and Internet in medicina. Guida alluso e applicazioni pratiche, published by the Pensiero Scientifico Editore, Rome) and of one section about Internet and medicine, included in one of the most important italian medical encyclopedia (Enciclopedia Medica Italiana, UTET 2007). He also collaborates to the publication of the Italian National Bioethics Committees guidelines about ethics, health, and the new information technologies.
Selected publications Santoro E. "Web 2.0 e Medicine: how social networks, podcasts, wikis and blogs are transforming communication, care, and education in health. Il Pensiero Scientifco Editore, Roma 2009. Santoro E., Tinazzi A.Clinical Trials Data Management. In Clinical Trials Handbook (Wiley 2009, Edited by Gad S.C.). Santoro E. Podcast, wiki e blog: il web 2.0 al servizio della formazione e dellaggiornamento del medico. Recenti Prog Med 2007;98:484-494. Santoro E, Rossi Valentina, Pandolfini C, Bonati M. DEC-NET: The development of the European register of clinical trials on medicines for children. Clin Trials 2006; 3: 366-375 Clivio L, Tinazzi A, Mangano S, Santoro E. The contribution of information technology: Towards a better clinical data management. Drug Dev Res 2006; 67: 245-250 Santoro E. Internet and information on breast cancer: an overview. Breast 2003; 12: 424-431

Cristina Bosetti got her degree in Mathematics in 1994 at the University of Milan, School of Mathematics, and the Post-Graduate Diploma in Pharmacological Research in 1999 at the Mario Negri Institute for Pharmacological Research in Milan.

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Work experiences: She is Head of the Unit of Cancer Epidemiology, Department of Epidemiology, Istituto di Ricerche Farmacologiche Mario Negri, Milan since 2005. Previous work experiences include: Visiting scientist at Life style and cancer group of the International Agency for Research on Cancer (IARC), Lyon, France (Oct 2009); Collaboration with the International Epidemiology Institute, Rockville, MD, USA (2002-2009); Visiting scientist at the Unit of Field and intervention studies, IARC, Lyon, France (Sept. 2000/June 2001); Visiting scientist at the Department of Epidemiology, Harvard School of Public Health, Boston, MA (Sept-Nov 1998); Researcher at the Laboratory of Epidemiology, Istituto di Ricerche Farmacologiche Mario Negri, Milan (1998-2005); Researcher at the Laboratory of Mother and Child Health, Istituto di Ricerche Farmacologiche Mario Negri, Milan (1996-1997). Areas of interest: Epidemiology of cancer, cardiovascular diseases and other chronic conditions. In particular case-control studies on cancers of the upper respiratory and digestive sites, thyroid, breast, hormone-related cancers, and on ischemic heart disease; analysis of risk related to diet, alcohol, tobacco, reproductive and hormonal factors, occupational and environmental exposure to toxic substances, through the application of generalized linear models; meta-analysis and systematic reviews of the epidemiologic evidence on cancer risk in relation to various (environmental) exposures . She is author/coauthor of more than 230 publications on peer-reviewed scientific journals publications on peer-reviewed scientific Journals cited in PubMed/MEDLINE. Mean I.F.: 3.8. H-index: 34 (on Google Scholar or SCOPUS).
Selected publications: Bosetti C, Scelo G, Chuang SC, Tonita JM, Tamaro S, Jonasson JG, Kliewer EV, Hemminki K, Weiderpass E, Pukkala E, Tracey E, Olsen JH, Pompe-Kirn V, Brewster DH, Martos C, Chia KS, Brennan P, Hashibe M, Levi F, La Vecchia C, Boffetta P. High constant incidence rates of second primary cancers of the head and neck: A pooled analysis of 13 cancer registries. Int J Cancer. 2010 Sep 7. [Epub ahead of print] Bosetti C, Bertuccio P, Chatenoud L, Negri E, Levi F, La Vecchia C. Childhood cancer mortality in Europe, 1970-2007. Eur J Cancer. 2010;46:384-94. Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C. Tobacco Smoking, Smoking Cessation, and Cumulative Risk of Upper Aerodigestive Tract Cancers.Am J Epidemiol. 2007; 167:468-73. Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, 19702000. Ann Oncol 2005; 16: 489-511. Bosetti C, Spertini L, Parpinel M T, Gnagnarella P, Lagiou P, Negri E, et al. Flavonoids and breast cancer risk in Italy. Cancer Epidemiol Biomarkers Prev 2005; 14: 805-808. Smith J S, Herrero R, Bosetti C, Munoz N, Bosch F X, Eluf-Neto J, et al. IARC Multicentric Cervical Cancer Study Group Herpes simplex virus-2 as a human papillomavirus cofactor in the etiology of invasive cervical cancer. J Natl Cancer Inst 2002; 94: 1604-1613.

Liliane Chatenoud Doctor in Science Biology, University of Milan (1987); Postgraduate Doctor in Health Statistics University of Milan (1995). Work experiences: Since Sept. 2005: Unit Head, Lifestyle and Prevention, Department of Epidemiology. 1993-2005: Researcher at the Laboratory of Epidemiology; 1988-1990: Staff Statistician Bracco S.p.A., Milan. Areas of interest: Epidemiological studies on obstetric diseases. Dermato-epidemiology. Cancer epidemiology (case-control studies on cancers of the breast, female genital tract). Analysis of temporal trends and geographical distribution of perinatal, infant mortality, cancer and other selected conditions (over 150 publications on these topics, 1993-2005).
Selected publications Chatenoud L, Malvezzi M, Pitrelli A, La Vecchia C, Bamfi F. Asthma mortality and long-acting beta2-agonists in five major European countries, 1994-2004. J Asthma 2009 46 : 546-551 Naldi L, Chatenoud L Registry research in dermatology. Dermatol Clin 2009 27 : 185-19 Naldi L, Chatenoud L, Belloni A, Peserico A, Balato N, Virgili A R, Bruni P L, Ingordo V, Lo Scocco G, Solaroli C, Schena D, Di Landro A, Pezzarossa E, Arcangeli F, Gianni C, Betti R, Carli P, Farris A, Barabino G F, La Vecchia C, Parazzini F Medical history, drug exposure and the risk of psoriasis. Evidence from an Italian case-control study Dermatology 2008 216 : 125-132 Valentini L G, Casali C, Chatenoud L, Chiaffarino F, Uberti Foppa C, Broggi G Surgical site infections after elective neurosurgery: a survey of 1747 patients. Neurosurgery 2008 62 : 88-95 Chatenoud L, Mosconi P, Malvezzi M, Colombo P, La Vecchia C, Apolone G. Impact of a major thermoelectric plant on self-perceived health status. Prev Med. 2005;41:328-33.

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Naldi L, Chatenoud L, Linder D, Belloni Fortina A, Peserico A, Virgili AR, et al. Cigarette smoking, body mass index, and stressful life events as risk factors for psoriasis: results from an Italian case-control study. J Invest Dermatol. 2005;125:61-7. Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, 19702000. Ann Oncol 2005; 16: 489-511.

Silvano Gallus was born in Milan on the 20th of November 1970, and got his degree in Computer Science in 1999 at the University of Milan. Work experiences: Chief of the Unit of Epidemiology for Clinical Research of the Department of Epidemiology (since 2006); computer analyst, graphic designer, and statistical and epidemiological consultant, Milan and Bergamo (since 2002); researcher at the Laboratory of Epidemiology (since 1997); creator, designer and webmaster of the website of one of the major Italian public hospital, Milano (19992002). Areas of interest: Design, data managing, and statistical analyses of case-control studies on the associations between several risk factors (including in particular tobacco smoking, alcohol drinking and Mediterranean diet) and risk of cancer, coronary heart disease and several other conditions. Analyses of occupational cohort studies. Monitoring of prevalence and trends of smoking habit and obesity in Italy and Europe. Author/coauthor of over 160 publications in peer-reviewed journals since 1998. Since 2008, Dr Gallus is Associate Editor (Deputy Section Editor since 2010) of the journal BMC Public Health, and is member of the editorial board of the following journals: The Open Obesity Journal (since 2008), The Open Demography Journal (since 2009), World Journal of Gastrointestinal Oncology (since 2009), World Journal of Dermatology (since 2010).
Selected publications Parente F, Molteni M, Marino B, Colli A, Ardizzone S, Greco S, Sampietro G, Foschi D, Gallus S. Are colonoscopy and bowel ultrasound useful for assessing response to short-term therapy and predicting disease outcome of moderate-tosevere forms of ulcerative colitis?: a prospective study. Am J Gastroenterol. 2010;105:1150-7. Gallus S, Naldi L, Carli P, La Vecchia C; Italian Group for Epidemiologic Research in Dermatology (GISED). Nevus count on specific anatomic sites as a predictor of total body count: a survey of 3,406 children from Italy. Am J Epidemiol. 2007;166:472-8. Gallus S, Scotti L, Negri E, Talamini R, Franceschi S, Montella M, Giacosa A, Dal Maso L, La Vecchia C. Artificial sweeteners and cancer risk in a network of case-control studies. Ann Oncol. 2007;18:40-4. Gallus S, Schiaffino A, La Vecchia C, Townsend J, Fernandez E. Price and cigarette consumption in Europe. Tob Control. 2006;15:114-9. Gallus S, Zuccaro P, Colombo P, Apolone G, Pacifici R, Garattini S, La Vecchia C. Effects of new smoking regulations in Italy. Ann Oncol. 2006;17:346-7. Clifford GM, Gallus S, Herrero R, Muoz N, Snijders PJ, Vaccarella S, Anh PT, Ferreccio C, Hieu NT, Matos E, Molano M, Rajkumar R, Ronco G, de Sanjos S, Shin HR, Sukvirach S, Thomas JO, Tunsakul S, Meijer CJ, Franceschi S; IARC HPV Prevalence Surveys Study Group. Worldwide distribution of human papillomavirus types in cytologically normal women in the International Agency for Research on Cancer HPV prevalence surveys: a pooled analysis. Lancet. 2005;366:991-8.

ACTIVITIES
The Department of Epidemiology is involved in the epidemiology of several common cancers (including cancers of the breast, female genital tract, respiratory and digestive sites, prostate and urinary organs, lymphoid malignancies, melanoma, etc.) and of cardiovascular diseases, both through a descriptive and an analytical approach. Among the activities of descriptive epidemiology are the analysis of temporal trends and geographical distribution of mortality from cancer, cardiovascular diseases, and other selected conditions, in Italy and Europe; the analysis of trends in tobacco consumption in the Italian population, and the corresponding effects on incidence and mortality from lung and other tobacco-related neoplasms. The analytic epidemiology activities include the conduction and analysis of case-control studies, aimed at identifying and better quantifying the association between genetic factors (family history), selected lifestyle habits (diet, tobacco, alcohol, etc.), use of exogenous hormones and exposure to various substances and the development of various forms of cancers and cardiovascular

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diseases. In particular, the Department works on the analysis of dietary correlates of cancer and cardiovascular disease risk; quantification of health effects of tobacco smoking, alcohol consumption and implications for prevention; epidemiological studies on the risk related to oral contraceptive and hormone replacement therapy use; evaluation of the impact of screening in the early diagnosis and prevention of cancer. Other activities include: the conduction of quantitative reviews and meta-analysis of published data; the re-analysis of original data from epidemiological studies of cancers of the oral cavity and pharynx, pancreas, thyroid, breast, ovary, and cervix; the analysis of historical cohort studies of occupational exposures to aromatic amines, asbestos, herbicides and other known or potential carcinogens; monitoring and prevention of injuries; epidemiological and clinical studies in dermatology, pediatrics and oncology in collaboration with other Italian groups. Other Departments activities are related to the development of medical websites, the study of the quality of medical information available on the Internet, and the training and research on issues related to medical informatics and those concerning the use in the medical field of the Internet, the social media, and the web 2.0 applications.

MAIN FINDINGS
Diets rich in foods of animal origin and animal fats were positively, and those rich in fruit, vegetables and vegetable fats inversely related to oral and pharyngeal cancer risk. Similarly, a diet rich in animal products and animal fats was directly related to laryngeal cancer risk, while one rich in fruit and vegetables was inversely associated. We found that a lower BMI enhanced smoking- and drinking-related odds ratios for oral cavity/pharyngeal cancer, but did not modify smoking and drinking odds ratios for laryngeal cancer. We found that quitting tobacco smoking and alcohol drinking resulted in a head and neck cancer risk reduction, mainly after 20 years of quitting. Intake of green tea is not associated with gastric cancer risk. However, the temperature of green tea may play a role in the etiology of the disease. We found that dietary proanthocyanidins have a favorable role on gastric cancer risk. Our findings do not support the inverse relation between aspirin use and gastric cancer reported in a recent meta-analysis. We found a direct association between metabolic syndrome and both colon and rectal cancers in men, but not in women. We observed an unfavorable role on colorectal cancer of a dietary pattern based on high intake of starch, and a protective role of dietary patterns characterized by vitamins and fiber, and unsaturated fats. We found an increased risk of colon cancer for higher intake of polyunsaturated fat, trans-fat, lactose, sucrose and cholesterol.

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Our case-control study of pancreatic cancer reported no association with dietary acrylamide intake. Occupational and recreational physical activity was not associated to a decreased risk of pancreatic cancer in our study. Alcohol consumption was associated to increased pancreatic cancer risk, but the ORs were significant only among heavy drinkers. The risk was over 4-fold higher in heavy smokers and heavy drinkers in comparison with never smokers who drunk no or low amounts. Our findings support a favorable role of vitamins E and C, selected carotenoids, and folate on pancreatic cancer risk. We confirmed that soft drink consumption is not materially related to pancreatic cancer risk. We observed no association between regular aspirin use and pancreatic cancer risk, although our results suggested a possible protective effect for long-term current users. We found that diabetes and smoking are related to pancreatic cancer risk, with a multiplicative effect between the two factors. There is an inverse association between fruit and vegetables and pancreatic cancer risk, and a direct one with meat. A diet poor in animal proteins and rich in sugars (mainly derived from fruit) appears to have a beneficial effect on pancreatic cancer risk. A case-control study of endometrial cancer showed a direct association with the metabolic syndrome. Our data suggest the potential importance of lignin intake in the prevention of endometrial cancer at Italian consumption levels. Our data support earlier findings of an increase in risk of endometrial cancer with duration of use of fertility drugs. We found a lack of association between alcohol consumption and endometrial cancer risk in our case-control study and in a meta-analysis including other 19 case-control and 7 cohort studies (more than 13,000 cases). We added relevant information on the absence of a consistent association between aspirin use and endometrial cancer risk, even in high-risk overweight and obese women. We found that a diet low in trans-fat could reduce prostate cancer risk. Adherence to the Mediterranean diet in an Italian population showed no significant change over the last 15 years in both men and women. Our case-control study indicated that the metabolic syndrome, as well as its components, are associated with an increased risk of cataract extraction in an Italian population.

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Glycemic index and glycemic load were inversely related to body mass index and not related to waist to hip ratio in an Italian population. A review on flavonoids and cancer risk provided evidence of a protective effect of flavanones on upper aerodigestive tract cancers, flavonols, anthocyanidins and proanthocyanidins on colorectal cancer, flavonols and flavones on breast cancer, isoflavones on ovarian cancer, and flavonols on renal cancer. Our findings indicated that citrus fruit has a protective role against cancers of the digestive and upper respiratory tract. Several aspects of the Mediterranean diet were favorable on risk of various cancers, particularly of the digestive tract. A meta-analysis of epidemiological studies on dietary acrylamide and human cancer reported a lack of association with most neoplasms. The main association which will require further monitoring involves kidney cancer. In a meta-analysis of observational studies, a significant reduction in the risk of oral and pharyngeal cancer of about 35% emerged for highest coffee drinkers as compared to lowest drinkers, while no relation was found between coffee drinking and laryngeal and esophageal cancer. In a meta-analysis of 31 observational studies on alcohol and cancers of the oral cavity and pharynx, we observed higher alcohol-related RRs for pharyngeal than for oral cancer, particularly at higher doses. A meta-analysis of casecontrol studies suggested a moderate favorable effect of coffee consumption on colorectal cancer risk. This may reflect a real protection, but can also partly or largely be due to reverse causation. A collaborative analysis provided evidence that cigar smoking is associated with an excess risk of pancreatic cancer, while no significant association emerged for pipe smoking and smokeless tobacco use. In a review of all studies of alcohol and oral and pharyngeal cancers, we observed a strong doseresponse effect on the intensity of alcohol use, but no apparent association for the duration of alcohol use. A significant positive association with non-Hodgkin lymphoma was found in higher socioeconomic status subjects. CHD and CVD mortality steadily declined in Europe, except in Russia, where rates were 10- to 15-fold higher than those of France, Italy or Sweden. Hungary and Poland, but also Scotland, where CHD trends were less favorable than in other western European countries, also emerge as priorities for preventive interventions. Though oral and pharyngeal mortality in Europe has declined in the last decade in men, there were still rises in a few central and eastern European countries, reaching exceedingly high rates in Hungary and Slovakia.

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In Brazil, total cancer mortality rates increased over the last decades in both sexes. Particularly high rates were registered in the last decade for head and neck cancers in men and cervix in women, cancers largely avoidable through prevention screening and early diagnosis. Overall, since 1970, rates for all childhood cancers dropped in both sexes in North America and Japan. In middle and low income countries, for leukemia, bone and kidney cancer, rates registered in 2005- 2007, are comparable to those registered in more developed areas in the early 1980s. High risk-populations in terms of risk factors related to cardiovascular mortality should give the highest priority to achieving favorable shifts in all modifiable risk factors. Analyzing cancer mortality in Europe during 2000-2004, we found a decline in overall cancer mortality from 185.2 to 168.0/100 000 (world standard, 29%) in men and from 104.8 to 96.9 (28%) in women. Although the reduction in postmenopausal hormone therapy use is a plausible explanation for the recent decline in breast cancer incidence observed in Europe and North America, the issue remains open to discussion. In an age-period-cohort analysis, we observed a leveling of the epidemic of oral cancer for men in most European countries, including Hungary and other central European countries where mortality from this cancer was exceedingly high. In another age-period-cohort analysis, we found that the favourable trends in gastric cancer mortality observed in recent years are likely to continue in the close future. In an analysis for breast cancer trends from Cordoba, Argentina, rates over most recent calendar years decreased, mainly in the most urbanized districts. Mortality from childhood cancer continued to decline over more recent years in most European countries. Some further improvement in childhood cancer mortality in eastern Europe is achievable through more widespread and better adoption of currently available treatments. The recent decreases in colorectal cancer mortality rates in several European countries are likely due to improvements in (early) diagnosis and treatment, with a consequent higher survival from the disease. Changes in breast cancer mortality after 1988 varied widely between European countries, and the UK is among the countries with the largest reductions. In South-eastern Europe, liver cirrhosis mortality was 10-20 times higher than in most other European states. Using data from two surveys on more than 3000 individuals representative of the Italian adult population, we found for the first time, after 5 decades, a significant increase in smoking prevalence. Countries of the European Union with a relatively stricter implementation of policies to control tobacco have lower smoking prevalence, lower self-reported exposure to SHS and higher support towards smoking bans in all workplaces.

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Supplementation with n-3 polyunsaturated fatty acids ameliorated depressive symptoms and quality of life in the treatment of depressed elderly female patients. Influenza vaccination among oncohematological children should be strongly recommended only for those who have been off therapy for less than 6 months. The direct involvement of parents significantly increased children acceptance of nasal swabs use to detect influenza virus, thus simplifying collection, without reducing the influenza virus detection rate. In a survey of Italian adolescents and parents, the likelihood of HPV infection was underestimated. This was associated with a lower propensity towards HPV vaccine. In a meta-analysis on the literature on risks factors for falls in older people we identified some indicators of disability that strongly predict the risk of falling. In a prospective study, we found that bowel ultrasound (US) may be used as a surrogate of colonoscopy in assessing the short-term response of severe forms of UC to therapy. Both US score and endoscopic score after 3 months of steroid therapy predict outcome of disease at 15 months. In Milan, the Emergency Medical System provides support to patients having an out-of-hospital cardiac arrest with characteristics comparable with those reported in other large urban areas, but the time from arrival-to-first CPR was longer than recommended by current guidelines. Serum amyloid A protein levels were predictive of an elevated risk of lung cancer, supporting the general view that inflammation is implicated in lung cancer development. Bronchial diverticula are a frequent finding in the major airways of smokers, and they are associated with other markers of smoking-related damage. Screening for lung cancer using airflow obstruction with FEV1% is a strategy worth future consideration. In a cohort of workers exposed to extensively high doses of aromatic amines, 56 deaths from bladder cancer were observed, compared with 3.4 expected. The incidence of second HN cancers does not increase with age, but remains constant, or if anything, decreases with advancing age. Preliminary results from 2 case-control studies conducted in Italy and Spain suggest a weak if any association between colorectal cancer and disinfection by-products. The excess risks of adverse pregnancy outcomes in relation to particulate matter, if any, are small, and it is unclear whether they are causal, due to misclassification of the exposure or some sources of bias/residual confounding. The evidence that human exposure to paraquat pesticide increases the risk for Parkinson Disease is rather limited.

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NATIONAL COLLABORATIONS
Associazione Italiana di Oncologia Medica (AIOM) Accademia Nazionale di Medicina, Genova Agenzia giornalismo scientifico Zadig, Milano Arcispedale S. Maria Nuova, Reggio Emilia Associazione Nazionale dei Medici Cardiologi Ospedalieri (ANMCO) Azienda Unit Sanitaria Locale di Ravenna Centro Cardiologico Monzino, Milano Centro Studi Comunicazione sul Farmaco, Milano Centro di Riferimento Oncologico, Servizio di Epidemiologia e Biostatistica, Aviano (PN) Comune di Milano, Direzione centrale salute, Settore politiche per la Salute Federazione Italiana delle Associazioni di Volontariato in Oncologia (FAVO) Fondazione LuVI Fondazione Politecnico di Milano Fondazione SmithKline, Milano Gruppo Italiano per lo Studio della Sopravivenza nellInfarto miocardico (GISSI) Gruppo Italiano Studi Epidemiologici in Dermatologia GISED, Bergamo Gruppo Italiano Documentalisti dellIndustria Farmaceutica e degli Istituti di Ricerca Biomedica Gruppo Studi Tumori Urologici (GSTU) International Centre for Pesticides and Health Risk Prevention, Milano Istituto Auxologico Italiano, Divisione Malattie Metaboliche III, IRCCS, Piancavallo (VB) Istituto Auxologico Italiano, Laboratorio Sperimentale di Ricerche Endocrinologiche (LSRE), IRCCS, Milano Istituto DOXA, Milano Istituto Europeo di Oncologia, Divisione di Epidemiologia e Biostatistica, Milano Istituto Europeo di Oncologia, Divisione di Chirurgia Cervico Facciale, Milano Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN), Roma Istituto Nazionale Neurologico Carlo Besta, Milano Istituto Nazionale per lo Studio e la Cura dei Tumori, Oncologia Sperimentale, Unit di Eredit Poligenica, Milano Istituto Superiore di Sanit, Osservatorio Fumo Alcol Droga, Roma Istituto Tumori Fondazione Pascale, Servizio di Epidemiologia, Napoli Novartis Vaccines SpA, Siena Ordine dei Medici della Provincia di Bari Ospedale Casa Sollievo della Sofferenza San Giovanni Rotondo Ospedale Niguarda Ca Granda, Dipartimento Trapianti di Fegato, Milano Ospedale Niguarda Ca Granda, Istituto di Fisiologia Clinica CNR, Sezione di Milano, Milano Ospedali Riuniti di Bergamo Ospedale Alessandro Manzoni, Unit di Gastroenterologia, Lecco (LC) Ospedale Luigi Sacco, Az Osp - Polo Universitario, Milano Policlinico di Monza, Unit Operativa di Endoscopia I, Monza (MB) Prima Clinica Ostetrico Ginecologica, Mangiagalli, Milano Regione Lombardia, U.O. Governo dei servizi sanitari territoriali e politiche di appropriatezza e controllo Struttura Sistemi di remunerazione e Osservatorio Epidemiologico Direzione Generale Sanit Societ Italiana Attivit Regolatorie Unione Nazionale dei Giornalisti Scientifici Italiani

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Universit Bocconi di Milano, Dipartimento di Analisi Istituzionale e Management Pubblico, Milano Universit Cattolica del Sacro Cuore, Unit di Epidemiologia genetica e Biologia Molecolare, Istituto di Igiene, Roma Universit degli Studi di Milano - Bicocca, Dipartimento di Statistica, Milano Universit degli Studi di Milano-Bicocca, I Clinica Otorinolaringoiatria, DNTB, Monza Universit di Milano, Clinica Pediatrica De Marchi, Milano Universit di Milano, Dipartimento di Medicina del Lavoro, Sezione di Statistica Medica e Biometria, Milano Universit di Milano, Prima Clinica Ostetrico Ginecologica, Milano Universit di Pavia, Azienda di Servizi alla Persona, Pavia Universit di Torino, Istituto di Medicina del Lavoro, CTO, Torino Universit di Verona, Clinica Ostetrico Ginecologica, Verona

INTERNATIONAL COLLABORATIONS
Catalan Institute of Oncology, Institut dInvestigaci Biomdica de Bellvitge (IDIBELL), Cancer Prevention and Control Unit, LHospitalet de Llobregat, Spain Center of Oncology, Dept. of Epidemiology and Cancer Prevention, Warsaw, Poland Centre for Research in Environmental Epidemiology (CREAL) and Municipal Institute of Medical Research (IMIM), Barcellona, Spain Evidence and Risk Assessment Division, Centre for Chronic Disease Prevention and Control, Public Health Agency of Canada, Ottawa, Ontario, Canada Harvard School of Public Health, Department of Epidemiology, Boston, USA Hellenic Health Foundation, Greece Hpital Necker - Enfants Malades, Centre of the Association Claude Bernard on Auto-immunes diseases, Parigi, France Institute de Academie des Sciences, Paris, France International Agency for Research on Cancer, Lione, France International Epidemiology Institute (IEI), Rockville, USA International Life Science Institute (ILSI), Bruxelles, Belgium International Prevention Research Institute (IPRI), Lyon, France Karolinska Institute, Department of Medical Epidemiology and Biostatistics, Stockholm, Sweden National Cancer Institute, Environmental Studies Section, Bethesda, USA National Cancer Institute, Radiation Epidemiology Branch, Bethesda, USA National School of Public Health, WHO, Atene, Greece Registre Vaudois des Tumeurs, Institut Universitaire de Mdecine Sociale et Prventive, Losanna, Switzerland Senologic International Society Society for Internet in Medicine The Tisch Cancer Institute and Institute for Translational Epidemiology, Mount Sinai School of Medicine, New York, NY, USA Tobacco Free Research Institute, Dublino, Ireland UNDP/UNFPA/WHO/WORLD Bank special programme of research development and research training in human reproduction, Ginevra, Svizzera Universitat Pompeu Fabra, Department of Experimental and Health Sciences, Barcellona, Spain

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University of Athens Medical School, Department of Hygiene and Epidemiology, Atene, Greece University of Cordoba, Faculty of Medical Diseases, Cordoba, Argentina University of Las Palmas de Gran Canaria, Department of Clinical Sciences, Las Palmas de Gran Canaria, Spain Vanderbilt University, Department of Medicine, School of Medicine, Nashville, TN, USA

EDITORIAL BOARD MEMBERSHIP

Advances in Therapy (Eva Negri) Alimentazione e Prevenzione (Carlo La Vecchia) Annals of Oncology (Carlo La Vecchia, Associate Editor) Archives of Medical Science (Carlo La Vecchia) BMC Public Health (Silvano Gallus, Associate Editor) Cancer Letter (Carlo La Vecchia, Associate Editor) Current Cancer Therapy Reviews (Carlo La Vecchia) Dermatology Research and Practice (Carlo La Vecchia) Digestive and Liver Disease (Carlo La Vecchia) Economia Politica del Farmaco (Carlo La Vecchia) European Journal of Cancer Prevention (Carlo La Vecchia, Associate Editor) European Journal of Clinical Nutrition (Carlo La Vecchia) European Journal of Nutrition (Carlo La Vecchia) Evidence Based Dermatology (Carlo La Vecchia, Liliane Chatenoud) In Scope Oncology & Haematology (Carlo La Vecchia) ISRN Cardiology (Eugenio Santoro) Maturitas (Carlo La Vecchia) Nutrition and Cancer (Carlo La Vecchia) Open Cancer Journal (Carlo La Vecchia) Oral Oncology (Carlo La Vecchia) Portale Partecipasalute.it http://www.partecipasalute.it (Eugenio Santoro) Revisiones en Ginecologa y Obstetricia (Carlo La Vecchia) Revista Espaola de Nutrici Comunitaria (Carlo La Vecchia) Revue dEpidmiologie et de Sant Publique (Carlo La Vecchia) Societ Italiana Attivit Regolatorie News, SIARNews (Eugenio Santoro) The Open Obesity Journal (Silvano Gallus) Tumori (Carlo La Vecchia) World Journal of Gastrointestinal Oncology (Silvano Gallus)

PEER REVIEW ACTIVITIES

Acta Dermato-Venereologica, Acta Psychiatrica Scandinavica, Alcologia, American Journal of Clinical Nutrition, American Journal of Epidemiology, Annals of Epidemiology, Annals of Oncology, Archives of Internal Medicine, BMC Public Health, British Journal of Cancer, British Journal of Nutrition, British Medical Journal, Bulletin of the World Health Organization, Canadian Journal of Physiology and Pharmacology, Cancer, Cancer Causes and Control, Cancer Detection and Prevention, Cancer Epidemiology Biomarkers and Prevention, Computer Methods and Programs in Biomedicine, Diabetes/Metabolism Research and Reviews, Digestive

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Liver Disease, Epidemiologia & Prevenzione, Epidemiology, Epidemiology & Biostatistic, European Heart Journal, European Journal of Cancer, European Journal of Cancer Prevention, European Journal of Clinical Nutrition, European Journal of Epidemiology, European Journal of Public Health, Evidence-Based Healthcare and Public Health, Gynecological Endocrinology, Gut, Hepatology, Human Reproduction, International Journal of Cancer, International Journal of Environmental Research and Public Health, International Journal of Epidemiology, International Journal of Obesity, JAMA, Journal of American College of Nutrition, Journal of Clinical Endocrinology and Metabolism, Journal of Clinical Epidemiology, Journal of Epidemiology and Community Health, Journal of Investigative Dermatology, Journal of Medical Economics, Journal of Medical Internet Research , Journal of the National Cancer Institute, Lancet Oncology, Lung Cancer, Maturitas, Melanoma Research, Nicotine & Tobacco Research, Nutrition and Cancer, Obstetric and Gynecology, Oncology, PLoS ONE, Preventive Medicine, Public Health, Public Health Nutrition, QJM, Radiation Research, Revue dEpidmiologie et de Sant Publique, The Breast, The Cancer Journal, The Lancet, The Open Obesity Journal, Tobacco Control, Tumori, World Journal of Gastroenterology.

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Advisory Committee of the Oxford Collaborative group on Aetiological Factors in Cancers of the Female Genital Tract Comitato Scientifico del Gruppo Italiano Studi Epidemiologici in Dermatologia Comitato Scientifico della Societ Italiana di Colposcopia e Patologia Cervico Vaginale Comitato Scientifico del portale www.familyhealth.it Data and Safety Monitoring Board of the Phase II therapeutic trial with a humanized nonmitogenic CD3 (ChAgly CD3) monoclonal antibody in recently diagnosed type I diabetic patients Executive Committee, International Head and Neck Cancer Epidemiology (INHANCE) consortium Ministero della Salute, Sottocomitato fumo Scientific Review Committee del UND/WHO/World Bank Human Reproduction Programme

EVENT ORGANIZATION
Istituto di Ricerche Farmacologiche Mario Negri, Milan; Conference: Presentazione trial sulleffetto del consumo di chewing-gum sullappetito e il consumo di snack. 15 January 2010. Corso La ricerca bibliografica su database biomedici, organizzato in collaborazione con lASL di Bergamo Dipartimento Cure Primarie e Continuit Assistenziale, Bergamo 11 February 2010 V edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 8 June 2010 V edizione del corso ECM "Internet e laggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 9 June 2010 V edizione del corso ECM "Il web 2.0 al servizio della formazione e dellaggiornamento del medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 10 June 2010

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V edizione del corso ECM "Twitter, Facebook, Youtube e i nuovi social media per laggiornamento del medico e delloperatore sanitario", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 11 June 2010 Corso Web 2.0 e social media in medicina organizzato presso il Centro Cardiologico Monzino di Milano, 5,19 October 2010 Corso di formazione e aggiornamento per i giornalisti Internet e Medicina: a caccia di salute sul web, organizzato in collaborazione con la Unione Nazionale dei Giornalisti Scientifici Italiani (UNAMSI), Istituto di Ricerche Farmacologiche Mario Negri, Milano, 16 October 2010 VI edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 2 November 2010 VI edizione del corso ECM "Internet e laggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 3 November 2010 VI edizione del corso ECM "Il web 2.0 al servizio della formazione e dellaggiornamento del medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 4 November 2010 VI edizione del corso ECM "Twitter, Facebook, Youtube e i nuovi social media per laggiornamento del medico e delloperatore sanitario", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 5 November 2010

Corso Aggiornarsi in oncologia con gli strumenti del web 2.0 organizzato dal personale del dipartimento in collaborazione con lAssociazione Italiana Oncologia Medica e svolto nellambito del XII Congresso Nazionale dellAssociazione Italiana Oncologia Medica, Roma 6 November 2010
Corso Il web 2.0 e i social media come strumento di comunicazione in sanit promosso dalla Scuola Umbra di Amministrazione Pubblica, Perugia 10 November 2010 Corso Il web 2.0 e i social media al servizio della formazione e dellaggiornamento del medico e delloperatore sanitario organizzato presso lAzienda Unit Sanitaria Locale di Ravenna, Ravenna 14-15 December 2010

CONFERENCE AND WORKSHOP CONTRIBUTIONS

Corso Laccademia del cittadino: valutare la qualit e la sicurezza dei servizi sanitari per fare scelte consapevoli e partecipare al miglioramento Modulo 3: Informazione e comunicazione sulla salute. Il ruolo della associazioni nella costruzione dellalleanza tra cittadino e sistema sanitario organizzato da Partecipasalute e Regione Toscana. Titolo della relazione: Facebook, blog, wiki: le informazioni di salute corrono sul web 2.0, Montecatini Terme 21 January 2010. Corso Le nuove tecnologie al servizio dellefficacia e dellefficienza in diabetologia, promosso da Roche Diagnostics, Buttrio (UD) 22-23 January 2010 Convegno. Cibo degli dei, cibo dei demoni alimentazione fra salute e malattia. Cibi contro il cancro e cibi che lo fanno venire. Cosa c di vero? Lugano, Svizzera 28 January 2010

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International scientific workshop. The truth about wine. Alcohol and health, with focus on moderate doses e Wine consumption and longevity. Castello di Grinzane Cavour (CN) 5-7 February 2010 Open day. Hiwate. Health impacts of long term exposure to disinfection by-products in drinking water. London, UK 23 February 2010 43rd Annual Meeting of the Italian Association for the Study of the Liver. Coffee and liver diseases. Roma, 24 February 2010 Convegno Alleati per la salute, Roma 5-6 March 2010. Titolo della relazioneComunicare il non profit: Internet e i Social Network Corso di aggiornamento. Gli studi caso-controllo: dal disegno agli strumenti epidemiologicostatistici. Lettura di un articolo scientifico ed esercitazione: disegnare uno studio casocontrolloCalcolo della numerosit campionaria in uno studio caso-controllo. Particolari tipi di studi caso-controllo. Roma 9 March 2010 Pancreatic cancer case-control (PANC4) consortium. Non-cigarette tobacco use and pancreatic cancer. Rockville, MD 18 March 2010 European Breast Cancer Conference. EBCC7. Impact of lifestyle on breast cancer. Barcellona, Spagna 24-27 March 2010 Corso, Siti, strumenti ed applicazioni web 2.0 in ambito medico promosso dalla Biblioteca Medica della AIL Biella-Fondazione Clelio Angelino onlus, Biella 14-15 aprile 2010. VII International Nutrition and dietetics Congress. Mediterranean diet and cancer. Istanbul, Turchia 14-18 April 2010 Corso Internet e laggiornamento professionale in ambito medico, promosso dal servizio formazione del personale medico della Provincia Autonoma di Bolzano, Bolzano 23 April 2010 Alimentazione, stili di vita e promozione della salute. Componenti della frutta e verdura e prevenzione del cancro. Bologna, 24-27 Aprile2010 Bocconi University, Milan; Master course: Epidemiology of tobacco in Italy, with a focus on the effects of the smoking ban legislation. Milano. 4 Maggio 2010. Master Universitario di I livello in Comunicazione e Salute nei Media Contemporanei, Universit degli Studi di Milano, facolt di Farmacia, anno accademico 2009-2010. Ruolo di docenze nel modulo Il Web 2.0 come strumento per la comunicazione della salute", Milano 7 May 2010 V Giornata nazionale del malato oncologico promossa dalla Federazione Italiana delle Associazioni di Volontariato in Oncologia (FAVO). Titolo della relazione: Le associazioni dei pazienti nellera dei social network e dei social media, Roma 15 May 2010 PPACTE Meeting: WP2: European Survey on economic aspects of smoking. IARC, Lyon, France. 16 May 2010.

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IARC Handbook Meeting: Chapter 5; tax, price and adult tobacco use. IARC, Lyon, France. 17 May 2010. Minimaster Come leggere un lavoro clinico Ricerca bibliografica online organizzato dallAssociazione Nazionale Medici Cardiologi Ospedalieri, Firenze 19 May 2010. Titolo della relazione Web 2.0 e banche dati: utilizzo di feed RSS, aggregatori di notizie e social bookmarks I edizione Convegno Nazionale Fondazione GSTU (Gruppo Studi Tumori Urologici). Loncologia dei nostri giorni: fra comunicazione e comunicabilit. Titolo della relazione Strumenti di informazione sanitaria disponibili su Internet: come orientarsi, Palermo 21 May 2010 XII Convegno nazionale tabagismo e servizio sanitario nazionale. Fumo e donne: nulla peggio per la loro salute. Roma, 31 May 2010 Convegno Medicina e Media: sfide per una comunicazione di qualit organizzato dallUniversit degli Studi di Padova, Padova 8 June 2010. Titolo della relazione: Internet e web 2.0: cosa cambia per la comunicazione in medicina. Convegno I nuovi canali del informazione scientifica tra e-detailing e TV digitale terrestre, Business International. Titolo della relazione Social Networking in Sanit: come scambiare esperienze e condividere conoscenze mediche e sanitarie attraverso la rete, Roma 16 June 2010. 1 European Focus Meeting. Excellence in young women breast care. Epidemiologia del carcinoma mammario (La Vecchia moderatore), Famigliarit, ereditariet e counseling genetico (Negri). Castel dellOvo, Napoli, 24-25 June 2010 IPRI-Dundee University Summer School of epidemiology. Exposure assessment, Introduction to biasDundee, UK, 5-7 July 2010 Meeting The International meta-analysis on lung cancer screening: recent data- Peter B. Bach. Fondazione IRCCS Istituto Nazionale dei Tumori. Milano, 22 July 2010 Prevention, from theory to policy. Lifestyle and cardiovascular risk. Losanna, Svizzera. 7 September 2010 ESBRA Nordmann Award & ISBRA Satellite Meeting Alcohol & Cancer. Epidemiology of alcohol cancer. Heidelberg 17-18 September 2010 I edizione del corso Gli strumenti del web 2.0 per la distribuzione e la condivisione dellinformatica medica, promosso dallArcispedale Santa Maria Nuova, Azienda Ospedaliera di Reggio Emilia, Reggio Emilia 22 September 2010 II World Congress of Public Health Nutrition. Application of low-calorie sweeteners, safety and contributions to health. Porto, Portogallo. 23-25 September 2010 Europa Donna Breast Cancer Advocacy Leader Conference: reducing health inequalities and fostering healthy ways of life. Epidemiology and facts about lifestyle and breast cancer prevention. Milano. 25 September 2010

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II edizione del corso Gli strumenti del web 2.0 per la distribuzione e la condivisione dellinformatica medica, promosso dallArcispedale Santa Maria Nuova, Azienda Ospedaliera di Reggio Emilia, Reggio Emilia 1 October 2010 Corso Le nuove tecnologie al servizio dellefficacia e dellefficienza in diabetologia, promosso da Roche Diagnostics, Cavenago 1-2 October 2010 Convegno. Alimentazione e tumori: dalla prevenzione al support nutrizionale. Componenti della dieta utili nella prevenzione dei t umori. Milano. 8 October 2010 Breast cancer conference. Swedish Cancer Society. Impact of lifestyle on breast cancer. Stoccolma, Svezia. 15 October 2010 Vivere in salute alla Bocconi. La prevenzione come strumento utile al fine di ridurre i rischi di malattie. Milano. 18 October 2010 Dental Lessons. Alcohol e colluttori: dati epidemiologici. Roma. 19 October 2010 3rd International Congress on Nutrition and Cancer. Overweight, diabetes and Cancer risk. Bodrum, Turkey. 20-24 October 2010 III edizione del corso Gli strumenti del web 2.0 per la distribuzione e la condivisione dellinformatica medica, promosso dallArcispedale Santa Maria Nuova, Azienda Ospedaliera di Reggio Emilia, Reggio Emilia 20 October 2010 II edizione Convegno Nazionale Fondazione GSTU (Gruppo Studi Tumori Urologici). Loncologia dei nostri giorni: fra comunicazione e comunicabilit. Titolo della relazione Strumenti di informazione sanitaria disponibili su Internet: come orientarsi, Catania 30 October 2010 Conference EUROEPI: Impact of cigarette price on demand for tobacco products. Florence, 8 November 2010. Conference EUROEPI: Colorectal cancer and disinfection by-products in Italy and Spain. Florence, 7 November 2010. XXXIV Congresso AIE 2010. Associazione Italiana Epidemiologia. Consumo di caff e t e rischio di tumore alla testa e collo: una pooled analysis di studi del consorzio INHANCE (The International Head and Neck Cancer Epidemiology). Firenze. 9 November 2010 Conferenza Da LightHouse al web 2.0: il futuro dellinformazione biomedica in Lombardia, conferenza annuale del Sistema Bibiotecario Biomedico Lombardo (SBBL). Titolo della relazione Il ruolo del web 2.0 e dei social media nella diffusione dellinformazione biomedica, Milano 16 November 2010. XVI Congress of International Federation of Health Records Organizations (IFHRO), Milano 15-19 November 2010. Titolo relazione: Personal Health Record and web 2.0: a new way for patients and citizens to collect, handle, and share their own health data Convegno Comunicazione sociale per la salute: salute 2.0 esperienze e interrogativi promosso dalla Agenzia Informazione e Ufficio Stampa della Giunta Regionale Emilia Romagna in collaborazione con lUniversit di Bologna e Fondazione Pubblicit e Progresso. Titolo della

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relazione Partecipazione, formazione e salute:lesperienza del portale partecipasalute.it, Bologna 26 November 2010 Seminario La registrazione dei trial clinici: come e perch effettuarla e come interrogare i database, Ospedale Maggiore di Bologna, Bologna 2 Dicember 2010 Convegno Questioni di cuore: linfarto miocardico nellera della comunicazione globale promosso dalla Azienda Unit Sanitaria locale di Forl. Titolo della relazione La comunicazione e gli ammalati (potenziali) di domani: web, iphone, ipad, centro di ascolto, condivisione delle immagini, second opinion, Forl 4 December 2010 Convegno Innovazioni tecnologiche in medicina sportiva, promosso dalla Federazione Medico Sportiva Italiana (FMSI) Puglia e Basilicata, Matera 3-4 December 2010. Titolo della relazione Il web 2.0: filosofia e strumenti. Master Universitario di I livello in Ricerca Clinica, Universit degli Studi di Milano, anno accademico 2010-2011. Ruolo di docenze nel modulo Internet e le nuove tecnologie per laggiornamento medico-scientifico, Milano 9 December 2010 EU-FP6 Project HiWATE Final Meeting. 22-24 February 2010 Londra

GRANTS AND CONTRACTS

AIFA Arcispedale Santa Maria Nuova, Azienda Ospedaliera di Reggio Emilia Associazione Italiana Oncologia Medica Associazione Italiana per la Ricerca sul Cancro (AIRC) Azienda Sanitaria Locale di Bergamo Azienda Unit Sanitaria Locale di Ravenna Centro Cardiologico Monzino Comune di Milano Lega Italiana Lotta contro i Tumori (LILT) European Commission (FP7) European Research Council (ERC) Fondazione Politecnico di Milano GISED Ministero della Salute Regione Lombardia Weber Shandwich Consorzio Assomela ISA Perfetti Van Melle Provincia Autonoma di Bolzano Roche Diagnostics Scuola Umbra di Amministrazione Pubblica Unione Nazionale dei Giornalisti Scientifici Italiani

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SCIENTIFIC PUBLICATIONS (2010)

Deandrea S, Foschi R, Galeone C, La Vecchia C, Negri E, Hu J. Is temperature an effect modifier of the association between green tea intake and gastric cancer risk? Eur. J. Cancer Prev 19:1822 (2010) Parazzini F, Ricci E, Chiaffarino F, Cipriani S, Tozzi L, Fedele L Induced abortion and risk of preterm birth. Gynecol Obstet Invest. 69:40-45 (2010). Tavani A, Scotti L, Bosetti C, Dal Maso L, Montella M, Ramazzotti V, Negri E, Franceschi S, La Vecchia C. Aspirin and risk of renal cell cancer in Italy. Eur. J. Cancer Prev., 19: 272-274 (2010) Bosetti C, Niewenhuijsen M, Gallus S, Cipriani S, La Vecchia C, Parazzini F. Ambient particulate matter and preterm birth or birth weight: review of the literature. Arch Toxicol. 2010;84:447-60. Pelucchi C, Galeone C, Negri E, La Vecchia C. Trends in adherence to the Mediterranean diet in an Italian population between 1991 and 2006. Eur J Clin Nutr, 64: 1052-1056 (2010) Powles J, Shroufi A, Mathers C, Zatonski W, La Vecchia C, Ezzati M National cardiovascular prevention should be based on absolute disease risks, not levels of risk factors. EJPH, 20: 103106 (2010) Bidoli E, Pelucchi C, Zucchetto A, Negri E, Dal Maso L, Polesel J, Montella M, Franceschi S, Serraino D, La Vecchia C, Talamini R. Fiber intake and endometrial cancer risk. Acta Oncol. 49: 441446 (2010) Bravi F, Edefonti V, Bosetti C, Talamini R, Montella M, Giacosa A, Franceschi S, Negri E, Ferraroni M, La Vecchia C, Decarli A. Nutrient dietary patterns and the risk of colorectal cancer: case-control study. Cancer Causes Control, 21: 1911-1918 (2010) Rossi M, Negri E, Parpinel M, Lagiou P, Bosetti C, Talamini R, Montella M, Giacosa A, Franceschi S, La Vecchia C. Proanthocyanidins and the risk of colorectal cancer in Italy. Cancer Causes Control, 21: 243-250 (2010) Gaudet MM, Olshan AF, Chuang SC, Berthiller J, Zhang ZF, Lissowska J, Zaridze D, Winn DM, Wei Q, Talamini R, Szeszenia-Dabrowska N, Sturgis EM, Schwartz SM, Rudnai P, ElufNeto J, Muscat J, Menezes A, Matos E, Bucur A, Levi F, Lazarus P, La Vecchia C, Koifman S, Kelsey K, Herrero R, Hayes RB, Franceschi S, Wunsch-Filho V, Fernandez V, Fabianova E, Daudt AW, Dal Maso L, Curado MP, Chen C, Castellsague X, Benhamou S, Boffetta P, Brennan P, Hashibe M.

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Body mass index and risk of head and neck cancer in a pooled analysis of case-control studies in the International Head and Neck Cancer Epidemiology (INHANCE) Consortium. Int J Epidemiol, 39: 1091-1102 (2010) Deandrea S, Lucenteforte E, Bravi F, Foschi R, La Vecchia C, Negri E. Risk factors for falls in community-dwelling older people. A systematic review and metaanalysis. Epidemiology, 21: 658-668 (2010) Foschi R, Pelucchi C, Dal Maso L, Rossi M, Levi F, Talamini R, Bosetti C, Negri E, Serraino D, Giacosa A, Franceschi S, La Vecchia C. Citrus fruit and cancer risk in a network of casecontrol studies Cancer Causes Control, 21:237242 (2010) Marron M, Boffetta P, Zhang Z-F, Zaridze D, Wunsch-Filho V, Winn DM, Wei Q, Talamini R, Szeszeia-Dabrowska N, Sturgis EM, Smith E, Schwartz SM, Rudnai P, Purdue M, Olshan AF, Eluf-neto J, Menezes A, Mclean M, Matos E Mates IN, Lissowska J, Levi F, Lazarus P, La Vecchia C, Koifman S, Kelsey K, Herrero R, Hayes RB, Franceschi S, Fernandez L, Fabianova E, Daudt AW, Da Maso L, Curado MP, Cadoni G, Chen C, Castellsague X, Boccia S, Benhamou S, Ferro G, Berthiller J, Brennan J, Moller H, Hashibe M. Cessation of alcohol drinking, tobacco smoking and the reversal of head and neck cancer risk. Int. J. Epidemiol, 39: 182-196 (2010) Talamini R, Polesel J, Gallus S, Dal Maso L, Zucchetto A, Negri E, Bosetti C, Lucenteforte E, Boz G, Franceschi S, Serraino D, La Vecchia C. Tobacco smoking, alcohol consumption and pancreatic cancer risk: a case-control study in Italy. Eur. J. Cancer, 4 6 : 3 7 0 3 7 6 (2010) La Vecchia C, Bosetti C, Lucchini F, Bertuccio P, Negri E, Boyle P, Levi F. Cancer mortality in Europe, 2000-2004, and an overview of trends since 1975. Ann. Oncol., 21: 1323-1360 (2010). Chatenoud L, Bertuccio P, Bosetti C, Levi F, Curado PM, Malvezzi M, Negri E, La Vecchia C. Trends in cancer mortality in Brazil, 1980-2004. Eur J Cancer Prev. 19:79-86. (2010) Rossi M, Rosato V, Bosetti C, Lagiou P, Parpinel M, Bertuccio P, Negri E, La Vecchia C. Flavonoids, proanthocyanidins and the risk of stomach cancer. Cancer Causes Control, 21: 1597-1604 (2010) Lucenteforte E, Talamini R, Bosetti C, Polesel J, Franceschi S, Serraino D, Negri E, La Vecchia C. Macronutrients, fatty acids, cholesterol and pancreatic cancer. Eur. J. Cancer, 46: 581-587 (2010) Lubin JH, Gaudet MM, Olshan AF, Kelsey K, Zhang Z-F, Winn D, Wei Q, Talamini R, Szeszenia-Dabrowska N, Sturgis EM, Smith E, Shangina O, Schwartz SM, Rudnai P, Neto JE, Muscat J, Morgenstern H, Menezes A, Matos E, Mates IN, Lissowska J, Levi F, Lazarus P, Vecchia C, Koifman S, Herrero R, Franceschi S, Wnsch-Filho V, Fernandez L, Fabianova E, Daudt AW, Dal Maso L, Curado MP, Chen C, Castellsague X, Brennan P, Boffetta P, Hashibe M, Hayes RB.

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Body mass index, cigarette smoking and alcohol consumption and risk of cancer of the larynx, pharynx and oral cavity: modeling risk in pooled case-control data. Am. J. Epidemiol, 171:12501261 (2010) Morici N, De Luca G, Lucenteforte E, Chatenoud L, Lorito F, Palgiosa A, La Vecchia C, Sesana G. Emergency Medical System response to out-of hospital cardiac arrest in Milan, Italy. Eur. J. Emergency Med., 17: 234-236 (2010) Bosetti C, Bertuccio P, Chatenoud L, Levi F, Negri E, La Vecchia C. Childhood cancer mortality in Europe, 1970-2007. Eur. J. Cancer, 4 6 : 3 8 4 3 9 4 (2010) Edefonti V, Bravi F, Garavello W, La Vecchia C, Parpinel M, Franceschi S, Dal Maso L, Hashibe M, Bosetti C, Boffetta P, Ferraroni M, Decarli A. Nutrient-based dietary patterns and laryngeal cancer: evidence from an exploratory factor analysis Cancer Epidemiol Biomarkers Prev, 19: 18-27 (2010) Sverzellati N, Ingegnoli A, Calabr E, Randi G, La Vecchia C, Marchian A, Kuhnigk JM, Hansell DM, Zompatori M, Pastorino U. Bronchial diverticula in smokers on thin-section CT. Eur. Radiol., 20: 88-94 (2010) Rondanelli M, Giacosa A, Opizzi A, Pelucchi C, La Vecchia C, Montorfano G, Negroni M, Berra B, Politi P, Rizzo AM. Effect of omega-3 fatty acids supplementation on depressive symptoms and on health-related quality of life in the treatment of elderly women with depression. A double-blind, placebocontrolled, randomized clinical trial. J. Am. College Nutr., 29: 55-64 (2010) Garavello W, Bertuccio P, Levi F, Lucchini F, Bosetti C, Negri E, La Vecchia C. The oral cancer epidemic in central and eastern Europe. Int. J. Cancer. 127: 160171 (2010) Polesel J, Talamini R, Negri E, Bosetti C, Boz G, Lucenteforte E, Franceschi S, Serraino D, La Vecchia C. Dietary habits and risk of pancreatic cancer: An Italian case-control study. Cancer Causes Control, 21:49350, (2010) Niclis C, del Pilar Diaz M, La Vecchia C. Breast cancer mortality trends and patterns in Cordoba, Argentina 1986-2006. Eur. J. Cancer Prev., 19: 94-99 (2010) Pelucchi C, Levi F, La Vecchia C. The rise and fall in menopausal hormone therapy and breast cancer incidence. The Breast, 19: 198-201 (2010) Tramacere I, Scotti L, Jenab M, Bagnardi V, Bellocco R, Rota M, Corrao G, Bravi F, Boffetta P, La Vecchia C. Alcohol drinking and pancreatic cancer risk: a meta-analysis of the dose-risk relation. Int. J. Cancer, 126: 14741486 (2010)

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Calabr E, Randi G, La Vecchia C, Sverzellati N, Marchian A, Villani M, Zompatori M, Cassandro R, Harari S, Pastorino U. Lung function predicts lung cancer risk in smokers: a tool for targeting screening programs. Eur. Respir. J., 35: 146-151 (2010) Galeone C, Turati F, La Vecchia C, Tavani A. Coffee consumption and risk of colorectal cancer: a meta-analysis of case-control studies. Cancer Causes Control, 21: 1949-1959 (2010) Edefonti V, Bravi F, La Vecchia C, Randi G, Ferraroni M, Garavello W, Franceschi S,Talamini R, Boffetta P, Decarli A. Nutrient-based dietary patterns and the risk of oral and pharyngeal cancer Oral Oncol, 46: 343-348 (2010) Galeone C, Petracci E, Pelucchi C, La Vecchia C, Tavani A. Metabolic syndrome, its components and risk of age-related cataract extraction: a case-control study in Italy. Ann. Epidemiol., 20:380384 (2010) Rossi M, Bosetti C, Negri E, Lagiou P, La Vecchia C. Flavonoids , proanthocyanidins and cancer risk: a network of case-control studies from Italy. Nutr Cancer, 62: 871-877 (2010) Rossi M, Lipworth L, Polesel J, Negri E, Bosetti C, Talamini R, McLaughlin J, La Vecchia C. Dietary glycemic index and glycemic load and risk of pancreatic cancer: a case-control study. Ann Epidemiol, 20: 460-465 (2010) Negri E. Sun exposure, vitamin D and risk of Hodgkin and non Hodgkin lymphoma. Nutr Cancer, 62: 878-882 (2010) Rota M, Bellocco R, Scotti L, Tramacere I, Jenab M, Corrao G, La Vecchia C, Boffetta P, Bagnardi V Random-effects meta-regression model for studying nonlinear dose-response relationship Stat Med, 29: 2679-2687 (2010) Goldstein BY, Chang S-C, Hashibe M, La Vecchia C, Zhang Z F Alcohol consumption and cancer of the oral cavity and pharynx from 1988 to 2009: an update Eur J Cancer Prev, 19: 431-465 (2010) Grulich A E, Vajdic C M , Falster M O, Kane E , Smedby K E, Bracci P M, De Sanjose S, Becker N, Turner J, Martinez-Maza O, Melbye M, Engels E A, Vineis P, Seniori Costantini A, Holly E A, Spinelli J J, La Vecchia C, Zheng T, Chiu B C H, Dal Maso L, Cocco P, Maynadie M, Foretova L, Staines A, Brennan P, Davis S, Severson R, Cerhan J R, Breen E C, Birmann B, Cozen W Birth order and risk of non-Hodgkin lymphoma true association or bias? Am J Epidemiol, 172: 621-630 (2010) Hu J, La Vecchia C, Gibbons L , Negri E, Mery L, Canadian Cancer Registries Epidemiology Research Group Nutrients and risk of prostate cancer

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Nutr Cancer, 62: 710-718 (2010) F. Parente, M. Molteni, B. Marino, A. Colli, S. Ardizzone, S. Greco, G. Sampietro, D. Foschi, S. Gallus. Are colonoscopy and bowel ultrasound useful to assess response to short-term therapy and predict disease outcome of moderate to severe forms of ulcerative colitis ? A prospective study Am J Gastroenterol, 105: 1150-1157 (2010) Pelucchi C, Negri N, Talamini R, Levi F, Giacosa A, Crispo A, Bidoli E, Montella M, Franceschi S, La Vecchia C Metabolic syndrome is associated with colorectal cancer in men Eur J Cancer, 46: 1866-1872 (2010) Hu J, La Vecchia C, Negri E, Mery L Nutrients and Risk of Colon Cancer Cancer, 2: 51-67 (2010) Bosetti C, Bravi F, Talamini R, Montella M, Negri E, La Vecchia C. Aspirin and risk of endometrial cancer: a case-control study from Italy. Eur J Cancer prev, 19: 401-403 (2010) Bonifazi M, Gallus S, Bosetti C, Polesel J, Serraino D, Talamini R, Negri E, La Vecchia C. Aspirin use and pancreatic cancer risk Eur J Cancer Prev, 19: 352-354 (2010) Martnez-Sncheza,JM, Fernndeza,E, Fua,M, Gallus S, Martneza,C, Sureda X, La Vecchia C, Clancy L Smoking behaviour, involuntary smoking, attitudes towards smoke-free legislation, and tobacco control activities in the European Union. PLoS ONE, 5: e13881 (2010) Esposito S, Cecinati V, Scicchitano B, Delvecchio GC, Santoro N, Amato D, Pelucchi C, Jankovic M, De Mattia D, Principi N. Impact of influenza-like illness and effectiveness of influenza vaccination in oncohematological children who have completed cancer therapy. Vaccine. 28: 1558-1565 (2010) Galeone C, Tavani A, Pelucchi C, Turati F , Winn DM, Levi F, Yu G-P, Morgenstern H, Kelsey K, Dal Maso L, Purdue MP, McClean M, Talamini R, Hayes RB, Franceschi A, Schantz S, Zhang Z-F, Ferro G, Chuang S-C, Boffetta P, La Vecchia C, Hashibe M Coffee and tea intake and risk of head and neck cancer: pooled analysis in the international head and neck cancer Epidemiology consortium. Cancer Epidemiol Bio Prev, 19: 1723-36 (2010) Tramacere I, Negri E, Bagnardi V, Garavello W, Rota M, Scotti L, Islami F, Corrao G, Boffetta P, La Vecchia C A meta-analysis of alcohol drinking and oral and pharyngeal cancers: 1. Overall results and dose-risk relation Oral Oncol, 46: 497503 (2010) Zatonski W, Sulkowska U, Manczuk M, Rehm J, Boffetta P, Lowenfels AB, La Vecchia C.

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Liver cirrhosis mortality in Europe, with special attention to Central and Eastern Europe. Eur Addiction Res, 16:193-201 (2010) Autier P; Boniol M; La Vecchia C; Vatten L; Gavin A; Hry C; Heanue M Disparities in breast cancer mortality trends between thirty European countries BMJ, 341: c3620 (2010) doi:10.1136/bmj.c3620 Chatenoud L, Bertuccio P, Bosetti C, Levi F, Negri E, La Vecchia C Childhood cancer mortality in America, Asia, and Oceania, 1970 through 2007 Cancer, 116: 5063-5074 (2010) Peleteiro B, Lunet N, Carrilho C, Dures C, Machado JC, La Vecchia C, Barros H. Association between cytokine gene polymorphisms and gastric precancerous lesions: systematic review and meta-analysis. Cancer Epidemiol Biomarkers Prev, 19: 762-776 (2010) Turati F, Gallus S, Tavani A, Tramacere I, Polesel J, Talamini R, Montella M, Scotti L, Franceschi S, La Vecchia C Alcohol and endometrial cancer risk: a case-control study and a meta-analysis Cancer Causes Control, 21: 1285-1296 (2010) Lpez-Miranda J, Prez-Jimnez F, Ros E, De Caterina R, Badimn L, Covas MI, Escrich E, Ordovs JM, Soriguer F, Abi R, Alarcn de la Lastra C, Battino M, Corella D, ChamorroQuirs J, Delgado-Lista J, Giugliano D, Esposito K, Estruch R, Fernandez-Real JM, Gaforio JJ, La Vecchia C, Lairon D, Lpez-Segura F, Mata P, Menndez JA, Muriana FJ, Osada J, Panagiotakos DB, Paniagua JA, Prez-Martinez P, Peinado MA, Pineda-Priego M, Poulsen HE, Quiles JL, Ramrez-Tortosa MC, Ruano J, Serra-Majem L, Sol R, Solanas M, Solfrizzi V, R de la Torre-Fornell, Trichopoulou A, Uceda M, Villalba-Montoro JM, Villar-Ortiz JR, Visioli F, Yiannakouris N. Olive oil and health: Summary of the II international conference on olive oil and health consensus report, Jaen and Cordoba (Spain) 2008 Nutr Metab Cardiovasc Dis., 20: 284-294 (2010) Malvezzi M, Bonifazi M, Bertuccio P, Levi F, La Vecchia C, Decarli A, Negri E An Age-Period-Cohort Analysis of Gastric Cancer Mortality from 1950 to 2007 in Europe. Ann Epidemiol, 20: 898-905 (2010) Peleteiro B, Lunet N, Barros R, La Vecchia C, Barros H. Factors contributing to the underestimation of Helicobacter pylori-associated gastric cancer risk in a high-prevalence population. Cancer Causes Control. 21:12571264 (2010) The ESHRE Capri Workshop Group (In appendix: E Negri and C La Vecchia) Bone fractures after menopause. Human Reproduction Update, 16: 761-773 (2010) Ricci E, Cipriani S, Chiaffarino F, Malvezzi M, Parazzini F. Effects of soy isoflavones and genistein on glucose metabolism in perimenopausal and postmenopausal non-Asian women: a meta-analysis of randomized controlled trials Menopause, 17: 1080-1086 (2010)

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Lucenteforte E, Garavello W, Bosetti C, La Vecchia C. Dietary factors. In: Epidemiology, pathogenesis, and prevention of head and neck cancer. (Olshan AF, editor), Springer, New York. pp 117-136 (2010). Parazzini F, Pelucchi C, Talamini R, Montella M, La Vecchia C Use of fertility drugs and risk of endometrial cancer in an Italian case-control study. Eur J Cancer Prev, 19: 428-430 (2010) Turati F, Garavello W, Tramacere I, Bagnardi V, Rota M, Scotti L, Islami F, Corrao G, Boffetta P, La Vecchia C, Negri E A meta-analysis of alcohol drinking and oral and pharyngeal cancers. Part 2: Results by subsites Oral Oncol, 46: 720-726 (2010) Berry Sir C, La Vecchia C, Nicotera P Paraquat and parkinsons disease Cell Death Differentiation, 17: 1115-1125 (2010) Randi G, Edefonti V, Ferraroni M, La Vecchia C, Decarli A. Dietary patterns and the risk of colorectal cancers and adenomas. Nutrit Review, 68:389-408 (2010) Bertuccio P, Bravi F, Bosetti C, Negri E, La Vecchia C Aspirin and gastric cancer risk Eur J Cancer Prev, 19: 426-427 (2010) Islami F, Tramacere I, Rota M, Bagnardi V, Fedirko V , Scotti L, Garavello W, Jenab M, Corrao G, Straif K, Negri E, Boffetta P, La Vecchia C Alcohol drinking and laryngeal cancer: overall and dose-risk relation- a systematic review and meta-analysis Oral Oncol, 46: 802-810 (2010) Cibula D, Gompel A, Mueck AO, La Vecchia C, Hannaford PC, Skouby SO, Zikan M, Dusek L, Crosignani P Hormonal contraception and risk of cancer. Hum Repr Update, 16: 631-650 (2010) Bosetti C, Rossi M, Pelucchi C, La Vecchia C. Mediterranean diet and cancer. CML Kidney Cancer, 2: 69-76 (2010) La Vecchia C. Intake of artificially sweetened soft drinks and risk of preterm delivery. Am J Clin Nutr 92: 1540 (2010) Ricci E, Cipriani S, Chiaffarino F, Malvezzi M, Parazzini F Soy isoflavones and bone mineral density in perimenopausal and postmenopausal western women: a systematic review and meta-analysis of randomized controlled trials. J Womens Health, 19: 1609-1617 (2010)

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McLaughlin JK, LaVecchia C, Tarone RE, Lipworth L, Blot WJ Re: False-positive results in cancer epidemiology: a plea for epistemological modesty (response to Cogliano). JNCI, 102: 134-135 (2010) McLaughlin JK, Lipworth L, Tarone RE, La Vecchia C, Blot WJ, Boffetta P Authors response: A further plea for adherence to the principles underlying science in general and the epidemiologic enterprise in particular (by Hauptmann and Ronckers) Int J Epid, 39: 1679-1680 (2010) Parazzini F, Chiaffarino F, Chatenoud L, Cipriani S, Ricci E, Chiantera V, Bortolus R, Maffioletti C. Exposure to Video DisplayTerminals and Risk of Small-for-Gestational-Age Birth J Environ Health 72: 24-27 (2010) Rossi M, Talamini R, Lagiou P, Franceschi S, La Vecchia C. Glycemic index and glycemic load in relation to body mass index and waist to hip ratio. Eur J Nutr, in press http://www.ncbi.nlm.nih.gov/pubmed/20390288 Gallus S, Tramacere I, Boffetta P, Fernandez E, Rossi S, Zuccaro P, Colombo P, La Vecchia C. Temporal changes of under-reporting of cigarette consumption in population-based studies. Tobacco Control, in press http://www.ncbi.nlm.nih.gov/pubmed/20861005 Cremona M, Calabr E, Randi G, De Bortoli M, Mondellini P, Sozzi G, Pierotti MA, La Vecchia C, Pastorino U, Bongarzone I. Elevated levels of the acute-phase serum amyloid A are associated with heightened lung cancer risk. Cancer, in press. http://www.ncbi.nlm.nih.gov/pubmed/20087959 Bagnardi V, Randi G, Lubin J, Consonni D, Lam TK, Subar AF, Goldstein A, Wacholder S, Goldin L, Bergen A, Tucker M, Decarli A, Caporaso N, Bertazzi PA, Landi MT. Alcohol consumption and lung cancer risk in the Environment and Genetics in Lung cancer Etiology (EAGLE) study. Am. J. Epidemiol., in press http://www.ncbi.nlm.nih.gov/pubmed/19933698 Ricci E, Chiaffarino F, Cipriani S, Malvezzi M, Parazzini F. Diet in pregnancy and risk of small for gestational age birth: results from a retrospective casecontrol study in Italy. Maternal Child Nutr., in press. http://www3.interscience.wiley.com/journal/122651373/abstract?CRETRY=1&SRETRY=0 Bravi F, Polesel J, Bosetti C, Talamini R, Negri E, Dal Maso L, Serraino D, La Vecchia C. Dietary intake of selected micronutrients and the risk of pancreatic cancer: an Italian casecontrol study. Ann Oncol, in press. http://www.ncbi.nlm.nih.gov/pubmed/20530201 Pira E, Piolatto G, Negri E, Romano C, Boffetta P, Lipworth L, McLaughlin JK, La Vecchia C

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Bladder cancer mortality of workers exposed to aromatic amines: a 58-year follow-up J Natl Cancer Inst, in press http://www.ncbi.nlm.nih.gov/sites/pubmed/20548022 Rossi M, Negri E, La Vecchia C, Campos H Smoking habits and the risk of nonfatal acute myocardial infarction in Costa Rica Eur J Card Prev Rehabil, in press Cecinati V, Esposito S, Scicchitano B, Dalvecchio G C , Amato D, Pelucchi C, Jankovic M, Mattia D D, Principi N Effectiveness of recall systems for improving influenza vaccination coverage in children with oncohematological malignancies. Human Vaccine, in press http://www.ncbi.nlm.nih.gov/sites/entrez/19946216 Rosato V, Zucchetto A, Bosetti C, Dal Maso L, Montella M, Pelucchi C, Negri E, Franceschi S, La Vecchia C Metabolic syndrome and endometrial cancer risk Ann Oncol, in press http://www.ncbi.nlm.nih.gov/pubmed/20937645 Pelucchi C, Esposito S, Galeone C, Semino M, Sabatini C, Picciolli I, Consolo S, Milani G, Principi N Knowledge of human papillomavirus infection and its prevention among adolescents and parents in Italy BMC Public Health, in press Bosetti C, Scelo G, Chuang S - C, Tonita J M , Tamaro S, Jonasson J G , Kliewer E V , Hemminki K, Weiderpass E , Pukkala E, Tracey E , Olsen J H , Pompe-Kirn V , Brewster D H , Martos C , Chia K - S , Brennan P, Hashibe M, Levi F, La Vecchia C, Boffetta P High constant incidence rates of second primary cancers of the head and neck: a pooled analysis of 13 cancer registries Int J Cancer, in press http://www.ncbi.nlm.nih.gov/pubmed/20824702 Pelucchi C, Galeone C, Talamini R, Negri E, Polesel J, Serraino D, La Vecchia C Dietary acrylamide and pancreatic cancer risk in an italian case-control study. Ann Oncol, in press Bertuccio P, La Vecchia C, Silverman D, Petersen G, Bracci P, Negri E, Donghui L, Risch H A , Olson S H , Gallinger S, Miller A B, Bueno-De-Mesquita H B, Talamini R, Polesel J, Ghadirian P, Baghurst P A, Zatonski W, Fontham E , Bamlet W R, Holly E A, Lucenteforte E, Hassan M, Yu H , Kurtz R C , Cotterchio M, Su J , Maisonneuve P, Duell E J, Bosetti C, Boffetta P Cigar and pipe smoking, smokeless tobacco use and pancreatic cancer: a re-analysis from the International Pancreatic Cancer Case-Control Consortium (PanC4) Ann Oncol, in press Esposito S, Molteni CG, Daleno C, Valzano A, Tagliabue C, Galeone C, Milani G, Fossali E, Marchisio P, Principi N. Collection by trained pediatricians or parents of mid-turbinate nasal flocked swabs for the detection of influenza viruses in childhood.

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Virology Journal 2010, 7:85 http://www.ncbi.nlm.nih.gov/pubmed/20433729 Bertuccio P, Levi L, Lucchini F, Chatenoud L, Bosetti C, Negri E, La Vecchia C Coronary heart and cerebrovascular disease mortality in young adults: recent trends in Europe Eur j cardiovascular prev, in press Bosetti C, Levi F, Rosato V, Bertuccio P, Lucchini F, Negri E, La Vecchia C Recent trends in colorectal cancer mortality in Europe Int J Cancer, in press http://www.ncbi.nlm.nih.gov/pubmed/20824701 Lipworth L, Zucchetto A, Bosetti C, Franceschi S, Talamini R, Serraino D, McLaughlin J K, La Vecchia C, Negri E Diabetes mellitus and other medical conditions and pancreatic cancer: a case-control study Diabetes Metab Res Rev, in press Bonifazi M, Malvezzi M, Bertuccio P, Edefonti V, Garavello W, Levi F, La Vecchia C, Negri E Age-Period-Cohort Analysis of Oral Cancer Mortality in Europe: the End of an Epidemic? Oral Oncol, in press Tramacere I, La Vecchia C, Negri E Tobacco smoking and esophageal and gastric cardia adenocarcinoma: A metaanalysis Epidemiology, in press Gallus S, Turati F, Polesel J, Talamini R, Franceschi S, La Vecchia C Soft drinks, sweetened beverages and risk of pancreatic cancer Cancer Causes and Control, in press http://www.ncbi.nlm.nih.gov/pubmed/20981481 Gallus S, Tramacere I, Pacifici R, Zuccaro P G, Colombo P, Ghislandi S, La Vecchia C Smoking in Italy 2008-2009: a rise in prevalence related to the economic crisis? Prev Med, in press http://www.ncbi.nlm.nih.gov/pubmed/21130111 Fedirko V , Tramacere I, Bagnardi V, Rota M, Scotti L, Islami F, Negri E, Straif K, Romieu I, La Vecchia C, Boffetta P, Jenab M A meta-analysis of alcohol drinking and colorectal cancer risk: a dose-response analysis of published studies Ann Oncol, in press Hu J, La Vecchia C, Morrison H, Negri E, Mery L, Canadian Cancer Registries Epidemiology Research Group Salt, processed meat and the risk of cancer. Eur J Cancer Prev, in press Islami F, Fedirko V, Tramacere I, Bagnardi V, Jenab M, Scotti L, Rota M, Corrao G, Garavello W, Schuz J, Straif K, Negri E, Boffetta P, La Vecchia C Alcohol drinking and esophageal squamous cell carcinoma with focus on light-drinkers and never-smokers A systematic review and meta-analysis Int J Cancer, in press

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Pelucchi C, Zucchetto A, Tavani A, Dal Maso L, Serraino D, La Vecchia C. Physical activity and pancreatic cancer risk. Int J Cancer, in press http://www.ncbi.nlm.nih.gov/pubmed/20617512 Randi G, Malvezzi M, Levi F, Ferlay J, Negri E, Franceschi S, La Vecchia C. Re: Classification of biliary tract cancer (BTC): evaluation of all entities. Ann. Oncol., in press. Turati F, Galeone C, La Vecchia C, Tavani A. Coffee and Cancers of the Upper Digestive and Respiratory Tracts: Meta-analyses of observational studies Ann Oncol, in press http://www.ncbi.nlm.nih.gov/pubmed/20943597 Pelucchi C, et al. Exposure to acrylamide and human cancer-a review and meta-analysis of epidemic studies. Ann Oncol, in press

LAY PRESS SELECTION (2010)

Santoro E. Dai social network ai social media. Ricerca & Pratica 2010 ; n.154 : 159-160 Santoro E. Medpedia: il Wikipedia della medicina. Ricerca & Pratica 2010 ; n.153 : 123-124 Santoro E. La medicina viaggia su YouTube. Ricerca & Pratica 2010 ; n.151 : 27-28 Santoro E. Wikipedia o wiki medici? Ricerca & Pratica 2010 ; n.152 : 69-70 Santoro E. La newsletter web 1.0 del Ministero della Salute. Ricerca & Pratica 2010 ; n. 155 : 202-203 Santoro E. E indispensabile il coinvolgimento dei medici. Avvenire Medico 2010;8:13-14 Santoro E. Web 2.0 e diabete: il nuovo web al servizio dellaggiornamento del medico. Giornale Italiano di Diabetologia e Metabolismo 2010; 30:45-48 Santoro E. Internet, web 2.0 e malati cronici: i risultati di unindagine. Partecipasalute 2010; http://www.partecipasalute.it/cms_2/node/1467 Santoro E. La cartella clinica delude le aspettative. Partecipasalute 2010; http://www.partecipasalute.it/cms_2/node/1520 Santoro E. Per Big Pharma il futuro del marketing su Facebook? Partecipasalute 2010; http://www.partecipasalute.it/cms_2/node/1660 Santoro E. Niente e-mail tra medico e paziente. Partecipasalute 2010; http://www.partecipasalute.it/cms_2/node/1645 Santoro E. La salute ai tempi del web 2.0. Il Sole 24 Ore Sanit 29 giugno 5 luglio 2010; 1617

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Santoro E. Fascicolo elettronico, il mezzo flop inglese. Il Sole 24 Ore Sanit 19-25 ottobre 2010; 12

OTHER PUBLICATIONS (2010)

Levi F, La Vecchia C. Cancer of the prostate In: Tobacco - Science, policy and public health, 2e. (Chapter 19 and Prelims) Rossi M, Negri E, Bosetti C, Pelucchi C, La Vecchia C. Epidemiology behind fruit and vegetable consumption and cancer risk with a focus on flavonoids. In: Plant phenolics and human health. Biochemistry, Nutrition and Pharmacology. (Cesar G. Fraga, ed.). John Wiley & Sons. Inc., Hoboken, New Jersey. pp. 471-487 (2010)

RESEARCH ACTIVITIES Laboratory of General Epidemiology CASE-CONTROL STUDIES ON CANCER Dietary patterns and upper aero-digestive tract cancers
In a case-control study of laryngeal cancer, we identified five major dietary patterns named animal products, starch-rich, vitamins and fiber, vegetable unsaturated fatty acids, and animal unsaturated fatty acids. The vitamins and fiber dietary pattern was inversely associated with laryngeal cancer, whereas the animal products and the animal unsaturated fatty acids patterns were directly associated with it. There was no significant association between the vegetable unsaturated fatty acids and the starch-rich patterns and laryngeal cancer risk. These findings suggest that diets rich in animal products and animal fats are directly related, and those rich in fruit and vegetables inversely related, to laryngeal cancer risk. A similar analysis of oral and pharyngeal cancer indicated that diets rich in animal origin and animal fats are positively, and those rich in fruit and vegetables, and vegetable fats inversely related to oral and pharyngeal cancer risk.

Green tea and gastric cancer

We considered the relationship between green tea and gastric cancer risk in Harbin, China, an area with high baseline risk of stomach cancer. No association emerged when tea consumption alone was considered. When tea consumption was further classified according to the temperature, however, the odds ratio (OR) was 0.19 for cold tea intake 750 g/year and 1.27 for hot tea intake as compared to non drinkers.

Flavonoids, proanthocyanidins and gastric cancer

Flavonoids have been suggested to be responsible for the potential beneficial properties of fruit and vegetables on stomach cancer risk. We analyzed data from a case-control study conducted

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in Italy including 230 cases with incident gastric cancer and 547 controls. Using logistic regression models, strong inverse relations were found for proanthocyanidins. The OR for the highest versus the lowest quintile was 0.44 for monomers and dimers combined, and 0.36 for polymers with three or more mers. Further adjustment for fruit and vegetables, or vitamin C, did not materially change these associations.

Aspirin and gastric cancer risk

Studies on the relation between aspirin and non-steroideal anti-inflammatory drugs (NSAIDs), and the risk of gastric cancer are inconsistent. We analyzed data of our case-control study conducted in Northern Italy in 1997-2007, based on 230 cases and 547 controls. Regular use of aspirin was reported by 21 (9.2%) cases and 46 (8.5%) controls, and was not related with risk of gastric cancer (OR=1.09 95% confidence interval, CI: 0.611.94). No associations were found with duration of use, age at first use, time since first use, and time since last use.

Nutrients and colon cancer

We were also involved in a nationwide case-control studies from 8 Canadian provinces. With reference to colon cancer, intake of polyunsaturated fat, trans-fat and cholesterol were significantly associated with the risk of colon cancer. The association was stronger with proximal colon cancer. An increased risk was also observed with increasing intake of sucrose for both proximal and distal colon cancers. An elevated risk of proximal colon cancer was also found with increased lactose intake.

Dietary patterns and colorectal cancer

In an Italian case-control study we identified five nutrient-based dietary patterns, and we investigated their role on colorectal cancer risk. Direct associations were observed between the Starch-rich pattern and both cancer of the colon (OR=1.68) and rectum (OR=1.74). Inverse relations were found between the Vitamins and fiber pattern and rectal cancer (OR=0.61), between the Unsaturated fats (animal source) and the Unsaturated fats (vegetable source) and cancer of the colon (OR=0.80 and OR=0.79, respectively). The Animal products pattern was not associated with colorectal cancer.

Proanthocyanidins and colorectal cancer

In vitro and animal studies suggest that proanthocyanidins decrease cancer risk, particularly colorectal cancer. We analyzed data from an Italian case-control study on 1953 incident cases of colorectal cancer and 4154 controls. Using logistic regression methods, we found a decreased risk of cancer with increasing intake of proanthocyanidins. The OR for the highest quintile compared to lowest was 0.88 for monomers, 0.75 for dimers, 0.84 for trimers, 0.80 for polymers with 4-6 meres, 0.79 for polymers with 7-10 meres, and 0.69 for polymers with more than 10 meres. The associations appeared stronger for the rectum than the colon. Our results may explain the protective effect of fruit and vegetables on these tumors.

Metabolic syndrome and colorectal cancer

We analysed data from a multicentre case-control study conducted in Italy and Switzerland, including 1378 cases of colon cancer, 878 cases of rectal cancer and 4661 controls to assess the relation between metabolic syndrome (MetS) and its components and colorectal cancer. MetS was defined according to the International Diabetes Federation criteria. With reference to each component of the MetS, the ORs of colorectal cancer were increased in men only. Accordingly, colorectal cancer risk was increased in men (OR=1.86), but not women (OR=1.13), with MetS. Results were similar for colon and rectal cancers. This study supported a direct association between MetS and both colon and rectal cancers in men, but not in women.

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Dietary habits and pancreatic cancer

In the case-control study on pancreatic cancer we investigated the role of dietary habits. Frequent meat consumption was associated to a two-fold increased risk of pancreatic cancer; the risk was significant for meat cooked by boiling/stewing or broiling/roasting. Added table sugar (OR = 2.23) and potatoes (OR = 1.79) were significantly related to pancreatic cancer. A significant inverse association emerged for non-citrus fruits (OR = 0.41), cooked vegetables (OR = 0.57), and, possibly, for pulses (OR = 0.59). The present study supports an inverse association between fruit and vegetables and pancreatic cancer risk, and it confirms a direct relation with meat.

Macronutrients, fatty acids, cholesterol and pancreatic cancer

A positive association was found between pancreatic cancer and animal proteins (OR=1.85 for the highest versus the lowest quintile of intake; p for trend=0.039), whereas a negative association was observed for sugars (OR=0.52; p for trend=0.003). Non-significant negative associations emerged for vegetable proteins (OR=0.69) and polyunsaturated fatty acids (OR=0.67). A diet poor in animal proteins and rich in sugars (mainly derived from fruit) appears to have a beneficial effect on pancreatic cancer risk.

Micronutrients and pancreatic cancer

Several studies have reported an inverse relation between fruit and vegetables and pancreatic cancer, but no specific component of these foods has been clearly identified as beneficial. We estimated the intakes of selected vitamins, carotenoids and minerals, using an Italian food composition database applied on information collected through a food frequency questionnaire. We observed inverse associations with vitamin E (OR=0.60), vitamin C (OR=0.44), folate (OR=0.56), and potassium (OR=0.57). Non significant protections were also observed for carotene, -carotene, and -cryptoxanthin, while no relation was found with retinol, lycopene, lutein/zeaxanthin, total carotenoids, vitamin D, thiamin, riboflavin, niacin, vitamin B6, calcium, phosphorus, iron, zinc, and sodium.

Dietary acrylamide and pancreatic cancer

In the same database of pancreatic cancer, we investigated the relation with dietary acrylamide exposure. The OR of pancreatic cancer for an increase in acrylamide intake of 10 g/day was 1.01. No meaningful difference between ORs was found in strata of smoking habit, alcohol drinking, body mass index (BMI) and other selected covariates. Thus, this study found no association between dietary acrylamide and pancreatic cancer.

Soft drinks, sweetened beverages and pancreatic cancer

We considered the association between carbonated drink consumption and pancreatic cancer risk in an Italian case-control study conducted on 326 pancreatic cancer cases and 652 matched controls. We also combined the results from all the studies on soft drinks or sweetened beverages and pancreatic cancer published before June 2010, using a meta-analytic approach. In the case-control study, compared with non-drinkers, the multivariate OR was 1.02 for carbonated drink consumers and 0.89 for regular consumers. From the literature search, we identified 4 other case-control (1919 cases) and 6 cohort studies (2367 cases). The pooled relative risks (RR) for soft drink consumers vs. non-consumers were 0.97 for case-control, 1.05 for cohort, and 1.02 for all studies.

Physical activity and pancreatic cancer

We also analyzed data on physical activity and pancreatic cancer risk. For the highest vs. lowest level of occupational physical activity, the ORs were 1.24 at age 15-19, 1.21 at age 30-39 and 1.41 at age 50-59, with no significant trend in risk at any age considered. The corresponding

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ORs for recreational physical activity were 1.44 at age 15-19, 1.52 at age 30-39 and 1.39 at age 50-59. Therefore, occupational and recreational physical activity were not associated to a decreased risk of pancreatic cancer in our study.

Tobacco, alcohol and pancreatic cancer

Tobacco smoking is the major established risk factor for pancreatic cancer, whereas the role of alcohol consumption is still open to debate. In our case-control study including 326 cases with pancreatic cancer and 652 controls, pancreatic cancer was associated to current smoking (OR=1.68), and the risk rose with increasing number of cigarettes/day (OR=2.04 for 20 cigarettes/day). No association emerged for former smokers. Alcohol consumption was associated to increased pancreatic cancer risk, but ORs were significant only among heavy drinkers. The risk was 4.3-fold higher in heavy smokers and heavy drinkers in comparison with never smokers who drunk <7 drinks/week.

Diabetes mellitus, other medical conditions and pancreatic cancer

We considered diabetes and other medical conditions in relation to pancreatic cancer risk among two datasets collected between 1983 and 2008 and including 683 cases. Diabetes was associated with pancreatic cancer risk, and the association was significant for diabetes diagnosed up to 10 years before pancreatic cancer. As compared to non-diabetic non-smokers, the OR was 1.85 among non-diabetic current smokers, 2.18 among diabetic never/former smokers, and rose to 4.67 among diabetic current smokers, indicating a multiplicative effect between these two risk factors. Pancreatic cancer was also significantly associated with pancreatitis, primarily among those diagnosed within 2 years.

Aspirin use and pancreatic cancer

We analyzed the role of aspirin use in pancreatic carcinogenesis using data from a hospitalbased case-control study (308 cases e 477 controls) conducted in Italy between 1991 and 2008. A total of 22 cases (7%) and 37 controls (8%) reported regular aspirin use, with a corresponding adjusted OR of 0.87. A slight protection, although not significant, was observed for duration of use > or =5 years (OR=0.53) and for time since first use > or =10 years (OR=0.69). The risk of pancreatic cancer was significantly below unity for current users of > or =5 years (OR=0.23).

Dietary fiber and endometrial cancer

The epidemiological evidence on the relation between dietary fiber intake and endometrial cancer is contradictory. We further investigated the issue in a case-control study of 454 women with incident, histologically confirmed, endometrial cancer and 908 controls. Information on diet was elicited using a validated food frequency questionnaire. Lignin intake was significantly inversely related to endometrial cancer (OR=0.6 for the highest vs. the lowest quintile of intake), whereas total fiber intake was not associated with risk. Data suggest the potential importance of lignin intake in the prevention of endometrial cancer at Italian consumption levels.

Metabolic syndrome and endometrial cancer

The same case-control study - conducted between 1992 and 2006 in the provinces of Milan, Pordenone and Naples - showed a significant association between the metabolic syndrome (a cluster of central obesity, diabetes, hyperlipidemia, hypertension) and risk of endometrial cancer. According to different definitions of the metabolic syndrome, women with at least three components of the syndrome were 3 to almost 9 times more likely to develop endometrial cancer than women with no component.

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Aspirin and risk of endometrial cancer

We provided additional information on the role of aspirin on endometrial cancer. Regular aspirin use was reported by 28 (6.3%) cases and 46 (6.8%) controls, corresponding to a non significant multivariate OR of 0.65. There was no consistent pattern of risk with duration of use, nor with age at first use, time since first use or time since last use. Further, there was no difference in risk estimates across strata of BMI. This study -the first one from a population outside North America- adds relevant information on the absence of a consistent association between aspirin use and endometrial cancer risk, even in high-risk overweight and obese women.

Use of fertility drugs and endometrial cancer

The same case-control study analyzed time-related aspects of the use of fertility drugs in association to the risk of endometrial cancer. The OR of endometrial cancer for ever use of fertility drugs was 3.26. The risk was higher for duration of use 12 months or more (OR= 6.10), time since last use 25 years or less before the interview (OR= 5.30), and for age at first use less than 30 years (OR= 5.14). Thus, our data support earlier findings of an increase in risk of endometrial cancer with duration of use of fertility drugs.

Nutrients and prostate cancer

In a companion study of prostate cancer, an increased risk was observed with increasing intake of sucrose and disaccharides. In contrast, men in the highest quartile of cholesterol intake were at lower risk of prostate cancer. No association was found with intake of total proteins, total fat, monounsaturated fats, polyunsaturated fats, monosaccharides, and total carbohydrates. The findings provide evidence that a diet low in trans-fat could reduce prostate cancer risk.

Citrus fruit and cancer

Our findings indicated that citrus fruit has a protective role against cancers of the digestive and upper respiratory tract. No association was found with breast, endometrial, ovarian, prostate and renal cell cancer (RCC).

Flavonoids, proanthocyanidins and cancer

We considered flavonoids and proanthocyanidins in a network of Italian case-control studies including about 10,000 incident cases of selected cancers, and over 16,000 controls. Using logistic regression analysis, these studies provide evidence of a protective role of flavanones on upper aerodigestive tract cancers, flavonols, anthocyanidins and proanthocyanidins (in particular, those with higher degree of polymerization) on colorectal cancer, flavonols and flavones on breast cancer, isoflavones on ovarian cancer, and flavonols on renal cancer. No association between flavonoids and prostate cancer emerged.

Glycemic index and glycemic load and to body mass index and waist to hip ratio
High-glycemic index (GI) diet has been associated with obesity, but epidemiological data are inconsistent. We investigated the relation between GI and glycemic load (GL) with BMI and waist-to-hip ratio (WHR), using data of 7,724 patients from the control group of a network of hospital-based case-control studies from Italy. Mean BMI decreased from the lowest to the highest tertile of GI from 26.59 to 26.18 kg/m2 in men (p ~ 0.005), and from 25.81 to 25.09 kg/m2 in women (p < 0.001). With respect to GL tertiles, the corresponding values were 26.41 and 26.25 kg/m2 in men (p ~ 0.51), and 26.01 and 24.93 kg/m2 in women (p <0.001). No consistent association was found with WHR.

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Trends in adherence to the Mediterranean diet

We investigated whether adherence to the Mediterranean diet changed during the period 19912006 in an Italian population. Subjects included in the study were 3247 adults with a wide spectrum of acute conditions unrelated to long term modifications of diet. For each subject, we computed a Mediterranean diet score on the basis of nine a priori defined peculiar characteristics of the Mediterranean dietary pattern. Adherence to the Mediterranean diet showed no significant change over the last 15 years in both sexes. Subjects aged 55-64 years, those with high education, and those born in central and southern Italy showed the highest adherence to the Mediterranean diet.

POOLED- AND META-ANALYSES AND REVIEW PAPERS Alcohol drinking and oral and pharyngeal cancers
We performed a review of all studies of alcohol, oral and pharyngeal cancers published between 1988 and 2009. A strong doseresponse effect on the intensity of alcohol use was reported in most of the studies. However, no apparent association was observed for the duration of alcohol use. A decreased risk of approximately 10 to 15 years was associated with alcohol cessation. Similar associations have been observed among nonsmokers in over 20 studies. In general, the dominant type of alcohol consumption in each population was associated with the greatest increase in risk. We also conducted a meta-analysis of available data on the association between alcohol drinking and oral and pharyngeal cancer (OPC) risk, published up to September 2009. We identified 43 case-control and 2 cohort studies, including a total of 17,085 OPC cases. The pooled RR was 1.21 (95% CI, 1.10-1.33) for 1 drink per day, and rose to 5.24 (95% CI, 4.366.30) for heavy alcohol drinking (4 drinks per day). The dose-risk analysis resulted in RR of 1.29 for 10 g ethanol/day, 3.24 for 50 g ethanol/day, 8.61 for 100 g ethanol/day, and 13.02 for 125 g ethanol/day. This meta-analysis provided more precise evidence of a gross excess of OPC risk for heavy alcohol drinkers. It also indicated an increased risk for moderate doses, i.e. 1 drink or 10 g ethanol/day.

Alcohol drinking and esophageal squamous cell carcinoma

We conducted a systematic review and meta-analysis using 40 case-control and 13 cohort studies that reported the risk of esophageal squamous cell carcinoma (ESCC) associated with alcohol drinking for at least 3 levels of consumption. In studies adjusted for age, sex, and tobacco smoking, the RR and 95% CI for the association between light alcohol drinking (12.5g/d) and risk of ESCC was 1.38 (1.14-1.67). The association was slightly stronger in Asian countries than in other populations. The adjusted RRs (95% CIs) were 2.62 (2.07-3.31) for moderate drinking (>12.5-<50g/d) and 5.54 (3.92-7.28) for high alcohol intake (50g/d); the RRs were slightly higher in non-Asian populations. In prospective studies, the RR (95% CI) was 1.35 (0.92-1.98) for light, 2.15 (1.55-2.98) for moderate, and 3.35 (2.06-5.46) for high alcohol intakes; light drinking showed an association with ESCC in Asia (5 studies) but not in other regions (3 studies). Among never-smokers (9 studies), the RR (95% CI) was 0.74 (0.471.16) for light, 1.54 (1.09-2.17) for moderate, and 3.09 (1.75-5.46) for high intakes. This metaanalysis further corroborated the association of moderate and high alcohol intake with risk of ESCC and provided risk estimates based on multiple prospective studies. Light alcohol intake appeared to be associated to ESCC mainly in studies in Asia, which suggests a possible role of genetic susceptibility factors.

Tobacco smoking and esophageal and gastric cardia adenocarcinoma

We conducted a meta-analysis of 33 studies published up to January 2010 on the association between tobacco smoking and esophageal and gastric cardia adenocarcinoma risk. Compared to

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never smokers, the pooled RR was 1.76 for ever, 2.32 for current, and 1.62 for ex-smokers. No significant difference emerged between esophageal (RR=1.85 for ever vs never smokers) and gastric cardia (RR=1.76) adenocarcinoma. We found a direct association with quantity (RR=2.48 for 20 cig/day) and duration (RR=2.32 for 40 years) of cigarette consumption. This meta-analysis showed a significant excess of esophageal and gastric cardia adenocarcinoma risk for smokers.

Alcohol drinking and laryngeal cancer

We conducted a meta-analysis on the association between alcohol drinking and laryngeal cancer risk. Forty studies (38 case-control, 2 cohort) reporting at least three levels of consumption were included. Overall, alcohol drinking versus non-drinking was associated with an approximately 2-fold increase in risk of laryngeal cancer (RR=1.90; 95% CI: 1.59-2.28). While light alcohol drinking (1 drink/day) did not show any significant association with risk of laryngeal cancer (12 studies. RR=0.88; 95% CI: 0.71-1.08), moderate drinking (>1 to <4 drinks/day) was associated with a 1.5-fold increase in risk (35 studies. RR=1.47; 95% CI: 1.25-1.72) and heavy drinking (4 drinks/day) was associated with a 2.5-fold increased risk (33 studies. RR=2.62; 95% CI: 2.13-3.23). Thus, this meta-analysis showed a significant and positive association between alcohol drinking and laryngeal cancer risk, even at moderate doses.

Alcohol drinking and colorectal cancer risk

A meta-analysis with dose-risk estimation was performed to evaluate the association between alcohol drinking and colorectal cancer risk from published epidemiological literature. We identified 27 cohort and 34 case-control studies. The summary RR was 1.21 (95% CI, 1.131.28) for moderate alcohol drinking (>1 to 3 drinks/day), and 1.52 (95% CI, 1.27-1.81) for heavy alcohol drinking (4 drinks/day). The RR for moderate drinkers, compared to non- or occasional drinkers, was stronger for men (RR=1.24, 95% CI, 1.13-1.37) than for women (RR=1.08, 95% CI, 1.03-1.13). For heavy drinkers, the association tended to be stronger in Asian studies (RR=1.81, 95% CI, 1.33-2.46) compared to studies conducted in other geographical regions, and in particular in Europe (RR=1.16, 95% CI, 0.95-1.43). The dose-risk analysis found RRs of 1.07 (95% CI, 1.04-1.10), 1.18 (95% CI, 1.12-1.25), 1.38 (95% CI, 1.281.50), and 1.82 (95% CI, 1.41-2.35) for 10, 25, 50, and 100 g/day of alcohol, respectively. This meta-analysis provided strong evidence for a positive association between colorectal cancer risk and alcohol drinking, even at relatively low doses.

Dietary patterns and colorectal adenomas and cancer

We conducted a systematic review of dietary patterns and colorectal disease and cancer. Dietary patterns named as healthy, prudent, fruit and vegetables, fat-reduced/diet foods, vegetable/fish/poultry, fruit/whole grain/dairy, and healthy eating index-2005, recommended food and Mediterranean diet scores were all associated with reduced risk of colorectal cancer. In contrast, diets named Western, pork-processed meat-potatoes, meat-eaters, meat and potatoes, traditional patterns, and dietary risk and life summary scores were associated with increased risk of colorectal cancer. Dietary patterns for adenomas were consistent with those identified for colorectal cancer.

Cigar and pipe smoking, smokeless tobacco use and pancreatic cancer in the PANC4 consortium
We conducted a pooled analysis of cigar and pipe smoking and smokeless tobacco use and risk of pancreatic cancer using data from 11 casecontrol studies (6,056 cases and 11,338 controls) within the International Pancreatic Cancer Case-Control Consortium (PanC4). Compared with never tobacco users, the OR for cigar-only smokers was 1.6 (95% CI: 1.22.3), i.e. comparable to that of cigarette-only smokers (OR 1.5; 95% CI: 1.41.6). The OR was 1.1 (95% CI: 0.69

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1.6) for pipe-only smokers. There was some evidence of increasing risk with increasing amount of cigar smoked per day (OR 1.82 for 10 grams of tobacco), although not with duration. The OR for ever smokeless tobacco users as compared with never tobacco users was 0.98 (95% CI: 0.751.3).

Selected aspects of Mediterranean diet and cancer

We reviewed the role of the Mediterranean diet on cancer risk in our series of case-control studies. Several aspects of the Mediterranean diet (i.e., high vegetable and fruit consumption, high fish, low meat intake, use of olive oil, whole grain foods consumption) were favourable on risk of various cancer, particularly those of the digestive tract.

Acrylamide exposure and human cancer

We performed a critical review and meta-analysis of studies considering exposure to acrylamide and cancer risk. We identified 25 publications providing relevant results. The summary RRs for an increase of 10 g/day of dietary acrylamide intake were close to unity for all the cancers considered, ranging from 0.98 for esophageal cancer to 1.01 for colon, endometrial, ovarian and kidney cancer. None of the estimates was significant. The combined standardized mortality ratios for high occupational acrylamide exposure were 1.67 for pancreatic cancer and 2.22 for kidney cancer. Available studies consistently suggest a lack of an increased risk of most types of cancer from exposure to acrylamide. The main association which requires further monitoring involves kidney cancer.

InterLymph study
We participated to a consortium study of non-Hodgkin lymphoma (NHL) worldwide. In this pooled analysis, these was no significant association between increasing birth order and risk of NHL. However, a significant association was present for a number of B- and T-cell NHL subtypes. A significant positive association was present in higher socioeconomic status participants only. Results were very similar for the related variable of sibship size. The known correlation of high birth order with low socioeconomic status suggests that selection bias related to socioeconomic status may be responsible for the association between birth order and NHL.

Ambient particulate matter and preterm birth or low birth weight

We reviewed epidemiologic evidence on maternal exposure to particulate matter (PM) and adverse pregnancy outcomes, up to June 2009. We identified a total of 30 papers, including 13 with information on preterm birth, 17 on low birth weight (LBW) or very low birth weight (VLBW), and 4 on small for gestational age (SGA). Eight studies on preterm birth, 11 studies on LBW/VLBW and two studies on SGA reported some increased risk (by about 10-20%) in relation to exposure to PM; no meaningful associations was found in the remaining studies. However, even in studies reporting some excess risk, this was inconsistent across exposure levels and pregnancy periods. Epidemiologic studies on maternal exposure to PM during pregnancy thus do not provide convincing evidence of an association with the risk of preterm birth and LBW/VLBW and SGA.

OTHER PROJECTS HI-WATE project - Colorectal cancer and disinfection by-products in Italy and Spain
Experimental data suggest that disinfection by-products (DBPs) are possible colorectal carcinogens, but epidemiological evidence is inconclusive. To evaluate colorectal cancer risk in relation to long-term DBP exposure we conducted a case-control study in the greater Milan area and the provinces of

Pordenone and Udine, Italy, and the metropolitan area of Barcelona, Spain. Cases are incident,

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histologically confirmed colon and rectal patients, aged 20-85 years and resident in the study areas. Controls are hospital-based (Italy) or population-based patients (Spain) matched to cases by age and sex. Besides data on known or potential risk factors of colorectal cancer, information is collected on residential and water source history, water consumption and use, including ingestion, showering, bathing, dishwashing and swimming pool attendance. Retrospective data on DPB (mainly trihalomethane, THM) levels in the study areas is collected through local water companies. Based on 400 cases and 363 controls from Italy, and 500 cases and 436 controls from Spain, 46% of Italian subjects and 56% of Spanish subjects drunk water from public water at the longest residence (mean duration 37 years in Italy and 35 years in Spain); the remaining consumed water from bottles or other sources. The multivariate OR for subjects drinking water from public supplies as compared to those drinking bottled water was 1.17 (95% CI, 0.87-1.58) in Italy and 1.18 (95% CI, 0.79-1.77) in Spain. Taking long compared to short shower yield an OR of 1.16 (95% CI, 0.83-1.63) in Italy and of 1.04 (95% CI, 0.80-1.40) in Spain. Mean THM levels in Italy were <10 g/l, and ranged between 17.6 and 134 g/l in Spain. No clear dose-risk relations between residential THM exposure and colorectal cancer risk were observed. Preliminary results suggest a weak if any association between colorectal cancer and DBPs.

The MILD trial

We have collaborated with the Multicentric Italian Lung Diagnosis (MILD) trial, a randomised trial of CT scan in lung cancer disease, plus smoking cessation assistance to both intervention and control subjects. In this study, serum amyloid A (SAA) protein was strongly related to lung cancer risk. Thus, SAA levels were predictive of an elevated risk of lung cancer, supporting the general view that inflammation is implicated in lung cancer development. In another analysis of the MILD trial, subjects with grade 2 bronchial diverticula were heavier smokers, reported a history of coughing more frequently, and showed more severe functional impairment, greater extent of emphysema and more severe bronchial wall thickening compared with subjects with grade 1 and those without bronchial diverticula. Thus, bronchial diverticula are a frequent finding in the major airways of smokers, and they are associated with other markers of smoking-related damage. In a third analysis, even a relatively small reduction in FEV1 % was found to be a significant predictor of increased lung cancer risk. Thus, screening for lung cancer using airflow obstruction with FEV1% is a strategy worth future consideration.

Influenza vaccination in children

In a study investigating the best recall system to improve influenza vaccination coverage in children with oncohematological malignancies, 205 subjects were randomised to one of three intervention groups: i) call by a known pediatrician from the Oncohematological Unit and vaccination in the same Unit; ii) call by a known pediatrician from the Oncohematological Unit and vaccination in a general pediatric clinic by an unknown paediatrician; iii) call by a pediatrician not previously involved in caring for the children and vaccination in a general pediatric clinic by an unknown paediatrician. The results showed that all of the recall strategies were useful in increasing influenza vaccination coverage, particularly the first one, but the efficacy was optimal only in the children who had interrupted chemotherapy for less than six months. In the same study, it was found that the clinical and socio-economic impact of influenza-like illness and the effectiveness of influenza vaccination in children with oncohematological disease were related to the length of the off therapy period, and seemed to be significantly greater in those who had been off therapy for less than 6 months in comparison with healthy controls. This suggests that the administration of influenza

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vaccination should be strongly recommended only among oncohematological children who have been off therapy for less than 6 months. Another investigation from the same study evaluated the efficiency of pediatric mid-turbinate nasal flocked swabs to detect influenza viruses used by parents in 203 children aged 6 months to 5 years with signs and symptoms of respiratory disease. Two nasal samples were collected from each child in a randomised sequence: one by a trained pediatrician and one by a parent. In comparison with the pediatrician-collected samples, the sensitivity and specificity of the parental collections were respectively 89.3% and 97.7%. The children were significantly more satisfied with the parental collections. This study showed that mid-turbinate nasal flocked swabs specifically designed for infants and children can be used by parents without reducing the influenza virus detection rate.

Mortality of workers exposed to aromatic amines

We have updated to 58 years the follow-up of a cohort of workers exposed to extensively high doses of aromatic amines. Overall, 56 deaths from bladder cancer were observed, compared with 3.4 expected (standardized mortality ratio, SMR = 16.5, 95% CI, 12.4 to 21.4). The SMR for bladder cancer increased with younger age at first exposure and increasing duration of exposure. Although the SMR for bladder cancer steadily decreased with time since exposure stopped, the absolute risk remained approximately constant at 3.5 deaths per 1000 man-years up to 29 years after exposure stopped. Excess risk was apparent 30 years or more after last exposure.

Incidence rates of second primary cancers of the head and neck

Scanty data are available on the incidence of second cancers of the head and neck (HN) and its pattern with age. We investigated this issue using data from a multicentric study of 13 population-based cancer registries from Europe, Canada, Australia and Singapore for the years 1943-2000. A total of 99,257 patients had a first primary HN cancer, contributing to 489,855 person-years of follow-up. A total of 1,294 of the patients (1.3%) were diagnosed with second HN cancers. Male incidence rates of first HN cancer steeply increased from 0.68/100,000 at age 30-34 to 46.2/100,000 at age 70-74, and leveled off at older age; female incidence increased from 0.50/100,000 at age 30-34 to 16.5/100,000 at age 80-84. However, age-specific incidence of second HN cancers after a first HN cancer in men was around 200-300/100,000 between age 40-44 and age 70-74 and tended to decline at subsequent ages (150/100,000 at age 80-84); in women, incidence of second HN cancers was around 200-300/100,000 between age 45-49 and 80-84. The incidence of second HN cancers does not increase with age, but remains constant, or if anything, decreases with advancing age.

Knowledge and prevention of HPV infection

A study was aimed to investigate the knowledge of Italian adolescents and parents concerning human papillomavirus (HPV) infection and its prevention. Both students and parents underestimated the likelihood of HPV infection, and this was associated with a lower propensity for vaccination. This is an important indication for future training programmes concerning HPV prevention designed to increase the acceptance of HPV vaccine in families.

Effect of omega-3 supplementation on depression in elderly women

In a randomized controlled trial, it was investigated whether a supplement containing long chain omega-3 polyunsaturated fatty acids improved depressive symptoms and health-related quality of life in depressed elderly patients. The mean Geriatric Depression Scale at 8-week was significantly lowered for the n-3 group. Supplementation with n-3 polyunsaturated fatty acids

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was found to be efficacious in the amelioration of depressive symptoms and quality of life in the treatment of depressed elderly female patients.

Menopausal hormone therapy and breast cancer incidence

Recent studies conducted in different areas of North America and Europe showed a 5-10% decline in the incidence of breast cancer. This followed reductions up to 70% in menopause hormone therapy (HT) use after 2002. The observation that the decline in breast cancer incidence was larger in (or limited to) women aged 50 years weighs in favour of an effect of reduced HT use. However, changes in screening are also likely to play a role in the decreasing incidence of breast cancer observed in several countries. Thus, the reasons of the falls in incidence of breast cancer remain open to discussion.

Cytokine gene polymorphisms and gastric precancerous lesions

In a meta-analysis of studies of cytokine gene polymorphisms and gastric cancer from Portugal and Mozambique, the IL1RN*22 genotype seems to consistently increase the risk of gastric precancerous lesions, supporting a role for this polymorphism in the early stages of gastric carcinogenesis.

Paraquat and Parkinsons disease

We reviewed data on Paraquat (PQ) and Parkinsons disease (PD). Several studies have suggested that pesticide exposure and life in rural areas are significant risks factors for PD. Among other pesticides, PQ has been linked to PD by epidemiological studies and experimental work in rodents, in which it causes lesions in the substantia nigra, pars compacta. However, the evidence that human exposure to the chemical results in an increased risk for PD is rather limited and based on insufficient epidemiological data.

Emergency Medical System response to out-of-hospital cardiac arrest in Milan, Italy

The objective of this study was to investigate how rapidly the Emergency Medical System provides life support to 1426 patients having an out-of-hospital cardiac arrest in Milan, Italy between January 2007 and October 2008. The mean time interval from collapse-to-first shock was 18.67 5.37 min. The mean Emergency Medical System unit response time interval was 7.07 3.14 min; time elapsed from arrival-to first CPR was 7.75 4.32 min. The dispatch to arrival and dispatch to CPR intervals are comparable with those reported in other large urban areas, but the time from arrival-to-first CPR was longer than recommended by current guidelines.

TOBACCO CONTROL Smoking in Italy 2008-2009

Two surveys on smoking were conducted in 2008 and 2009, i.e., before and during the economic crisis. Each survey was based on a sample of more than 3000 individuals, representative of the Italian population aged 15 years or over. Compared to 2008, there was a significant increase in smoking prevalence overall (25.4% in 2009 vs 22.0% in 2008; <0.01), among men (28.9% in 2009 vs 26.4%, p=0.13), and among women (22.3% in 2009 vs 17.9% in 2008; <0.01). The proportion of ever smokers did not change, whereas that of ex-smokers decreased (18% in 2008 vs 15% in 2009). For the first time after several decades, in 2009 smoking prevalence increased in both sexes in Italy. This may be at least in part due to relapses of former smokers, because of stress related to the economic crisis.

Smoking behavior, involuntary smoking, attitudes towards smoke-free legislations, and tobacco control activities in the European Union

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We estimated the correlation between the Tobacco Control Scale (TCS; a scale that measures the six most important cost-effective policies on tobacco control in each country) and selected smoking patterns in the 27 countries of the European Union (EU27) in 2008. The correlation between smoking prevalence and TCS score was negative (r = -0.42); the correlation between TCS score and support to smoking bans in all workplaces was positive. The correlation between TCS score and self-reported exposure to SHS was negative, but statistically non-significant.

Temporal changes of under-reporting of cigarette consumption in population-based studies

We monitored trends in under-reporting of smoking in Italy over the last two decades, using 9 representative population-based surveys on smoking conducted in Italy in 1990 and annually between 2001 and 2008, covering 26,397 individuals. The difference in cigarette consumption between legal sale and self-reported data has substantially increased in Italy, from approximately 1% in 1990, 25% in 2001 and up to 35% in 2008. This reflects increasing underreporting of cigarette consumption mainly due to a decreasing social acceptability of smoking. The difference in cigarette consumption between legal sale and self-reported data has substantially increased over the last two decades in Italy, reflecting increasing under-reporting of cigarette consumption mainly due to a decreasing social acceptability of smoking.

PUBLIC HEALTH PREVENTION AND INFORMATION

The major products of our activity have also been published in the lay press, in order to increase the project impact on prevention and public health.

Laboratory of Epidemiological Methods MONITORING OF CANCER MORTALITY Update of the systematic analysis of cancer mortality in Europe for 20002004
We analysed cancer mortality in 34 European countries during 20002004, with an overview of trends in 19752004 using data from the World Health Organization (WHO). From 19901994 to 20002004, overall cancer mortality in the EU declined from 185.2 to 168.0/100 000 (world standard, 29%) in men and from 104.8 to 96.9 (28%) in women, with larger falls in middle age. Total cancer mortality trends were favourable, though to a variable degree, in all major European countries. The major determinants of these favourable trends were the decline of lung (216%) and other tobacco-related cancers in men, together with the persistent falls in gastric cancer, and the recent appreciable falls in colorectal cancer. In women, relevant contributions came from the persistent decline in cervical cancer and the recent falls in breast cancer mortality, particularly in northern and western Europe. Favourable trends were also observed for testicular cancer, Hodgkin lymphomas, leukaemias, and other neoplasms amenable to treatment, though the reductions were still appreciably smaller in eastern Europe.

Age-period-cohort analysis of oral cancer mortality in Europe

To update trends in oral cancer mortality and analyse the recent epidemic in central Europe, official death certifications for oral and pharyngeal cancer for 37 European countries were derived over the period 1970-2007, and an age-period-cohort model was fitted for selected countries. Male oral cancer mortality continued to decline in most European countries, and, more importantly, it also started to decline in some of the countries with the highest male rates, i.e. Hungary (18.9/100 000) and Slovakia (15.8/100 000); persisting rises were, however,

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observed in Belarus, Bulgaria and Romania. In women rates were lower than in men and showed no appreciable trend over the recent period in the EU. The estimated effects of age, period of death and cohort of birth for most selected countries showed a fall in effects from the cohorts born after the 1950s. For the period effect there was a rise for the earlier periods, an inversion in the 1990's and a continuous fall up to the last studied period. Only some former non-market economy countries, like Romania, Ukraine and Lithuania, had rising cohort effect trends up to most recent generations. The major finding of analysis is the leveling of the epidemic for men in most European countries, including Hungary and other central European countries where mortality from this cancer was exceedingly high. This essentially reflects the changes in alcohol and tobacco consumption patterns in various populations.

The oral cancer epidemic in central and eastern Europe

We analyzed oral and pharyngeal cancer mortality in 38 European countries over the period 19752004. Joinpoint analysis was used to identify significant changes in trends. In the EU, male mortality rates rose by 2.1% per year between 1975 and 1984, by 1.0% between 1984 and 1993, and declined by 1.3% between 1993 and 2004, to reach an overall age-standardized rate of 6.1/100,000 in 20002004. Mortality rates were much lower in women, and the rate in the EU rose by 0.9% per year up to 2000, and leveled off to 1.1/100,000 in 20002004. In France and Italy, which had the highest rates in the past, male rates have steadily declined during the last two decades (annual percent change, APC=-4.8% in 19982004 in France and -2.6% in 19862003 in Italy). Persisting rises were, however, observed in several central and eastern European countries, with exceedingly high rates in Hungary (21.1/100,000; APC=6.9% in 19751993 and 1.4% in 19932004) and Slovakia (16.9/100,000; APC=0.1% in 19922004). The highest rates for women were in Hungary (3.3/100,000; APC=4.7% in 19752004) and Denmark (1.6/100,000; APC=1.3% in 19752001). Oral and pharyngeal cancer mortality essentially reflects the different patterns in tobacco smoking and alcohol drinking, including drinking patterns and type of alcohol in central Europe.

Age-period-cohort analysis of gastric cancer mortality in Europe

In order to analyse the favourable trends in gastric cancer mortality in Europe, we applied an age-period-cohort regression model to official, certified gastric cancer mortality data from 42 countries of the European region and the EU as a whole, from 1950 to 2007. We found that Central and northern European countries, such as France, the UK, Sweden and Switzerland that had very low mortality rates in the 2005-2007 period (between 4.5 and 5.5 / 100,000 men and 1.5 and 2.5 /100,000 women), showed steep descending period effects and cohorts that fell steeply from the earliest cohorts to those born after the 1940s where they stabilised. Conversely, many former non-market economy countries had rates greater than 20/100,000 in men and 10/100,000 in women. In these countries cohort effect falls were only seen later but the fall gradient remained stable throughout recent cohorts. Mortality rates were also found to be high in some countries of Southern and eastern Europe. The observed falls in gastric cancer mortality are driven by downward trends in the effects of both cohorts and periods, although these are stronger in the former, indicating that these favourable trends should continue in the foreseeable future, particularly in those countries that still had high mortality rates, while in some countries with very low rates there appears to be an asymptote in cohort effects born after the 1940s.

Colorectal cancer mortality in Europe

We updated to 2007 colorectal cancer mortality trends in Europe using data from the WHO. In the EU, between 1997 and 2007 mortality from colorectal cancer declined by around 2% per year, from 19.7 to 17.4/100,000 men (world standardized rates) and from 12.5 to 10.5/100,000 women. Persisting favourable trends were observed in countries of western and northern

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Europe, while there were more recent declines in several countries of eastern Europe particularly in women (but not in Romania and the Russian Federation). In 2007, a substantial excess in colorectal cancer mortality was still observed in Slovakia, Hungary, Croatia, the Czech Republic and Slovenia in men (rates over 25/100,000), and in Hungary, Norway, Denmark and Slovakia in women (rates over 14/100,000). Colorectal mortality trends were more favourable in the young (30-49 years) from most European countries, with a decline of 2% per year since the early 1990s in both men and women from the EU.

Disparities in breast cancer mortality trends in Europe

We also separately considered mortality from breast cancer. Changes in breast cancer mortality after 1988 varied widely between European countries, and the UK is among the countries with the largest reductions. Women aged <50 years showed the greatest reductions in mortality, also in countries where screening at that age is uncommon. The increasing mortality in some central European countries reflects avoidable mortality.

Liver cirrhosis mortality in Europe

With reference to liver cirrhosis, an increase of the burden of mortality appeared in Eastern Europe in two specific areas: South-eastern Europe and North-eastern Europe. In the first group of countries, liver cirrhosis mortality was 10-20 times higher than in most other European states, levels never before observed in Europe.

Childhood cancer mortality in Europe

We analyzed mortality data derived from the WHO for all childhood neoplasms and selected cancers in 30 European countries up to 2007. Between 1990-1994 and 2005-2007, mortality from all neoplasms steadily declined in most European countries (from 5.2 to 3.5/100,000 boys and from 4.3 to 2.8/100,000 girls in the EU). In 2005-2007, however, mortality rates from childhood cancers were still higher in countries from Eastern (4.9/100,000 boys and 3.9/100,000 girls) and Southern (4.0/100,000 boys and 3.1/100,000 girls) Europe than in those from Western (3.1/100,000 boys and 2.5/100,000 girls) and Northern (3.2/100,000 boys and 2.5/100,000 girls) Europe. Similar temporal trends and geographic patterns were observed for leukaemias, with declines from 1.7 to 0.9/100,000 boys and from 1.3 to 0.7/100,000 girls between 1990-1994 and 2005-2007 in the EU. For kidney cancer and NHL mortality rates were low and have been declining in larger European countries over the last 15 years. The pattern of trends was less clear for bone cancer, with no systematic downward trends at age 0-14, though some fall was evident at age 15-19. Thus, mortality from childhood cancer continued to decline over more recent years in most European countries. However, the mortality rates in Eastern - but also Southern - European countries in the mid 2000's were similar to those in the Western and Northern European ones in the early 1990's.

Coronary heart and cerebrovascular disease mortality in young adults, in Europe

We analyzed trends in mortality from coronary heart diseases (CHD) and cerebrovascular diseases (CVD) at age 35-44 in the EU as a whole and in 12 selected European countries, over the period 1980-2007. With joinpoint regression analysis we identified significant changes in trends and estimated average annual percent changes (AAPC). CHD mortality rates at ages 3544 years have decreased in both sexes since the 1980s for most countries, except for Russia (130/100,000 men and 24/100,000 women, in 2005-07). The lowest rates (around 9/100,000 men and 2/100,000 women) were in France, Italy and Sweden. In men, the steepest declines in mortality were in the Czech Republic (AAPC=-6.1%), the Netherlands (-5.2%), Poland (-4.5%) and England and Wales (-4.5%). Patterns were similar in women, though with appreciably lower rates. The AAPC in the EU was -3.3% for men (rate=16.6/100,000 in 2005-07) and -2.1%

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for women (rate=3.5/100,000). For CVD, Russian rates in 2005-07 were 40/100,000 men and 16/100,000 women, 5- to 10-fold higher than in most western European countries. The steepest declines were in the Czech Republic and Italy for men, in Sweden and the Czech Republic for women. The AAPC in the EU was -2.5% in both sexes, with steeper declines after the mid-late 1990's (rates=6.4/100,000 men and 4.3/100,000 women in 2005-07).

Trends in cancer mortality in Brazil, 1980-2004

We have also considered trends in cancer mortality in Brazil since 1980. Age-standardized mortality rates between 1980 and 2004, derived from the WHO database, were computed for all cancers and 24 major cancer sites in Brazil. Total cancer mortality rates increased over the last decade in men to reach 101.2/100,000, and in women 71.3/100,000. In men, upward trends were observed for cancers of the oral cavity and pharynx with a rate of 5.9/100,000 in 20002004, intestines (whose rate, however was low, i.e. 7.6), prostate (12.2), and leukemias (3.4). Breast cancer mortality leveled off at around 10/100,000 in the last decade, whereas declines were observed for cancers of the uterus, whose rate (8.3) however, remained comparatively high. Declines were observed for stomach cancer in both sexes, with rates of 11.1 in men and 4.6 in women. In conclusion, most rates were comparatively low, but key issues were the high

rates of head and neck cancers in men and (cervix) uterine cancer in women, i.e., in principle cancers that are largely avoidable through prevention, screening and early diagnosis. Trends in breast cancer mortality in Argentina
In an analysis for breast cancer trends from Cordoba, Argentina, rates over most recent calendar years decreased, mainly in the most urbanized districts. Ageperiodcohort models for Cordoba province and Cordoba Capital showed a favorable cohort effect for generations born after 1955, in the absence of a clear interpretation.

Childhood cancer mortality in America, Asia, and Oceania

In this article we analyzed and compared patterns in childhood cancer mortality in 24 developed and middle-income countries in America, Asia, and Oceania between 1970 and 2007. Childhood age-standardized mortality rates were derived from the WHO database for all neoplasms, bone and kidney cancer, NHL, and leukemias. Since 1970, rates for all childhood cancers dropped from approximately 8 to 3 per 100,000 boys and from 6 to 2 per 100,000 girls in North America and Japan. Latin American countries registered rates of about 5 and 4 per 100,000 boys and girls respectively for 2005 through 2007, similar to the rates registered in more developed areas in the early 1980s. Similar patterns were observed for leukemias. Highest bone cancer rates for 2005 through 2007 were registered in Argentina, in Mexico for kidney cancer and in Colombia for NHL, whereas the lowest rates were registered by Japan for kidney and by Japan and the United States for NHL. Improvements in the adoption of current integrated treatment protocols in Latin American and other lower- and middle-income countries worldwide would avoid a substantial proportion of childhood cancer deaths.

OTHER PROJECTS Risk factors for falls in community-dwelling older people

We performed a systematic review and meta-analysis on the literature on risk factors for falls in community-dwelling older people. We identified a total of 74 studies, considering several risk factors including sociodemographic, mobility, sensory, psychologic, and medical factors, and medication use. The strongest associations were found for history of falls (OR=2.8 for all fallers; OR=3.5 for recurrent fallers), gait problems (OR =2.1 and 2.2), walking aids use

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(OR=2.2 and 3.1), vertigo (OR=1.8 and 2.3), Parkinson disease (OR=2.7 and 2.8), and antiepileptic drug use (OR=1.9 and 2.7).

Follow-up of subjects enrolled in case-control studies

The main aim of this project is to collect follow-up information on vital status and, when needed, cause of death for subjects included in the case-control studies, in order to obtain a prospective study from retrospectively collected data and to analyse factors influencing survival from several cancers. We thus collected information on vital status of the subjects with pancreatic cancer included in our database, by contacting the competent institutional sources (Municipalities, Local Health Units, General Registry Offices).

Random-effects meta-regression models for studying nonlinear doseresponse relationship

A fundamental challenge in meta-analyses of published epidemiological dose-response data is the estimate of the function describing how the risk of disease varies across different levels of a given exposure. We developed a method, based on a two-step process, that addresses simultaneously the following issues: within studies variability, between studies heterogeneity, and nonlinear trend components. First, two-term fractional polynomial models are fitted within each study included in the meta-analysis, taking into account the correlation between the reported estimates for different exposure levels. Second, the pooled dose-response relationship is estimated considering the between studies heterogeneity, using a bivariate random-effects model.

Vitamin D and cancer risk

The epidemiological literature on sun exposure, vitamin D and lymphomas has been reviewed. Similarities in the epidemiology of melanoma and NHL led to the hypothesis that UV exposure increases the risk of NHL. Epidemiologic studies, however, have not confirmed this hypothesis. If anything, an inverse association with recreational sun exposure emerged. Sun exposure is a major determinant of vitamin D status. Studies investigating the association of dietary or serum vitamin D with NHL are scanty and inconsistent, and the vitamin D-NHL relation remains therefore largely undefined.

Strategies for prevention of cardiovascular mortality

An analysis of prevalence of risk factor and national cardiovascular mortality in Europe indicated that high-risk populations should give the highest priority to achieving favourable shifts in all modifiable risk factors. Irrespective of the level of any particular risk factor, the rewards will be greatest in these populations.

Laboratory of Epidemiology of Chronic Diseases CASE-CONTROL STUDIES Organization for data and biological sample collection for case-control studies
Data collection of epidemiological data is going on and it includes: 1) interview and interviewer management and training activity for new interviewers; 2) contacts with hospital department and ethical committee for study approval and conduction; 3) check and codification of patient questionnaires; 4) diagnosis and histological exam check; 5) organization and management of biological sample collection; 6) data input management.

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Ongoing case-control studies include: cancer of the oral cavity, pharynx, larynx, esophaguscardias, biliary tract, colorectum, bladder, lymphomas, myelomas and sarcomas. The overall updated dataset include about: 1250 cases of cancers of oral cavity and pharynx, 700 of the esophagus, 1100 of the stomach, 6500 of the colorectum, 600 of the liver, 120 of the biliary tract, 600 of the pancreas, 850 of the larynx, 500 cutaneous malignant melanoma, 7000 of the breast, 1000 of the cervix, 1000 of the endometrium, 200 of trophoblastic gestational disease, 200 of the vulva, 2000 of the ovary, 1300 of the prostate, 700 of the bladder, 800 of the kidney and renal pelvis, 600 of the thyroid, 200 of Hodgkin disease, 500 of non-Hodgkin disease, 200 of sarcomas, 300 of myelomas and about 18,000 controls. Biological sample collection, aimed to study genetic polymorphisms, includes cancers of the oral cavity, pharynx, larynx, bladder and colorectum.

Aspirin use and renal cell cancer

Aspirin has been related to decreased risk of several cancers, but studies on the relation with the risk of renal cell cancer (RCC) are inconsistent. We analyzed data of our case-control study. Regular use of aspirin for at least six months was reported by 67 cases and 99 controls and was not related with risk of RCC (OR = 0.98). The lack of association was consistent in both sexes, in two strata of age, in users for less than 3 years or 3 years or longer, and among users of aspirin as analgesic or for cardiovascular disease prevention.

Metabolic syndrome and cataract extraction

We explored the relationship between age-related cataract extraction and the metabolic syndrome or its various components separately and in various combinations in an Italian casecontrol study, including 761 cases and 1522 hospital controls. The ORs were 1.41 for a history of central obesity, 1.42 for hypertension, 1.25 for hyperlipidemia, and 1.16 for diabetes. Patients with the metabolic syndrome had an increased risk of cataract, with an OR of 2.01 (95% CI, 1.432.83).

META-ANALYSIS Coffee and cancers of the upper aero-digestive tract

We performed a meta-analysis on coffee consumption and risk of cancers of the upper aerodigestive tract, comparing the highest vs the lowest categories of coffee intake. For oral and pharyngeal cancer, laryngeal cancer and esophageal squamous cell carcinoma, we selected only studies with age, sex and smoking adjustments; for esophageal adenocarcinoma those with age, sex and BMI (or energy) adjustments. We found a significant reduction in the risk of oral and pharyngeal cancer of about 35% in highest coffee drinkers as compared to lowest drinkers, but no relation of coffee drinking with laryngeal and esophageal cancer.

Alcohol drinking and oral and pharyngeal cancer

We conducted a meta-analysis to provide a quantification of the association of alcohol consumption with oral and pharyngeal cancer separately, as well as with their subsites. We found higher alcohol-related RRs for pharyngeal than for oral cancer, particularly at higher doses: compared to non- or occasional drinkers, the pooled RRs for 4 drinks/day were 4.64 for oral (17 studies) and 6.62 for pharyngeal (17 studies) cancer. The association with cancer of the tongue was similar to that of oral cancer and weaker than that with hypopharyngeal and oropharyngeal cancers.

Coffee consumption and colorectal cancer

A meta-analysis of casecontrol studies on coffee consumption and colorectal cancer risk was conducted. Twenty-four eligible studies published before May 2010 were identified, including a

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total of 14,846 cases of colorectal, colon or rectal cancer. Compared to non/occasional drinkers, the ORs for drinkers were 0.83 (95% CI, 0.730.95) for colorectal, 0.93 (95% CI, 0.811.07) for colon and 0.98 (95% CI, 0.851.13) for rectal cancer, with significant heterogeneity among studies. The results of this meta-analysis of casecontrol studies suggested a moderate favorable effect of coffee consumption on colorectal cancer risk.

Alcohol drinking and pancreatic cancer

We performed a meta-analysis of relevant dose-risk results on the association between alcohol consumption and pancreatic cancer risk. The pooled RR was 0.92 for <3 drinks/day and 1.22 for 3 drinks/day. The increased risk for heavy drinking was similar in women and men, but apparently stronger in cohort studies (RR=1.29), in studies with high quality index (RR=1.30), and did not appear to be explained by residual confounding by either history of pancreatitis or tobacco smoking. Given the moderate increase in risk and the low prevalence of heavy drinkers in most populations, alcohol appears to be responsible only for a small fraction of all pancreatic cancers.

Alcohol and endometrial cancer

We analyzed data from our case-control study including 454 endometrial cancer cases and 908 controls, finding that alcohol drinking was not associated with the risk of this cancer, even for relatively high doses (OR=1.19 for 15 drinks/week, compared with never drinking). A lack of association was also observed for any type of alcoholic beverages (wine, beer and spirits). We also performed a meta-analysis on alcohol and endometrial cancer, including other 19 casecontrol and 7 cohort studies (for a total of 13,120 cases). Compared to non/low drinkers, the pooled RRs were 0.95 (95% CI 0.88-1.03) for drinkers and 1.12 (95% CI 0.87-1.45) for heavy drinkers.

OTHER PROJECTS INHANCE study

We were also actively involved in the International Head and Neck Cancer Epidemiology (INHANCE) consortium, which includes data from 33 head and neck cancer studies worldwide, and a total of about 25,000 cases of 33,000 controls. In an analysis for the INHANCE consortium, including over 8.000 cases, a lower BMI enhanced smoking- and drinking-related ORs for oral cavity/pharyngeal cancer (P < 0.01), while BMI did not modify smoking and drinking ORs for laryngeal cancer. Likewise, in the INHANCE dataset, quitting tobacco smoking for 14 years resulted in a head and neck cancer risk reduction (OR=0.70), with the risk reduction due to smoking cessation after 20 years, reaching the level of never smokers. For alcohol use, a beneficial effect on the risk of head and neck cancer was only observed after 20 years of quitting (OR=0.60, 95% CI, 0.400.89 compared with current drinking). Only a few studies explored the relation between coffee and tea intake and head and neck cancers, with inconsistent results. In the INHANCE consortium, including 5139 cases and 9028 controls with information on the issue, we found that coffee intake was inversely related with the risk of cancer of the oral cavity and pharynx: the ORs were 0.96 (95% CI, 0.940.98) for an increment of 1 cup per day and 0.61 (95% CI, 0.470.80) in drinkers of >4 cups per day versus nondrinkers. No association of coffee drinking was found with laryngeal cancer. Data on decaffeinated coffee were too sparse for detailed analysis, but indicated no increased risk. Tea intake was not associated with head and neck cancer risk.

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Definition and management of a cohort of residents in Asti for the cardiovascular risk evaluation
The study is conducted with the collaboration of the ALMA association and the Ordine dei Medici di Asti and includes data collection and first analysis of follow-up.

Laboratory of Medical Informatics Development of a medical collaborative platform

In collaboration with the Arcispedale S. Maria Nuova of Reggio Emilia a project with the following three aims has been started: the training of the medical staff working at the Arcispedale hospital in using web 2.0 tools and social media for professional education and update; the designing of tools to allow physicians to share medical knowledge; the development of a collaborative platform to allow physicians to share evidence-based medical practices, practice management information and other relevant medical information, and to discuss clinical cases. As of today, more than 80 physicians, health professionals and medical librarians have been trained on these issues. In addition, in collaboration with the Medical Library of the Arcispedale S. Maria Nuova of Reggio Emilia, we have developed a blog where physicians may access to and discuss on selected evidence-based medical literature. A collaborative platform for sharing medical slides, images and videos is planned to be developed during 2011.

Maintenance of the CARDIO.CARE, ONCO.CARE, GASTRO.CARE, NEURO.CARE, PNEUMO.CARE, BPCO.CARE, PAIN.CARE and DERMA.CARE websites
These indexes have been developed by the Laboratory of Medical Informatics in order to collect, classify, evaluate, and describe the most useful medical information on the web, and to provide Internet users with an easy means to surf the net. Several medical areas are covered including oncology (http://www.oncocare.it), neurology (http://www.neurocare.it), gastroenterology (http://www.gastrocare.it), cardiology (http://www.cardiocare.it), pulmonology (http://www.pneumocare.it, http://www.bpcocare.it ), the pain care and management (http://www.paincare.it), and dermatology (http://www.dermacare.it). The project is in collaboration with intramural departments (Department of Oncology, Laboratory of Neurological Disorders and Department of Cardiovascular Research, Laboratory of General Practice Research, Laboratory of Translational and Outcome Research in Oncology) and extramural research groups (Italian Group for Epidemiologic Research in Dermatology, GISED). During 2010, information about the use of web 2.0 tools and technologies (such as podcast, RSS feeds, blogs, webcasts, and webinars) by the indexed websites has been collected. RSS feeds and podcasting services have been activated on the CARE websites in order to allow the users to access to the corresponding applications available on the classified websites.

Studies on the typology of the web 2.0 applications in medicine

The Laboratory of Medical Informatics is involved in studies and surveys which aim is to describe the typology of the web 2.0 applications and tools (including social networks, podcasts, feed RSS, blogs, and wikis) in medicine available on the net, and how these are perceived by the medical community.

Internet as a research and formative tool on the chronic pain in cancer patient
This research project was born in the framework of the project "Pain in the patient with cancer", in collaboration with the Laboratory of Medical Research and Consumer Involvement and the

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Laboratory of Translational and Outcome Research in Oncology. Its aim is to make available for doctors, patients and families correct information about therapies for cancer pain and to produce greater tests on the effectiveness of therapies based on the use of analgesic drugs. This project is articulated in two main activities: Paincare, set-up a catalogue of selected web pages dedicated to the cancer pain and relative periodical up-to-date, http://www.paincare.it; Adaptation of the methods and instruments of Evidence Based Medicine to the resources available on the Internet about chronic pain in patients with cancer. This is done through a specific questionnaire. We are also considering the opportunity to set up a new Italian cancer pain website.

Training activities
In 2010, the Laboratory of Medical Informatics continued its training activity on issues related to the use of the Internet in medicine, and extended it to the use of the recent social media and web 2.0 technologies and tools in the medicine area. The members of the laboratory staff activated (or attended as invited teachers) a number of training courses, workshops, and master courses. Onsite CME courses for the Italian physicians have also been organized using the training/educational facilities and equipment available at the Mario Negri Institute.

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DEPARTMENT OF PUBLIC HEALTH

STAFF

Head Department

Maurizio BONATI, MD.

"Angelo & Angela Valenti" Centre for Health Economics (CESAV)

Head of Laboratory Livio GARATTINI, Econ.D.

Laboratory of Clinical Epidemiology

Head of Laboratory Guido BERTOLINI, MD.

Clinical Knowledge Engineering Unit

Head Unit Davide LUCIANI, MD.

Computer Methods and Programs for Clinical Research Unit

Head Unit Abramo ANGHILERI, IT.

Laboratory for Mother and Child Health

Head Laboratory Maurizio BONATI, MD.

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CURRICULA
Maurizio Bonati has a Medical School degree at the University of Milan. Areas of interest: Monitoring and epidemiological evaluation of drug utilisation and effects of drugs and vaccines in motherhood and childhood. Research methodology in general hospital and paediatric community practice. Transfer of information to the community. Epidemiology of paediatric and perinatal care. Past and present roles both at the Mario Negri Institute and in other institutions: 1973-77 Research Fellow at the IRFMN, within the Neurochemistry Lab.; 1977-85 Research Assistant at the IRFMN, within the Clinical Pharmacology Lab.; 1986-93 Chief of the Perinatal Clinical Pharmacology Unit at the IRFMN; Advisor to WHO for the Drug Utilization Research Group (pregnancy, paediatrics and breastfeeding); 1987-92 coordinator of the International Cooperative Study of Drug Use in Pregnancy, under the auspices of WHO and the support of EEC; 1992-93 co-editor of The Kangaroo; 2000-05 coordinator of the European Cooperative Study: Development of the European register of clinical trials on medicines for children (DEC-net), under the 5th Framework Programmes Quality of life and Management of Living Resources; since 1989 he has been director of the Centre for Drug Information; since 1993 head of the Lab. for Mother and Child Health; since 1997 teacher for the Lombardy regions professional training courses; since 2000 teacher for the Lombardy regions professional training courses; since 2002 Editor of the Ricerca & Pratica scientific journal; since 2003 professor of the School of Specialisation in Paediatrics - University of Milan Bicocca; teacher at the annual European course Evaluation of Medicinal Products in Children (promoted by ESDPPP and Eudipharm); from May 2008 Head of Department Public Health at the "Mario Negri" Institute for Pharmacology Research; since 2010 coordinator of the European Cooperative Study COHEMI-Coordination resources to Assess and Improve health status of migrants from Latin America, under the 7th Framework Programme for Research and Technological Development (Programme Cooperation- Health).
Selected publications Santoro E, Rossi V, Pandolfini C, Bonati M. DEC-net: the development of the European Register of Clinical Trials on Medicines for Children. Clinical Trials 2006;3:366-375. Clavenna A, Rossi E, De Rosa M, Bonati M. Use of Psychotropic Medications in Italian Children and adolescents. Eur J Pediatr 2007;166:339-47. Maschi S, Clavenna A, Schiavetti B, Campi R, Bernat M, Bonati M. Neonatal outcome following pregnancy exposure to antidepressants: a prospective controlled cohort study. BJOG 2008;115:283-289. Usala T, Clavenna A, Zuddas A, Bonati M. Randomised controlled trials of Selective Serotonin Reuptake Inhibitors in treating depression in children and adolescents: a systematic review and meta-analysis. European Neuropsychopharmacology 2008;18:62-73. Fortinguerra F, Clavenna A, Bonati M. Psychotropic drug use during breastfeeding: a review of the evidence. Pediatrics 2009;124:e547-e556. Fortinguerra F, Maschi S, Clavenna A, Bonati M. Pain management in the paediatric population. The regulatory situation in Europe. Arch Dis Child 2010;95:749-753.

Livio Garattini: got his degree in Economics in March 1983 at the Bocconi University in Milan. Educational activities: Kings Fund College, London: courses of health care management; Centre for Health Economics, York: review of publications on the English NHS; Ecole Nationale de la Sant Publique, Rennes: courses of health policy. Areas of interest: Health Economics and Health Policy Analysis. At present he is the Director of CESAV (Centre of Health Economics A. e A. Valenti - M. Negri Institute); 1981-1983: researcher at M. Negri Institute; 1983-1984: clerk at Banca Commerciale Italiana in Milan; 1984- 1985: junior consultant at Sogess srl in Milan; 1985-1990: researcher at Bocconi University in Milan.
Selected publications: Cornago D, Li Bassi L, De Compadri P, Garattini L. Pharmacoeconomic studies in Italy: a critical review of the literature. The European Journal of Health Economics 2007;8(2):89-95. Garattini L, Cornago D, De Compadri P. Pricing and reimbursement of in-patent drugs in seven European countries: A comparative analysis. Health Policy 2007;82:330-339. Garattini L, Motterlini N, Cornago D. Prices and distribution margins of in-patent drugs in pharmacy: A comparison in seven European countries. Health Policy 2008;85(3):305-313.

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Garattini L, Casadei G. Health technology assessment: for whom the bell tolls? The European Journal of Health Economics 2008;9(4):311-312. Garattini L, Casadei G, Freemantle N. Continuing medical education funding and management in Europe: room for improvement? (Editorial) JME 2009;12(1):56-59. Garattini L, Gritti S, De Compadri P, Casadei G. Continuing Medical Education in six European countries: A comparative analysis. Health Policy 2010;94(3):246-54.

Guido Bertolini got his Medical degree in 1989 at the University of Bologna, and the specialization in Pharmacological Research in 1993 at the Mario Negri Institute and in Gastroenterology in 1994 at the University of Pavia. He founded and chaired from 1997 to 2000 the School of Clinical Methodology and Quality of Care Improvement at the Ospedali Riuniti di Bergamo and the Istituto di Ricerche Farmacologiche Mario Negri. From 1999 to 2003 he has been contract professor at the post-doctoral schools in Anaesthesia and Intensive Care, University of Brescia and Milano; from 2002 to 2005 he has been contract professor of Educational Science at the Faculty of Lettere e Filosofia, University of Bergamo. Current research interests: Clinical Research Methodology, Continuous Quality of Care Assessment and Improvement, Health services research and outcome, Medical decision making, Medical Education. These interests are mainly developed within the fields of Intensive Care Medicine and Rare Diseases. Since 1997 he chairs the GiViTI Coordinating Center for research in intensive care medicine. He has been Head of the Unit of Epidemiology and Education for Clinical Practice at the Mario Negri Institute and since 2001 he is the Head of the Laboratory of Clinical Epidemiology. From 2001 to 2005 he has been Vice-chairman of the Research Group on Cost-effectiveness, Section on Health Services Research and Outcomes European Society of Intensive Care Medicine and, from 2001 to 2005, he has been President of the Scientific Committee of the Ospedale maggiore in Crema.
Selected publications Bertolini G, Rossi C, Anghileri A, Livigni S, Addis A, Poole D. Use of drotrecogin alfa (activated) in Italian intensive care units: the results of a nationwide survey. Intensive Care Med 2007;33:426-434. Malacarne P, Langer M, Nascimben E, Moro ML, Giudici D, Lampati L, Bertolini G, GiViTI. Building a continuous multicenter infection surveillance system in the intensive care unit: findings from the initial data set of 9,493 patients from 71 Italian intensive care units. Crit Care Med 2008;36:1105-1113. Poole D, Bertolini G, Garattini S. Errors in the approval process and post-marketing evaluation of drotrecogin alfa (activated) for the treatment of severe sepsis. Lancet Infect Dis 2009;9:67-72. Guido Bertolini, Simona Boffelli, Paolo Malacarne, Mario Peta, Mariano Marchesi, Camillo Barbisan, Stefano Tomelleri, Simonetta Spada, Roberto Satolli, Bruno Gridelli, Ivo Lizzola, Davide Mazzon. End-of-Life Decision-Making and Quality of ICU Performance: An Observational Study in 84 Italian Units. Intensive Care Med. 2010 Sep;36(9):1495504. J.C. Marchall, K. Reinhart, D. Angus, A. Argent, G. Bernard, G. Bertolini, S. Bhagwanjee, J.P. Cobb, D.J. cook, D. Fedson, S. Finfer, R. Fowler, C. Gomersall, E. Jimenez, N. Kissoon, D. McAuley, S. Opal, J.L. Vincent, S. Webb. InFACT: a global vritical care clinical research reponse to severe pendemic H1N1. Lancet. 2010 Jan 2;375(9708):11-3.

Marchall JC, Reinhart K, Angus D, Argent A, Bernard G, Bertolini G, Bhagwanjee S, Cobb JP, Cook, Fedson D, Finfer S, Fowler R, Gomersall C, Jimenez E, Kissoon N, McAuleyD, Opal S, Vincent JL, Webb S. InFACT: a global vritical care clinical research reponse to severe pendemic H1N1. Lancet 2010 Jan 2;375(9708):11-3.

Davide Luciani got his Medical Degree at the University of Bologna in 1995, and the post-doctoral certificate in "Tropical Medicine and Hygiene" at the University of Liverpool in 1997. In 2001, he spent one year at the Department of Statistical Science (University College London). Bayesian probabilistic applications, decision theory and the graphical approach to pathophysiological modelling represent his main interests. Within his research activity, these skills are meant as the main methodological ingredients in the formalization of clinical reasoning, in order to improve its effectiveness and to exploit its educational value. Since 2005 he is responsible of the Unit of Clinical Knowledge Engineering.
Selected publications Luciani D, Marchesi M, Bertolini G. The role of Bayesian Network in the diagnosis of pulmunary embolism. J Thromb Haemost 2003;1:698-707. Galli M, Luciani D, Bertolini G, Barbui T. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of thrombosis in the antiphospholipid syndrome. Blood 2003;102 (8):2717-23. Luciani D, Cavuto S, Antiga L, Miniati M, Monti S, Pistolesi M, Bertolini G. Bayes pulmonary embolism assisted diagnosis: a new expert system for clinical use. Em Med J 2007;24:157-164.

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M.Cesana, R.Cerutti, E.Grossi, E.Fagiuoli, M.Stabilini, F.Stella, D Luciani. Bayesian Data Mining Techniques: The Evidence Provided by Signals Detected in Single-Company Spontaneous Reports Databases. Drug Information Journal 2007;41:11-21. Latronico N, Bertolini G, Guarneri B, Botteri M, Peli E, Andreoletti S, Bera P, Luciani D, Nardella A, Vittorielli E, Simini B, Candiani A Simplified electrophysiological evaluation of peripheral nerves in critically ill patients: the Italian multi-centre CRIMYNE study. Crit Care 2007;11(1):R11. Bertolini G, Luciani D, Biolo G Immunonutrition in septic patients: A philosophical view of the current situation. Clin Nutr 2007;26:25-29.

Abramo Anghileri got his high-school certificate at the ITIS P. Paleocapa of Bergamo in 1997 and attended several courses in information technology (IT) applied to health care. Since 2001 he coordinates the IT activities of the Laboratory of Clinical Epidemiology at the Mario Negri Institute for Pharmacological Research. His main area of interest is informatics applied to clinical research. Since 2009, he is the head of the Unit of Computer Methods and Programs for Clinical Research of the Laboratory of Clinical Epidemiology.
Boffelli S, Rossi C, Anghileri A, Giardino M, Carnevale L, Messina M, Neri M, Langer M, Bertolini G. Continuous quality improvement in intensive care medicine. The GiViTI Margherita project - Report 2005. Minerva Anestesiol 2006; 72: 419-432. Bertolini G, Rossi C, Anghileri A, Livigni S, Addis A, Poole D. Response to the letter by Williams et al. Intensive Care Med 2007; 33(8): 1490-1 Bertolini G, Rossi C, Anghileri A, Livigni S, Addis A, Poole D. Use of drotrecogin alfa (activated) in Italian intensive care units: the results of a nationwide survey. Intensive Care Med 2007; 33(3): 426-34 Di Bartolomeo S, Valent F, Rossi C, Beltrame F, Anghileri A, Barbone F. Geographical differences in mortality of severely injured patients in Italy. Eur J Epidemiol 2008 Poole D, Rossi C, Anghileri A, Giardino M, Latronico N, Radrizzani D, Langer M, Bertolini G. External validation of the simplified acute physiology score (SAPS3) in a cohort of 28.357 patients from 147 Italian intensive care units. Intensive Care Medicine (2009)35:1916-1924

ACTIVITIES
The main aim of the Public Health Department is to understand which factors affect the health of individuals or entire populations and to define effective interventions for responding to their health needs. Special emphasis is therefore placed on prevention, so that the risks of contracting illness are lowered, and on the dissemination of independent, evidence-based information. The departments effort cannot disregard the National Health System, however, which must guarantee access to, and quality of, care that is based on principles of equity and appropriateness and must guarantee it especially to the more vulnerable patient groups. It is in this context that the Public Health Department carries out its activities. In addition to its formal research activity, the department participates in, and organises, initiatives involving information dissemination, training, and debate aimed at healthcare operators and social care workers, but also at the general population. These activities are also supported by the publication of the departments two journals: Ricerca&Pratica and Quaderni di Farmaco Economia. During 2009, 34 people worked in the Department

"A. and A. Valenti" Centre for Health Economics (CESAV)

The "Angelo e Angela Valenti" Centre for Health Economics (CESAV) was established in 1992 at the "M. Negri Institute" and based at Villa Camozzi- Ranica (Bergamo)-Italy. CESAV is primarily a research centre, but also does educational work. The centre is involved in health economics and health policy research. The main areas of research are: Economic Evaluation of Health Care Programs (i.e. assessment of costs and benefits of alternative health care treatments and services) and Comparative Health Policy Analysis (i.e. study of domestic and foreign health care systems, in particular aimed at identifying possible innovations for European countries).

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Laboratory of Clinical Epidemiology

The general aim of the Laboratory of Clinical Epidemiology is to contribute to the improvement of health care in different medical fields. The guiding principles are mainly two: to help physicians in using the available knowledge and resources at their best, and to contribute to the growth of applied knowledge for clinical practice. The Laboratory operates in the field of Intensive Care Medicine and Rare Diseases. Within the Laboratory, the Unit of Clinical Knowledge Engineering aims to bring the value of clinical reasoning out, through the implementation of probabilistic models for its formalization, thus favouring the evaluation and the continuous improvement of complex clinical activities. The main area of activity of the Unit of Computer Methods and Programs for Clinical Research is the development of an electronic health record for the ICU that would be able to conjugate the needs of clinical practice with those of clinical research, with the aim of closing the gap between the two.

Laboratory for Mother and Child Health

The main objective of the Laboratory for Mother and Child Health is to ensure a better mother and child well-being by undertaking interdisciplinary and collaborative work in the field. Four broad areas, or spheres, of research have been selected: - monitoring and epidemiological evaluation of utilisation and effects of drugs and vaccines; - research methodology in general hospital and paediatric community practice; - public health determinants of childrens well-being; - transfer of health information to the community. Special attention is given to activities involving countries in the north and south of the world. In addition to the formal research activities, the Laboratory promotes initiatives in the public health field, in particular those involving mother and child health care. The initiatives involve the participation in, and the organisation of, educational, training, and information-dissemination activities. The critical and active transfer of scientific knowledge is a continuous, daily stimulus to the Laboratorys activity.

NATIONAL COLLABORATIONS "A. and A. Valenti" Centre for Health Economics (CESAV)
Public and private institutions, other health care organizations (Ministry of Health, Regional and Local Health Authorities, Hospital Trusts).

Laboratory of Clinical Epidemiology

Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo Ospedale A. Manzoni, U.O. Anestesia e Rianimazione 1, Lecco. Ospedale Regionale S. Maria dei Battuti C Foncello, Dipartimento di Neurologia, Treviso Ospedale San Giovanni Bosco, Servizio Anestesia e Rianimazione, Torino Universit degli Studi di Brescia, Dipartimento di Specialit Chirurgiche, Scienze Radiologiche e Medico Forensi, Cattedra di Anestesia. Universit di Firenze, Facolt di Medicina e Chirurgia,Cattedra di Fisica e Informatica Medica.

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Universit di Milano Comunicazione.

Bicocca,

Dipartimento

di

Informatica

Sistemistica

Laboratory for Mother and Child Health

Centre for Child Health, (CSB) Cultural Paediatric Association, (ACP) Federfarma Lombardia Hospital of Bergamo Ospedali Riuniti, Poison Control Centre, Unit Clinical Toxicology Italian Drug Agency, (AIFA) Italian Global Health Watch (OISG) Italian National Institute of Health (ISS) Italian Society of Preparers Pharmacists (SIFAP) Italian Society of Clinical Pharmacy (SIFO) Il Pensiero Scientifico Editore Lombardy Region, Ministry of Health Operating unity of Neuropsychiatry of the childhood and of the adolescence, Foundation Policlinico di Milano, (UONPIA) University of Cagliari, Department of Neuroscience, Clinic of Child and Adolescent Neuropsychiatry University of Milan-Bicocca, Faculty Medicine, Paediatric Clinic

INTERNATIONAL COLLABORATIONS "A. and A. Valenti" Centre for Health Economics (CESAV)
CES (Collge des Economistes de la Sant) of Paris Corvinus University of Budapest Global Fund of Geneva WidO of Bonn Servicio Canario de la Salud, S/C de Tenerife University of Birmingham University of Hannover University of York University Pompeu Fabra of Barcelona University Erasmus of Rotterdam

Laboratory of Clinical Epidemiology

Centre for Intensive Care Medicines, Bloomsbury Institute, London, UK Clinic of Anesthesiology and Intensive Therapy, Jena, Germany Machine Intelligence Group, University di Aalborg, Denmark Institute of Anesthesia and Intensive Cares, Semmelweis University, Budapest, Hungary Department of Anesthesiology and Intensive Cares, University of Warsaw, Poland Department of Intensive Care, Hospital Generale di Novo Mesto, Slovenia Department of Pneumologia and Intensive Cares, Hospital Generale di Nicosia, Cyprus Laboratory of Experimental Physiopathology, Academic Unity of Sciences of the

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Health, University of the Studies Extremo Sul Catarinense, Cricima, SC, Brasil Department of Anesthesiology and Intensive Cares, University of Toronto, Canada Intensive Medicine, Regional Hospital of Lugano, Switzerland Intensive Medicine, Regional Hospital Beata Vergine, Mendrisio - Ticino, Switzerland Terapia Intensiva Pediatrica, Soroka University Medical Center, Beer-Sheva, Israele

Laboratory for Mother and Child Health

Centre for Tropical Diseases (CECOMET), Ecuador Clnica Infantil Colsubsidio, Bogot, Colombia European Medicines Agency (EMA) European Society for Developmental, Perinatal and Paediatric Pharmacology. (ESDPPP) European Union (EU) Hospital Robert Debr, Paris, France International Society of Drug Bulletins (ISDB) World Health Organization (WHO) University of Nottingham, Derbyshire Childrens Hospital, Derby, United Kingdom

EDITORIAL BOARD MEMBERSHIP "A. and A. Valenti" Centre for Health Economics (CESAV)
INTERNATIONAL: Acta Bio Medica; Applied Health Economics and Health Policy; Biomedical Statistics and Clinical Epidemiology; BMC-Health Services Research; Health Policy; Journal of Medical Economics; The European Journal of Health Economics. NATIONAL: FarmacoEconomia News; Farmeconomia e Percorsi Terapeutici; L'Internista; PharmacoEconomics Italian Research Articles; Quaderni di FarmacoEconomia.

Laboratory of Clinical Epidemiology

INTERNATIONAL: Intensive Care Medicine NATIONAL: Ricerca & Pratica; Dedalo. Gestire i sistemi complessi in sanit.

Laboratory for Mother and Child Health

INTERNATIONAL: European Journal Clinical Pharmacology; Journal of Clinical Pharmacology & Pharmacoepidemiology; Saludarte. NATIONAL: Dialogo sui Farmaci; Disturbi dAttenzione e Iperattivit; Quaderni ACP; Quaderni di Farmacoeconomia; Ricerca & Pratica.

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PEER REVIEW ACTIVITIES "A. and A. Valenti" Centre for Health Economics (CESAV)
Applied Health Economics and Health Policy; BMC-Health Services Research; Health Policy; PharmacoEconomics; The European Journal of Health Economics.

Laboratory of Clinical Epidemiology

INTERNATIONAL: British Medical Journal; Intensive Care Medicine; Critical Care Medicine; American Journal of respiratory and Critical Care Medicine. NATIONAL:

Ricerca & Pratica Laboratory for Mother and Child Health

INTERNATIONAL: Acta Paediatrica; African Health Sciences; Annals of African Medicine; Annals of African Medicine; Antimicrobial Agents and Chemoterapy; BMC Public Health; BMC Pulmonary Medicine; British Journal of Clinical Pharmacology; Current Drug Metabolism; Drug Metabolism Letters; European Journal of Clinical Pharmacology; Health and Quality of Life Outcome; Journal of Infection and Public Health; Pharmaceuticals; Pediatrics; Swiss National Foundation. NATIONAL: Dialogo sui Farmaci; Medico e Bambino; Quaderni ACP.

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP Laboratory of Clinical Epidemiology Ethical committee A.O. Bolognini in Seriate (Bergamo) National Health Plan Research Commission, Trent Autonomous Province Scientific Council of the Health and Illness Interdisciplinary Research Centre Scientific Commitee of the Lombardy Region on cancer research Laboratory for Mother and Child Health Ethical committee A.O. "Ospedale Maggiore" of Crema Technical Commission for the management, update, and elaboration of the Regional Therapeutic Formulary, Valle d'Aosta Autonomous Region Breastfeeding Promotion Work Group, Lombardy Region Scientific Advisory Board (PRIOMEDCHILD)

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EVENT ORGANIZATION "A. and A. Valenti" Centre for Health Economics (CESAV)
Regional Congress of Pharmacoeconomics: Economia del farmaco: fra soluzioni tecniche e decisioni politiche, May 25-26, Ranica (BG). Regional Congress of Pharmacoeconomics Politiche vaccinazione HPV regionali: esperienze a confronto, October 22, Milan.

Laboratory of Clinical Epidemiology

Congress, Meeting annuale GiViTI, October 27-29, Pesaro. Workshop, Startup Meeting Crimyne2, April 16, Milan. Workshop, Meeting Compact, May 12, Milan. Workshop, Meeting MargheritaTre, May 18-19, Milan. Course, Statistica applicata alla Ricerca biomedica, from September to December, Ranica (BG).

Laboratory for Mother and Child Health

Congress Modelli innovativi di intervento nella crisi acuta in adolescenza Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico e Laboratory for Mother and Child Health, IRFMN, Milan. Congress Bisogni comunicativi complessi e partecipazione nei contesti di vita Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico e Laboratory for Mother and Child Health, IRFMN, Milan.

CONFERENCE AND WORKSHOP CONTRIBUTIONS "A. and A. Valenti" Centre for Health Economics (CESAV)
May Congress, Societal impact of pain, Bruxelles. Congress, Le vaccinazioni alla luce delle pi recent innovazioni, Ranica. July Congress, Adozione del PDT HIV e determinanti costo-efficaci nella terapia antiretrovirale, Monza. September Congress, Risk sharing: il prezzo dellinnovazione, Milan. October Course, 5 Corso di aggiornamento: novit e criticit nellattivit regolatoria dei farmaci, Rome. Congress, Bioequivalenza e pregiudizio-Unesperienza di informazione condivisa sul farmaco equivalente, Comano Terme (TN). November Congress, ISPOR 13 Annual European Congress, Prague. Congress, Farmaci a brevetto scaduto: tecnologia farmaceutica, appropriatezza e strumenti di governo, Legnago (VR).

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Congress, Economia, Politica del Farmaco e HTA, Udine.

Laboratory of Clinical Epidemiology

January Meeting, MargheritaTre; Palermo. Congress, MargheritaDue; Lugano. February Congress, National meeting PROSAFE; Lubiana. Congress, Formazione del medico; Bolzano. March Congress, Progetto StART Piemonte; Torino. Congress, National meeting PROSAFE; Budapest. Meeting, Presentazione Margheirta Tre; Carrara. April Workshop: Startup Crimyne 2; Ranica (BG). Meeting: Presentazione Margheirta Tre; Tortona. Meeting: Presentazione Margheirta Tre; Massa. May Congress, Progetto StART Lombardia; Milano. Workshop, Meeting Compact; Milano. Workshop, Meeting MargheritaTre; Milano. Congress, Progetto StART Toscana; Firenze. Congress, Congresso Nazionale SIARED ; Cagliari. June Congress, Convegno CPFA; Vicenza. August Meeting: Presentazione Margheirta Tre; Massa. September Seminar, Scuola specializzazione; Milano. October Congress, Terzo Simposio Nazionale sulle decisioni di fine vita: Le testimonianze; Sottomarina di Chioggia Master Nefrologia; Bergamo. Congress, Appropriatezza dei ricoveri in Terapia Intensiva, Tuscany; Lucca. Congress, Meeting GiViTI 2010; Pesaro. November Congress, National meeting PROSAFE; Varsavia. Congress, Tecnical meeting PROSAFE; Lubiana.

Laboratory for Mother and Child Health

January Le prospettive della ricerca in campo pediatrico. Congress Dove va la pediatria?. Associazione Pediatria di Comunit (APeC), Cesena (FC). March Come superare il problema delloff-label. Course: La prescrizione off-label in et pediatrica: evidenze scientifiche, aspetti medico-legali e comunicazione efficace. OCM Comunicazioni; Verona.

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Il profilo epidemiologico del bambino con spettro autistico. National Congress: Attualit sullautismo: dai modelli interpretativi allefficacia del trattamenti. ULSS 10 Veneto Orientale, Comune di San Don di Piave; San Don di Piave (VE). April Farmaci in gravidanza e durante lallattamento. Congress Allattamento Materno e Ospedale Amici dei Bambini: Dieci Passi Insieme per la qualit del percorso nascita. UNICEF, SIGO (Societ Italiana di Ginecologia e Ostetricia); Palermo. Lappropriatezza (efficacia, sicurezza, costo) dellutilizzo dei farmaci in et pediatrica. Farmaci off-label in ambito pediatrico. Course Il medico di continuit Assistenziale e lappropriato uso dei farmaci. ASL provincia di Bergamo Dipartimento Cure Primarie e Continuit Assistenziale; Bergamo. Il ruolo del mediatore culturale in neuropsichiatria infantile. National Course of Mediazione Transculturale. INMP (Istituto Nazionale Salute, Migrazione e Povert); Rome. Sfide nel trattamento della malaria nei bambini. II International Congress Lotta alla malaria in Africa e infuso di Artemisia annua L.. ICEI (Istituto Cooperazione Economica Internazionale); Rome. Educazione e istituzione: fattori essenziali per lo sviluppo e il benessere dei bambini. Congress Conclusione della campagna di prevenzione incidenti in casa, a scuola, negli ambienti di svago, sulla strada, sulluso improprio di psicofarmaci, videogiochi, Personal Computer, alimentazione. MOICA (Movimento Italiano Casalinghe); Treviso. The TINN survey in european neonatal intensive care units (NICUs). Ciprofloxacin safety in infants newborns. General Meeting TINN Period 1. TINN (Treat Infections In Neonates); Paris (F). Vietato ammalarsi: povert e discriminazione nellaccesso delle cure sanitarie. XVI Course multidisciplinare di educazione ai diritti. UNICEF; Milan. May Costo/efficacia: parametro efficace? Come si misura la sostenibilit dellintervento nei paesi poveri? Round Table. Convegno Gratuit e qualit delle cure: il Progetto EMERGENCY. Commissione Straordinaria per i Diritti Umani del Senato, EMERGENCY; Rome. Crisi acuta in adolescenza: lo scenario italiano ed esperienze internazionali. Congress Modelli innovativi di intervento nella crisi acuta in adolescenza. Fondazione IRCCS Ca Granda (Ospedale Maggioer Policlinico, ASL Milano; Milan. I farmaci: quali evidenze. Workshop Opzioni chirurgiche per il bambino e ladolescente obeso. ICP (Istituti Clinici di Perfezionamento), Ospedale dei Bambini Buzzi; Milan. Reazioni avverse in gravidanza e pediatria. Master and Course Perfezionamento in Farmacovigilanza Gestione del farmaco sul territorio e farmacosorveglianza. SEFAP (Servizio di Epidemiologia e Farmacologia Preventiva) Universit degli Studi di Milano Dipartimento di Scienze Farmacologiche; Milan. Farmaci essenziali, politiche ed uso nei paesi poveri. VIII Course In memoria di Antonio Terrizzi. Medici in Africa; Genoa. Lettura: implicazioni cliniche e generalizzazione dei risultati degli RCTs. 7 National Congress SIARED; Villasimius (CA). Farmaci, test diagnostici e screening: i rischi nelle decisioni mediche. Incontro Formazione Giornalisti. Ordine dei Giornalisti di Milano; Milan. June Determinants of the drug utilization profile in the pediatric population in Italys Lombardy Region. Course: Indagine sul consenso prescrittivo nella pediatria di famiglia: considerazioni e proposte. Regione Lombardia ASL Monza e Brianza; Monza (MB). Luso razionale degli psicofarmaci in et evolutiva. GCP e buona pratica clinica. Il monitoraggio delle reazioni avverse da farmaci. Seminari del luned. Fondazione Istituto Neurologico Nazionale C. Mondino, Dipartimento di Scienze Neurologiche Universit di Pavia; Pavia, (PV).

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La ricerca clinica nella pediatria di famiglia. Course La sperimentazione clinica in pediatria ASL Roma E, Ospedale S.Spirito in Sassia; Rome. July Paediatric Clinical Pharmacology in Italy. Congress PAEDIATRIC Clinical Pharmacology and Child Health. The Medical School Royal Derby Hospital, NHS Fondation Trust; Derby (UK). September Round Table. Regional Congress Prescrizioni di antibiotici in pediatria: si pu migliorare? Valutazione di unesperienza veneta di percorsi diagnostico terapeutici. Regione del Veneto, Unit di Informazione sul Farmaco, Azienda ULSS 20 Verona; San Bonifacio (VR). La prescrizione dei farmaci in et pediatrica. Il profilo prescrittivo in et pediatrica in Regione Lombardia. Course ASL Milano Due Una lista di farmaci essenziali basata sulle attitudini prescrittive dei pediatri. Azienda Sanitaria Locale della Provincia di Milano 2; Milan. Il network e la ricerca. Congress Network Neonatale Italiano: cure, esiti e ricerca per i neonati pretermine. SIN (Societ Italiana Neonatologia), Network Neonatale Italiano; Roma. La realt dei centri di riferimento a due anni dallavvio del Registro Nazionale. Registro regionale dellADHD. Giornate di studio e approfondimento Lavorare insieme nellADHD. Esperienze dei centri di riferimento della Lombardia. Regione Lombardia, ASL di Brescia, UONPIA Ospedale dei Bambini e Ospedale Civile di Brescia; Iseo (BS). October La valutazione degli esiti nelle situazioni complesse. Course Bisogni comunicativi complessi e partecipazione nei contesti di vita. ASL Milano, Fondazione RCCS Ca Granda Ospedale Maggiore Policlinico, Azienda Ospedaliera Treviglio; Milan (MI). November I farmaci off-label in et pediatrica. Congress Progetto MEAP. La terapia farmacologica in pediatria: dallempirismo alla razionalizzazione. Regione Lombardia Sanit; Milan. December Le caratteristiche della farmacocinetica in et pediatrica. Sperimentazione clinica in pediatria. Course Utilizzo appropriato dei farmaci nelle patologie oncologiche dellet pediatrica e adolescenziale. CRO Aviano (Centro di Riferimento Oncologico) Istituto Nazionale Tumori; Aviano (PN).

GRANTS AND CONTRACTS "A. and A. Valenti" Centre for Health Economics (CESAV) Abbott AIFA Grunenthal-Prodotti Formenti Merck Serono Sanofi Aventis Sanofi Pasteur MSD Schering Plough Vivisol

Laboratory of Clinical Epidemiology ARESS Piemonte ASL TO-2 Piemonte

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Azienda ULSS 16, Padova Italia Bellco SpA Draeger Italia Regione Lombardia Regione Toscana Commissione Europea DG SANCO Hill-Rom Brahms Astellas

Laboratory for Mother and Child Health A.O. Spedali Civili of Brescia
Boehringer Ingelheim European Union

IRCCS Burlo Garofolo, Trieste IRCCS C Granda Hospital Maggiore Policlinico, Milan
Italian Drug Agency, (AIFA) Province of Milan Regional Health Ministry - Lombardy Region Regional Health Ministry - Valle dAosta Region

SCIENTIFIC PUBLICATIONS (2010)

"A. and A. Valenti" Centre for Health Economics (CESAV)

Garattini L, Gritti S, De Compadri P, Casadei G. Continuing Medical Education in six European countries: A comparative analysis. Health Policy 2010; 94(3):246-254. Garattini L, Casadei G. Letter to the Editor. (2nd) Vaccine 2010; 28:880. Koleva D, Cortelazzo S, Toldo C, Garattini L. Health Care Costs of Multiple Myeloma: an Italian Study. European Journal of Cancer Care (e-pub); 2010. Garattini L, Koleva D, Casadei G. Modeling in pharmacoeconomic studies: Funding sources and outcomes. International Journal of Technology Assessment in Health Care 2010;26(3):330-333. Virgili G, Koleva D, Garattini L, Banzi R, Gensini GF. Utilities and QALYs in health economic evaluations: glossary and introduction. Internal and Emergency Medicine 2010;5:349-352.

Laboratory of Clinical Epidemiology

Csomos A, Varga S, Bertolini G, Hibbert C, Sandor J, Capuzzo M, Guidet BR. Intensive care reimbursement practices: results from the ICUFUND survey. Intensive Care Med 2010 Oct;36(10):1759-64. Guido Bertolini, Simona Boffelli, Paolo Malacarne, Mario Peta, Mariano Marchesi, Camillo Barbisan, Stefano Tomelleri, Simonetta Spada, Roberto Satolli, Bruno Gridelli, Ivo Lizzola, Davide Mazzon. End-of-Life Decision-Making and Quality of ICU Performance: An Observational Study in 84 Italian Units. Intensive Care Med 2010 Sep;36(9):1495-504. Cogo PE, Poole D, Codazzi D, Boniotti C, Capretta A, Langer M, Luciani D, Rossi C, Bertolini G.Outcome of children admitted to adult intensive care units in Italy between 2003 and 2007. Intensive Care Med 2010Aug;36(8):1403-9. Corona A, Bertolini G, Lipman J, Wilson P, Singer M. Use and impact of antibiotics on outcome in bacteraemic critical

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illness: the BActeraemia study in intensive care (BASIC). J Antimicrob Chemother 2010Jun;65(6):1276-85. Bruno Giometto; Wolfgang Grisold; Roberta Vitaliani; Francesc Graus; Jrme Honnorat; Guido Bertolini; for the PNS Euronetwork. Paraneoplastic Neurologic Syndrome in the PNS Euronetwork Database: A European Study From 20 Centers. Arch Neurol 2010;67(3):330-335. P. Malacarne, D. Boccalatte, A. Acquarolo, F. Agostini, A. Anghileri, M.Giardino, D. Giudici, M. Langer, S. Livigni, E. Nascimben, C. Rossi, G. Bertolini. Epidemiology of Nosocomial Infection in 125 Italian Intensive Care Units. Minerva Anestesiologica 2010 Gennaio;76(1):13-23. J.C. Marchall, K. Reinhart, D. Angus, A. Argent, G. Bernard, G. Bertolini, S. Bhagwanjee, J.P. Cobb, D.J. Cook, D. Fedson, S. Finfer, R. Fowler, C. Gomersall, E. Jimenez, N. Kissoon, D. McAuley, S. Opal, J.L. Vincent, S. Webb. InFACT: a global vritical care clinical research reponse to severe pendemic H1N1. Lancet 2010 Jan 2;375(9708):11-3.

Laboratory for Mother and Child Health

Bianchi M, Clavenna A, Bonati M. Inter-country variations in anti-asthmatic drug prescriptions for children. Systematic review of studies published during the 20002009 period. Eur J Clin Pharmacol 2010;66: 929-36. Bonati M. Commentary. Evidence-Based Mental Health 2010;13:43. Bonati M, Pandolfini C. Safety of ciprofloxacin in neonates with sepsis. Adverse Drug Reactions Bulletin 2010:265:1019-1022. Clavenna A, Pandolfini C, Bianchi M, Bonati M. Do children really need more research on respiratory drugs? Acta Paeditr 2010;99:1445-46. Clavenna A, Sequi M, Bonati M. Drug prescribing by Italian family paediatricians: an exception? Acta Paediatr 2010;99:754-757. Clavenna A, Sequi M, Bonati M. Differences in the drug prescriptions to children by Italian paediatricians and general practitioners. Eur J Clin Pharmacol 2010;66:519-524. Fortinguerra F, Maschi S, Clavenna A, Bonati M. Pain management in the paediatric population. The regulatory situation in Europe. Arch Dis Child 2010;95:749-753. Panei P, Arcieri R, Bonati M, Bugarini M, Didoni A, Germinario E. Safety of psychotropic drug prescribed for attention-deficit/hyperactivity disorder in Italy. Adverse Drug Reaction Bulletin 2010;260:999-1002.

LAY PRESS SELECTION (2010) "A. and A. Valenti" Centre for Health Economics (CESAV)
Clavenna A, Gangemi M, Casadei G, Garattini L, Bonati M. Efficacia del beclometasone versus placebo nella profilassi del wheezing virale in et prescolare. Ricerca & Pratica 2010; 26: 19-25. Padula A, Casadei G, Motterlini N, Garattini L . Vaccinazione antinfluenzale in ambiente di lavoro: una valutazione economica. Quaderni di Farmaco Economia 2010; 11:9-15. Garattini L, Koleva D, Casadei G . Lapplicazione dei modelli di Markov in farmacoeconomia. Quaderni di Farmaco Economia 2010; 12:7-12. Gallus S, Apolone G, Padula A, Casadei G, Motterlini N, Garattini L . Quaderni di Farmacoeconomia risponde ai lettori. Quaderni di farmaco economia 2010; 12:24-31. Garattini L, Casadei G . Cure H: far bene la gara fa bene alla spesa. Sanit del Sole 24Ore 26 Gennaio 2010 pag. 11. Garattini L . Finanziaria 2010. Dialogo sui Farmaci 2010; 1:25-26. Garattini L, Casadei G . Cure H: far bene la gara fa bene alla spesa. Ricerca & Pratica 2010; 26: 75-80.

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Garattini L . Riunificare i tetti di spesa per responsabilizzare chi fa i prezzi. Sanit del Sole 24Ore 1-7 Giugno 2010 pag.6-7. Garattini L, Casadei G . No alla riserva indiana per gli equivalenti. Sanit del Sole 24Ore 15-21 Giugno 2010 pag. 7. Garattini L, Casadei G . Ma attenti alla spesa ospedaliera. LEspresso 3 Settembre 2010 pagg.137-138. Casadei G, Garattini L . Contratti desito, puntualizzare le verifiche. Sanit del Sole 24Ore 7-13 Settembre 2010 pag. 19. Clavenna A, Gangemi M, Casadei G, Garattini L, Bonati M . Efficacia del beclometasone versus placebo nella profilassi del wheezing virale in et prescolare. Ricerca & Pratica 2010; 151:21. Casadei G, Garattini L . Spesa farmaceutica e manovra finanziaria: riflessioni. Ricerca & Pratica 2010; 26:135-140. Garattini L . Mercato, Salute e Povert. Club3-Vivere in armonia Dicembre 2010 pag. 129. De Compadri P, Koleva D . Costi nosocomiali del condiloma acuminato, con particolare riferimento alla Regione Lombardia. Farmacoeconomia News 2010; 1:23-30. De Compadri P . Corticosteroidi Inalati somministrati a bambini in et pre-scolare con manifestazioni asmatiche durante infezione delle vie respiratorie superiori: valutazione economica budesonide. Quaderni di Farmaco Economia 2010; 13:7-14. Casadei G . Stato, Regioni e Concorrenza. Quaderni di Farmaco Economia 2010; 11:5-7. Casadei G . Spesa, federalismo e varie. Quaderni di Farmaco Economia 2010; 12:4-6. Gritti S . Valutazione economica della vaccinazione antinfluenzale in ambiente di lavoro. Ricerca & Pratica 2010; 26(3):130-131.

Laboratory of Clinical Epidemiology

Guido Bertolini. Per un medico nuovo: saper collaborare, saper apprendere, saper sapere. Recenti Progressi in Medicina, Luglio-Agosto 2010:303-304. Guido Bertolini. La fibrosi cistica e levoluzione della specie. R&P 2010;26:161-173. Abramo Anghileri, Michele Giardino, Guido Bertolini. Margherita Due: lassistenza informatica. R&P 2010; 26: 3839. Bertolini G. Imparare dallerrore. Appunti per una riflessione in medicina. Medico e bambino 2010; 29:566-568.

Laboratory for Mother and Child Health

Bianchi M. Variabilit dellinosina monofosfato deidrogenasi nei pazienti con trapianto di rene in terapia cronica con micofenolato mofetile. R&P 2010;152:67. Bonaccorsi A. Farmaci in prima pagina. R&P 2010;152:78-80. Bonaccorsi A. Displasia broncopolmonare e danno cerebrale nei neonati prematuri: una relazione complessa. R&P 2010;152:66. Bonaccorsi A. Farmaci in prima pagina (traduzione). R&P 2010;152:75-80. Bonaccorsi A. Innovazione e brevetti nella synthetic biology. R&P 2010;26:207-209. Bonati M. Uso e abuso degli pasicofarmaci nella ricerca e nellassistenza ai minori e agli adulti. In: 1978-2008 Trentanni di sanit tra bioetica e prassi quotidiana. Regione Toscana, Firenze 100-103; 2010. Bonati M. Il 90 per cento dei lettori legge solo labstract. R&P 2010;26:43-44.

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Bonati M, Severino F, Bagnati R, Carr A, Roberto F. Sfide nel trattamento della malaria nei bambini. R&P 2010;152:163-164. Campi R, Bonati M. I bambini ci guardano e ... aspettano ancora. R&P 2010;152:47-49. Clavenna A. Utilit clinical dei criteri di Roma per la gestione dei disturbi funzionali gastrointestinali nelle cure primarie pediatriche. R&P 2010;152:66. Clavenna A, Bonati M. La ricerca indipendente; faranno apposta? Quaderni acp 2010;17:191-192. Clavenna A, Bonati M, Sequi M, Nobili A, Franchi C, Tettamanti M, Bortolotti A, Merlino L, Fortino I. La prescrizione di farmaci per i bambini e gli anziani in Regione Lombardia. R&P 2010;151:9-18. Clavenna A, Bonati M. Troppe inadempienze ritardano la sperimentazione clinica nel territorio. Quaderni acp 2010;17:49. Clavenna A, Fortinguerra F. Psicofarmaci in allattamento: per un terzo dei medicinali non ci sono evidenze. Quaderni ACP 2010;17:35. Clavenna A, Fortinguerra F. Lista dei farmaci cardiovascolari per i bambini: un passo per superare gli off label. Quaderni acp 2010;17:82. Clavenna A, Fortinguerra F. Beta-agonisti a lunga durata dazione: mai pi da soli nella terapia dellasma. Quaderni acp 2010;17:131. Clavenna A, Fortinguerra F. Mucolitici e propiltiouracile: aggiornamenti sulla sicurezza di impiego nei bambini. Quaderni acp 2010;17:181. Clavenna A, Fortinguerra F. Quando il farmaco somministrato per far male: un problema in aumento negli Stati Uniti. Quaderni acp P 2010;17:227. Clavenna A, Gangemi M, Casadei G, Garattini L, Bonati M. Efficacia del beclometasone versus placebo nella profilassi del wheezing virale in et prescolare. R&P 2010;151:21. Clavenna A, Sequi M, Bortolotti A, Fortino I, Merlino L, Bonati M. Farmaci essenziali e attitudini prescrittive dei pediatri. M&B 2010;29:565-569. De Compadri P, Clavenna A. Corticosteroidi Inalati somministrati per manifestazioni asmatiche in et prescolare: valutazione economica budesonide. Quaderni di Farmacoeconomia 2010;13:7-14. De Fiore L, Bonati M. La fiera dei congressi. R&P 2010;151:3-8. Didoni A. A colloquio. R&P 2010;151:32. Didoni A. Una madre e una figlia alla ricerca delle proprie identit. R&P 2010;152: 74. Fortinguerra F. Pittogrammi gratuiti per farmacie e ospedali. R&P 2010;151:28-29. Fortinguerra F. Prescribe nel regno di Sua Maest. R&P 2010;151:201. Fortinguerra F. Nuovi farmaci sottoposti a studi clinici pediatrici in UE: a che punto siamo? Giornale italiano di Farmaci Clinica 2010;24:55-56. Fortinguerra F. LEMA presenta una sezione pediatrica rinnovata allinterno del suo sito web. Giornale italiano di Farmaci Clinica 2010;24:116-117. Fortinguerra F. Uso di formulazioni orali non appropriate negli studi clinici pediatrici Giornale italiano di Farmaci Clinica 2010;24: Fusco F, Sambugaro D, Clavenna A. Linvermictina per os nella pediculosi di difficile trattamento. M&B 2010; http://www.medicoebambino.com/?id=AP1005_10.html.

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Marchetti F, Ronfani L, Maestro A, Rovere F, Zanon D, Arrighini A, Bertolani P, Biban P, Da Dalt L, Di pietro P, Renna S, Guala A, Mannelli F, Pazzaglia A, Messi G, Perri F, Reale A, Tondelli MT, Urbino AF, Valletta E, Vitale A, Zangardi T, Clavenna A, Bonati M. Trial controllato randomizzato multicentrico di valutazione comparativa dellondansetron verso domperidone per il trattamento sintomatico del vomito acuto da gastroenterite nel bambino: protocollo di studio. M&B 2010; http://www.medicoebambino.com/?id=PST1009_10.html. Severino F, Bonati M. Migranti e salute: tra diritto (alle cure) e reato (di clandestinit). R&P 2010;152:50-61. Working Group Pediatrico dellAIFA, Bonati M. La prima lista dei farmaci cardiovascolari autorizzati per un uso pediatrico. M&B 2010;29:172-176. Working Group Pediatrico dellaAIFA, Bonati M. Farmaci cardiovascolari: la prima lista di farmaci autorizzati per un uso pediatrico. Clinica e management 2010;2:9-12.

OTHER PUBLICATIONS (2010) Laboratory of Clinical Epidemiology

Rossi C, Di Gangi S, Bertolini G. Progetto Margherita - Promuovere la ricerca e la valutazione in Terapia Intensiva RAPPORTO 2009. Bergamo: Edizioni Sestante, 2010. [report]

Laboratory for Mother and Child Health

Bonati M, Garattini S. Malattie Tropicali dimenticate. In: rapporto di Medici Senza Frontiere. Le crisi umanitarie dimenticate dai media 2009. Marsilio Editori spa, Venezia, 2010;94-100. [chapter book] Bonati M. Lo specialista risponde. In: Il grande libro italiano della gravidanza. Rizzoli, Milano, 2010;252-253. [chapter book] Clavenna A, Braguglia A. Peculiarit della farmacocinetica nel neonato. In: Farmacoterapia Neonatale. Guida pratica con supporto interattivo. I edizione 2009. Editore Biomedia, Milano 2009;4-6. [chapter book] Bonati M, Campi R. Quale futuro per chi nasce e cresce oggi nel mezzogiorno? Saluteinternazionale.info 17 febbraio 2010; http://saluteinternazionale.info/2010/02/quale-futuro-per-chi-nasce-e-cresce-oggi-nel-mezzogiorno/. [Information in the Media] Fattore E, Davoli E, Bonati M. Il fantasma della discarica. Il Sole 24 Ore Sanit 30 novembre - 6 dicembre 2010; pag. 14-15 [Information in the Media] Per un uso razionale dei farmaci. Controparte: Assessorato alla Sanit, Regione Lombardia. [report] Registro Nazionale ADHD. Controparte : AIFA. [report] Formazione farmaco epidemiologia pediatrica. Controparte: Boehringer Ingelheim S.p.A. [report] During 2009 the laboratorys activities were covered by the national media 40 times

RESEARCH ACTIVITIES "A. and A. Valenti" Centre for Health Economics (CESAV) Educational activity
Educational activities are developed only if related to research studies, in order to offer original contributions which naturally reinforce the research aims.

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Economic Evaluation of Health Care Programs

The aim of this research area is to assess the costs of pathologies and the cost-effectiveness ratios of the diagnostic/therapeutic existing alternatives. In general, analyses can be classified into two groups: partial economic evaluations (e.g. cost of illness analysis) and full economic evaluations (e.g. cost-effectiveness analyses).

Comparative Health Policy Analysis

The aim of this research area is to study the organization of health care systems, in order to draw lessons from international comparisons. This is particularly important in a "market" like health care where economic competition lacks by definition and therefore public regulation plays a crucial role.

Laboratory of Clinical Epidemiology Quality of care in the Intensive Care Units

The main purpose of these research projects is the assessment and improvement of the quality of care in Italian Intensive Care Units (ICUs). It is a multi-annual project promoted on behalf of GiViTI, a collaborative network composed by more than half of the Italian ICUs and coordinated by the Laboratory. The main focus is the Project Margherita. Its aim is the continuous evaluation of the quality of care and it is based on a free software developed by the Laboratory and distributed to all the ICUs adhering to the GiViTI group. The software has been realized on a modular structure, which enables to easily integrate the basic data collection (the core of Margherita) with the data collection of specific research projects (the petals of Margherita). Since January 2011, Margherita became an international project. Thanks to funding from the European Union have in fact been able to develop new software and to distribute the project to six countries: Slovenia, Hungary, Poland, Cyprus, Brazil and Israel.

Appropriateness of the Intensive Care Units

ICU is a high technology environment, that requires a high number of high-level personnel. Hence, the cost of these units is extremely important and a special attention not to waste resources is mandatory. In this field, the Laboratory launched a study to assess the level of appropriateness of the use of ICU beds, in four Italian regions: Lombardia, Piemonte, Toscana e Campania. Such an evaluation is based on the understanding that the level of care provided by an ICU should correspond to the level of care it can theoretically provide, given the available resources. In this framework, patients are classified as requiring high-, low-, or ordinary-care, and beds are independently classified are high- or low-level. The appropriateness evaluation protocol adopted verify the concordance between these two separate classifications.

Influenza A/H1N1 in ICU

On 11 June 2009 the World Health Organization confirmed the presence of a new pandemic influenza A, caused by a H1N1 virus. The first experiences showed that this influenza was characterized by a higher incidence of severe viral pneumonia in children, middle-aged patients, and pregnant women and, apparently, by a higher mortality than expected in a normal influenza season. This caused a generalized alarm all over the world. On October 2009 the GiViTI group, coordinated by the Laboratory of Clinical Epidemiology, set up the H1N1 registry, with the aim to assess the characteristics and outcome of affected patients, the impact of the pandemic influenza on the activity of Italian ICUs from September 2009 to April 2010, and the correspondence between the generalized alarm on this phenomenon and what really took place in Italian ICUs.

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Studies on Multiple Organ Failure pathological processes

The Laboratory of Clinical Epidemiology has lead several investigations to clarify which pathophysiological mechanisms induce multiple organ failure, a condition still burdened with high mortality. Among these, the investigation on the neuromuscular impairment in critical patients (the observational study 'Crimyne'), the study on the impact of enteral feeding, and the new treatments proposed for severe sepsis (Xigris and the removal of inflammation mediators through specific filter applied to circuit of plasma-filtration). The value of a strict glycemic control of critical patients has been recently emphasized, given its connection to a drug like insulin that, in spite of its large availability and low cost, induces a relevant reduction of mortality in ICU. The Unit of Clinical Knowledge Engineering has developed a model based on a differential equations system whose aim is to support the physician in dosing both insulin and glucose infusions, in order to extend the possibility of a strict control even to patients with a high risk of hypoglycaemia. This model represents a precious opportunity to investigate the pathophysiological mechanisms behind the benefits of insulin already demonstrated at an empirical level, allowing the explanation of the dynamic behaviour of glycemic fluctuations on the basis of the patient's metabolic profile.

The reconstruction of clinical reasoning in the medical practice and education

This area represents the main concern of the Unit of Clinical Knowledge Engineering, whose objective is the valorization of clinical reasoning in solving complex clinical problems. The diagnosis of pulmonary embolism still represents a relevant clinical challenge, due to the complexity of the patient's clinical presentation and the variability of diagnostic resources among Centres. In this regards, we are conducting an Italian multicenter study, involving mainly Emergency Units, with the aim of prospectively validating the diagnostic software BayPAD (Bayes Pulmonary embolism Assisted Diagnosis). Such a tool, relying on a probabilistic model covering 72 clinical variables and doing without the need to input all the contemplated observations, would overcome the main reasons which prevented ordinary clinical guidelines to be largely accepted. Moreover, the results of the retrospective validation of the system have been obtained. The Unit started a project for the realization of a software assisting the physician in tracing back the basis of his clinical decisions before the description provided by clinical reports, among those that are typical of particular medical specialty. The software has the double target to create specific applications based on probabilistic models representing complex clinical decision problems, and to involve physicians in their construction. The last target is achievable given the strong analogy between the causal structure of the exploited models (bayesian networks) and the pathophysiological structure of medical knowledge. By this, it will be given the chance to adopt this system within medical training projects, with a special attention to e-learning programs.

Clinical Epidemiology of rare diseases and orphan medicine

Our purpose is to find out and describe clinical and research problems related either to rare diseases or to neglected aspects of well-known diseases. We also focus on the needs of the patients with rare diseases. A specific project is connected with this research activity: PNS Euronetwork. It is a European project on paraneoplastic neurological syndromes that is financed by the Fifth and Sixth Framework Program of the European Community. Its purposes are various: to develop a network of reference centers for these pathologies all sharing a common database; to organize a sample bank of biological fluids and cerebrospinal

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liquid to point out the best antibody indicators for the diagnosis and prognosis of these patients; to realize some research projects on the treatment of this syndrome.

An electronic health record to promote research in Intensive Care Medicine

The main aim of this project is to develop an electronic health record (EHR) the allows the assessment of indicators of the process of care in the ICU. A multidisciplinary team of intensivists, ICU nurses, epidemiologists, statisticians, and IT specialists, had the responsibility of planning the HER, that is now already shared by 15 Italian ICUs.

Laboratory for Mother and Child Health Efficacy of Nebulised Beclometasone versus placebo in preventing viral wheezing in pre-school children. (ENBe)
The Laboratory for Mother and Child Health coordinates the Efficacy Of Nebulised Beclometasone Versus Placebo In Preventing Viral Wheezing In Pre-School Children (ENBe) randomised controlled trial. The study is funded by the Italian Medicines Agency (Agenzia Italiana del Farmaco, AIFA) with a grant for independent research on drugs and it is performed in collaboration with the Associazione Culturale Pediatri and the "Angelo and Angela Valenti" Centre for Health Economics (CESAV). The ENBe study involves 36 family paediatricians in 9 Italian local health units (LHU) representative of geographical distribution and setting. The aim of the study is to evaluate the safety and effectiveness of beclometasone in preventing wheezing in viral upper respiratory tract infection (URTI). A total of 576 children 1-5 years old, with at least one episode of viral wheezing in the previous 12 months, will be enrolled. Children with steroid hypersensitivity, chronic respiratory disease (e.g. cystic fibrosis, broncho-pulmonary dysplasia), presence of wheezing at the entry visit or using inhaled and/or oral corticosteroid use in the preceding month will be excluded. Patients will be randomly allocated to receive beclometasone suspension 400 mcg b.i.d or placebo b.i.d. Active drug and placebo will be administered through a nebuliser, in the morning and in the evening. The duration of the treatment will be 10 days, and a 6-month follow up period will be performed to monitor the recurrence of respiratory tract infections and viral wheezing. The percentage of children with wheezing (diagnosed by the paediatrician) during the URTI episode will be the primary outcome measure. The bureaucratic procedures for obtaining the authorization by each LHU ethics committee lasted a total of 19 months and only in October 2010 it was possible to start the patient enrollment. On 9 January 2011, 13 weeks after the beginning of the study, 615 eligible children were visited, 218 of which (35%) were enrolled: 137 were males and 81 females, with an average age of 2.7 years.

Pharmacoepidemiology in the Lombardy Region

The Laboratory for Mother and Child Health is involved in the analysis of the drug prescription profile in children and adolescents in the EPIFARM (Epidemiologia del farmaco) project funded by the Lombardy Region. The main results of the project are summarised below: During 2008, 826,727 children and adolescents < 18 years old (51% of the population) received at least one drug prescription.

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The highest prevalence of drug prescription was observed in the 1-5 year old range (average value 68%), and then decreased to 40% in the 12-17 year range. Each treated child received an average of 3 prescriptions and 4 packages. Antibiotics were the most commonly prescribed therapeutic class (41% of the population), followed by anti-asthmatics (15%) and anti-histamines (5%). Amoxicillin+clavulanic acid was the most prescribed drug (21% of children) , followed by amoxicillin (12%) and inhaled beclometasone (9%). Only 42 out of 746 drugs were prescribed by at least 50% of the paediatricians. Six drugs were prescribed by all the paediatricians (amoxicillin+clavulanic acid, amoxicillin, beclometasone, clarithromycin, salbutamol, and cefaclor) Quantitative and qualitative differences in the prescription profile were found between different local health units (LHU). The prevalence ranged between 41% in Milan and 56% in Valle Camonica. In general, the LHUs with a greater prevalence of drug prescriptions were also characterised by a lower degree of appropriate prescriptions This is particularly evident in the case of antibiotic: the average ratio of children treated with penicillins/children treated with cephalosporins by paediatrician in Milan LHU was 4.7 (median 4.1, interquartile range: 3.0-5.8), while the average ratio penicillin/cephalosporin treated children by paediatrician in Brescia was 1.9 (median 1.7, interquartile range 1.2-2.2). The quantitative and qualitative differences observed in a quite homogeneous context like Lombardy region suggest that educational interventions for physicians and parents with the aim to improve the rational use of drugs should be adapted to the local setting.

Prescription, efficacy, and safety of psychotropic drugs in the Italian paediatric population
Diagnostic and therapeutic approaches in children and adolescents with neuropsychiatric disorders in the Local Health Unit of Verona, Italy. Aims. The safety and effectiveness of psychotropic drug use in the paediatric population are widely debated, in particular because of the lack of data concerning the long term effects. In Italy the prevalence of psychotropic drug prescriptions increased in the 2000-2002 period and decreased afterwards. In such a context, a study with the aim to estimate the prevalence of psychotropic drug prescription in the paediatric population and to describe diagnostic and therapeutic approaches was performed. Methods. The study population was composed of 76,000 youths <18 years of age living in an area covered by the Local Health Unit of Verona, Italy. Children and adolescents receiving psychotropic drug prescriptions were identified. Questionnaires were sent to the prescribers with the aim to collect data concerning diagnostic and therapeutic approaches, and care strategies. Results. Between 1 January 2005 and 31 December 2006, 111 youths (0.8 per 1,000) received at least one psychotropic drug prescription. Nintyone youths received antidepressants and 25 received antipsychotics. Only 29 patients were under the care of child and adolescent outpatient psychiatric services. Information concerning diagnostic and therapeutic approaches, and care strategies, were collected for 52 patients (47%). Anxietydepressive syndrome and attention disorders were the diseases for which psychotropic drugs were most commonly prescribed. In all, 85% youths received also psychological support and 20% were prescribed drugs for 2 or more years. Child psychiatrists made the first diagnosis in 50% of the cases, but in 1/3 of the cases the parents met with 2 or more physicians before the diagnostic procedure was completed. Conclusions. Despite the low prevalence of drug prescriptions, the finding that most children were not cared for by child and adolescent psychiatric services is of concern and calls for a systematic continuous monitoring of psychopharmacological treatments.

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Migration and psychological difficulties in growth and adulthood

The project involves seven Childhood and Adolescence Neuropsychiatry Units (UONPIAs) of Milan, 2 psychiatric services and the family, childhood and school-age service of Milans local health unit, Milans social and educational services, the Cesare Beccaria Penal Institute for Minors, 7 third-sector organizations, the Provincial School Office and the Mario Negri Institute. The project aims to improve the response to the specific problems of the school-aged migrant population, with particular attention to adolescents and minors who arrived in Italy unaccompanied by family members. During 2010 data concerning 727 patients were collected and analyzed, most of whom are males, aged between 0 and 23 years (mode: 7 years). 64, 9% were born in Italy. In 40.2% of the cases the school sent the children to the services, most commonly for academic difficulties, behavioral problems, and language or communication problems. In 53% of cases, migrant patients, who are not Italian citizens, have regular residence permits. Patients generally have a good knowledge of Italian. The mother tongue is Italian only in 10.7% of patients. The patients parents were generally born abroad, migrated without the family of origin (50%) in order to search for a job, and both hold a residence permit in 40% of cases.

Complex communication needs and participation

(CAA: Augmentative Alternative Communication)

Children who do not speak or who can only say a few words, children who have difficulty in understanding anothers words... There are about 8000 people between 0 and 18 years in the Lombardy Region that suffer from communication disorders, some for a few months, others for years, some forever. In Italy there are almost 50,000. Some also have mobility problems, others a genetic syndrome or autistic disorder. For all, growing up without the ability to communicate is very tough, and it is essential to find ways of making it possible to communicate without use of the voice, through measures of support (augmentative communication). Symbol-based systems, as well as images, can be used, in which all the figures are written above the word or the verb they represent. The child is therefore able to recognize the images and the interlocutor to read the words. Digital tools programmed to lend a voice when necessary can be used, as well as information technology and computer technology adapted to personal needs and systems that allow people who cannot use the alphabet or cannot use a pen, to read and write. Anyone who needs to communicate with a child must use a language (written, spoken, visual) appropriate to the childs ability and this should be implemented and guaranteed forever. For this reason, it is not enough to train specific operators, but it is necessary to also involve parents and teachers. To date, only a few patients can access the required support interventions for communication, and this generally occurs very late for the shortage of adequately trained personnel within departments of Child and Adolescent Psychiatry. To meet these unaddressed needs, Milans Local Health Unit (ASL) has launched a specific project, financed by the Lombardy Region for the years 2010-2012, involving 15 Child and Adolescent Psychiatry Units of the region. In this context, a conference was organized on October 21st by the the IRCCS Ca Granda Ospedale Maggiore Policlinico di Milano Foundations Centro Sovrazonale di Comunicazione Aumentativa (Supranational Centre for Augmentative Communication), in collaboration with Milans Local Health Unit (ASL), Treviglios UONPIA (Infant and Adolescent Neuropsychiatry Operational Unit), and the Laboratory for Mother and Child Health of the Mario Negri Institute for Pharmacological Research. The conference addressed the complex communication needs in children, described what represents an augmentative communication intervention, and how it can be applied to certain situations in order to become a possible resource for all children. The project was presented

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in detail and a series of experiences already in place that led to the structuring of the project were described.

Fp7 Projects 1) TINN Treat Infections in NeoNates

The TINN project, Treat Infections in Neonates, is part of the European Unions Seventh Framework Project and is aimed at gathering the experience in the neonatal research field of numerous centres across Europe in order to produce detailed evidence on the safety and efficacy of ciprofloxacin and fluconazole use in neonatal sepsis. The final goal of the project is to obtain a Pediatric Use Marketing Authorization (PUMA) for the two drugs. A survey on the use of ciprofloxacin and fluconazole by neonatal intensive care units (NICU) in Europe was conducted in the first phase of the project, from December 2009 to June 2010. A total of 200 NICUs participated, representing 32 countries. The countries represented by the greatest number of NICUs (disregarding country size), were Italy, the United Kingdom, and France. The survey results highlight a heterogeneous picture of the therapeutic schemes and indications for use of the two drugs, both between and within countries. Significant doubts on the part of clinicians concerning safety and efficacy issues were also revealed, highlighting a need for additional evaluation and information on the optimal use of the drugs. The survey, however, also found a notable interest on the part of NICUs across Europe in participating in studies on the two drugs in order to obtain the necessary, lacking information.

2) COHEMI
Coordinating resources to assess and improve health status of migrants from Latin AmericaCOHEMI Project- HEALTH.2010.3.4-5 European health systems are committed to meeting the challenge of understanding the needs of migrant populations and adapting their services to meet these needs. The difficulties inextricably linked to this challenge are caused by the complexity of migration patterns and the differences between migrant population across EU countries. At present, the limited available data show that attempts to incorporate migrants health needs, in particular those of migrants from non-EU countries, into EU health systems have remained scattered and uncoordinated. In January 2011 started the project COHEMI-Coordination resources to Assess and Improve health status of migrants from Latin America, a three-year project, funded under the 7th Framework Programme for Research and Technological Development (Programme Cooperation- Health), which sees the Mario Negri Institute coordinator of a major consortium of 10 partners located between Europe and Latin America. COHEMIs general objective is to coordinate referral centres dealing with specific Latin American (LA) diseases in order to: provide an analysis of health systems and legislation on migrants and assess the determinants of health by evaluating 3 types of diseases typical of LA countries in order to produce new procedures and suggestions shared at migration policies level, that take into account the specific cultural needs of migrants.

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NEONATOLOGY Part of the Laboratorys effort is currently dedicated specifically to the neonatological area. In addition to the TINN project, Treat Infections in Neonates (see dedicated section), there are three projects underway, two of which related to developing countries: A literature review on the use of drugs in neonatal intensive care units in low to middle income countries. An assessment of mortality and duration of stay of neonates admitted to the largest neonatal intensive care unit in Nicaragua, after modification of the strategy used to employ mechanical ventilation. A literature review on the use of moderate cerebral hypothermia for prevention of brain injury, with particular attention to neonatal hypoxic-ischaemic encephalopathy. Io e lAsma Project Under an agreement with the Hospital of Brescia, the data collected during the "Io e lasma" project, performed by the Bronchopneumology Unit of the Hospital, were evaluated. The aim of the project is to create multidisciplinary network of different figures (physicians, nurses, parents, teachers, educators) who share common educational and therapeutic approaches for the management of asthma. During the September 2007-September 2010 period a total of 515 children (64% male) have made at least three visits to the Bronchopneumology Unit. 40% of the children had adequate control of asthma at the baseline visit, a figure that increased to 76% at the time of the third visit (after 12-16 weeks). The degree of asthma control improved in 47% of children and was adequately maintained in 34% of cases. The rate of hospitalization and of school absences due to asthma in children followed at the clinic were reduced (from 7.6 to 2.7%, and from 13.4 to 6.1%, respectively). 30% of children did not need any controller medication, while 62% of children receiving drug therapy were treated with an inhaled steroid (fluticasone) at low doses. After attending the unit all children used spacer device appropriate for the age, and in 2/3 of the cases the symptoms were recorded on a daily diary. This pilot study documented that the multidisciplinary approach used within the "Io e lasma" project was effective in improving and maintaining an adequate control of asthma symptoms in more than 80% of children.

National ADHD Register

The National Register for ADHD in children has been active since June 2007. The register monitors the diagnostic, therapeutic, and health care approaches and evaluates the adverse effects of two drugs; methylphenidate and atomoxetine. These two drugs are marketed in Italy for the treatment of ADHD in children and adolescents. The register was initially supposed to be active for two years (2008-9), but its duration was extended to three (to the end of June 2010). The register is a tool with great potential for responding, in an appropriate manner and at the national level, to specific, unmet health needs in youth with ADHD (and their families). The Laboratory for Mother and Child health, as part of the scientific committee, participated in writing the protocol that defines the National ADHD Registers structure and activities and in monitoring it. The Laboratory is currently involved in monitoring the data present in the register and in creating the data reports for the health operators working in the services involved. The Lab is especially involved in supporting the creation of a functional network of referral centers. The Lombardy Region, with 300 patients, is the first region in number of patients monitored by the national register. There are 229 patients with a first methylphenidate and atomoxetine prescription (76%), reported by 15 of 22 referral centres (68%) indicated by the Lombardy Region: 57 were present in 2007, 82 in 2008, 42 in 2009, and 39 in 2010. In all, 88% of

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patients are male, aged between 5 and 17 years (median 11 years, mode 12). For 129 patients (56,3%) at least one familiarity was found, in particular: 12 with ADHD, 24 with ADHD and learning disabilities, and 17 with learning disabilities. In agreement with the protocols indications, 66 patients were evaluated with a multidimensional approach (28,8% of the total, with K-SADS, CPRS, CTRS e WISC). The most common diagnosis was complex ADHD (79,5%), compared to the disorder characterized prevalently by attention deficit (14,4%), or hyperactivity (4,4%). In 54 cases (23,6% of the total) no comorbidities were found, while in 100 cases one was found and in 1 case five were found. The most common comorbidities were learning disorders (49,8%) and oppositional defiant disorder (34,5%). Both were diagnosed in 34 patients (14,8% of total). In all, 149 patients began treatment with atomoxetine (65%) and 80 with methylphenidate. Adverse effects were found in 109 patients (47,6% of the total). The laboratory set up ADHDNews, a newsletter aimed at providing interested health operators with a monthly bibliographic update of the recent scientific literature via email. This initiative is part of the Italian Medicines Agencys (AIFA) independent research project implemented in collaboration with the Istituto Superiore di Sanit and called Long term safety of drugs used to treat school-aged children with Attention Deficit Hyperactivity Disorder and epidemiology of the disorder in the Italian population. A total of 24 issues of ADHDNews have been produced so far and 277 people have registered to receive the update (125 psychiatrists and neuropsychiatrists, 48 family physicians, 36 psychologists and speech therapists, 32 pediatricians, 7 pharmacists, 7 school workers, and 22 other). The newsletter identified more than 1010 scientific articles.

The Lombardy Regions ADHD Register

Thanks to the network created after the abovementioned project, it was possible for the referral centres to actively put forward a three-year project Sharing diagnostic-therapeutic approaches for ADHD in Lombardy. The project, whose coordinator is the UONPIA A.O. Spedali Civili of Brescia, began in January 2010, involves 18 referral centres, and is funded by the Lombardy Region. In this project, the laboratorys role is to build a new registry, that will be more practical than the national one, in order to increase the centers compliance in providing information, with the aim to improve data quality. The Registry will also be extended not only to patients with a diagnosis of ADHD in drug treatment with methylphenidate or atomoxetine, but to all those who belong to the center with suspected ADHD. The registry will then permit the:

Quantification of the workload Monitoring of diagnostic paths Outlining of the prevalence of the disorder Monitoring of also non-drug treatment programs Maintenance of pharmacovigilance by extending the monitoring on the use of the drugs, including drugs other than atomoxetine and methylphenidate.

The registry will most likely be accessible from the early months of 2011.

Workgroup on the Convention for child and adolescent rights. The Laboratory for Mother and Child Health is part of the Working Group for the Convention on the Rights of the Child (CRC) in Italy. This year the CRC Group has published the 2nd Supplementary Report in English and sent it to the 'NGO Group for the CRC, which will forward it to the UN Committee that will consider Italy in 2011.

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The English version of the 2nd supplementary report can be downloaded at: http://gruppocrc.net/IMG/pdf/2_Rapporto_CRC_2010_StC.pdf. The 2nd Supplementary Report on the Rights of the Child in Italy in Italian was presented November 18, 2009, on the occasion of the twentieth anniversary of the CRC. CRC Group, as happened in 2003 and 2006, will attend the pre-session (closed-door meeting between the members of the UN and representatives of NGOs) and the sessions will be held in 2011. EAHC (Executive Agency for Health and Consumers).
The Laboratory for Mother and Child Health collaborated in writing the Health and social care chapter of the First European child health report for the age group of 0-12 years in European countries, funded by the Executive Agency for Health and Consumers of the European Union. In particular, the Laboratory was involved in evaluating the policies concerning the access to health care for migrant people developed by European and EFTA countries, and the drug prescription profile in paediatric population.

Ricerca & Pratica

Ricerca & Pratica was born in January, 1985, as a manifestation of the . Mario Negri. Institute for Pharmacological Research. Today, the journal is part of the International Society of Drug Bulletins (ISDB), which represents independent journals. For more than twenty years, the journal has represented an arena for all those professionals who collect data and carry out studies in general practice with the aim to increase their knowledge and to improve their practice. Ricerca & Pratica is also appreciated for its ability to go beyond the merely clinical aspect of medicine, without, however, forgetting that it is to this aspect that the readers dedicate most of their time and effort. Through its activity, Ricerca & Pratica can therefore represent an exclusive, independent observation point. It is also an area that promotes contemplation, evaluation, and information by applying . tools. such as data trustworthiness and importance, the balance between benefits and risks and between benefits and costs, independence from conflicts of interest, and the realistic objective to contribute to a progressive, equally distributed improvement in the populations health.

Co-operation with countries with limited resources

As an expression, test, and original method of expression of the choice to make the Laboratorys research transferable and accessible to all populations, the Laboratory promoted and provided assistance to projects in, and for, the . South of the world. , also in collaboration with the World Health Organization. The technical and organisational support for local groups and international non-governmental organisations for carrying out sociosanitary projects in countries with limited resources, especially Colombia, Ecuador, Brazil, Bolivia, Cuba, Vietnam and the Balkans, continues.

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LABORATORY OF REGULATORY POLICIES

STAFF

Head Senior resercher Research fellow Visiting scientist

Vittorio BERTELE, M.D. Rita BANZI, Chem Pharm. D, PhD Brian GODMAN, BSc Roberta Joppi, Chem Pharm. D

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CURRICULUM
Vittorio Bertele is a clinical pharmacologist. He got his MD degree in 1977 and the specialization in Internal Medicine in 1982, both at the Milan University Medical School. He was research fellow at the Harvard Medical School and then worked at the Milan University and the Mario Negri Institute. His main areas of interest have been clinical pharmacology of drugs active on the haemostatic and vascular system1,2, epidemiology of interventions in the cardiovascular area, and clinical trials and drug utilization studies in the cardiovascular area3,4. He was CPMP expert at the EMEA, member of the Committee for Drug Price Negotiation at the Italian Ministry of Health, and member of the TechnicalScientific Committee at the Italian Drug Agency. At present he is head of the Regulatory Policies Laboratory at the "Mario Negri" Institute, secretariat of the ECRIN Scientific Board, and member of the Italian Horizon Scanning Center5-10. Selected publications 1. Bertele' V., Falanga A., Tomasiak M., Dejana E., Cerletti C., De Gaetano G. Platelet thromboxane synthetase inhibitors with low doses of aspirin: Possible resolution of the "aspirin dilemma". Science 1983; 220: 517-519 2. Bertele' V., Falanga A., Tomasiak M., Chiabrando C., Cerletti C., De Gaetano G. Pharmacological inhibition of thromboxane synthetase and platelet aggregation: Modulatory role of cyclooxygenase products. Blood 1984; 63: 1460-1466 (1984). 3. The i.c.a.i. Group (Gruppo di studio dell'Ischemia cronica Critica degli Arti Inferiori). Prostanoids for chronic critical leg ischemia: A randomized, controlled, open-label trial with prostaglandin E1. Ann Int Med 1999; 130: 412-421 4. Garattini S, Bertele V. Adjusting regulatory rules to public health needs. Lancet 2001; 358: 6467 5. Garattini S, Bertele' V. Efficacy, safety, and cost of new anticancer drugs. BMJ 2002; 325: 269271 6. Garattini S, Bertele V, Li Bassi L. How can research ethics committees protect patients better? BMJ 2003; 326:1199201 7. Joppi R, Bertele' V, Garattini S. Disappointing biotech. BMJ 2005; 331: 895-897 8. Garattini S, Bertele' V. Non-inferiority trials are unethical because they disregard patients' interests. Lancet 2007; 370 : 1875-1877 9. Garattini S, Bertele' V. Homoeopathy: not a matter for drug-regulatory authorities. Lancet 2009; 374 : 1578-1580 10. Garattini S, Bertele' V. Europe's opportunity to open up drug regulation. BMJ 2010; 340:c1578

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ACTIVITIES
Critical appraisal of clinical methodology Cooperation to the development and functioning of the pan-European Infrastructure for clinical trials (ECRIN, European Clinical Research Infrastructure Network) achieved by the ECRIN Scientific Board Secretariat and the European Correspondent for Italy Coordination of the Task 9.3 Identification of key steps on monitoring activities within the WP9 of the ECRIN, European Clinical Research Infrastructure Network Critical evaluation of the benefit-risk profile of drugs Assessment of emerging technologies Optimisation of drug use and healthcare fund stewardship including potential reforms and initiatives to achieve this Critical appraisal and recommendations for European Pricing and Reimbursement systems including generics, interchangeable products within a class and new innovative medicines Cooperation to the design and conduct of pharmacovigilance and pharmacoepidemiology studies in Europe Evaluation of the appropriateness of drug legislation, institutions, and regulatory procedures with respect to public health needs. Cooperation to the development and to the solution of regulatory issues in developing countries.

MAIN FINDINGS
Critical appraisal of clinical research methodological aspects as the adoption of equivalence/non-inferiority design in clinical trials Critical evaluation of transnational clinical research projects to be conducted with the methodological and operating support of ECRIN (European Clinical Research Infrastructure Network). Development of Pan-European strategies for the rational use of new and existing drugs including policies to enhance the managed entry of new drugs as well as reduce prescribing of more expensive interchangeable single sourced products in a class once generics are available: establishment of the Piperska Group Development of new models and strategies to optimise the managed entry of new drugs including suggestions for risk sharing arrangements in the future given current concerns. This co-ordinated via the Piperska group Recommendations for Pan-European pricing policies for generics as well as interchangeable brands in a class once generics are available

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Assessment of emerging technologies in the frame of the Italian Horizon Scanning Project which provides decision makers with timely information on the potential clinical impact and cost-effectiveness of new health technologies Critical review of drug documentation at the basis of marketing authorizations. Critical review of the criteria to assess pharmaceutical innovation and include new drugs in the national reimbursement schemes. Participation in the ENCePP (European Network of Centres of Pharmacovigilance and Pharmacoepidemiology) and the PROTECT consortium which under the coordination of the European Medicines Agency (EMA) aims at improving design and conduct of pharmacovigilance activities and pharmacoepidemiological studies in Europe. The PROTECT consortium is also developing and testing innovative methods to integrate and present information on drug-related benefits and risks Raising awareness among interested parties about the deficiencies of the present EU pharmaceutical legislation and about our proposals to improve it in the public health interest.

NATIONAL COLLABORATIONS
Italian Drug Agency (AIFA) Istituto Superiore di Sanit Department of Health Lombardy Region Italian Horizon Scanning Project

INTERNATIONAL COLLABORATIONS
European Medicine Agency (EMA) European Clinical Research Infrastructure Network (ECRIN) European Network of Centres in Pharmacoepidemiology and Pharmacovigilance (ENCePP) Piperska network involving health authority and health insurance personnel from acoss Europe to enhance the rational use of new and existing drugs Karolinska Institutet, Division of Clinical Pharmacology, Department of Laboratory Medicine, and Centre for Pharmacoepidemiology; Department of Drug Management and Informatics, SE University of Liverpool Management School, Prescribing Research Group, UK Cochrane Collaboration

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International Information Network on New and Emerging Health Technologies (EuroScan) World Health Organisation (Department of Essential Drugs and Medicines Policy) Association of South East Asian Nations (ASEAN)

EDITORIAL BOARD MEMBERSHIP

Ricerca & Pratica Dialogo sui Farmaci Frontiers in Clinical Trials and Pharmacotherapy (associate editor)

Frontiers in Pharmacology: Pharmacoeconomics and Outcomes Research (Associate Editor)

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

European Clinical Research Infrastructure Network (ECRIN) Scientific Board, Secretariat Scientific Committee of the Italian Horizon Scanning Project

SCIENTIFIC PUBLICATIONS (2010)

Garattini S, Bertele' V. Bevacizumab and ranibizumab. A matter of public interest. BMJ 2010 341 : c3721 Garattini S, Bertele' V. Dutasteride and prostate cancer. Reply to. N Engl J Med 2010 363 : 794 Garattini S, Bertele' V. Europe's opportunity to open up drug regulation. BMJ 2010 340 : c1578 Garattini S, Bertele' V. Alternative medical practices: flashbacks from the dark ages. Eur J Intern Med 2010 21 : 245-246 Garattini S, Bertele' V. Rosiglitazone and the need for a new drug safety agency. BMJ 2010 341 : c5506 Garattini S, Bertele' V. Health authorities should drive clinical research. Public Service Review European Union 2010 20 : 162-163 Adamski J, Godman B, Ofierska-Sujkowska G, Osinska B, Herholz H, Wendykowska K,Laius O, Jan S, Sermet C, Zara C, Kalaba M, Gustafsson R, Garuoliene K, Haycox A, Garattini S, Gustafsson LL. Review of risk sharing schemes for pharmaceuticals: considerations, critical evaluation and

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recommendations for European payers. BMC Health Services Research 2010, 10:153 doi:10.1186/1472-6963-10-153 Godman B, Shrank W, Wettermark B, Andersen M, Bishop I, Burkhardt T, Garuoliene K, Kalaba M, Laius O, Joppi R, Sermet C, Schwabe U, Teixeira I, Tulunay FC, Wendykowska K, Zara C, Gustafsson LL. Use of generics a critical cost containment measure for all healthcare professionals in Europe? Pharmaceuticals 2010; 3:2470-94 doi 10.3390/ph/3082470 ISSN 1424-8247 Wettermark B, Godman B, Eriksson C, van Ganse E, Garattini S, Joppi R, Malmstrm RE, Paterson K, Gustafsson LL. Einfhrung neuer Arzneimittel in europische Gesundheitssysteme. GGW 2010;10(3):2434 (Introduction of new medicines into European healthcare systems) Godman B, Wettermark B. Commentry Cost awareness when prescribing treatment. British Journal of Healthcare Management 2010;16( 2); 72 Wettermark B, Elseviers M, Godman B, Ronning M, Tilson L, Vlahovic-Palcevski V. Methodological challenges in cross-national comparisons in drug utilisation. Pharmacoepidemiology and Drug Safety 2010:19:S16.DOI: 10.1002/pds P. K. Cheema, S. Gavura, B. Godman, L. Yeung, M. E. Trudeau. Global variations in reimbursement of new cancer therapeutics: Improving access through risk-sharing agreements. J Clin Oncol 28:15s, 2010 (suppl; abstr 6050). Godman B, Shrank W, Andersen M, Berg C, Bishop I, T. Burkhardt T, Garuolien K, Herholz H, Joppi R, Kalaba M, Laius O, McGinn D, Samaluk V, Sermet C, Schwabe U,Teixeira I, Tilson L, Tulunay FC, Vlahovi-Palevski V, Wendykowska K, Wettermark B, Zara C, Gustafsson LL. Comparing policies to enhance prescribing efficiency in Europe through increasing generic utilisation: changes seen and global implications. Expert Rev. Pharmacoeconomics Outcomes Res 2010; 10: 707722 Godman B, Buscics A, Burkhardt T, Schmitzer M, Wettermark B, Wieninger P. Initiatives to enhance renin-angiotensin prescribing efficiency in Austria; impact and implications for other countries. Expert Rev Pharmacoeconomics and Outcomes Research 2010;10: 199-207 Sermet C, Andrieu V, Godman B, Van Ganse E, Haycox A, Reynier JP. Ongoing pharmaceutical reforms in France; implications for key stakeholder groups. Applied Health Economics and Health Policy 2010;8:7-24 Wettermark B, Persson M, Wilking N, Kalin M, Korkmaz S, Hjemdahl P, Godman B, Petzold M Gustafsson LL for the Regional Drug Expert Consortium. Forecasting drug utilization and expenditure in a metropolitan health region. BMC Health Services Research 2010, 10:128 McGinn D, Godman B, Lonsdale J, Way R, Wettermark B, Haycox A. Initiatives to enhance the efficiency of statin and proton pump inhibitor prescribing in the UK; impact and implications. Expert Rev Pharmacoeconomics and Outcomes Res 2010; 10: 73-85 Moon J, Flett A, Godman B, Grosso A, Wierzbicki A. Getting better value from the NHS drug budget. BMJ 341:c6449 2010.

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RESEARCH ACTIVITIES Critical appraisal of clinical methodology

Raising awareness about potential biases in clinical research

Critical evaluation of the EU pharmaceutical legislation

Raising awareness among interested parties about the deficiencies of the present EU pharmaceutical legislation and about our proposals to improve it in the public health interest

Development of a Pan-European Infrastructure for clinical trials

Participation in a distributed infrastructure linking national networks of clinical research centres and clinical trials units (ECRIN, European Clinical Research Infrastructure Network) which provides integrated one-stop shop services to investigators and sponsors in multinational studies.

Critical appraisal of ongoing reforms including pricing reforms in major European countries
Evaluation of ongoing reforms across Europe to enhance generic prescribing rates, drive down generic prices and corresponding originator brands, as well as potential prices of interchangeable brands once standards become available as generics, and the potential for cross cultural learnings to release valuable resources to fund increased volumes and new innovative drugs in the future without prohibitive increases in general taxation or health insurances to continue to provide equitable and comprehensive healthcare in Europe

Development of Pan-European strategies for rational use of drugs

Enhancing rational use in line with an approach that has become known as the four Es, namely: economics; enforcement; education and engineering to further fund increased volumes and new valuable innovative drugs. In addition, development of new models to optimize the managed entry of new drugs including horizon scanning and critical drug evaluation pre-launch (below) and post launch activities

Development of Pan-European strategies for pharmacovigilance

Developing and testing innovative methods to integrate and present information on benefits and risks in order to provide all stakeholders (patients, prescribers, regulators and pharmaceutical companies) with accurate and useful information on drug-related risks and benefits

Assessment of emerging technologies

Collecting information on emerging medicines with respect to their potential clinical impact and their cost effectiveness and ranking the new products according to their possible marketing authorization date, their potential innovation grade, therapeutic and economic impact, possible price and NHS sustainability with the aim to provide decision makers with timely information on the potential clinical impact and cost effectiveness of new health technologies

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CENTRE OF COMPUTER SCIENCE ENGINEERING

STAFF

Research and Communication Informatics Head of Division Division of I.C.T. Services and Management Head of Division BAZZI Davide ROSSI Lorenzo Marco

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CURRICULA Lorenzo Marco Rossi graduated in Biomedical Engineering with specialization in Hospital Clinical Instrumentation at Politecnico of Milan. He has been working with the Institute Mario Negri since 1998. Main areas of interest are: 1.Planning and realization of software for clinical research 2.Planning and realization of software system for in-plant automatization 3.Planning and realization of products for multimedia divulgation Davide Bazzi graduated in Informatics with ABACUS specialization at Istituto Tecnico Industriale Statale of Corsico. He has been working with the Institute of Mario Negri since 1997. Main areas of interest are: 1. Planning, realization and management of communication Network and Data Center 2. Definition and management of technological innovation for ICT systems 4. Planning and realization of technological innovation for ICT systems 3. Definition and application of organizations methodologies and processes for the Informatics Security Management ACTIVITIES In order to fulfill even more specialization needs in informatics development, the Centre of Computer Science Engineering is organized, considering the acquired skills, in three distinct division bound each other by a strong collaborative relationship. The Centre of Computer Science Engineering gathering informatics multidisciplinary aspects promotes and propose itself to coordinate and harmonize the development of the tools for the management information, improving the integration between informative procedures making more efficacious communication process and management of scientific and administrative datas, in order to support and fasten decisional, management, clinical trials and scientific processes. RESEARCH ACTIVITIES Implementation of Clinical Trials gathering forms (E-CRF) Lab. Translational and Outcome Research in Oncology (Dep. Oncology) Trial CERP Lab. Clinical Trials (Dep. Oncology) Amendment trial FOLFOX Trial ITACAS 2 Lab. New Drug Development Strategies (Dep. Oncology)

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Trial MUCOSITIS Maintenance and management of data gathering forms for the following clinical trials Lab. Neurological Disorders (Dip. Neuroscience): Registro Europeo SLA Trial L-ACETYLCARNITINE Trial ANTIEPILETTICI Trial EPILESSIA E STROKE Trial EPO VS MP IN SPINAL SHOCK Trial VALPROATO Trial THEOREM Trial ANTIEPILETTICI Trial ADONE Trial EDU-COM Lab. Clinical Trials (Dip. Oncology) Trial FOLFOX Trial TOP Trial COMETS Trial TAILOR Trial HEAD & NECK Trial GLAUCOMA PEDIATRICO Amendment to Trial TAILOR Lab. New Drug Development Strategies (Dip. Oncology) Trial MAPS Trial STARPAN Trial TRIAC Dip. Epidemiology Trial CADASIL Lab. Quality Assessment of Geriatric Therapies and Services Trial GISAS Patients registry for Polipathologies and Politherapies SIMI web

Web based applications connected to the projects

Design and realization of Monitoring Instruments for the current Trials Gathering Data Program Order Handling System (Statistics Module) Management of the Database hostings datas about recovers, prescriptions and examinations provided from Regione Lombardia for covenant data analysis. Support to data processing in recipes analysis for Regione Lombardia Creation of utility for the software of Interaction between Drugs

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Other projects Web based applications connected to the projects New database for the Institute Donators Updates of statistics for the Institute Distributed Publications Management of the database and credits of ECM Clinical Trial Register for the Institute Administration of the management system for the entrance exams of the Regione Lombardia school Administration of the management system for the entrance exams for PhD Utility for School Administration

Multimedia Communication Management of multimedia contents for Institutional website Video editing and DVD realization for Presentations

Websites

Management of the Official Institutes website Management of linked websites

Informatics Infrastructures

Realization of physical-to-virtual migration and stabilization of server on virtual platform Realization of support infrastructure (server and database) for Valhidate project Migration of the new e-mail system on virtualized platform Coordination of activities for setup and starting of the new Km Rosso sites Informatics and Telecommunications Activation completed for new linking infrastructures between sites on MPLS network Realization of a new backup system (disk-to-disk) in secondary server farm destined to a disaster recovery site Ordinary activities of helpdesk and Informatics Infrastructures and Telecommunications management

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ITALIAN COCHRANE CENTRE

STAFF

Head

Alessandro LIBERATI, M.D.

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CURRICULUM
Alessandro Liberati obtained his MD Degree in 1978 at the University of Milano and his post doctoral degree in Hygiene and Preventive Medicine in 1981 at the same university. Teaching activities: Primary responsibility of several academic and non academic training courses on the methodology of clinical research and systematic reviews/metanalyses. He is Director of the advanced Master Evidence based medicine e metodologia della ricerca sanitaria, at the Universit degli Studi di Modena e Reggio Emilia. Areas of scientific expertise: methodology of clinical research with particular reference to controlled clinical trials, epidemiological methods for research synthesis (systematic reviews and metanalyses); methods of practice guidelines production and implementation, evaluation of ethical implications of clinical research. Past and current roles at the Mario Negri Institute: 1980-1986 Junior researcher at the Laboratory of Clinical pharmacology; 1987-1989 Head, Unit of Clinical Epidemiology and Health Services Research; 1990-1998 Head Laboratory of Clinical Epidemiology; since 1994 he is Director of the Italian Cochrane Centre; since 1998 he is Associate Professor of Clinical Biostatistics and Epidemiology at the University of Modena and Reggio Emilia; since 1997 he is President of the Associazione per la Ricerca sulla Efficacia dellAssistenza Sanitaria - Centro Cochrane Italiano (AREAS-CCI); he collaborates since 2004 with the Social and Health Regional Agency of Emilia Romagna; since 2004 he is Member of the Commissione Nazionale Ricerca Sanitaria; since 2005 he is Member of the Commissione Ricerca e Sviluppo dellAgenzia Italiana del Farmaco (AIFA).

Selected publications Liberati A et al (Italian Medicines Agency (AIFA) Research & Development Working group). Feasibility and challenges of independent research on drugs: the Italian medicines agency (AIFA) experience Eur J Clin Invest 2010; 40 (1): 69-86 Banzi R, Liberati A, Moschetti I, Tagliabue L, Moja L, A review of online evidence-based practice point of care information summary providers. J Med Internet Res 2010; 7:12 (3) Parmelli E, Papini D, Moja L, Bandieri E, Belfiglio M, Ciccone G, De Palma R, Leoni M, Longo G, Magrini N, Moschetti I, Liberati A, Updating clinical recommendations for breast, colorectal and lung cancer treatments: an opportunity to improve methodology and clinical relevance Ann Oncol 2010 Moher D, Liberati A Reporting systematic reviews and meta-analyses: Asking authors, peer reviewers, editors and funders to do better Med Clin 2010 Colombo C, Mosconi P, Buratti MG, Liberati A, Donati S, Mele A, Satolli R, Press coverage of hormone replacement therapy and menopause. Eur J Obstet Gynecol Reprod Biol 2010 Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA GROUP Preferrred reporting items for systematic reviews and meta-analysis. The PRISMA Statement. Int J Surg 2010; 8: 336-41 Liberati A, D'Amico R, Commentary: The dabate on non inferiority trials: "when meta-analysis alone is not helpful" Int J Epidemiol 2010; 39: 1582-3 Colombo C, Satolli R, Liberati A, Mosconi P, Giurie dei cittadini Working Group. Treating prehypertension. Citizens' juries in health care. BMJ 2010; 22: 341 Traversa G, Sagliocca L, Liberati A, Martini N, The need to promote independent research on drugs. Ann Oncol 2010; 21: 2295 Liberati A, So many questions, so few answers. Inerview by Les Olson. Bull World Health Organ 2010; 88: 568 Moher D, Liberati A, Reporting systematic reviews and meta-analysies: asking authors, peer reviewers, editors and funders to do better. Med Clin (Barc) 2010; 135: 505

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INTRODUCTION TO THE CENTRE'S ACTIVITIES

The Italian Cochrane Centre (ICC) (http://www.cochrane.it) was founded in 1994 and is affiliated to the Cochrane Collaboration (CC). The CC is an international non profit organization that prepares, maintains and promotes systematic reviews of the effects of health care interventions. The main product of the Cochrane Collaboration is the Cochrane Library, a monthly publication containing Cochrane systematic reviews and other relevant databases of other siblings international organizations. The objectives of the ICC are centered around supporting various activities of the Cochrane Collaboration within Italy. In particular: a) to disseminate the knowledge of CC and CC activities throughout Italy; b) to provide methodological and practical support to all individuals and groups who are interested in collaborating with the CC; c) to develop methodological projects to progress scientific knowledge on science of research synthesis d) to contribute to the adoption and dissemination of Evidence-based Medicine (EBM) and Evidence based Health Care (EBHC) in Italy. e) to collaborate with NHS to improve the quality of clinical and epidemiological research.

FINDINGS/MAIN RESULTS
The ICC has created a national network with researchers and health care providers who are producing systematic reviews for the Cochrane Collaboration and are actively involved in other activities related to the dissemination of evidence based medicine. Collaboration with Agenzia Italiana del Farmaco (AIFA) in information projects Collaboration with Agenzia Sanitaria e Sociale Regione Emilia Romagna to production guidelines and raccomendations in clinical oncology.

NATIONAL COLLABORATIONS
Istituto Neurologico "Carlo Besta", Milano Agenzia Sanitaria Regionale, Regione Emilia Romagna, Bologna Universit degli Studi di Modena e Reggio Emilia, Modena Universit degli Studi di Milano, Milano Dipartimento di Epidemiologia della Regione Lazio, Roma Osservatorio epidemiologico della Direzione sanit e servizi sociali della Regione Umbria, Perugia Istituto Ortopedico Galeazzi, Milano Istituti Ortopedici Rizzoli, Bologna Istituto Superiore di Sanit, Roma

INTERNATIONAL COLLABORATIONS
The Cochrane Collaboration, Oxford, UK Centre for Reviews and Dissemination, University of York, York, UK British Medical Journal Publishing Group, London, UK Centre for Statistics in Medicine, Oxford, UK

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Thomas Chalmers Centre for Systematic Reviews, Ottawa, Canada The Campbell Collaboration, Philadelphia, USA Centre for Evidence-Based Medicine, Oxford, UK Institute of Health and Society, Newcastle University, Newcastle, UK Clinical Epidemiology, Ottawa Hospital Research Institute, Ottawa, Canada Department of Epidemiology and Community Medicine, Faculty of Medicine, University of Ottawa, Ottawa, Canada

EDITORIAL BOARD MEMBERSHIP

Evidence Based Health Policy and Research, Journal of Clinical Epidemiology, Journal of Health Services Research, BMJ Evidence Centre (BMJ Publishing Group), European Journal of Clinical Investigation

PEER REVIEW ACTIVITIES

Annals of Internal Medicine, British Medical Journal, JAMA, Evidence Based Health Policy and Research, Canadian Medical Association Journal, PLoS Medicine. International Journal of Epidemiology.

EVENT ORGANIZATION
Corso di perfezionamento avanzato in Revisioni sistematiche e meta-analisi metodologia Cochrane November 2010- March 2011 Master in Promozione e Governo della Ricerca nella Aziende Sanitarie March December 2010, Modena Workshop: November 14 2010, Modena - Cochrane Collaboration e miglioramento della pratica clinica - Assistenza pazienti con ictus XIV Annual Meeting of the Italian Cochrane Network "Orientare e informare le decisioni nel campo degli interventi complessi" November 15 2010, Modena

PARTICIPATION IN EVENTS IN WHICH THE CENTRE WAS INVOLVED Clinical Trial Day May 19 2010, Milano Course Dalle evidenze alle raccomandazioni per la pratica clinica: istruzioni per luso

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May 30 31 2010, Ischia Annual Continental European Cochrane Entities Meeting (CECEM) June 2 4 2010, Perugia Annual Croatian Cochrane Simposium June 26 27, Spalato XII Cochrane Colloquium October 18 22 2010, Keystone, Colorado GRANTS AND CONTRACTS
Istituto di Ricerche Farmacologiche Mario Negri, Milano Universit degli studi di Modena e Reggio Emilia Agenzia sanitaria e sociale regionale, Regione Emilia-Romagna, Bologna Universit degli Studi di Milano, Milano

SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2010

All publications of the Italian Cochrane Center are available on web site www.cochrane.it. Spagnolo P, Del Giovane C, Luppi F, Cerri S, Balduzzi S, Walters EH, D'Amico R, Richeldi L. Non-steroid agents for idiopathic pulmonary fibrosis. Cochrane Database of Systematic Reviews 2010; Issue 9: CD003134 Iorio A, Matino D, D'Amico R, Makris M, Recombinant Factor VIIa concentrate versus plasma derived concentrates for the treatment of acute bleeding episodes in people with haemophilia and inhibitors. Cochrane Database of Systematic Reviews 2010; Issue 8: CD004449 Liberati A, D'Amico R, Commentary: The dabate on non inferiority trials: "when metaanalysis alone is not helpful" Int J Epidemiol 2010; 39: 1582-3 Colombo C, Satolli R, Liberati A, Mosconi P, Giurie dei cittadini Working Group. Treating prehypertension. Citizens' juries in health care. BMJ 2010; 22: 341 Traversa G, Sagliocca L, Liberati A, Martini N, The need to promote independent research on drugs. Ann Oncol 2010; 21: 2295 Liberati A, So many questions, so few answers. Inerview by Les Olson. Bull World Health Organ 2010; 88: 568 Moher D, Liberati A, Reporting systematic reviews and meta-analysies: asking authors, peer reviewers, editors and funders to do better. Med Clin (Barc) 2010; 135: 505

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Corbetta D, Sirtori V, Moja L, Gatti R, Constraint - induced movement therapy in stroke patients: systematic review and meta-analysis. Eur J Phy Rehabil Med 2010; 46: 537-44 Dall'Olmo L, Moja L, Treatment for non-dysplastic Barrett's oesophagus: a well informed, demanding patient. Int Emerg Med 2010; 5: 433-5 Macura A, Abraha I, Kirkham J, Gensini GF, Moja L, Iorio A, Selective outcome reporting: telling and detecting true lies. The state of the science. Int Emerg Med 2010; 5: 151-5 Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA GROUP Preferrred reporting items for systematic reviews and meta-analysis. The PRISMA Statement. Int J Surg 2010; 8: 336-41 Moja L, Clinicla trials: trial registration cannot alone trasform scientific conduct. Nat Rev Urol 2010; 7: 7-8 Colombo C, Mosconi P, Buratti MG, Liberati A, Donati S, Mele A, Satolli R, Press coverage of hormone replacement therapy and menopause. Eur J Obstet Gynecol Reprod Biol 2010 Virgili G, Koleva D, Garattini L, Banzi R, Gensini GF, Utilities and QALYs in health economic evaluations: glossary and introduction. Internal Emerg Med 2010; 5: 349-352 Banzi R, Liberati A, Moschetti I, Tagliabue L, Moja L, A review of online evidence-based practice point of care information summary providers. J Med Internet Res 2010; 7:12 (3) Parmelli E, Papini D, Moja L, Bandieri E, Belfiglio M, Ciccone G, De Palma R, Leoni M, Longo G, Magrini N, Moschetti I, Liberati A, Updating clinical recommendations for breast, colorectal and lung cancer treatments: an opportunity to improve methodology and clinical relevance Ann Oncol 2010 Moher D, Liberati A Reporting systematic reviews and meta-analyses: Asking authors, peer reviewers, editors and funders to do better Med Clin 2010 Italian Medicines Agency (AIFA) Research & Development Working group. Feasibility and challenges of indipendent research on drugs: the Italian medicines agency (AIFA) experience Eur J Clin Invest 2010; 40 (1): 69-86 Moja L. Trial registration cannot alone transform scientific conduct. Nature reviews. Urology 2010;7(1):7-8 Sirtori V, Corbetta D, Moja L, Gatti R. Constraint-induced movement therapy for upper extremities in patients with stroke. Stroke 2010;47:e57-e58

RESEARCH ACTIVITIES Educational and dissemination activities

The ICC is a partner of the Thomas C. Chalmers Interuniversity Center together with eight Italian schools of medicine. This networks main objective is to disseminate systematic reviews

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knowledge, production and use within the academia activities. In particular, many post-graduate courses were organized on EBM, clinical trial and systematic review methodology, and the GRADE approach to develop clinical guideline. In the context of the Thomas C. Chalmers activities, the ICC has organized a post-graduate advanced course on systematic reviews, Corso di Perfezionamento avanzato in revisioni sistematiche e meta-analisi per la produzione di linee guida metodologia Cochrane, second edition. In collaboration with PartecipaSalute project, the ICC translates the Cochrane Library press releases and disseminates them to scientific journalists, doctors and consumers (through the PartecipaSalute website).

Research activities
ICC researchers are involved in methodological projects focused on the study of the quality of Systematic Reviews. The ICC is involved in many multidisciplinary projects: - Point-of-Care information tools: to describe and evaluate information tools used by clinicians during their practice. Specifically the project focuses on editorial quality, evidence-based methodology and updating of Point-of-Care products. - GRADE: international collaboration with the GRADE working Group to develop and transparent methodology on evidence quality and strength of recommendation assessment, used in the production of the clinical guideline. - PRISMA: publication of a reporting guideline to evaluate the completeness of systematic review reporting. - E-learning: two systematic reviews on efficacy of e-learning to improve the knowledge of students and health professionals in health care.

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THE CATULLO AND DANIELA BORGOMAINERIO CENTER

One of the buildings on the Mario Negri Institute campus is The Catullo and Daniela Borgomainerio Center built in 1987 thanks to a donation from Mrs. Angela Marchegiano Borgomainerio. This is a Center for the study of rare childhood diseases and even today some of the laboratories housed in the building still conduct this research. For example, the study of new therapies used to treat a very rare form of acute myeloid leukemia, know as acute promyelocytic leukemia. A number of new studies are being done to identify new drugs having different mechanisms able to synergize with trans retinoic acid. Research on epidemiological childhood leukemia is also done at the Borgomainerio and a similar line of research involves testicular cancer in adolescents and young adults. We also do research aimed at finding evidence based therapies for children. Paediatric research activities done at the Borgomainerio Center are also performed in collaboration with groups located at other Institute locations including, The Aldo and Cele Dacc Center for Clinical Research on Rare Diseases at Ranica in Bergamo, the Regional Centre for Drug Information (CRIF) and the Laboratory for Mother and Child Health (Department of Public Health) which are both located in Milan.

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THE LIBRARY
STAFF

Head Librarian

Vanna Pistotti

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The Library, specialised in pharmacology and clinical epidemiology, was founded in 1963 thanks to a generous donation from the Gustavus and Louise Pfeiffer Research Foundation, in Denville, New Jersey, USA. Numerous public and private organisations help keep it operative, through donations in money or books, and subscriptions to periodicals.

STAFF
One Head and two Assistants plus a clerical worker

WHAT THE LIBRARY OFFERS

The library has a collection of about 5000 textbooks, monographs and congressional proceedings, and 150 periodicals of which a major part are in an electronic format. The books are classified according to the US National Library of Medicine Classification and the Medical Subject headings of Medline (MeSH). Besides the internal collection, the Library has access to the National Periodical Catalogue and to other Library systems (SBBL, GIDIF-RBM).

DATABESES AND ELECTRONIC JOURNALS

From every computer in the Institute it is now possible to have access to more than 2000 electronic journals and to three of the most important databases, PubMed, the Cochrane Library and Embase.

SPECIAL PROJECTS
The Library cooperates to the realisation of the Italian Information Specialists (GIDIF, RBM) journal catalog which is updated annually and to the catalog of the Lombardy Biomedical Library Consortium, a network that serves, through Internet, the scientific community in this District.

It collaborates to the Institute web site, particularly taking care of the Publications section, both scientific and lay press.

TRAINING
Every year courses on the use of the database and electronic journals are organised. These courses are designed for use by those working at the Institute but outsiders who are interested may attend.

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CONTRACTS
Since 1994 the library has been part of the Lombard Biomedical Library System. 14 university and research organisation libraries in Lombardy take part in this project, which allows easy, free access to scientific information to over 140 centres and institutions the Lombardy Region.

EVENTS AND COURSES Corso La ricerca bibliografica su database biomedici, organizzato in collaborazione con lASL di Bergamo Dipartimento Cure Primarie e Continuit Assistenziale, Bergamo 11 febbraio 2010 V edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 8 giugno 2010 V edizione del corso ECM "Internet e laggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 9 giugno 2010 Corso di formazione e aggiornamento per i giornalisti Internet e Medicina: a caccia di salute sul web, organizzato in collaborazione con la Unione Nazionale dei Giornalisti Scientifici Italiani (UNAMSI), Istituto di Ricerche Farmacologiche Mario Negri, Milano, 16 ottobre 2010 VI edizione del corso ECM "La ricerca bibliografica su database biomedici", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 2 novembre 2010 VI edizione del corso ECM "Internet e laggiornamento professionale in ambito medico", Istituto di Ricerche Farmacologiche Mario Negri, Milano, 3 novembre 2010 Minimaster Come leggere un lavoro clinico Ricerca bibliografica online organizzato dallAssociazione Nazionale Medici Cardiologi Ospedalieri, Firenze 19 maggio 2010. Titolo della relazione La ricerca bibliografica su database biomedici"

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Negri Bergamo Laboratories

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departments and laboratories

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DEPARTMENT OF MOLECULAR MEDICINE

STAFF

Head

Ariela BENIGNI, Biol.Sci.D., Ph.D.

Laboratory of Cell Biology and Xenotransplantation*

Head Marina MORIGI, Biol.Sci.D., Ph.D.

Unit of Platelet-Endothelial Cell Interaction

Head Miriam GALBUSERA, Biol.Sci.D.

Laboratory of Immunology and Genetics of Organ Transplantation and Rare Diseases

Head Marina NORIS, Chem.Farm.D., Ph.D.

Unit of Cellular biology of Autoimmunity and Transplant Rejection Head Sistiana AIELLO, Biol.Sci.D

Unit of Cellular and Molecular Biology of Transplantation Tolerance Head Federica CASIRAGHI, Chemist

Laboratory of Experimental Models of Kidney Diseases*

Head Unit of Pathology and Immunophatology Head Mauro ABBATE, M.D. Carla ZOJA, Biol.Sci.D., Ph.D.

Laboratory of Cell and Gene Therapy*

Head Susanna TOMASONI, Biol.Sci.D., Ph.D.

Unit of Advanced Microscopy Head Elena GAGLIARDINI, Biol.Sci.D., Ph.D.

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*Since November 1 2010, the following Laboratories have changed

st

name:
Laboratory of Cell Biology and Xenotransplantation is now entitled Laboratory of Cell Biology and Regenerative Medicine Laboratory of Experimental Models of Kidney Diseases is now entitled Laboratory of Pathophysiology of Experimental Renal Disease and Interaction with other Organ systems Laboratory of Cell and Gene Therapy is now entitled Laboratory of Gene Therapy and Cellular Reprogramming

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CURRICULA
Ariela Benigni got the Biol.Sci. degree in 1979 at the University of Milano, Italy, and the Ph.D. at Maastricht University, Netherlands, in 2001. Educational training: in 1979 Post Doctoral Fellow, Istituto di Ricerche Farmacologiche Mario Negri (IRFMN), Laboratory of Cancer Chemotherapy, Milan, Italy; in 1980-1981 Post Doctoral Fellow, Associazione Bergamasca per lo Studio delle Malattie Renali, Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Italy; in 1982 Post Doctoral Fellow, Centre Regional de Transfusion Sanguigne de Strasbourg, France. Areas of interest: vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on endothelin-1; combined treatment of antipertensive and renoprotective drugs to halt progressive renal injury; use of stem cells for tissue regeneration in acute renal failure in vivo e in vitro gene transfer; prevention of acute graft rejection through gene therapy; induction of kidney transplant tolerance by gene therapy; correction of genetic deficiency in rare diseases. Employement: in 1983 Scientist, IRFMN, Laboratory of Kidney Disease, Bergamo, Italy; in 19901994 Head Laboratory of Prostaglandin and Leukotriene Metabolism, IRFMN, Bergamo, Italy; from January 1991 Scientific Secretary, IRFMN, Bergamo, Italy; in 1994-1999 Head Laboratory of Vasoactive and Inflammatory Mediators of Tissue damage, IRFMN, Bergamo, Italy; from January 2000 Head, Department of Molecular Medicine, IRFMN, Bergamo, Italy; from March 2007 Consultant World Health Organization (WHO) for the multicentre observational study Screening for Pre-eclampsia: evaluation of the predictive ability of angiogenic factors for Pre-eclampsia; during 2007 Senior Fellow at the University of Oxford, Nuffield Department of Obstetrics & Gynaecology.
Selected publications: Gagliardini E, Conti S, Benigni A, Remuzzi G, Remuzzi A. Imaging of the porous ultrastructure of the glomerular epithelial filtration slit. J Am Soc Nephrol. 2010;21:2081-2089. H. Mischak, G. Allmaier, R. Apweiler, T. Attwood, M. Baumann, A. Benigni, S.E. Bennett, R. Bischoff, E. BongcamRudloff, G. Capasso, J.J. Coon, P. D'Haese, A.F. Dominiczak, M. Dakna, H. Dihazi, J.H. Ehrich, P. Fernandez-Llama, D. Fliser, J. Frokiaer, J. Garin, M. Girolami, W.S. Hancock, M. Haubitz, D. Hochstrasser, R.R. Holman, J.P. Ioannidis, J. Jankowski, B.A. Julian, J.B. Klein, W. Kolch, T. Luider, Z. Massy, W.B. Mattes, F. Molina, B. Monsarrat, J. Novak, K. Peter, P. Rossing, M. Snchez-Carbayo, J.P. Schanstra, O.J. Semmes, G. Spasovski, D. Theodorescu, V. Thongboonkerd, R. Vanholder, T.D. Veenstra, E. Weissinger, T. Yamamoto, A. Vlahou. Recommendations for biomarker identification and qualification in clinical proteomics. Sci Transl Med. 2010;2:46ps42 Benigni, P. Cassis, G. Remuzzi. Angiotensin II revisited: new roles in inflammation, immunology and aging. EMBO Mol Med. 2010; 2;247-257. Review Benigni, M. Morigi, G. Remuzzi. Kidney regeneration. Lancet. 2010;375:1310-1317. F. Prefumo, G. Pagani, N. Fratelli, A. Benigni, T. Frusca.Increased concentrations of antiangiogenic factors in mirror syndrome complicating twin-to-twin transfusion syndrome. Prenat Diagn. 2010;30:378-379. B. Smeets, ML. Angelotti, P. Rizzo, H. Dijkman, E. Lazzeri, F. Mooren, L. Ballerini, E. Parente, C. Sagrinati, B. Mazzinghi, E. Ronconi, F. Becherucci, A. Benigni, E. Steenbergen, L. Lasagni, G. Remuzzi, J. Wetzels, P. Romagnani. Renal progenitor cells contribute to hyperplastic lesions of podocytopathies and crescentic glomerulonephritis. J Am Soc Nephrol. 2009;20:2593-2603.

Marina Morigi got her Biol.Sci. degree in 1987 at the University of Milano, Milano, Italy and the Ph.D. at Maastricht University, Netherlands, in 2005. Educational training: in 1984-1987 Research training, IRFMN, Bergamo, Italy; in 1987-1995 Post Doctoral Fellow, IRFMN, Bergamo, Italy; in 1991 Stage at Brigham and Womens Hospital, Laboratory of Dr. P. Marsden, Boston, USA. Employement: since 1995 Scientist, IRFMN, Bergamo, Italy; in 1996-1999 Head, Unit of Renal and Endothelial Cell Biology; since 2000 Head, Laboratory of Cell Biology and Xenotransplantation, IRFMN, Bergamo, Italy. Areas of interest: Stem cell therapy and tissue regeneration: the potential of adult stem cells of different origin, and renal progenitor cells to differentiate and to regenerate renal tissue in acute and chronic experimental models of renal disease. Stem cell therapy with embryonic stem cells to cure acute and chronic renal diseases and to correct genetic defects in experimental mouse models. Kidney Organogenesis. Role of Shigatoxin in the pathogenesis of endothelial dysfunction and microvascular thrombosis in Hemolytic Uremic Syndrome. In vitro model of hyperacute xenograft rejection (porcine endothelium exposed to human serum as a source of xenoreactive natural antibodies and complement).

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Renal toxicity of the proteins filtered through the capillary barrier: in vitro model to study intracellular signals, gene expression and production of inflammatory mediators in cultured proximal tubular cells and glomerular epithelial cells.
Selected publications Morigi M, Buelli S, Zanchi C, Longaretti L, Macconi D, Benigni A, Moioli D, Remuzzi G, Zoja C. Shigatoxin-induced endothelin-1 expression in cultured podocytes autocrinally mediates actin remodeling. Am J Pathol. 2006 Dec;

169(6):1965-75.
Morigi M, Benigni A, Remuzzi G, Imberti B. The regenerative potential of stem cells in acute renal failure. Cell Transplant. 2006;15 Suppl 1:S111-7. Review. Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol. 2008 Sep 15;181(6):3933-46. Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A, Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice. Stem Cells. 2008 Aug;26(8):2075-82. Epub 2008 May 22. Figliuzzi M, Cornolti R, Perico N, Rota C, Morigi M, Remuzzi G, Remuzzi A, Benigni A. Bone marrow-derived mesenchymal stem cells improve islet graft function in diabetic rats. Transplant Proc. 2009 Jun;41(5):1797-800. Benigni A, Corna D, Zoja C, Sonzogni A, Latini R, Salio M, Conti S, Rottoli D, Longaretti L, Cassis P, Morigi M, Coffman TM, Remuzzi G. Disruption of the Ang II type 1 receptor promotes longevity in mice. J Clin Invest. 2009 Mar;119(3):524-30. Zoja C, Buelli S, Morigi M. Shiga toxin-associated hemolytic uremic syndrome: pathophysiology of endothelial dysfunction. Pediatr Nephrol. 2010 Nov;25(11):2231-40. Epub 2010 Apr 28. Benigni A, Morigi M, Remuzzi G. Kidney regeneration. Lancet. 2010 Apr 10;375(9722):1310-7. Morigi M, Rota C, Montemurro T, Montelatici E, Lo Cicero V, Imberti B, Abbate M, Zoja C, Cassis P, Longaretti L, Rebulla P, Introna M, Capelli C, Benigni A, Remuzzi G, Lazzari L. Life-sparing effect of human cord bloodmesenchymal stem cells in experimental acute kidney injury. Stem Cells. 2010 Mar 31; 28(3):513-22.

Marina Noris got her degree in Pharmaceutical Chemistry and Technologies in 1986 at the University of Rome La Sapienza) and the Ph.D. at Maastricht University, Netherlands, in 2005. Educational training: in 1984-1986 Fellow, Istituto di Chimica Farmaceutica e Tossicologica, University of Rome, Italy; in 1986-1987 Post Doctoral Fellow, Istituto di Chimica Farmaceutica e Tossicologica, University of Rome, Italy; in September 1987-March 1994 Post Doctoral Fellow, IRFMN, Unit of Mediators of Inflammation and Tissue Damage, Laboratory of Kidney Disease, Bergamo, Italy. Areas of interest: immunology of transplantation, tolerance induction; genetics of hemolytic uremic syndrome, thrombotic thrombocytopenic purpura, focal segmental glomerulosclerosis, diabetic nephropathy, role of nitric oxide and arginine dysfunctions in uremia and in pre-eclampsia. Employment: in 1994-1996 Head, Unit of Endothelial Cell Pathophysiology, IRFMN, Bergamo, Italy; 1996-1999 Head, Laboratory of Cellular and Molecular Biology of the immune response and autoimmunity, IRFMN, Italy; from January 2000: Head, Laboratory of Immunology and Genetics of Rare Diseases and Organ Transplantation, Department of Molecular Medicine, IRFMN, Bergamo, Italy.
Selected publications Noris M, Remuzzi G. Atypical Hemolytic Uremic Syndrome N Engl J Med. 2009 Oct 22;361(17):1676-87. Noris M, Cassis P, Azzollini N, Cavinato R, Cugini D, Casiraghi F, Aiello S, Solini S, Cassis L, Mister M, Todeschini M, Abbate M, Benigni A, Trionfini P, Tomasoni S, Mele C, Garlanda C, Polentarutti N, Mantovani A, Remuzzi G. The Toll-IL1R member Tir8/SIGIRR negatively regulates adaptive immunity against kidney grafts. J Immunol. 2009 Oct 1;183(7):424960. Delvaeye M, Noris M, De Vriese A, Esmon CT, Esmon NL, Ferrell G, Del-Favero J, Plaisance S, Claes B, Lambrechts D, Zoja C, Remuzzi G, Conway EM. Thrombomodulin mutations in atypical hemolytic-uremic syndrome. N Engl J Med. 2009 Jul 23;361(4):345-57. Castelletti F, Donadelli R, Banterla F, Hildebrandt F, Zipfel PF, Bresin E, Otto E, Skerka C, Renieri A, Todeschini M, Caprioli J, Caruso RM, Artuso R, Remuzzi G, Noris M. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Proc Natl Acad Sci U S A. 2008;105(7):2538-43. Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P, Mele C, Bresin E, Cassis L, Gamba S, Porrati F, Bucchioni S, Monteferrante G, Fang CJ, Liszewski MK, Kavanagh D, Atkinson JP, Remuzzi G. Genetics of HUS: the impact of MCP, CFH and IF mutations on clinical presentation, response to treatment, and outcome. Blood. 2006; 108(4):1267-79. Noris M, Brioschi S, Caprioli J, Todeschini M, Bresin E, Porrati F, Gamba S, Remuzzi G, on behalf of the International Registry of Familial and Recurrent HUS/TTP. Familial haemolytic uraemic syndrome and an MCP mutation. Lancet. 2003; 362:1542-47.

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Susanna Tomasoni got her Biological Science degree in 1991 at the University of Milan and the Ph.D. at the University of Bologna in 1995. Educational training: in 1989-1991 Graduate student, University of Milan; in 1991-1994 PhD student, University of Milan; in 1994 Research Fellow, Renal Division, Brigham & Womens Hospital, Harvard Medical School, Boston, USA; 1995-1998: Post Doctoral Fellow, IRFMN, Bergamo, Italy. Areas of interest: construction of adenoviral vectors for gene therapy; in vitro and in vivo gene transfer techniques; use of adenoviral and adeno-associated viral vectors to prevent acute and chronic allograft rejection; induction of kidney transplant tolerance by cell and gene therapy; correction of genetic deficiency in rare diseases by gene therapy; involvement of microRNAs in the progression of renal disease; generation of induced-pluripotent stem cell from adult somatic cells; mesenchymal stem cellderived exosomes as mediators of cell-to-cell communication. Employment: in 1998-2000 Scientist; from 2000 Head, Unit of Gene Therapy; from 2010 Head, Laboratory of Cell and Gene Therapy; IRFMN, Bergamo, Italy.
Selected publications Trionfini P, Tomasoni S, Galbusera M, Motto D, Longaretti L, Corna D, Remuzzi G, Benigni A. Adenoviral-mediated gene transfer restores plasma ADAMTS13 antigen and activity in ADAMTS13 knockout mice. Gene Therapy. 2009; 16: 1379-85. Tomasoni S, Remuzzi G, Benigni A. Allograft rejection: acute and chronic studies. Contrib Nephrol. 2008; 159:122-34. Review. Imberti B, Morigi M, Tomasoni S, Rota C, Corna D, Longaretti L, Rottoli D, Valsecchi F, Benigni A, Wang J, Abbate M, Zoja C, Remuzzi G. Insulin-like growth factor-1 sustains stem cell mediated renal repair. J Am Soc Nephrol. 2007; 18: 2921-8. Benigni A, Tomasoni S, Turka LA, Longaretti L, Zentilin L, Mister M, Pezzotta A, Azzollini N, Noris M, Conti S, Abbate M, Giacca M, Remuzzi G. Adeno-associated virus-mediated CTLA4Ig gene transfer protects MHC-mismatched renal allografts from chronic rejection. J Am Soc Nephrol. 2006, 17: 1665-72. Tomasoni S, Aiello S, Cassis L, Noris M, Longaretti L, Cavinato R, Azzollini N, Pezzotta A, Remuzzi G, Benigni A. Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent CD4+ Tregulatory cells. Transplantation. 2005; 79: 1056-61. Tomasoni S, Azzollini N, Casiraghi F, Capogrossi M C, Remuzzi G, Benigni A. CTLA4Ig gene transfer prolongs survival and induces donor-specific tolerance in a rat renal allograft. J Am Soc Nephrol. 2000; 11: 747-52.

Carlamaria Zoja got her Biol.Sci. degree at the University of Milano, Italy, in 1979 and the Ph.D. at the University of Maastricht, The Netherlands in 2001. Educational Training: in 1979-1981 Post Doctoral Fellow, Associazione Bergamasca per lo studio delle Malattie Renali, Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Italy; in 1981-1983 Post Doctoral Fellow, Center for Thrombosis and Vascular Research, Department of Research Katholieke Universiteit, Leuven, Belgium; in 1983-1985: Post Doctoral Fellow, IRFMN, Laboratory of Kidney Disease, Bergamo, Italy; in 1988 stage at Case Western Reserve University, Cleveland, Ohio, USA; in 1989 stage at Brigham and Womens Hospital, Boston, USA. Areas of interest: experimental models of kidney diseases of immunological and non immunological origin; vasoactive and inflammatory mediators of renal disease progression; role of proteinuria in progressive kidney damage; protection of renal disease progression by a multidrug approach; novel immunosuppressive and anti-inflammatory strategies for the treatment of lupus nephritis; role of Shigatoxin in the pathogenesis of endothelial dysfunction in Hemolytic Uremic Syndrome. Employement: since 1985 Scientist, IRFMN, Bergamo, Italy; in 1990-1994: Head, Unit of Experimental Modelling for Human Renal Diseases, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; since 1995: Head, Laboratory of Experimental Models of Kidney Diseases, IRFMN, Bergamo, Italy. In November 2010 Lab denomination changed to Laboratory of Physiopathology of Experimental Renal Disease and Interaction with other Organ Systems. In 2004-2007 member Editorial Board, Journal of the American Society of Nephrology. Since January 2010 Leader WP5.2, SysKid collaborative project (FP7.
Selected publications: C. Zoja, D. Corna, D. Camozzi, D. Cattaneo, D. Rottoli, C. Batani, C. Zanchi, M. Abbate, G. Remuzzi. How to fully protect the kidney in a severe model of progressive nephropathy: a multidrug approach. J Am Soc Nephrol 2002;13:2898-2908. R. Donadelli, C. Zanchi, M. Morigi, S. Buelli, C. Batani, S. Tomasoni, D. Corna, D. Rottoli, A. Benigni, M. Abbate, G. Remuzzi, C. Zoja. Protein overload induces fractalkine upregulation in proximal tubular cells through NF-kB and p38 MAPK dependent pathways. J Am Soc Nephrol 2003; 14:2436-2446.

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C.Zoja, F.Casiraghi, S.Conti, D.Corna, D.Rottoli, R.A.Cavinato, G.Remuzzi, A.Benigni. Cyclin-Dependent kinase inhibition limits glomerulonephritis and extends lifespan of mice with systemic lupus. Arthritis & Rheumatism 2007; 56; 1629-1637. C.Zanchi, C.Zoja, M. Morigi, F.Valsecchi, XY Liu, D. Rottoli, M. Locatelli, S. Buelli, A.Pezzotta, P.Mapelli, J. Geelen, G.Remuzzi, J.Hawiger J. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to Shiga toxin. J Immunol. 2008; 181:1460-9. N.Perico, C.Zoja, D.Corna, D. Rottoli, F.Gaspari, L.Haskell, G.Remuzzi.V1/V2 Vasopressin receptor antagonism potentiates the renoprotection of renin-angiotensin system inhibition in rats with renal mass reduction. Kidney Int. 2009; 76:960-7. C.Zoja, D.Corna, E.Gagliardini, S.Conti, L.Arnaboldi, A. Benigni, G.Remuzzi G. Adding a statin to a combination of ACE inhibitor and ARB normalizes proteinuria in experimental diabetes, which translates into full renoprotection. Am J Physiol Renal Physiol. 2010; 299:F1203-11.

Mauro Abbate obtained his M.D. degree in 1988 at the University of Brescia, Italy. Educational training: in 1984-1988 Graduate Student, IRFMN, Bergamo, Italy; in 1988-1992 Post Doctoral Fellow, IRFMN, Bergamo, Italy; in 1992-1994 Research Fellow, The Renal Unit, Massachusetts General Hospital, HMS, Boston, USA. Areas of interest: renal disease progression: the role of proteinuria, complement, and mediators of injury in progressive kidney damage; mechanisms of glomerular injury; anti-GBM glomerulonephritis; mechanisms of tubular injury; kidney fibrosis; the renal biopsy; membranous nephropathy. Employement: in 1996 - 2000: Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Renal Pathology and Immunopathology, IRFMN, Bergamo, Italy.
Selected publications Buelli S, Abbate M, Morigi M, Moioli D, Zanchi C, Noris M, Zoja C, Pusey CD, Zipfel PF, Remuzzi G. Protein load impairs factor H binding promoting complement-dependent dysfunction of proximal tubular cells. Kidney Int. 2009 May;75(10):1050-9. Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol. 2008 Nov;3(6):1652-9. Abbate M, Zoja C, Corna D, Rottoli D, Zanchi C, Azzollini N, Tomasoni S, Berlingeri S, Noris M, Morigi M, Remuzzi G. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J Am Soc Nephrol. 2008 Jun;19(6):1158-67. Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A, Remuzzi G. Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target for renoprotective intervention. Am J Pathol. 2006 Apr;168(4):1073-85. Abbate M, Zoja C, Remuzzi G. How does proteinuria cause progressive renal damage? J Am Soc Nephrol. 2006 Nov;17(11):2974-84. Review Abbate M, Zoja C, Morigi M, Rottoli D, Angioletti S, Tomasoni S, Zanchi C, Longaretti L, Donadelli R, Remuzzi G. Transforming Growth Factor-beta 1 Is Up-Regulated by Podocytes in Response to Excess Intraglomerular Passage of Proteins: A Central Pathway in Progressive Glomerulosclerosis. Am J Pathol. 2002;161,2179-93.

Sistiana Aiello got the Biol.Sci. degree in 1993 at the University of Milano, Italy, and the Specialization in Pharmacology Research in 1996, at IRFMN, Bergamo, Italy. Educational training: in 1990-1993 research training, IRFMN, Bergamo; in 1993-2000 post doctoral fellow, IRFMN, Bergamo. Areas of interest: transplant immunology with a particular interest on dendritic cell biology and mechanisms by which T regulatory cells arise and work; in vitro and in vivo studies on new compounds with immunosuppressive capacity or capable to prevent ischemia/reperfusion tissue injury; vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on platelet activating factor (PAF) and nitric oxide (NO). Employement: since 2000 Scientist within Laboratory of Immunology and Genetics of Rare disease and Organ Transplantation; IRFMN, Bergamo; since 2006 Head, Unit of Cellular Biology of Autoimmunity and Transplant Rejection, IRFMN, Transplant Research Center Chiara Cucchi de Alessandri e Gilberto Crespi, Ranica.
Selected publications: Noris M, Cassis P, Azzollini N, Cavinato R, Cugini D, Casiraghi F, Aiello S, Solini S, Cassis L, Mister M, Todeschini M, Abbate M, Benigni A, Trionfini P, Tomasoni S, Mele C, Garlanda C, Polentarutti N, Mantovani A, Remuzzi G. The Toll-IL1R Member Tir8/SIGIRR Negatively Regulates Adaptive Immunity against Kidney Grafts. J Immunol. 2009, 183(7): 4249-60.

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Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol. 2009;20(1):123-30. Pezzotta A, Mister M, Monteferrante G, Cassis L, Azzollini N, Aiello S, Satta M, Benigni A, Remuzzi G, Noris M. Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat. Transplantation. 2008; 85(10):1476-82. Aiello S, Cassis P, Cassis L, Tomasoni S, Benigni A, Pezzotta A, Cavinato RA, Cugini D, Azzollini N, Mister M, Longaretti L, Thomson AW, Remuzzi G, Noris M. DnIKK2-transfected dendritic cells induce a novel population of iNOS-expressing CD4+CD25- cells with tolerogenic properties. Transplantation. 2007; 83:474-84. Tomasoni S, Aiello S, Cassis L, Noris M, Longaretti L, Cavinato RA, Azzollini N, Pezzotta A, Remuzzi G, Benigni A. Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent CD4+ T regulatory cells. Transplantation. 2005;79:1056-61.

Federica Casiraghi has obtained his degree in Industrial Chemistry in 1988, and the degree in Clinical Monitoring and in Biochemical Research in 1993-1994 at IRFMN, Bergamo, Italy. Educational Training: 1989-1994 research fellow, IRFMN, Bergamo. Areas of interest: Transplant immunology with particular focus on pharmacological and cellular therapies for induction and maintenance of transplantation tolerance. Characterization of regulatory T cells in renal transplant patients and in experimental models of allograft tolerance. Impact of different immunosuppressive drugs on T cell function in renal transplant patients. Vasoactive and inflammatory mediators of progressive renal injury with a particular emphasis on arachidonic acid metabolites. Employment: since 1994 Scientist within Laboratory of Immunology and Genetics of Rare Disease and Organ Transplantation, IRFM, Bergamo; since 2006 Head, Unit of Cellular and Molecolar Biology of Transplantation Tolerance, Transplant Research Center Chiara Cucchi de Alessandri e Gilberto Crespi, Ranica.
Selected Publications: Perico N, Casiraghi F, Introna M, Gotti E, Todeschini M, Cavinato RA, Capelli C, Rambaldi A, Cassis P, Rizzo P, Cortinovis M, Maras M, Golay J, Noris M, Remuzzi G. Autologous Mesenchymal Stromal Cells and Kidney Transplantation: A Pilot Study of Safety and Clinical Feasibility. Clin J Am Soc Nephrol. 2010 Oct 7. [Epub ahead of print Noris M, Cassis P, Azzollini N, Cavinato R, Cugini D, Casiraghi F, Aiello S, Solini S, Cassis L, Mister M, Todeschini M, Abbate M, Benigni A, Trionfini P, Tomasoni S, Mele C, Garlanda C, Polentarutti N, Mantovani A, Remuzzi G. The Toll-IL-1R Member Tir8/SIGIRR Negatively Regulates Adaptive Immunity against Kidney Grafts. J Immunol. 2009 183(7): 4249-60. Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol. 2008;181(6):3933-46. Noris M, Casiraghi F, Todeschini M, Cravedi P, Cugini D, Monteferrante G, Aiello S, Cassis L, Gotti E, Gaspari F, Cattaneo D, Perico N, Remuzzi G. Regulatory T Cells and T Cell Depletion: Role of Immunosuppressive Drugs. J Am Soc Nephrol. 2007; 18 (3):1007-18. Ruggenenti P, Perico N, Gotti E, Cravedi P, D'Agati V, Gagliardini E, Abbate M, Gaspari F, Cattaneo D, Noris M, Casiraghi F, Todeschini M, Cugini D, Conti S, Remuzzi G. Sirolimus versus cyclosporine therapy increases circulating regulatory T cells, but does not protect renal transplant patients given alemtuzumab induction from chronic allograft injury. Transplantation. 2007, 84(8):956-64.

Elena Gagliardini got her Biological Science degree in 1998 at the University of Milan and the Ph.D. at the Open University of London, UK, in 2006. Educational training: in 1996-1998 graduate student, IRFMN, Bergamo, Italy; in 1998-2006 Research Fellow, IRFMN, Bergamo, Italy. Areas of interest: vasoactive and inflammatory mediators of progressive renal injury; combined treatment of antipertensive and renoprotective drugs to halt and also regress progressive renal injury; mechanisms underlying tissue regeneration; ultrastucture and function of glomerular filter in physiological or pathological conditions. Employment: from 1996 Scientist, IRFMN, Bergamo, Italy; from 2010 Head, Unit of Advanced Microscopy, IRFMN, Bergamo, Italy
Selected publications Imaging of the porous ultrastructure of the glomerular epithelial filtration slit. Gagliardini E, Conti S, Benigni A, Remuzzi G, Remuzzi A. J Am Soc Nephrol. 2010 Dec;21(12):2081-9. Adding a statin to a combination of ACE inhibitor and ARB normalizes proteinuria in experimental diabetes, which

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translates into full renoprotection. Zoja C, Corna D, Gagliardini E, Conti S, Arnaboldi L, Benigni A, Remuzzi G. Am J Physiol Renal Physiol. 2010 Nov;299(5):F1203-11. Podocyte repopulation contributes to regression of glomerular injury induced by ACE inhibition. Macconi D, Sangalli F, Bonomelli M, Conti S, Condorelli L, Gagliardini E, Remuzzi G, Remuzzi A. Am J Pathol. 2009 Mar;174(3):797-807 ACE inhibition reduces glomerulosclerosis and regenerates glomerular tissue in a model of progressive renal disease. Remuzzi A, Gagliardini E, Sangalli F, Bonomelli M, Piccinelli M, Benigni A, Remuzzi G. Kidney Int. 2006 Apr;69(7):1124-1130. Selective impairment of gene expression and assembly of nephrin in human diabetic nephropathy. Benigni A, Gagliardini E, Tomasoni S, Abbate M, Ruggenenti P, Kalluri R, Remuzzi G. Kidney Int. 2004 Jun;65(6):2193-200. Changes in glomerular perm-selectivity induced by angiotensin II imply podocyte dysfunction and slit diaphragm protein rearrangement. Benigni A, Gagliardini E, Remuzzi G. Semin Nephrol. 2004 Mar;24(2):131-40. Review.

Miriam Galbusera got her Biol.Sci. degree in 1981 at the Universit degli Studi di Milano. Educational training: in 1981-1983 Post Doctoral Fellow, Istituto di Patologia Speciale Medica dell'Universit degli Studi di Milano, Italy; in 1985 - 1989 Post Doctoral Fellow, IRFMN, Bergamo, Italy; in 1989-1991 Post Doctoral Fellow at Scripps Clinic and Research Foundation, Laboratory of Thrombosis and Hemostasis, La Jolla, CA, USA; in 1991-1995 Post Doctoral Fellow, IRFMN, Bergamo, Italy. Areas of interest: ADAMTS-13 and VWF in thrombotic microangiopathies, VWF biochemistry, xenotransplantation, platelet-endothelial cell interaction under flow condition, platelet pathophysiology in uremia, receptor studies in kidney and platelets. Employement: 1995 - 1999: Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of PlateletEndothelial Cell Interaction, IRFMN, Bergamo, Italy.
Selected publications Trionfini, P, Tomasoni S, Galbusera M, Motto D, Longaretti L, Corna D, Remuzzi G, Benigni A. Adenoviral-mediated gene transfer restores ADAMTS13 plasma levels and activity in knockout mice. Gene Therapy. 2009; 16:1373-9. Galbusera M, Remuzzi G, Boccardo P. Treatment of bleeding in the dialysis patients. Semin Dial. 2009; 22:279-86. Galbusera M, Noris M, Remuzzi G. Inherited thrombotic thrombocytopenic purpura. Haematologica. 2009; 94:166-70. Bresin E, Gastoldi S, Daina E, Belotti D, Pogliani E, Perseghin P, Scalzulli PR, Paolini R, Marcen R, Remuzzi G, Galbusera M. Rituximab to prevent relapses in patients with thrombotic thrombocytopenic purpura and evidence of anti-ADAMTS13 autoantibodies. Thromb Haemost. 2009; 101:233-8. Benigni A, Caroli C, Longaretti L, Gagliardini E, Zoja C, Galbusera M, Moioli D, Romagnani P, Tincani A, Andreoli L, Remuzzi G. Renal tubular TLR9 is involved in the development of tubulointerstitial injury of systemic lupus. Arthritis Rheum. 2007; 56:1569-78. Donadelli R, Banterla F, Galbusera M, Capoferri C, Bucchioni S, Gastoldi S, Ruggeri ZM, Bresin E, Scheiflinger F, Rossi E, Remuzzi G, Noris M. In vitro and in vivo consequences of mutations in the von Willebrand factor cleaving protease ADAMTS13 in thrombotic microangiopathies. Thromb Haemost. 2006; 96:454-64.

INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES

The Department of Molecular Medicine was established in 1999 at the Negri Bergamo laboratories to coordinate the work of four laboratories and five units. The activities of the Department of Molecular Medicine are strictly interrelated with those of the Department of Renal Medicine of the Clinical Research Center for Rare Diseases Aldo e Cele Dacc. The following major objectives have been pursued: 1) identification of mediators and mechanisms responsible for the relentless decline of renal function in kidney diseases and development of therapeutic interventions to slow or even halt the disease progression to end-stage renal failure; 2) understanding the mechanisms underlying endothelial cell dysfunction in thrombotic microangiopathies and hyperacute rejection of xenograft 3) finding new strategies for modulating the immune response and preventing acute and chronic rejection of kidney allograft as well as exploration of immunological pathways leading to donor specific unresponsiveness and tolerance of the graft;

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4) investigation of the molecular and genetic basis of rare diseases such as hemolytic uremic syndrome/thrombotic thrombocytopenic purpura and pre-eclampsia and search for diseasesusceptibility genes or gene polymorphisms predicting the patient's response to drug therapy in more common and complex polygenic disorders. Such goals have been pursued using various approaches: 1) experimental models of kidney diseases of immunological and non-immunological origin mimicking human renal diseases to study vasoactive and inflammatory mediators and to test novel antiproteinuric and renoprotective drugs; 2) in vitro cultures of renal cells to address the toxicity of protein overload reproducing the condition of exaggerated protein traffic of proteinuric progressive nephropathies; 3) in vitro models to assess the interaction of vascular endothelial cells with leukocytes and platelets under controlled flow conditions; 4) experimental models of kidney allotransplant to study immunological processes responsible for acute and chronic rejection, the nephrotoxicity of immunosuppressor drugs as well as to explore pathways responsible for accomodation; 5) gene transfer of viral constructs carrying genes encoding immunomodulatory molecules to overcome acute rejection of allotransplantation avoiding immunosuppression; 6) identification of candidate genes with linkage analysis and search for mutations as well as assessment of gene polymorphisms.

FINDINGS/MAIN RESULTS
Mesenchymal stromal cell infusion inhibits memory favouring regulatory cell expansion in kidney transplant recipients. lymphocytes while

Identified the ultrastructure of the glomerular filtration pores whose alterations might be responsible for the increased filtration of plasma proteins in glomerular disease. A triple therapy to protect the diabetic kidney. Alterations in thrombomodulin a coagulation protein Hemolytic Uremic Syndrome. worsens Shigatoxin-associated

Human amniotic fluid stem cell therapy ameliorates the outcome of experimental acute kidney injury and improves animal survival. Embryonic kidneys transplanted in rats with chronic renal injury develop new renal structures and contribute to regeneration of damaged tissue. Inhibiting ACE promotes renal repair by limiting progenitor cell proliferation and restoring the glomerular niche

NATIONAL COLLABORATIONS
Consorzio per la Ricerca sul Trapianto di Organi, Tessuti, Cellule e Medicina Rigenerativa CORIT, Padova Laboratorio di Biologia dello Sviluppo, Dipartimento di Biologia Animale, Universit degli Studi di Pavia, Pavia Stem Cell Processing Laboratory, Clinic of Paediatric Oncohematology, University of Padova

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Sezione di Istologia e Embriologia, Dipartimento di Medicina Sperimentale, Universit degli Studi di Parma, Parma Fondazione I.R.C.C.S. Policlinico San Matteo, Pavia Laboratorio di Terapia genica e cellulare, G. Lanzani, Divisione di Ematologia, Ospedali Riuniti di Bergamo Laboratorio di Tecnologie della Riproduzione, AVANTEA Srl, Cremona
Laboratorio di Virologia, Istituto Nazionale per le Malattie Infettive L. Spallanzani, Roma

Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano

Dipartimento di Biotecnologie, Universit di Verona

Dipartimento di Istologia Microbiologia e Biotecnologie Mediche, Universit di Padova Dipartimento di Patofisiologia Clinica, Sezione di Nefrologia, Universit di Firenze International Centre for Genetic Engineering and Biotechnology, Molecular Medicine Group, Trieste U.O. di Ostetricia e Ginecologia, Azienda Ospedaliera Spedali Civili di Brescia I.R.C.C.S. Policlinico San Matteo, Pavia Istituto di Medicina Interna e Geriatria e Centro di Ricerca Emostasi, Universit Cattolica, Roma Dipartimento di Fisiologia e Patologia, Universit di Trieste Centro Regionale Indicatori biochimici di tumori, Associazione ABO, AULSS 12, Ospedale Civile, Venezia

INTERNATIONAL COLLABORATIONS
Academisch Ziekenhuis Maastricht, Interne Geneeskunde, Maastricht, Olanda Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA Children's Hospital and Regional Medical Center, University of Washington, Seattle, USA Department of Cell and Developmental Biology, SUNY Upstate Medical University, Syracuse, NY, USA Departments of Pediatrics and Human Genetics, University of Michigan, Ann Arbor, USA Deparment of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, USA Duke University Medical Center and Durham Veterans Affairs Medical Center, Durham, North Carolina, USA Erasmus University of Rotterdam, Olanda Hans-Knoll Institute for Natural Products Research, Jena, Germania Hospital of Bellvitge, Barcelona, Spagna Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Palo Alto, USA Klinikum der Ludwig Maximillians Universitat Munchen, Germania Max Delbruck Center for Molecular Medicine, Berlin, Germania Max-Plank Gesellschaft zur Forderung der Wissenshaften, Hpi of experimental endocrinology, Hannover, Germania Medical University of Innsbruck, Austria MISOT (Mesenchymal Stem Cells in Solid Organ Transplantation) study group Pediatric Nephrology Division, Center for Pediatrics and Adolescence Medicine, Heidelberg, Germany Rosalind Franklin University of Medicine and Science, Chicago, USA Surgery Unit, Great Ormond Street Hospital and Institute of Child Health, University College London, UK UCD Conway Institute, University College Dublin, Ireland

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University of British Columbia, Vancouver, Canada University of Colorado Cardiovascular Institute, Denver, USA University of Groningen, Olanda University of Liverpool, School of Biological Sciences, UK University of Pittsburgh School of Medicine, Pittsburgh, USA Wake Forest Institute of Regenerative Medicine, Wake Forest University of School of Medicine, Winston-Salem, NC, USA Weizmann Institute of Science, Rehovot, Israele World Health Organization, Geneva, Svizzera

EDITORIAL BOARD MEMBERSHIP

International Journal of Artificial Organs

PEER REVIEW ACTIVITIES

American Journal of Kidney Diseases American Journal of Pathology American Journal of Physiology American Journal of Transplantation Archives of Medical Science Cell transplantation Cytotherapy Diabetologia Drug Discovery Today Experimental Gerontology Kidney International Journal of Clinical Investigation Journal of Immunology Journal of Nephrology Journal of the American Society of Nephrology Laboratory Investigation Molecular Medicine Nature Reviews Nephrology Nephrology, Dialysis and Transplantation New England Journal of Medicine Pediatric Nephrology Plos One Proteomics Stem Cells and Development The Lancet Tissue and Cell Transplant Immunology Transplantation

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PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED

Star-T Rek Annual Meeting, Firenze, February 16 2010 EVOLVA meeting, Zurigo, March 2, 2010

SysKid Meeting, FP7, Vienna, March 8-9 2010

International Society of Nephrology, ISN Nexus Symposium; Kyoto, Japan, April 15-18, 2010 American Transplant Congress, San Diego, CA, May 1-5, 2010 2nd Meeting of the Forum of Italian Researchers on Mesenchymal and Stromal Stem Cells, Modena, May 3 2010 Curarsi con le cellule, Bergamo, May 22, 2010 Tavola Rotonda su Next Generation sequencing, Bergamo, May 24, 2010 Stem Cells and the Kidney, Genova, June 18-19, 2010

SysKid Meeting , FP7, Munich, Germany, June 24, 2010

XLVII ERA-EDTA Congress, Munich, Germany June 25-28, 2010 Renal Biopsy in Medical Diseases of the Kidneys, 33rd Annual Postgraduate Medicine Course, August 4- 7, 2010, Columbia University, New York Grandangolo 2010 in Nefrologia, Dialisi e Trapianto, Bologna, September 15, 2010 II Meeting Internazionale: Modern trends in Andrologia e Riproduzione Assistita - Taranto September, 17-18, 2010 12th Cambridge Massachusetts Biennial meeting of the International society for applied cardiovascular biology, Boston, USA, September 22-25, 2010 International Stem Cell Symposium, Seoul, Korea, October 2, 2010 51 Congresso Nazionale Societ Italiana di Nefrologia, Rimini, October 6-9, 2010 Renal Master Class, Ranica, Bergamo, October 7-8, 2010 PodoNet Symposium on Podocyte Disorders at the Meeting of the Turkish Society of Pediatric Nephrology Ankara , Turkey, October 8, 2010 New paradigma in hypertension research, American Heart association HBPR, Washington, USA, October 13, 2010

43rd Annual Meeting, American Society of Nephrology, Denver, November 16-21, 2010

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First European Conference on Mesenchymal Stem Cells, Toulouse, France, November 18-20, 2010 Second International Symposium on Albuminuria, Amsterdam, December 12-14, 2010

GRANTS AND CONTRACTS

Comitato Telethon Fondazione ONLUS Commissione Europea FP6 e FP7, Programma E-RARE Fondazione Aiuti per la Ricerca sulle Malattie Rare (ARMR) Fondazione ART per la Ricerca sui Trapianti ONLUS Fondazione Cariplo Fondazione Compagnia di San Paolo Fondazione Chiesi F. Hoffman La Roche Ltd Istituto Superiore di Sanit Ablynx NV ACRAF (Aziende Chimiche Riunite Angelini Francesco Spa) Alexion Pharmaceuticals Evolva AG Genzyme Corporation (Genzyme Renal Innovations Program) Genzyme Europe B.V. LFB Biotechnologies Reata Pharmaceuticals Sigma-Tau SpA

SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2010

Zoja C, Corna D, Gagliardini E, Conti S, Arnaboldi L, Benigni A, Remuzzi G. Adding a statin to a combination of ACE inhibitor and ARB normalizes proteinuria in experimental diabetes, which translates into full renoprotection. Am J Physiol Renal Physiol. 2010 Nov;299(5):F120311. Zoja C, Buelli S, Morigi M. Shiga toxin-associated hemolytic uremic syndrome: pathophysiology of endothelial dysfunction. Pediatr Nephrol. 2010 Nov;25(11):2231-40.
Perico N, Casiraghi F, Introna M, Gotti E, Todeschini M, Cavinato RA, Capelli C, Rambaldi A, Cassis P, Rizzo P, Cortinovis M, Maras M, Golay J, Noris M, Remuzzi G. Autologous Mesenchymal Stromal Cells and Kidney Transplantation: A Pilot Study of Safety and Clinical Feasibility. Clin J Am Soc Nephrol. 2010 Oct 7. [Epub ahead of print] Cugini D and Noris M. Toward a B-cell signature of tolerance? Commentary on Kidney Int. 2010 Sep;78(5):435-7.

Aiello S, Noris M. Klotho in acute kidney injury: biomarker, therapy, or a bit of both? Kidney Int. 2010 Dec;78(12):1208-10

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Benigni A, Morigi M, Remuzzi G. Kidney Regeneration. Lancet 2010;375:1310-7. Cassis P, Conti S, Remuzzi G, Benigni A. Angiotensin receptors as determinants of life span. Pflugers Arch. 2010 Jan;459(2):325-32. Review R. Schiffmann, S. Waldek, A. Benigni, C. Auray-Blais. Biomarkers of Fabry disease nephropathy. Clin J Am Soc Nephrol. 2010;5:360-364. A. Benigni, M. Morigi, G. Remuzzi. Kidney regeneration. Lancet. 2010;375:1310-1317. F. Prefumo, G. Pagani, N. Fratelli, A. Benigni, T. Frusca.Increased concentrations of antiangiogenic factors in mirror syndrome complicating twin-to-twin transfusion syndrome. Prenat Diagn. 2010;30:378-379. M. Morigi, C. Rota, T. Montemurro, E. Montelatici, V. Lo Cicero, B. Imberti, M. Abbate, C. Zoja, P. Cassis, L. Longaretti, P. Rebulla, M. Introna, C. Capelli, A. Benigni, G. Remuzzi, L. Lazzari. Life-sparing effect of human cord blood-mesenchymal stem cells in experimental acute kidney injury. Stem Cells. 2010;28:513-522. A. Benigni, P. Cassis, G. Remuzzi. Angiotensin II revisited: new roles in inflammation, immunology and aging. EMBO Mol Med. 2010; 2;247-257. Review H. Mischak, G. Allmaier, R. Apweiler, T. Attwood, M. Baumann, A. Benigni, S.E. Bennett, R. Bischoff, E. Bongcam-Rudloff, G. Capasso, J.J. Coon, P. D'Haese, A.F. Dominiczak, M. Dakna, H. Dihazi, J.H. Ehrich, P. Fernandez-Llama, D. Fliser, J. Frokiaer, J. Garin, M. Girolami, W.S. Hancock, M. Haubitz, D. Hochstrasser, R.R. Holman, J.P. Ioannidis, J. Jankowski, B.A. Julian, J.B. Klein, W. Kolch, T. Luider, Z. Massy, W.B. Mattes, F. Molina, B. Monsarrat, J. Novak, K. Peter, P. Rossing, M. Snchez-Carbayo, J.P. Schanstra, O.J. Semmes, G. Spasovski, D. Theodorescu, V. Thongboonkerd, R. Vanholder, T.D. Veenstra, E. Weissinger, T. Yamamoto, A. Vlahou. Recommendations for biomarker identification and qualification in clinical proteomics. Sci Transl Med. 2010;2:46ps42. M. Sandovici, L.E. Deelman, A. Benigni, R.H. Henning. Towards graft-specific immune suppression: Gene therapy of the transplanted kidney. Adv Drug Deliv Rev. 2010;62:13581368. Gagliardini E, Conti S, Benigni A, Remuzzi G, Remuzzi A. Imaging of the porous ultrastructure of the glomerular epithelial filtration slit. J Am Soc Nephrol. 2010;21:2081-2089.

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RESEARCH ACTIVITIES
The kidney's capability to regenerate by forming new nephrons, a characteristic of teleost and elasmobranch fish, was lost during the evolution of species. In humans, the kidney has been considered a terminally differentiated organ with low proliferative potential. However, as we reported in a recent review ( Lancet, 375: 1310-1317, 2010), studies in rodents suggest that, after an acute kidney injury, progenitors present in renal tissue or derived from bone marrow are able to activate molecular and cellular processes of regeneration. In this context, our research focused on studying whether and how adult stem cells of different origin or embryonic kidney tissues are able to regenerate and protect the kidney in models of acute and chronic kidney injury. Identification of renal progenitors in the adult tissue will allow to find new potential targets for regenerative therapies.

Laboratory of Cell Biology and Xenotransplantation Human amniotic fluid stem cells ameliorate the outcome of experimental acute kidney injury and animal survival.
Transplantation of bone marrow or cord blood mesenchymal stem cells (MSCs) has been indicated as a powerful tool for cell-based treatment of acute kidney injury (AKI). However, bone marrow collection is associated with patients discomfort and isolation of MSCs from cord blood is not invariably successful. By contrast, broadly multipotent stem cells, sharing characteristics with embryonic stem cells, can be easily obtained from amniotic fluid. Here we studied the regenerative potential of human amniotic fluid stem (hAFS) cells in mice with cisplatin-induced AKI. Infusion of hAFS cells in cisplatin-mice limited tubular damage and improved renal function, although not to control level, and prolonged animal survival. Human AFS cells engrafted injured kidney predominantly in peritubular region without acquiring tubular epithelial markers. Human AFS cells exerted antiapoptotic effect, activated Akt and stimulated proliferation of tubular cells possibly via local release of factors that we documented in vitro to be produced by hAFS cells. The therapeutic potential of hAFS cells was enhanced by cell pre-treatment with GDNF which, markedly ameliorated tubular injury and renal function by increasing stem cell homing to tubulo-interstitial area. By in vitro studies, GDNF increased hAFS cell production of growth factors, motility and expression of receptors involved in cell homing and survival. These findings indicate that hAFS cells can promote functional recovery and contribute to renal regeneration in AKI mice via local production of mitogenic and prosurvival factors. The effects of hAFS cells can be remarkably enhanced by GDNF preconditioning.

Kidney Organogenesis Activates Regeneration in a Rat Model of Progressive Renal Disease

Transplanted embryonic kidneys, metanephroi, were shown to develop renal structures in healthy animals. Here, in the context of chronic renal injury, we investigated the developmental capacity and the regenerative potential of metanephroi transplanted under the kidney capsule of rats with progressive renal disease. Six weeks post-transplantation, metanephroi developed tubuli and glomeruli, and produced diluted urine in cyst-like structures. In renal tissues, adjacent to the graft, proliferation and vascular density augmented together with a decrease of apoptosis and oxidative stress. Grafted animals showed tubuli positive to molecules normally expressed in immature renal tissue or in maturing nephrons and regenerating tubuli. Co-transplanted mesenchymal stem cells enhanced metanephros-induced cell proliferation, vascular density and graft filtering activity. Our data highlight that nephrogenesis occurres in chronically injured

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animals and activates regenerative processes. Moreover, mesenchymal stem cells revealed the capacity to speed-up and support renal recovery.

Inhibiting ACE promotes renal repair by limiting progenitor cell proliferation and restoring the glomerular niche
In collaboration with the Unit of Advanced Microscopy In the present study we have identified in rat kidneys a renal progenitor cell population localized in the inner surface of the Bowmans capsule. Accordingly with previous observation in human biopsies, the Bowmans capsule of healthy glomeruli is constituted by three different cell populations: immature renal progenitors, visceral epithelial cells or podocytes, and transitional cells which co-express markers of both the other cell populations. Under physiological condition renal progenitor cells may differentiate into podocytes and may be responsible for the generation of new podocytes. By using an experimental model of progressive nephropathy, we have observed that during the course of the disease renal progenitor cells undergo excessive proliferation, detach from the Bowmans capsule and invade the glomerular capillary tuft thus contributing to the formation of hyperplastic lesions. Treatment with the anti-hypertensive drug ACE inhibitor induces renal repair by significantly reducing the number and the extension of hyperplastic glomerular lesions. ACE inhibitor induces renoprotection by limiting renal progenitor cell activation and restoring the glomerular niche. Accordingly, progenitor cells in treated animals were no longer detectable within glomerular lesions but were localized along the Bowmans capsule to a pattern similar to controls. These results shed light on the mechanisms of glomerular injury and repair, and indicate the inhibition of angiotensin II synthesis as a potential selective way to enhance the intrinsic ability of the kidney to regenerate.

Laboratory of Experimental Models of Kidney Diseases Adding a statin to a combination of ACE inhibitor and Angiotensin II receptor blocker normalizes proteinuria in experimental diabetes which translates into full renoprotection
In collaboration with the Unit of Advanced Microscopy The activation of renin angiotensin system (RAS) plays a key role in favoring mechanisms leading to kidney function loss in several nephropathies and in diabetes. The capacity of RAS inhibitors to delay progression of diabetic nephropathy depends on the time at which therapy is started. A multimodal intervention is required to afford renoprotection in overt diabetic nephropathy. We assessed the effects of maximal RAS inhibition by angiotensin converting enzyme (ACE) inhibitor plus angiotensin II type I receptor blocker (ARB) in combination with statin in rats with overt diabetic nephropathy. Uninephrectomized rats made diabetic by streptozotocin were orally treated from 4 (when proteinuria and renal lesions had developed) to 8 months with vehicle, lisinopril plus candesartan, lisinopril plus candesartan plus rosuvastatin or rosuvastatin alone. Results showed that combined therapies were more renoprotective than single therapies. Systolic blood pressure increased in diabetic rats and was significantly lowered by combined therapies. Dual RAS blockade significantly reduced proteinuria with respect to vehicle. Addition of statin further lowered proteinuria to control levels. Glomerulosclerosis was ameliorated by RAS inhibitors or statin, and regression was achieved by the addition of statin. Loss of podocytes of diabetic rats was limited by ACE inhibitor plus ARB, while normalized by the three drugs. Defective nephrin expression of diabetes was increased by dual RAS blockade or statin, and restored by the triple therapy. Tubular damage, interstitial inflammation, and the expression of the fibrotic markers TGF and phosphorylated Smad 2/3 in tubuli were significantly reduced by the triple regimen. These data suggest a strategy to target proteinuria to

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try to achieve regression of renal disease in diabetic patients who do not fully benefit RAS inhibition alone.

Lack of the lectin-like domain of thrombomodulin worsens Shigatoxin (Stx)-induced HUS in mice.
In collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation Mutations of thrombomodulin (TM), a transmembrane endothelial cell glycoprotein with anticoagulant, anti-inflammatory and cytoprotective properties, have been recently implicated in the development of atypical HUS. We and others have found that in vitro TM expression was decreased in microvascular endothelial cells exposed to Stx, the causative agent of diarrheaassociated HUS (D+HUS), suggesting that impaired TM activity/levels may play a role also in D+HUS. In a murine model of Stx-HUS we studied the effect on disease course and severity of deletion of the lectin-like domain of TM that is critical for its anti-inflammatory and cytoprotective properties. Disease was induced by co-injection of Stx2 and LPS in mice lacking the lectin-like domain (TMLed/Led mice) or in wild type mice (TMwt/wt ). TMLed/Led mice had higher mortality after Stx2/LPS than TMwt/wt mice. TMLed/Led mice exhibited more severe thrombocytopenia and renal failure compared to TMwt/wt. In TMLed/Led mice, intraglomerular fibrinogen deposits were detected earlier (at 3h) than in TMwt/wt mice (at 6h). More abundant fibrinogen deposits were also found in the brain of TMLed/Led mice. Lack of the lectin-like domain of TM resulted in a stronger inflammatory reaction in the kidney after Stx2/LPS, with a higher number of neutrophils infiltrating glomeruli and renal interstitium. Interstitial accumulation of monocytes/macrophages was also more abundant. These data document that TMLed/Led mice exhibited earlier onset of HUS that, over the follow-up, was worse than in the TMwt/wt mice, suggesting that loss of TM function may increase susceptibility to the development of thrombotic microangiopathic lesions after Stx infection. These findings suggest that TM may be a therapeutic target for D+HUS.

Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation

Autologous Mesenchymal Stromal Cells and Kidney Transplantation: A pilot study of safety and clinical feasibility Multipotent mesenchymal stromal cells (MSCs) possess powerful immunomodulatory activity highlighting the potential for their clinical translation as a novel cell-based approach in solid organ transplantation. A safety and clinical feasibility study (ClinicalTrials.gov, NCT00752479) of autologous MSC infusion in recipients of kidneys from living-related donors is ongoing at our Centre. We reported on the first two patients. Patients were given T cell-depleting induction therapy and maintenance immunosuppression with cyclosporine and mycophenolate mofetil. On day 7 posttransplant, MSCs were administered intravenously. In both MSC-treated patients serum creatinine levels increased 7 to 14 days after cell infusion. A graft biopsy in patient 2 excluded acute graft rejection, but showed a focal inflammatory infiltrate, mostly granulocytes. In patient 1 protocol biopsy at 1-year posttransplant showed a normal graft. Both MSC-treated patients are in good health with stable graft function. A progressive increase of the percentage of CD4+CD25highFoxP3+CD127- Treg and a marked inhibition of memory CD45RO+RA-CD8+ T cell expansion were observed posttransplant. Patient T cells showed a profound reduction of CD8+ T cell activity.

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Findings from this study in the two patients show that MSC infusion in kidney transplant recipients is feasible, allows enlargement of Treg in the peripheral blood, and controls memory CD8+ T cell function. Future clinical trials with MSCs to look with the greatest care for unwanted side effects is advised.

Relative role of genetic complement abnormalities in sporadic and familial aHUS and their impact on clinical phenotype
Hemolytic uremic syndrome (HUS) is characterized by microangiopathic hemolytic anemia, thrombocytopenia, and renal impairment. Most childhood cases are caused by Shiga toxinproducing bacteria. The other form, atypical HUS (aHUS), accounts for 10% of cases and has a poor prognosis. Genetic complement abnormalities have been found in aHUS. We screened 273 consecutive patients with aHUS for complement abnormalities and studied their role in predicting clinical phenotype and response to treatment. We compared mutation frequencies and localization and clinical outcome in familial (82) and sporadic (191) cases. In >70% of sporadic and familial cases, gene mutations, disease-associated factor H (CFH) polymorphisms, or anti-CFH autoantibodies were found. Either mutations or CFH polymorphisms were also found in the majority of patients with secondary aHUS, suggesting a genetic predisposition. Familial cases showed a higher prevalence of mutations in SCR20 of CFH and more severe disease than sporadic cases. Patients with CFH or THBD (thrombomodulin) mutations had the earliest onset and highest mortality. Membrane-cofactor protein (MCP) mutations were associated with the best prognosis. Plasma therapy induced remission in 55 to 80% of episodes in patients with CFH, C3, or THBD mutations or autoantibodies, whereas patients with CFI (factor I) mutations were poor responders. aHUS recurred frequently after kidney transplantation except for patients with MCP mutations. Results underline the need of genetic screening for all susceptibility factors as part of clinical management of aHUS and for identification of patients who could safely benefit from kidney transplant.

Laboratory of Cell and Gene Therapy Mechanism of cell-to-cell communication between MSc and damaged tubular cells
In collaboration with the Laboratory of Cell Biology and Xenotransplantation Recent studies demonstrated that exogenous bone marrow-mesenchymal stem cells (BM-MSCs) may contribute to the recovery of tissue injury in several organs. We demonstrated that administration of mouse BM-MSC in a mouse model of cisplatin-induced acute kidney injury (AKI) ameliorates renal dysfunction and repairs tubular damage. This effect is possibly due to the ability of MSCs to promote renal tubular cell proliferation. The low number of MSC found within the renal tissue after injury together with the increased proliferation of tubular cells suggest that MSCs may exert their effects by a paracrine action on resident cells. One of the projects of the Department is to characterize the signalling mediators involved in the mechanism of cell-to-cell communication between MSCs and damaged tubular cells (growth factors, exosomes and microvesicles). We demonstrated that mouse and human BM-MSC release microvesicles (MV) and exosomes in the microenvironment. MV/exosomes were isolated from conditioned medium by serial ultracentrifugation and their identity was confirmed by electron microscopy. MV/exosomes are enriched in microRNA, mRNA and proteins. By in vitro experiments, we demonstrated that MV/exosomes horizontally transfer their mRNAs to damaged tubular cells, increasing their proliferation. We are now evaluating the possible pathways involved in such an effect and which pathways are modulated in targeted cells exposed to MSC-derived MV/exosomes.

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Induction of pluripotent stem cells from somatic cells

In collaboration with the Laboratory of Cell Biology and Xenotransplantation The therapeutic effects of human embryonic stem cells (hES cells) have been reported in several animal models of degenerative diseases, but only in January 2009 the US FDA approved the first clinical trial for treating patients affected by spinal cord injury with hES cells. The clinical applications of hES cells are associated with some obstacles: the immune rejection after transplantation and ethical concerns about the use of embryos. The induced pluripotent stem cells (iPS) technology could overcome the obstacles associated with hES cells. The iPS cells refer to pluripotent stem cells that are artificially induced from differentiated cells by introducing four transcription factors (OCT4, KLF4, SOX2 and cMyc) highly expressed in hES cells. Several methods have been used to deliver the four transcription factors into the somatic cells, but the plasmid-based transfection seems to be safer because it can avoid the integration into the host genome. On the basis of these evidences, our group cloned the four transcription factors in a single plasmid vector that ensures the same expression level for all factors into the differentiated cells. We have chosen to reprogram human adult dermal fibroblast cells because they are easy to obtain from patients. Preliminary experiments led to only partially reprogrammed iPS-like colonies. To overcome this problem we are now trying to improve transfection efficiency and the cell culture protocols. Recently, it has been reported that transfection with the RNA for the four transcription factors can reprogram adult cells with a high efficiency. This method seems to be safer then plasmid vector because the RNA cant integrate in the host genome. Given these data, our group is trying to optimize the in vitro RNA transcription technique in order to generate a unique RNA with all the four reprogramming factors. The final goal of our research will be to obtain iPS cells from patients affected from rare genetic diseases in order to get an in vitro model of the disease to study and correct the genetic defect.

MicroRNAs Involvement in the progression of the renal disease In collaboration with the Laboratory of Renal Biophysics (Department of Bioengineering) MicroRNAs are small, endogenously expressed non-coding RNA molecules that regulate target gene expression through translation repression or messenger RNA degradation. MicroRNAs are involved in a number of physiological processes like development, cellular proliferation, and apoptosis but also in pathological conditions such as in cardiovascular diseases and in cancer. One of the research lines of the Department is the identification of possible microRNAs modulated during the progression of renal disease in order to find gene and mechanisms predisposing to the renal damage. Munich Wistar Fromter (MWF) rats represent a genetic experimental model of spontaneous glomerulosclerosis, characterized by superficial glomeruli, reduced number of nephrons, increased glomerular volume and proteinuria. The male MWF rats develop hypertension, massive proteinuria and glomerulosclerosis with age. The reduction in podocytes number is an important determinant of podocytes dysfunction and progressive impairment of the glomerular permselectivity that leads to renal scarring. Taking advantage of this model, we performed a microRNA expression profile in isolated glomeruli and identified microRNAs differentially expressed in MWF rats in respect to control rats. We focused our attention on the most upregulated microRNA and, using different algorithms, we computationally predicted possible microRNA targets.

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Gene therapy to correct Thrombotic Thrombocytopenic Purpura In collaboration with the Department of Bioengineering
Thrombotic Thrombocytopenic Purpura is a rare thrombotic microangiopathy characterized by anemia and thrombocytopenia. The disease manifests prevalent, but not exclusive, neurologic symptoms and renal dysfunction. TTP is caused by deficiency of ADAMTS13 activity. ADAMTS13 is a plasma protein that regulates thrombi formation by cleaving von Willebrand (vWF) factor multimers secreted from endothelial cells. The treatment of choice for patients with familial TTP is plasma infusion that, however, exposes patients to high risk of infections, fluid volume overload and allergies. These limitations prompted us to look for an alternative therapy. Recently, we demonstrated that gene therapy restores ADAMTS13 plasma levels and activity in knockout mice. Indeed, intravenous administration of a recombinant adenoviral vector encoding for human ADAMTS13 into ADAMTS13-/- mice induces the release into the plasma of large amounts of functionally active ADAMTS13 protein, able to reduce the thrombi area in respect to control mice. Despite these promising results the use of an adenoviral vector as a gene transfer tool causes the production of antibodies against both the adenovirus and the transgene reducing the expression and activity of the recombinant protein in the long-term. In order to ameliorate our gene therapy approach we are now evaluating alternative gene delivery strategies based on the use of transposons. We are going to use the Sleeping Beauty (SB) system consisting of a plasmid containing ADAMTS13 cDNA flanked by inverted terminal repeats (ITR) typical of the SB transposon and a plasmid encoding for the transposase, a protein able to cut DNA at the level of the ITRs and to paste it into the genome. Of note, no dominant adverse effects associated with SB vectors integration have been observed in experimental animal and this system allows long-term expression of the transgene. Moreover, in order to overcome transgene expression in dendritic cells responsible for T cells activation, we inserted the target sequence for microRNA (miR)142-3p in the 3UTR of the ADAMTS13 cDNA. The miR142-3p is present in dendritic cells but not in hepatocytes. The perfect complementarity between the microRNA and its target sequence will suppress the expression of ADAMTS13 in dendritic cells, preserving ADAMTS13 into the hepatocytes, the constitutive site of ADAMTS13 production.

Imaging of the porous ultrastructure of the glomerular epithelial filtration slit

The ultrastructure of the glomerular filtration pores is still controversial. In the last 30 years, observations from transmission electron microscopy (TEM) and theoretical analysis of solute clearance produced conflicting results. In this study, the employment of a sophisticated technological approach permitted to observe details never detected in the past. Here, we used scanning electron microscopy (SEM) of last generation (Cross-Beam Supra con colonna Gemini, Carl Zeiss, Oberkochen, Germania) with a high-sensitivity detector to image the deepest regions of the filtration pores. In contrast to previous TEM imaging, we observed, in kidney samples of control animals, circular and ellipsoidal pores mainly located in the central region of the podocyte junctions. The normal mean pore radius estimated by digital morphometric analysis had a log-normal distribution, with an average value of 12.1 nm. Of note, in the kidney of proteinuric rats, the mean pore radius values were also log-normally distributed with the presence, however, of some very large pores, exceeding the sizes observed in normal conditions. These exceptionally large pores may be responsible for the increased filtration of plasma proteins in glomerular disease.

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DEPARTMENT OF BIOMEDICAL ENGINEERING

STAFF

Head

Andrea REMUZZI, Eng. D.

Laboratory of Renal Biophysics

Head Daniela MACCONI, Biol.Sci.D.

Laboratory of Biomedical Technologies

Head Unit of Tissue Engineering Head Marina FIGLIUZZI, Biol.Sci.D. Bogdan ENE-IORDACHE, Eng.D.

Unit of Medical Imaging Head Luca ANTIGA, Ph.D.

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CURRICULA
Andrea Remuzzi got his degree in Mechanical (Biomedical) Engineering in 1979, Politecnico di Milano. Research experience: 1980 Politecnico di Milano, Dipartimento di Ingegneria Biomedica; 1981 Istituto Mario Negri (Milano), Laboratorio di Farmacologia Cardiovascolare; 1982-83 Massachusetts Institute of Technology, Mechanical Engineering Department, Cambridge, USA. Areas of interest: biological transport phenomena, mathematical models, renal pathophysiology, cellular response to mechanical stimulation, tissue engineering, pancreatic islet transplantation, clinical databases, computational fluid dynamics. Chronology of appointment: From 1984 to 1986 Ricercatore Istituto Mario Negri (Bergamo), Laboratorio di malattie renali, 1986-1989 Head, Unit di Bioingegneria, Istituto Mario Negri, 1989-1993 Head, Laboratorio di Bioingegneria, Istituto Mario Negri, 1993-1999 Head, Dipartimento di Ricerca Renale, Istituto Mario Negri, from 2000 Head, Dipartimento di Bioingegneria, Istituto Mario Negri. From 1998 to 2007 contract professor of Bioengineering, Politecnico di Milano. For 2007 Professor of Bioengineering, University of Bergamo. Selected publications
Davies PF, Remuzzi A, Gordon EJ, Dewey CF Jr, Gimbrone MA Jr. Turbulent fluid shear stress induces vascular endothelial cell turnover in vitro. Proc Natl Acad Sci U S A. 1986 Apr;83(7): 2114-7. PMID: 3457378 Remuzzi A, Puntorieri S, Battaglia C, Bertani T, Remuzzi G. Angiotensin converting enzyme inhibition ameliorates glomerular filtration of macromolecules and water and lessens glomerular injury in the rat. J Clin Invest. 1990 Feb;85(2):541-9. PMID: 1688888 Noris M, Morigi M, Donadelli R, Aiello S, Foppolo M, Todeschini M, Orisio S, Remuzzi G, Remuzzi A. Nitric oxide synthesis by cultured endothelial cells is modulated by flow conditions. Circ Res. 1995 Apr;76(4):536-43. PMID: 7534657 Giavazzi R, Foppolo M, Dossi R, Remuzzi A. Rolling and adhesion of human tumor cells on vascular endothelium under physiological flow conditions. J Clin Invest. 1993 Dec;92(6):3038-44. PMID: 7504697 Antiga L, Ene-Iordache B, Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of blood vessels from MR and CT angiography. IEEE Trans Med Imaging. 2003 May;22(5):674-84. PMID: 12846436 Remuzzi A, Gagliardini E, Sangalli F, Bonomelli M, Piccinelli M, Benigni A, Remuzzi G. ACE inhibition reduces glomerulosclerosis and regenerates glomerular tissue in a model of progressive renal disease. Kidney Int. 2006 Apr;69(7):1124-30. Ruggenenti P, Remuzzi A, Remuzzi G. Decision time for pancreatic islet-cell transplantation. Lancet. 2008 371:883-4. Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Bruno S, Remuzzi G, Remuzzi A. Computed tomography evaluation of autosomal dominant polycystic kidney disease progression: a progress report. CJASN 2006 1:754-60. Cornolti R, Figliuzzi M, Remuzzi A. Effect of micro-and macroencapsulation on oxygen consumption by pancreatic islets. Cell Transplant. 2009; 18(2): 195-201. Remuzzi A, Cornolti R, Bianchi R, Figliuzzi M, Porretta-Serapiglia C, Oggioni N, Carozzi V, Crippa L, Avezza F, Fiordaliso F, Salio M, Lauria G, Lombardi R, Cavaletti G. Regression of diabetic complications by islet transplantation in the rat. Diabetologia 2009 sep 30; 52: 2653-2661. Cornolti R, Cattaneo I, Trudu M, Figliuzzi M, Remuzzi A. Effect of islet transplantation on metabolic glucose control in rats with diabetes. Diabetes Technol Ther. 2009 Dec; 11(12):805-11 Gagliardini E, Conti S, Benigni A, Remuzzi G, Remuzzi A. Imaging of the Porous ultrastructure of the glomerular epithelial filtration Slit. Journal American Society Nephrology 21: 20812089, 2010.

Daniela Macconi got her Biol.Sci.D. degree in Milan in the 1983. Research experience: 1977-81 CNR Institute of Neuroscience - Cell Mol Pharmacology - and Department of Medical Pharmacology, University of Milan, Milan, Italy; 1982-83 Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Bergamo, Italy; 1984-85 University of Michigan, Medical School, Department of Pathology, Medical Science I, Ann Arbor Michigan, USA; 1985-89 Mario Negri Institute for Pharmacological Research, Laboratory of Kidney Disease, Bergamo, Italy. Areas of interest: glomerular permeability, renal disease progression, podocytes, angiotensin II, reactive oxygen species Chronology of appointment: From 2000 Head Laboratory of Renal Biophysics, Department of Biomedical Engineering; 1994-2000 Head, Unit of Inflammatory Mediator of Leukocyte Origin; 198994 Scientist, 1985-89 post-doctoral fellow Mario Negri Institute for Pharmacological Research, Bergamo,

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Italy; 1982-83 fellow Laboratory of the Division of Nephrology e Dialysis, Ospedali Riuniti di Bergamo, Bergamo, Italy Selected publications
Macconi D. Targeting the renin angiotensin system for remission/regression of chronic kidney disease. Histol Histopathol 25:655-68, 2010. Review Macconi D, Sangalli F, Bonomelli M, Conti S, Condorelli L, Gagliardini E, Remuzzi G, Remuzzi A. Podocyte repopulation contributes to regression of glomerular injury induced by ACE inhibition. Am J Pathol 174:797-807, 2009 Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic peptides. J Am Soc Nephrol 20:123-30, 2009. Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A, Remuzzi G: Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target for renoprotective intervention. Am J Pathol 168:1073-85, 2006. Macconi D, Bonomelli M, Benigni A, Plati T, Sangalli F, Longaretti L, Conti S, Kawachi H, Hill P, Remuzzi G, Remuzzi A. Pathophysiologic implications of reduced podocyte number in a rat model of progressive glomerular injury. Am J Pathol 68:42-54, 2006. Morigi M, Buelli S, Zanchi C, Longaretti L, Macconi D, Benigni A, Moioli D, Remuzzi G, Zoja C. Shigatoxin induced endothelin-1 expression in cultured podocytes autocrinally mediates actin remodeling. Am J Pathol 169:1965-75, 2006 Morigi M, Macconi D, Zoja C, Donadelli R, Buelli S, Zanchi C, Ghilardi M, Remuzzi G: Protein overload-induced NFkappaB activation in proximal tubular cells requires H(2)O(2) through a PKC-dependent pathway. J Am Soc Nephrol 13:1179-89, 2002 Macconi D, Ghilardi M, Bonassi ME, Mohamed EI, Abbate M, Colombi F, Remuzzi G, Remuzzi A: Effect of angiotensin-converting enzyme inhibition on glomerular basement membrane permeability and distribution of zonula occludens-1 in MWF rats. J Am Soc Nephrol 11:477-89, 2000.

Bogdan Ene-Iordache got his M.Sc. in Mechanical Engineering in 1990 at the Petroleum & Gas University in Ploiesti (Romania). In 1992 he joined the Bioengineering Laboratory at NegriBERGAMO Laboratories. Main interests: renal research (hemodynamics and remodeling of the arteriovenous fistula for vascular access, morfometry of renal glomeruli) and controlled clinical trials (data management and data analysis). Other research interests include clinical research informatics (web-based applications) and applied clinical informatics (development of Electronic Health Record - EHR). Roles: since January 2000 is the Head of the Biomedical Technologies Laboratory, Department of Biomedical Engineering. He is coordinating the IT activities in the Clinical Research Center for Rare Diseases Aldo e Cele Dacc. Selected publications Ene-Iordache B, Imberti O, Foglieni O, Remuzzi G, Bertani T and Remuzzi A. Effects of angiotensin-converting enzyme inhibition on glomerular capillary wall ultrastructure in MWF/Ztm rats. J Am Soc Nephrol 5: 1378-1384, 1994. Ene-Iordache B and Remuzzi A. Numerical analysis of blood flow in reconstructed glomerular capillary segments. Microvasc Res 49: 1-11, 1995. Ene-Iordache B, Mosconi L, Remuzzi G, Remuzzi A. Computational fluid dynamics of a vascular access case for hemodialysis. J Biomech Eng 123(3): 284-292, 2001. Ene-Iordache B, Mosconi L, Antiga L, Bruno S, Anghileri A, Remuzzi G, Remuzzi A. Radial artery remodeling in response to shear stress increase within arteriovenous fistula for hemodialysis access. Endothelium 10(2): 95-102, 2003. Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. NEJM 351(19): 1941-1951, 2004. Ene-Iordache B, Carminati S, Antiga L, Rubis N, Ruggenenti P, Remuzzi G and Remuzzi A. Developing regulatory-compliant electronic case report forms for clinical trials: experience with the DEMAND trial. J Am Med Inform Assoc, 16(3):404-408, 2009.

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Marina Figliuzzi got her Biol.Sci.D. degree in Milan in the 1991. Research experience :1991-94 Mario Negri Institute for Pharmacological Research, Bergamo, Italy. Areas of interest: techniques of kidney decellularization, isolation of pancreatic islets from human, bovine, pig and rat pancreas, cell culture, immunoisolation devices for pancreatic islets, differentiation of progenitor pancreatic cells in insulin containing cells, immunhistochemistry. Chronology of appointment: From 2000 Head Unit of Tissue Engineering, Department of Biomedical Engineering; 1991-2000 fellow laboratory of Renal research, Mario Negri Institute for Pharmacological Research, Bergamo, Italy. Selected publications Figliuzzi M, Bonandrini B, Cattaneo I, Remuzzi G, Remuzzi A. Effect of inborn pancreatic islet deficit in the Munich Wister Frmter rat. Islets. 2010 Sep 1;2(5):318-22. Cornolti R, Cattaneo I, Trudu M, Figliuzzi M, Remuzzi A.Effect of islet transplantation on metabolic glucose control in rats with diabetes.Diabetes Technol Ther. 2009 Dec;11(12):805-11. Remuzzi A, Cornolti R, Bianchi R, Figliuzzi M, Porretta-Serapiglia C, Oggioni N, Carozzi V, Crippa L, Avezza F, Fiordaliso F, Salio M, Lauria G, Lombardi R, Cavaletti G. Regression of diabetic complications by islet transplantation in the rat. Diabetologia. 2009 Dec;52(12):26532661. Figliuzzi M, Cornolti R, Perico N, Rota C, Morigi M, Remuzzi G, Remuzzi A, Benigni A. Bone marrow-derived mesenchymal stem cells improve islet graft function in diabetic rats. Transplant Proc. 2009 Jun;41(5):1797-800. Cornolti R, Figliuzzi M, Remuzzi A. Effect of micro- and macroencapsulation on oxygen consumption by pancreatic islets.Cell Transplant. 2009;18(2):195-201. Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A.Assessment of in vitro differentiation of bovine pancreatic tissue in insulin-expressing cells. JOP 9(5):601-11, 2008. Figliuzzi M, Plati T, Cornolti R, Adobati F, Fagiani A, Rossi L, Remuzzi G, Remuzzi A. Biocompatibility and function of microencapsulated pancreatic islets. Acta Biomater. 2006 Mar;2(2):221-7. Figliuzzi M, Cornolti R, Plati T, Rajan N, Adobati F, Remuzzi G, Remuzzi A: Subcutaneous xenotransplantation of bovine pancreatic islets. Biomaterials. 26:5640-47, 2005. Figliuzzi M, Zappella S, Morigi M, Rossi P, Marchetti P, Remuzzi A: Influence of donor age on bovine pancreatic islet isolation. Transplantation. 70:1032-37, 2000

Luke Antiga graduated in 1999 in Biomedical Engineering and got his PhD in Bioengineering in 2003, Politecnico di Milano, having worked at the research laboratory of Biomedical Technology, Department of Bioengineering Institute Mario Negri. Training activities: 2003 Post-doctoral fellow at Imaging Research Laboratories, Robarts Research Institute, London, Ontario. Areas of interest: acquisition and image processing for medical and microscopy applications, numerical modeling of transport phenomena. Roles: from 2000 to 2002 Doctoral Student at the Laboratory of Biomedical Technology, Department of Bioengineering, from 2002 to 2003 visiting scientist Robarts Institute for medical Imaging, London, Ontario, Canada, from 2004 to 2006 resercher at the Laboratory of Biomedical Technology, Department of Bioengineering, from 2007 Head Unit of Medical Imaging, Department of Bioengineering. Selected publications
Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Bravo RF, Carminati S, De Leon FR, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A and Remuzzi G. Sirolimus therapy to halt the progression of Autosomal Dominant Polycystic Kidney Disease: The SIRENA study. JASN, 21(6):1031-1040, 2010. Caroli A, Antiga L, Cafaro M, Fasolini G, Remuzzi A, Remuzzi G and Ruggenenti P. Reducing polycystic liver volume in ADPKD: effects of the extended release somatostatin analogue octreotide. CJASN, 5(5): 783-789, 2010. Botti L, Piccinelli M, Ene-Iordache B, Remuzzi A and Antiga L. An adaptive mesh refinement solver for largescale simulation of biological flows. Communications in Numerical Methods in Engineering, 26(1): 86-100, 2010. Antiga L, and Steinman DA. Rethinking turbulence in blood. Biorheology, 46(2): 77-81, 2009. Piccinelli M, Veneziani A, Steinman DA, Remuzzi A and Antiga L. A framework for geometric analysis of vascular structures: application to cerebral aneurysms. IEEE Transactions on Medical Imaging, 28(8): 1141-55, 2009.

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Botti L, Piccinelli M, Ene-Iordache B, Remuzzi A and Antiga L. An adaptive mesh refinement solver for large-scale simulation of biological flows. Communications in Numerical Methods in Engineering, 26(1): 86-100, 2010. Ene-Iordache B, Carminati S, Antiga L, Rubis N, Ruggenenti P, Remuzzi G, Remuzzi A. Developing regulatorycompliant electronic case report forms for clinical trials: the DEMAND trial. JAMIA, 16(3): 404-408, May-Jun 2009. Lee SW, Antiga L and Steinman DA. Correlations among indicators of disturbed flow at the normal carotid bifurcation. Journal of Biomechanical Engineering, 131(6): , Jun 2009. Antiga L, Piccinelli M, Botti L, Ene-Iordache B, Remuzzi A and Steinman DA. An image-based modeling framework for patient-specific computational hemodynamics. Medical and Biological Engineering and Computing, 46: 1097-1112, Nov 2008. Antiga L, Wasserman B, Steinman D. On the overestimation of early wall thickening at the carotid bulb by black blood MRI, with implications for coronary and vulnerable plaque imaging. Magnetic Resonance in Medicine, 60(5): 10201028, Nov 2008. Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Ruggenenti P, Remuzzi G and Remuzzi A. Computed tomography evaluation of ADPKD progression: a progress report. Clinical Journal of the American Society of Nephrology (CJASN), 1(4): 754-760, Jul 2006. Thomas JB, Antiga L, Che S, Milner JS, Hangan Steinman DA, Spence JD, Rutt BK and Steinman DA. Variation in the carotid bifurcation geometry of young vs. older adults: Implications for "geometric risk" of atherosclerosis. Stroke, 36(11): 2450-2456, Nov 2005. Antiga L, Steinman DA. Robust and objective decomposition and mapping of bifurcating vessels. IEEE Transactions on Medical Imaging, 23(6): 704-713, June 2004. Antiga L, Ene-Iordache B and Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of blood vessels from MR and CT angiography. IEEE Transactions on Medical Imaging, 22(5): 674-684, May 2003. Antiga L, Ene-Iordache B, Remuzzi G and Remuzzi A. Automatic generation of glomerular capillary topological organization. Microvascular Research, 62: 346-354, June 2001.

INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES

The Department of Bioengineering conducts research activities in biomedicine, both at experimental and clinical level. Physiopathological processes are studied using engineering techniques with the aim to develop innovative treatment strategies. Several lines of research in basic, applied clinical research are currently active within the Department. The main tools used for this research consist of theoretical models, diagnostic imaging, histological measures, physical and chemical parameters in both experimental and clinical studies, cell culture techniques, biomaterials, computational technologies for archiving and processing clinical data. The ongoing studies relate to four main areas: 1) study of the mechanisms involved in the progression of chronic nephropathy; 2) studies on the role of hemodynamics in the development of vascular diseases; 3) development of laboratory techniques for tissue engineering; 4) development of information systems to manage clinical data and digital images generated in the context of controlled clinical trials and in routine clinical practice.

FINDINGS/MAIN RESULTS
We produced evidence demonstrating a possible relationship between the geometry of the cerebral arterial vessels, location of cerebral aneurysms. These findings are opening new perspectives in the understanding of the role of hemodynamic conditions as responsible for this disease. Demonstration of a significant relationship between morphometric parameters of the renal parenchyma, quantified through numerical analysis of CT images, and the loss of renal function in patients with polycystic kidney disease. Demonstration that the mechanism responsible for regeneration of the glomerular capillary, induced by drugs that inhibit the angiotensin II, depends on proliferation of parietal cells of the Bowmans capsule and their subsequent migration on to the glomerular capillary loops.

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Implementation of a paper less web-based system for the management, of clinical data generated in controlled clinical trials, in compliance with GCP. Set up a new network of specialists in Nephrology for long-term data collection aimed at monitoring the quality of treatment of chronic progressive nephropathy in current clinical practice.

NATIONAL COLLABORATIONS
Dipartimento di Bioingegneria, Politecnico di Milano, Milano. Unit di Diabetologia, Ospedali Riuniti, Bergamo. STMicroelectronics, Agrate Brianza, Milano Matematici, Universit di Bergamo Dipartimento di scienze Neurologiche e della visione, Universit di Verona. Dipartimento di Ingegneria Industriale e Dipartimento di Ingegneria dellinformazione e metodi Matematici Facolt di Medicina e Chirurgia, Universit degli studi di Milano

INTERNATIONAL COLLABORATIONS
Massachussetts Institute of Technology, Cambridge MA, USA. National Alliance for Medical Imaging Computing, USA. Harvard Medical School, Cambridge MA, USA. Department of Mathematics and Computer Science, Emory University, Atlanta, Georgia, US. Simula Laboratories, Oslo, Norway Academisch Medisch Centrum, Amsterdam, the Netherlands University of Toronto, Ontario, Canada. Ghent University, Ghent, Belgium. Technical University, Eindhoven, The Netherlands. University Hospital, Maastricht, The Netherlands. Universzitetni Klinikni Center Ljubjana, Ljubljana, Slovenia. The University of Sheffield, Sheffield, United Kingdom.

ESAOTE, Maastricht, The Nederland. EDITORIAL BOARD MEMBERSHIP

International Journal of Artificial Organs, (Andrea Remuzzi)

PEER REVIEW ACTIVITIES

Annals of Biomedical Engineering ASME Journal of Biomechanical Engineering

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Journal of Vascular Research Magnetic Resonance in Medicine Stroke Journal of the American Society of Nephrology Kidney International American Journal of Kidney Diseases American Journal of Pathology American Journal of Physiology Medical & Biological Engineering & Computing New England Journal of Medicine IEEE Transactions on Medical Imaging IEEE Transactions on Biomedical Engineering Medical Physics Journal of Biomechanics Medical Engineering and Physics Artificial Organs International Journal of Artificial Organs Biomaterials Contemporary Clinical Trials Journal of Endocrinological Investigation Pediatric Nephrology Nephrology Dialysis Translantation Acta Diabetologica

EVENT ORGANIZATION
Seminar: "Sviluppo di un sistema per l'elaborazione di immagini digitali al microscopio per la quantificazione di componenti cellulari in tre dimensioni, Benedetta Nodari (Universit degli studi di Bergamo, Facolt di Ingegneria Informatica), Sala Riunioni Centro Anna Maria Astori Km Rosso Bergamo. Seminar: "Hemodynamic factors in cardiovascular disease: Bridging the gap between Engineering and Medicine" Prof. David Steinman (Professor of Mechanical and Biomedical Engineering at the University of Toronto), October, Sala Conferenze Centro Anna Maria Astori Km Rosso - Bergamo. Seminar: " Biomedical Engineering: from basic research to clinical application. The Maastricht University Medical Center Experience", Dr. Frans Van de Vosse (Prof. of Cardiovascular Biomechanics, Eindhoven University of Technology, Netherlands), 10 Anni del Dipartimento di BIOINGEGNERIA, November, Sala Consiliare - Ospedali Riuniti di Bergamo. Seminar: "Evaluation of different DW-MRI Protocols on ex vivo mouse spinal cord", Jeire Steinbuch (Eindhoven University of Technology, Medical Engineering - The Netherlands), July, Sala Conferenze Centro Dacc Bergamo.

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Seminar: "Computational modelling and CFD models for hemodynamic behaviour in AVF", Raf Van Hoof (Eindhoven University of Technology, Biomedical technology - The Netherlands), July, Sala Conferenze Centro Dacc Bergamo. Seminar: "Sviluppo e testing di software web-based per il progetto COMGAN", Lorenzo Fugazza (Universit degli Studi di Bergamo, Facolt di Ingegneria), July, Sala Conferenze Centro Dacc Bergamo. Seminar: "Ingegneria del tessuto renale progetto e sperimentazione di un sistema per la decellularizzazione del rene", Paola Cucchetti (Dipartimento di Bioingegneria, Politecnico di Milano POLIMI), July, Sala Conferenze Centro Dacc Bergamo. Seminar: "Studio spettroscopico e morfologico della interazioni tra sistemi biologici e materiali organici ed inorganici", Prof. Loredana Latterini (Dipartimento di Chimica, Universit di Perugia), March, Sala Conferenze Centro Dacc Bergamo. Seminar: "Studio numerico dell'emodinamica nelle fistole artero-venose di tipo end-to-side utilizzate come accesso vascolare nell'emodialisi", Cristina Semperboni, February, Sala Conferenze Centro Dacc Bergamo.

PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED

VPH NoE WP2/WP3/VPH-I Meeting, 4th-5th February 2010, Laboratory of Anatomy, Biomechanics, Organogenesis, Universite Libre de Bruxelles (ULB) Erasme Campus, Bruxelles(The VPH ToolKit A Focus on Sustainability and Cooperation). 14th European Vascular Course(EVC2010), February 25-27, 2010 Maastricht, The Netherlands. CO.R.TE 2010: Conferenza Italiana per lo Studio e la Ricerca sulle Ulcere, Piaghe, Ferite e la Riparazione Tessutale. 4-6 Marzo 2010, Roma Cavalieri, Via Cadlolo, 101, Roma (www.romecavalieri.it) 46th Annual Meeting of the European Association for the Study of Diabetes (EASD), Stoccolma, Sweden Secondo Congresso Nazionale di Bioingegneria, GNB2010, 5-7 Luglio 2010, Torino ESB2010, 17th Congress of the European Society of Biomechanics, 5 - 8 July 2010, University of Edinburgh, UK. Call for an VPH FET Flagship All Hands meeting, Barcelona July 22nd, 2010. Meeting VPH FET Flagship All Hands, Zurigo 10 September 2010.

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VPH Conference 2010, Virtual Physiological Human Network of Excellence, 29-01 September 2010, Brussels. SIAPAV - XXXII Congresso Nazionale, Societ Italiana di Angiologia e Patologia Vascolare, Padova, 17-20 Novembre 2010 - Sheraton Hotel. Renal Master Class Mario Negri Institute, October 7-9 2010 "La Remission Clinic nella pratica clinica: nuove prospettive di prevenzione e trattamento per le nefropatie croniche progressive". Villa Camozzi, Bergamo, sabato18 December 2010. Corso Teorico-Pratico avanzato di eco color Doppler carotideo e vertebrale, Milano 11-13 February 2010.

GRANTS AND CONTRACTS

Research grants AIFA - trial clinici controllati (VARIETY, VALID, ATHENA, ARCADIA, NEMO). Research grant PKD foundation - ALADIN trial Effect of long-acting somatostatin on disease progression in ADPKD: a long-term three year follow-up study. Research grant Baxter ASAP trial Acute Start Access Programme. Research grant ISN per il Kidney Disease Data Center (KDDC ) del COMGAN. Contributo Regione Lombardia per data management del Centro di Coordinamento della Rete Regionale per le Malattie Rare. Project - FP7 UE - ARCH "Patient specific image-based computational modelling for improvement of acute and long-term outcomes of vascular access for hemodialysis FP7-224390 - Project Coordination. VASCOSILK, Fondazione Cariplo - N.536/5314 - Protesi vascolari in fibroina elettrofilata per la rigenerazione in vivo di arterie di piccolo calibro. LIGASILK, Regione Lombardia - N.534/5287 - Bioingegnerizzazione di tendini e legamenti: impiego combinato di supporti tessili in seta e cellule staminali adulte.

SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2010

1. A. Caroli, M. Lorenzi, C. Geroldi, F. Nobili, B. Paghera, M. Bonetti, M. Cotelli, G.B. Frisoni. Metabolic Compensation and Depression in Alzheimers Disease. Dement Geriatr Cogn Disord 2010;29:3745.

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2. Botti L, Piccinelli M, Ene-Iordache B, Remuzzi A, Antiga L. An adaptive mesh refinement solver for large-scale simulation of biological flows. Communications in Numerical Methods in Engineering 2010; 26(1): 86-100. 3. Caroli A, Antiga L, Cafaro M, Fasolini G, Remuzzi A, Ruggenenti P. Reducing polycystic liver in ADPKD: effects of somatostatin analogue octreotide. Clin J Am Nephrol. 2010 Feb 25. 4. Macconi D. Targeting the renin angiotensin system for remission/regression of chronic Kidney disease. Histol Histopathol. 2010 May;25(5):655-68. 5. Steinman DA, Antiga L, Wasserman BA. Overestimation of cerebral aneurysm wall thickness by black blood MRI? Magn Reson Imaging. 2010 Mar;31(3):766. 6. Boccardi M, Almici M, Bresciani L, Caroli A, Bonetti M, Monchieri S, Gennarelli M, Frisoni GB. Clinical and medial temporal features in a family with mood disorders. Neurosci Lett. 2010 Jan 4; 468(2): 93-97. 7. Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Bravo RF, Carminati S, De Leon FR, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A, Remuzzi G. Sirolimus therapy to halt the progression of ADPKD. J Am Soc Nephrol. 2010 Jun;21(6):1031-40. 8. Caroli A, Frisoni GB. The dynamics of Alzheimer's disease biomarkers in the Alzheimer's Disease Neuroimaging Initiative cohort. Neurobiol Aging. 2010 Jun 8. 9. Ruggenenti P, Iliev I, Filipponi M, Tadini S, Perna A, Ganeva M, Ene-Iordache B, Cravedi P, Trevisan R, Bossi A and Remuzzi G. Effect of trandolapril on regression of retinopathy in hypertensive patients with type 2 diabetes: a prespecified analysis of the BENEDICT Trial. J Ophthalmology, 2010. 10. Ruggenenti P, Cattaneo D, Rota S, Iliev I, Parvanova A, Perna A, Diadei O, Ene-Iordache B, Ferrari S, Bossi A, Trevisan R, Belviso A and Remuzzi G. Effects of combined ezetimibe and simvastatin therapy as compared to simvastatin alone in patients with type 2 diabetes: a prospective, randomized, double-blind clinical trial. Diabetes Care, 2010. 11. Ruggenenti P, Perna A, Tonelli M, Loriga G, Motterlini N, Rubis N, Ledda F, Rota S, Satta A, Granata A, Battaglia G, Cambareri F, David S, Gaspari f, Stucchi N, Carminati S, EneIordache B, Cravedi P and Remuzzi G. Effects of add-on fluvastatin therapy in patients with chronic proteinuric nephropathy on dual RAS blockade: the ESPLANADE trial. Clin J Am Soc Nephrol, 2010. 12. Cravedi P, Remuzzi A and Remuzzi G. Comment on: Robertson (2010) Islet Transplantation a Decade Later and Strategies for Filling a Half-Full Glass. Diabetes;59:12851291. Diabetes. 2010 Sep;59(9):e13. 13. Figliuzzi M, Bonandrini B, Cattaneo I, Remuzzi G and Remuzzi A. Effect of inborn pancreatic islet deficit in the Munich Wistar Fromter rat. Islets 2010. 14. Piccinelli M, Bacigaluppi S, Boccardi E, Ene-Iordache B, Remuzzi A, Veneziani A, Antiga L. Geometry of the ICA and recurrent patterns in location, orientation and rupture status of lateral aneurysms: an image-based computational study. Neurosurgery, 2010.

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15. Sharma KS, Hequn Z, Togtokh A, Ene-Iordache B, Carminati S, Remuzzi A, Wiebe N, Ayyalasomayajula B, Perico N, Remuzzi G and Tonelli M. Burden of CKD, proteinuria, and cardiovascular risk among Chinese, Mongolian, and Nepalese participants in the International Society of Nephrology screening programs. Am J Kidney Dis, 2010. 16. Morbiducci U, Gallo D, Ponzini R, Massai D, Antiga L, Montevecchi FM, Redaelli A. Quantitative Analysis of Bulk Flow in Image-Based Hemodynamic Models of the Carotid Bifurcation: The Influence of Outflow Conditions as Test Case. Annals of Biomedical Engineering, 2010. 17. Morbiducci U, Gallo D, Massai D, Consolo F, Ponzini R, Antiga L, Bignardi C, Deriu MA, Redaelli A. Outflow Conditions for Image-Based Hemodynamic Models of the Carotid Bifurcation: Implications for Indicators of Abnormal Flow. Journal of Biomechanical Engineering 2010 Sep;132(9):091005. 18. Gagliardini E, Conti S, Benigni A, Remuzzi G, Remuzzi A. Imaging of the Porous ultrastructure of the glomerular epithelial filtration Slit. Journal American Society Nephrology 21: 2081-2089, 2010.

RESEARCH ACTIVITIES Laboratory of Renal Biophysics

Three dimensional reconstruction of the glomerular capillary network

We have recently documented that angiotensin II blockade not only retards the progression of renal diseases, but also induces regression of the glomerular lesions. Quantification of the extent of the regression of sclerotic lesions and the potential regeneration of the glomerular capillary, induced by a therapy with angiotensin converting enzyme (ACE) inhibitors, can be achieved by a technology based on three dimensional (3D) reconstruction of tissues. A new approach is the vascular corrosion casting that produces replicas of normal and abnormal vasculature and microvasculature of various tissues and organs that can be viewed at the ultrastructural level. We applied this technique to the kidney that, after exanguination, is perfused with a rapidlyhardening polyurethane resin. Once hardening of the resin is complete, the kidney is excised and placed in an alkaline solution for maceration of the soft tissue leaving an intact cast of the renal vasculature to be analyzed by scanning electron microscopy. This allows the morphological analysis of the vascular architecture of the glomerular capillary and the estimation, by morphometry, of the geometrical parameters of the capillary network. Another electron microscopy technique, based on a combination of an electron beam with an ionic one, allows sectioning and acquisition of serial images of the whole glomerulus for 3D reconstruction of the glomerular capillary ultrastructure by mathematical algorithms.

Identification of immunogenic albumin peptides in the kidney of rats with proteinuric nephropathy
(In collaboration with the Department of Molecular Medicine and with the Protein Chemistry/Proteomics Unit, Institute of Biomedicine, Biomedicum, Helsinki, Finland) In proteinuric nephropathies abnormal filtration of proteins, mainly albumin, across the glomerular membrane, causes distress of the proximal tubular cell inducing the release into the interstitium of chemokines responsible for the recruitment and activation of inflammatory cells. The interstitial infiltrates are commonly characterized by immune-competent cells including

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dendritic cells (DCs) and CD8+ T cells that accumulate within renal parenchyma even in the absence of an immune insult. Within the kidney, DCs are in close contact with the tubular epithelium and work as immune sentinels to probe renal interstitium in search for foreign antigens. Upon injury, inflammatory stimuli released from the tubular cell enable DC to become immunogenic towards normally ignored self-antigens. We have recently documented that albumin is processed, through the concerted action of proximal tubular cells and DCs, in immunogenic peptides, thus triggering an immune response (JASN 20:223,2009). Proximal tubular cells in culture exposed to excess autologous albumin, as in case of proteinuric conditions, cleave albumin in the N terminal fragments the most abundant being the 24aminoacid peptide (Alb1-24). This peptide is taken up by DCs and digested, by a proteasomedependent pathway, to antigenic peptides that bear binding motifs for MHC class I and activate syngeneic CD8+ T cells. The functional role of Alb1-24 processing in inducing immune response was further confirmed in a rat model of proteinuria secondary to renal mass reduction (RMR) by 5/6 nephrectomy. Four weeks after surgery, DCs decreased in the kidney and increased in the renal lymph node, suggesting migration of DCs from the renal interstitium. CD8+ T cells isolated from renal lymph nodes from RMR rats were already activated at the first encounter, when incubated with Alb1-24 pulsed DC, suggesting that they were primed by albumin peptide in vivo during the course of the disease. By contrast, in vivo treatment with the proteasome inhibitor bortezomib prevented T cell activation. As follow-up of the study, we sought to evaluate the presence of Alb1-24 in the kidney of rats with overt proteinuria using a new technique, the imaging mass spectrometry that allows simultaneous analysis in the tissue of hundreds of different molecules including peptides/proteins. To this end, we started a collaboration with the Protein Chemistry/Proteomics Unit, Institute of Biomedicine, Biomedicum in Helsinki within the EUROKup (Urine and Kidney Proteomics), a multidisciplinary network from 26 European countries, who is focused on facilitating translational proteomic research in kidney diseases, by promoting interactions between basic scientists and clinicians. Kidney sections from RMR rats analyzed by MALDI (matrix-assisted laser desorption/ionization) imaging mass spectrometry showed the presence of a peptide with an average monoisotopic molecular mass corresponding with the predicted mass of Alb1-24 within and in proximity of the tubular epithelium.

Laboratory of Biomedical Technologies Remission Clinic Network

Many forms of chronic kidney diseases progress with a constant rate of renal function loss towards the end stage renal disease (ESRD). These forms of kidney disease are frequently associated with arterial hypertension and urine proteins, known as aggravating factors in the progression of the disease. Controlled clinical trials have demonstrated that specific treatments of hypertension with drugs that reduce the urinary excretion of proteins (ACE-inhibitors) are effective in reducing the rate of decline of GFR and even in obtaining stabilization or recovery of renal function allowing to delay the start of dialysis or the need of kidney transplant in subjects with chronic kidney disease. In collaboration with the Renal Department we have started a quality control and monitoring programme of patients affected by proteinuric nephropathies (Remission Clinic protocol) aimed at verifying whether GFR improvement might be obtained in routine clinical practice as well. Our laboratory developed a web-based application and established a network of specialists involved in the treatment of chronic progressive nephropathies distributed nationally (http://clinicalweb.marionegri.it/remission). Our tool offers computer support to medical specialists from all participating centers to gather, extract and analyze real time clinical data for patients with chronic kidney diseases treated according to the guidelines of Remission Clinic protocol. In addition, our tool allows real time analyses and quality controls of this clinical

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activity to assess in what extent adherence to the protocol may itself slow the progression of nephropathy in time.

KDDC a centre for data collection and surveillance of prevention programs on non-communicable chronic diseases in emerging countries
Chronic kidney diseases are emerging as a global threat to human health. Prevalence and incidence of renal diseases in developing countries are not known, and this is an obstacle to the adoption of preventive measures. Prevention is the only hope for these countries where treatment options for end stage renal failure are simply not available to the vast majority of the population because of their costs. The International Society of Nephrology (ISN), through the Commission for Global Advancement of Nephrology (COMGAN), has established a research committee in order to face the problem of prevention of kidney diseases in developing countries. The coordination of the team and intervention programs was committed to the Mario Negri Institute for Pharmacological Research at the Clinical Research Centre Aldo e Cele Dacc. The general aim of the project is to define programs in developing countries to identify those subjects who are at risk of developing a renal disease later in life, in order to design a prevention strategy on national basis by means of interventions of the local ministries of health to governmental and financial level. The Kidney Disease Data Centre (KDDC) established in our Laboratory, is dedicated to data management for the prevention programs underway in emerging countries. We have set up an a tool to collect clinical data from different centres located world-wide (http://comgan.marionegri.it). Data are stored in a dedicated server in our Laboratory. Results of our epidemiological analyses, shared also with medical staff of the centre, allow us to have a general overview on the health of population under study. The prevention programme has started in 2006 and today KDDC owns more than 45,000 records of subjects from 15 countries located all over the world. Recently we have published the results of a screening on over 10.000 subjects from Nepal, India and Mongolia showing the burden of chronic diseases in these countries and demonstrating the feasibility of the ISN prevention programme. Through the activity of KDDC it is thus possible to monitor the course of the actual screening projects, to tailor them to the specific needs of each participating country, and even more important to commence follow-up programs in low income countries.

Development of computerized systems for controlled clinical trials

Numerous clinical trials are conducted in the Clinical Research Center for Rare Diseases Aldo e Cel Dacc. These studies must be carried out in accordance with all regulatory requirements (GCP, EMEA, FDA). Every clinical study requires a paper case report form (CRF) for collection of patients clinical observations. These data must be verified for inconsistency by dedicated monitoring staff, and then recorded electronically. In our Lab we have developed applications tailored for data management of clinical studies using relational databases systems (RDBMS) and specific programs aimed to data elaboration, validation and extraction for subsequent statistic analyses. For the DEMAND study we have developed an innovative electronic CRF based on laptop computers. We have implemented also a web-based framework for electronic data capture and clinical data management for clinical trials. Thus, recently we have set-up a dedicated portal (http://clintrials.marionegri.it) as a show-case for the clinical trials conducted at the Clinical Research Center Aldo e Cele Dacc and a platform for the new web-based e-CRFs developed in our Lab.

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Data Management for the Registry for Rare Disease - Lombardy

Our laboratory contributes to the management of the Regional Network for Rare Diseases of Lombardy. We are directly involved for the development and maintenance of the web site of the centre and management of a regional Registry for Rare Diseases. We have set-up the databases and developed related web-pages for centres, rare diseases archive, patient associations and congenital rare diseases. These are published on the homepage of the web-site (http://malattierare.marionegri.it/). The Registry for Rare Diseases was born in 2007 as a collaboration between Mario Negri Institute, Lombardia Informatica (LI) and Regione Lombardia. The aim was to create a regional registry for rare diseases where all medical staff from Lombardy could register information regarding rare diseases. The application (Sistema Malattie Rare - SMR) is actually in use in almost all centres dedicated for rare diseases in Lombardy and can be used jointly with the patient health card.

Hemodynamics and vascular pathology

During the last ten years the presence of a close relationship between hemodynamics and vascular pathology has been pointed out both in the biological and in the clinical field. Typical fields of application are the investigation of the formation of atherosclerotic lesions in the arterial circulation, of intracranial and abdominal aneurysm disease, and of the effect of surgical and endovascular procedures. Thanks to innovative technologies developed during the last few years in the medical imaging and mathematical modeling fields, also within the Biomedical Engineering Department, it is now possible to accurately reproduce patient-specific hemodynamic force distribution from computed tomography (CT) or magnetic resonance (MR) acquisitions. Within the Department of Biomedical Engineering such technologies have three main applications: atherosclerosis (carotid bifurcation and renal artery level), intracranial aneurysm disease and complications of vascular access in hemodialysis. In particular, the Department is currently involved in an international collaborative project (ARCH) funded by the European Commission within the Seventh Framework Programme, aimed at improving the functionality of vascular access in hemodialysis patients, for which the Department is the coordinator centre. Within this project, computational tools for predicting post-operative from pre-operative scenario, aimed to help clinicians to plan for each patient the most suitable vascular access, are under development and validation.

Theoretical and experimental study of filtration of 'albumin in the kidney

Since years the Department have been studied the mechanisms responsible for glomerular filtration of proteins, and in particular, albumin. Recently, we have been dedicated to the theoretical modelling of filtration of albumin and of its concentration in the renal tubule by combining the experimental approach to the theoretical modelling. In an experimental model of nephropathy, we studied the selective properties of the glomerular membrane through infusion of specific fluorescent tracers and changes in albumin concentration in the glomerular filtration. Applying some mathematical models we evaluated the effect of the administration of an ACE inhibitor on the distribution of the membrane pores and the reabsorption of albumin in the tubules. The combination of the experimental approach with the theoretical modelling allowed us to elucidate an aspect of the renal pathophysiology which is still not highlighted such as the quantification of albumin concentration along the nephron either in normal and in pathological conditions.

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Imaging of glomerular filtration slit ultrastructure

In collaboration with the Department of Molecular Medicine. The glomerular capillary wall allows an important filtration of pleasure water but efficiently retains circulating proteins and macromolecules. This selective function is mainly due to the specific ultrastructure of the intercellular junction of the visceral epithelial cells, the podocytes, that allow water passage but retain macromolecules with radius greater than 4nm. The study on the morphology of this cellular junction are based essentially on very few observations of TEM done in the 70s, who suggested a zipper-like structure of the filtration slits, with openings smaller than 4nm. The inconsistency of these observations and the use of a new setup of a scanning electron microscope (SEM) allowed us to study in detail the ultrastructure of the filtration slits in physiological and in pathological conditions. The results are interesting since they disclosed a completely new ultrastructure of the pores of the epithelial filtration slits, and their dimensions.

Imaging and quantification in renal physiopathology

The use of imaging techniques such as CT, MR, and echography, and the application of advanced image processing tools make it possible to perform non-invasive in-vivo quantitative analysis of biological phenomena. Within the Department of Biomedical Engineering, this approach is applied to the investigation of renal physiopathology. Through CT and MR imagebased quantification, new therapies for autosomal dominant polycystic kidney disease (ADPKD) are currently being evaluated. To this purpose, the Medical Imaging Unit has been involved in several clinical trials, some of which still ongoing, one funded by the Polycystic Kidney Foundation, aimed at reducing the overall kidney cyst volume with the use of novel therapies. Using specific automatic algorithms, it has been possible to quantify on contrastenhanced CT images the volume of individual tissue components (cysts and residual parenchyma), beyond total kidney volume only. Moreover, CT image quantification has recently led to the discovery of a fibrotic tissue component (named intermediate volume), highly correlated with both renal function and disease progression rate, showing for the first time a likely direct relationship between structure and function, thus opening the way to new therapeutic targets. Beyond ADPKD studies, new methodologies for noninvasive characterization of renal functionality from diffusion-weighted MR images are currently under study. Preliminary studies on normal control subjects showed high resolution of the images and high reproducibility of the measures, indicative of both renal perfusion and diffusion mechanisms, pointing out the potential of DWI for the investigation of renal physiopathology.

Development of biomedical image analysis and computational modelling tools

The employment of quantitative imaging and mathematical modeling techniques aimed at the study of physiopathological processes is directly linked to medical image management and processing methodologies. Within the Department, research activity related to the development of new image processing algorithms and mathematical models for the numerical simulation of biological phenomena, both theoretical and in terms of software development, is actively performed. The department contributes to the development of three main open-source projects in the biomedical imaging field: Vascular Modeling Toolkit (www.vmtk.org), as main developer, Insight Toolkit (www.itk.org), a state-of-the-art library for medical image analysis, and Slicer 3D (www.slicer.org), one of the main medical imaging applications. The Department is directly involved in the development activities carried out within the National Alliance for Medical Image Computing, an inter-university consortium gathering the main academic institutions of the United States.

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Development of devices for the transplantation of immunoisolated islets

The projects main objective is to develop an immunisolation device for pancreatic islets or insulin producing cells that can be implanted in diabetic rats and to test the efficacy and in vivo resistance of the device. We have defined the most suitable geometry for an immunoisolation device providing large surface area in a small volume. We have developed a device made using parallel arrays of hollow fibers or a membrane device in the form of small coins. The device that we are developing can be implanted with minimally invasive surgical procedures and easy to retry. We have also developed a method for subcutaneous implantation as alternative site for transplantation. We are tested different types of material to be used as polysulfone hollow fiber or membranes of polyvinyl alcohol. The aims of our studies in the next months will be to improve functionality using nanotechnologies for materials characterization. Moreover we will develop new kinds of device and we will test new implantation sites.

Effect of pancreatic islet transplantation on diabetic complications

Diabetes type 1 is taking place as one of the most important worldwide public health problems with a high morbidity and mortality. Diabetes is associated with increased risk of a number of microvascular, neurologic and macrovascular complications due to poor glycemic control. The aim of this project is to evaluate whether the transplantation of pancreatic islets can induce regression of diabetic complications in a model of allotransplantation in rats with chemically induced diabetes. For this study, we used four groups of rats: healthy controls, diabetics, diabetics with transplanted islet four months after induction of diabetes and diabetics treated with insulin to adjust glycemia under 200 mg/dl. Transplantation of islet induces a lowering of blood glucose within a few days after transplantation, accompanied by an increase in body weight. All the neurological parameters, such as the behavioral tests for determination of thermal and mechanical nociceptive threshold and the measurement of nerve conduction velocity in the nerves of the tail (NCV) observed in diabetic rats significantly ameliorate in transplanted rats. Insulin treated rat group show similar improvement of the above described parameters. In conclusion, the transplantation of pancreatic islets not only normalizes blood glycemia levels in diabetic animals, but also reduce the neuropathy getting worse

Development of a protocol for kidney decellularization

Kidney tissue engineering applies the principles and methods of engineering to biological sciences in an attempt to completely regenerate damaged renal tissues affected by chronic nephropathy. This innovative strategy could solve problems related to dialysis therapies and overcome toxicity of immunosuppressive drugs for organ transplantation. The aim of this project is is to generate a biological and active support with the inherent ability to promote the growth, the proliferation and the differentiation of multipotent embryonic stem cells into glomerul, tubular and vascular renal cells. To this purpose we have developed a device includes a roller pump, an hydraulic closed circuit and a perfusion cylindric chamber, and provides the organ with constant physiological flows and pressures to obtain optimal cellular removal and to maintain the 3D architecture of renal ECM matrix. It represents a helpful operative tool whose features were experimented with success on rat kidneys. The proposed method consists in the use of different solutions such as ionic and non-ionic detergents (SDS, Triton X-100), saline, DNase and final washing with antibiotics and antimicrobials. Decellularization of kidney and the presence of the extracellular matrix was confirmed by histological and histochemical analysis. (In collaboration with the Department of Molecular Medicine).

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Vascular tissue engineering

The synthetic vascular grafts are studied for the replacement of large caliber vessels damaged by pathological events. Currently available synthetic vascular grafts are limited to large internal diameter grafts because of frequent thrombosis and occlusion. The alternative is represented from the use of autologous vascular graft, but they are not always available in patients affected from vascular pathology. Vascular tissue engineering has the objective to generate cellularized vascular prosthesis made of biodegradable materials that can be colonized by endothelial cells. For this purpose (in collaboration with Stazione Sperimentale della Seta in Milano and Politecnico di Milano), we have studied an innovative strategy for the vascular prosthesis realization using biodegradable material, the fibroin of the silk. Tubular structures were produced through elettrospinning of the fibrin of the silk, and studies of biocompatibility and biodegradation in vivo were performed. Tubular matrices (=1.8 mm) were implanted in situ in the abdominal aorta of Lewis rats by end-to-end anastomosis in order to evaluate the functionality of the graft. The histological analysis, performed on samples explanted after subcutaneous implantation, showed a thin fibrotic membrane overgrowth around the implant with cell infiltration in the construct. Immuno-fluorescence analysis demonstrated the presence of a low number of macrophages and the absence of T lymphocytes. At 5 days after implantation in abdominal aorta, histological analysis showed vascular tissue neoformation in the luminal side. This tissue is characterized by the presence of smooth muscolar cells and organized structures of neoformed elastin reach extracellular matrix. These data confirmed the regeneration of vascular intima with the presence of smooth muscular cells and elastin, resembling native arteries. These results indicate the great potential of this artificial structure to allow the regeneration of the arterial wall intima, with characteristics of biocompatibility requests to a surface in contact with blood.

The effect of flow on renal tubular cells

ADPKD (Autosomal Dominant Policystic Kidney Disease) is one of the major genetic renal pathologies with an incidence of 1 in 1000 and it represents the main genetic cause of renal insufficiency in adult. It is characterized by cysts growth in the tubular segment, which increase in size and in number throughout an individual's lifetime, that leads to renal failure requiring dialysis or transplant. This pathology is due to the mutation of two genes:PKD1 and PKD2. The mutation of PKD1 gene is the most common, and is responsible of the 85% of cases, the gene encoding for a protein, a membrane receptor, the polycystin 1, which is involved in the mainteinace of cell/cell or cell/matrix interactions while the PKD2 gene product, polycystin 2, is a ionic channel.Both the protein are localized in the cilia of renal tubular epithelial cells and act as a mechanosensory organs. The bending of the cilium, caused by tubular flow, leads to the activation of the polycystin 1 and to the generation of a peak of intracellular calcium concentration. This increase in intracellular calcium activates signalling pathways that modify cellular proliferation and other cellular functions. In pathologic conditions, polycystin modification impairs the mechanosensitive function of cilia, altering tubular cellular functions. The aim of this project is the in vitro study of renal tubular cells (MDCK2) in laminar flow conditions, to investigate the role of mechanical stimulation in the pathology development. Preliminary results showed that the application of a constant laminar flow to MDCK2 causes a different tridimensional organization of the cellular layer, as compared to static control. Furthermore, the inhibition of intracellular calcium increase impairs this reorganization when flow is applied.

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LABORATORY OF BIOLOGY AND THERAPY OF METASTASIS*

STAFF

Head

Raffaella GIAVAZZI, Biol.Sci.D., Ph.D.

Giulia TARABOLETTI, Biol.Sci.D.

Head

* Research activities of this Laboratory are listed in the Department of Oncology section (pag. 7)

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Aldo and Cele Dacc Center

Ranica (Bg)

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departments and laboratories

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DEPARTMENT OF RENAL MEDICINE

STAFF

Head Laboratory of Biostatistics

Head

Piero RUGGENENTI, M.D.

Annalisa PERNA, Stat.Sci.D.

Laboratory of Coordination and Conduction of Controlled Clinical Trials

Head Unit of Drug Monitoring Head Nadia RUBIS, Res.N. Giulia GHERARDI, Res.N.

Laboratory of Pharmacokinetics and Clinical Chemistry

Head Flavio GASPARI, Chem.D.

Laboratory of Advanced Development of Drugs

Head Unit of Early Clinical Evaluation of Drugs Head Aneliya ILIEVA PARVANOVA, M.D. Norberto PERICO, M.D.

Laboratory of Clinical Pathophysiology of Renal Disease and Transplantation

Head Paolo CRAVEDI, M.D., PhD.

Laboratory of Regulatory Affairs for Clinical Studies

Head Paola BOCCARDO, Bio.Sci.D.

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CURRICULA
Piero Ruggenenti got his Medicine degree in 1983 at the University of Milan, Italy; he got his specialization in Cardiology in 1985 and in Clinical Nephrology in 1989 at the same University; he specialized in Pharmacological Research in 1988 at IRFMN. Educational training: in 1980-1983 researcher at "Centro di Fisiologia Clinica e Ipertensione, Clinica Medica IV", Universit degli Studi di Milano; in 1984 Researcher at IRFMN, Bergamo, Italy in 19871988 Honorary Registrar of the Unit for Metabolic Medicine, Division of Medicine (University of London) of Guy's and St. Thomas's Hospitals, London; in 1988-1989 Assistant Professor of the Division of Nephrology and Dialysis of the Ospedali Riuniti di Bergamo. Areas of interest: mechanisms of chronic renal disease progression, diabetes and diabetic complications, clinical transplantation, thrombotic microangiopathies, cardiovascular complications of chronic renal disease, clinical trials, clinical pharmacology. Employment: from 1990 Assistant Professor of the Division of Nephrology and Dialysis of the Ospedali Riuniti di Bergamo; in 1994-1999 Head, Unit of Advanced Development of Drugs, Dacc Center, Ranica, Bergamo, Italy; since 2000 Head, Department of Renal Medicine, Dacc Center, Bergamo, Italy.
Selected publications: Ruggenenti P, Fassi A, Parvanova Ilieva A, Petrov Iliev I, Chiurchiu C, Rubis R, Gherardi G, Ene-Iordache B, Gaspari G, Perna A, Cravedi P, Bossi A, Trevisan R, Motterlini N, Remuzzi G, for the BENEDICT-B Study Investigators. Effects of verapamil added-on trandolapril therapy in hypertensive type 2 diabetes patients with microalbuminuria: the BENEDICT-B randomized trial. J Hypert. In press. Ruggenenti P, Cattaneo D, Loriga G, Ledda F, Motterlini N, Gherardi G, Orisio S, Remuzzi G. Ameliorating hypertension and insulin resistance in subjects at increased cardiovascular risk: effects of acetyl-L-carnitine therapy. Hypertension, 2009 Sep; 54(3): 567-74. Remuzzi G, Cravedi P, Perna A, Dimitrov BD, Turturro M, Locatelli G, Rigotti P, Baldan N, Beatini M, Valente U, Scalamogna M, Ruggenenti P Dual Kidney Transplant Group. Long-term outcome of renal transplantation from older donors. N Engl J Med, 2006; 354: 343-352. Ruggenenti P, Fassi A, Parvanova A, Bruno S, Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med, 2004; 351: 1941-1951. Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S, Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P. Mycophenolate mofetil versus azathioprine for prevention of acute rejection in renal transplantation (MYSS): a randomised trial. Lancet, 2004 Aug 7-13; 364 (9433): 503-12. Remuzzi G, Chiurchiu C, Abbate M, Brusegan V, Bontempelli M, Ruggenenti P. Rituximab for idiopathic membranous nephropathy. Research Letter. Lancet, 2002; 360: 923-924. Ruggenenti P, Perna A, Gherardi G, Garini G, Zoccali C, Salvadori M, Scolari F, Schena FP, Remuzzi G. Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet ,1999; 354: 359-364.

Paola Boccardo got her classic High School Diploma in 1979 and the Biol. Sci. Degree at the University of Pisa in 1985. In 1987 she passed the qualifying examination and got the license of Biologist. Educational training: she performed her training first at Mutagenesis and Differentiation Institute, CNR, of Pisa, and then at Mario Negri Institute for Pharmacological Research, where in 1990, got her diploma of Specialist in Pharmacological Research. Since 1987 has been working as full-time researcher at Mario Negri Institute, till 1995 at Molecular Medicine Department and then at Renal Medicine Department. Area of interest: since 1995 she is in charge of Regulatory Affairs and attends to the planning, organizing and conducting of clinical studies in accordance with the principles of Good Clinical Practice and with the laws in force. Employment: since June 2006 to October 2009 Responsible of Clinical Trials Office; since November 2009 Head, Laboratory of Regulatory Affairs for Clinical Studies. Member of Internal Staff for Security, since May 2008 she is Security Manager at Clinical Research Center for Rare Diseases Aldo e Cele Dacc.
Selected publications: Perico N, Delaini F, Lupini C, Benigni A, Galbusera M, Boccardo P, Remuzzi G. Blunted excretory response to atrial natriuretic peptide in experimental nephrosis. Kidney Int, 1989; 36: 57-64.

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Benigni A, Perico N, Dadan J, Gabanelli M, Galbusera M, Boccardo P, Mennini T, Remuzzi G. Functional implications of decreased renal cortical ANP binding in experimental diabetes. Circ Res, 1990; 66: 1453-60. Benigni A, Boccardo P, Noris M, Remuzzi G, Siegler RL. Urinary excretion of platelet-activating factor in hemolytic uremic syndrome. Lancet, 1992; 339: 835-6. Noris M, Benigni A, Boccardo P, Aiello S, Gaspari F, Todeschini M, Figliuzzi M, Remuzzi G. Enhanced nitric oxide synthesis in uremia: implications for platelet dysfunction and dialysis hypotension. Kidney Int, 1993; 44: 445-450. Benigni A, Boccardo P, Galbusera M, Monteagudo J, De Marco L, Remuzzi G, Ruggeri ZM. Reversible activation defect of the platelet glycoprotein IIb-IIIa complex in patients with uremia. Am J Kidney Dis, 1993; 22: 668-676. Boccardo P, Noris M, Remuzzi G. Prevention and therapeutic management of bleeding in dialysis patients. In: Dialysis Therapy, 3rd edition. Edited by Nissenson A.R., Fine R.N. Hanley & Belfus, Inc., Philadelphia 2001; 3: 190-194. Remuzzi G, Galbusera M, Boccardo P. Disorders of hemostasis in dialysis patients. In: Heinrich, W.L. Ed. Principles and Practice of Dialysis, 4th ed. Philadelphia, PA: Lippincot W & W, 2009.

Paolo Cravedi got his Medicine degree (cum laude) in 1999 at the University of Milan, Italy; he got his specialization in Nephrology (cum laude) in 2004 at the University of Parma. In 2009 got a Ph.D. degree from the Open University of London. Educational training: in 2005 Master on Organ Transplant at the University of Milano Bicocca; in 2006 researcher at the Mario Negri Institute, Bergamo; since 2007 to 2008 Research Fellow at the Transplant Branch of the National Institutes of Health (NIH) (Mentor Dr. Roslyn Mannon); since 2009 researcher at the Mario Negri Institute, Bergamo. Areas of interest: mechanisms of chronic renal disease progression, diabetes and diabetic complications, clinical transplantation, membranous nephropathy, clinical pharmacology. Employment: since 2010, Head Laboratory of Clinical Pathophysiology of Renal Disease and Transplantation.
Selected publications: Remuzzi G, Cravedi P, Perna A, Dimitrov BD, Turturro M, Locatelli G, Rigotti P, Baldan N, Beatini M, Valente U, Scalamogna M, Ruggenenti P Dual Kidney Transplant Group. Long-term outcome of renal transplantation from older donors. N Engl J Med, 2006; 354: 343-352. Cravedi P, Ruggenenti P, Remuzzi G. Sirolimus to replace calcineurin inhibitors? Too early yet. Lancet, 2009; 373:1235-6. Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol, 2008; 3: 1652-9. Cravedi P, Ruggenenti P, Sghirlanzoni MC, Remuzzi G. Titrating rituximab to circulating B cells to optimize lymphocytolytic therapy in idiopathic membranous nephropathy. Clin J Am Soc Nephrol, 2007; 2: 932-7. Remuzzi G, Cravedi P, Costantini M, Lesti M, Ganeva M, Gherardi G, Ene-Iordache B, Gotti E, Donati D, Salvadori M, Sandrini S, Segoloni G, Federico S, Rigotti P, Sparacino V, Ruggenenti P. Mycophenolate mofetil versus azathioprine for prevention of chronic allograft dysfunction in renal transplantation: the MYSS follow-up randomized, controlled clinical trial. J Am Soc Nephrol, 2007; 18: 1973-85.

Flavio Gaspari got his Chemistry degree in 1977 at the University of Milano, Italy, and the specialization in the same University in 1979. Educational training: in 1981-1985 Fellow and Researcher at IRFMN, Milan; in 1985-1991 at IRFMN, Bergamo, Italy. Areas of interest: pharmacokinetics and the metabolism of xanthines in different animal species; drug pharmacokinetics in uremic patients and in subjects with different degrees of renal function; analytical methods to measure the most important immunosuppressive drugs to determine their pharmacokinetics in kidney, heart, and liver transplant recipients; evaluation of the renal function by using different approaches, in the study of renal disease progression, and in the comparison of different methods for albuminuria determination. Employement: he is Head of Laboratory of Pharmacokinetics and Clinical Chemistry since January 2000 and he was Head of this Unit since 1991.
Selected publications: Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Flores Bravo R, Carminati S, Rodriguez De Leon F, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A, Remuzzi G. Sirolimus therapy to halt the progression of ADPKD. JASN 2010; 21: 1031-1040. Gaspari F, Cravedi P, Mandal M, Perico N, de Leon FR, Stucchi N, Ferrari S, Labianca, R, Remuzzi G, Ruggenenti P. Predicting Cisplatin-Induced Acute Kidney Injury by Urinary Neutrophil Gelatinase-Associated Lipocalin Excretion: A Pilot Prospective Case-Control Study. Nephron Clin Pract 2010;115:c154-c160. Ruggenenti P, Perna A, Tonelli M, Loriga G, Motterlini N, Rubis N, Ledda F, Rota S Jr, Satta A, Granata A, Battaglia G, Cambareri F, David S, Gaspari F, Stucchi N, Carminati S, Ene-Iordache B, Cravedi P, Remuzzi G; for the ESPLANADE Study Group. Effects of Add-on Fluvastatin Therapy in Patients with Chronic Proteinuric Nephropathy on Dual ReninAngiotensin System Blockade: The ESPLANADE Trial. Clin J Am Soc Nephrol. 2010; 5:1928-38.

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Perico N, Zoja C, Corna D, Rottoli D, Gaspari F, Haskell L, Remuzzi G. V1/V2 Vasopressin receptor antagonism potentiates the renoprotection of renin-angiotensin system inhibition in rats with renal mass reduction. Kidney Int, 2009 Nov; 76(9): 9607. Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol, 2008 Nov; 3(6): 1652-9. Gotti E, Perico N, Gaspari F, Cattaneo D, Lesti MD, Ruggenenti P, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Federico S, Sparacino V, Mourad G, Bosmans JL, Dimitrov BD, Iordache BE, Remuzzi G. Blood cyclosporine level soon after kidney transplantation is a major determinant of rejection: insights from the Mycophenolate Steroid-Sparing Trial. Transplant Proc, 2005 Jun; 37(5): 2037-40. Perico N, Gaspari F, Remuzzi G. Assessing renal function by GFR prediction equations in kidney transplantation. Am J Transplant, 2005 Jun; 5(6): 1175-6. D. Cattaneo, F. Gaspari, S. Zanoni, S. Baldelli, E.Gotti, A. Perna, N. Perico, G. Remuzzi. Two-hour post-dose cyclosporine monitoring does not fit all in kidney transplantation. Therapy, 2005; 2: 95-105. Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G; Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med, 2004 Nov 4; 351 (19): 1941-51. Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M, Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G, Perico N; MY.S.S. Study Investigators. Performance of different prediction equations for estimating renal function in kidney transplantation. Am J Transplant, 2004 Nov; 4 (11): 1826-35.

Giulia Gherardi got her Scientific High School Diploma in 1989 at the Liceo Scientifico Marie Curie in Zogno (Bergamo), the Nurse Diploma in 1995 at the Scuola per Infermieri Professionali, Ospedali Riuniti, Bergamo and the 1st Level Master in Clinical Research in 2008 at the Medicine and Surgery Faculty of the University in Milan. Educational training: Clinical Research Nurse Diploma on 1997 at IRFMN Dacc Center. Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology, and diabetology; the coordination, conduction and monitoring of controlled clinical trials. Employement: in 1997-2003 involved as co-organizing, speaker, co-speaker and tutor for the Clinical Research Course for Nurse at IRFMN Dacc Center (Ranica Bergamo). Several training activities for Nurses in Clinical Research area. In 1997-1999, Clinical Research Monitor at IRFMN Dacc Center; in 2000-2008 Head of the Monitoring Drug Unit at IRFMN Dacc Center. Since 2009 Head of the Laboratory of Coordination and Conduction of Controlled Clinical Trials at IRFMN Dacc Center.
Selected publications: Ruggenenti P, Fassi A, Parvanova Ilieva A, Petrov Iliev I, Chiurchiu C, Rubis R, Gherardi G, Ene-Iordache B, Gaspari G, Perna A, Cravedi P, Bossi A, Trevisan R, Motterlini N, Remuzzi G, for the BENEDICT-B Study Investigators. Effects of verapamil added-on trandolapril therapy in hypertensive type 2 diabetes patients with microalbuminuria: the BENEDICT-B randomized trial. J Hypert. In press. Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Bravo RF, Carminati S, De Leon FR, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A, Remuzzi G. Sirolimus therapy to halt the progression of ADPKD. J Am Soc Nephrol. 2010 Jun; 21(6): 1031-40. Ruggenenti P, Cattaneo D, Loriga G, Ledda F, Motterlini N, Gherardi G, Orisio S, Remuzzi G. Ameliorating hypertension and insulin resistance in subjects at increased cardiovascular risk: effects of acetyl-L-carnitine therapy. Hypertension, 2009 Sep; 54 (3): 567-74. Remuzzi G, Cravedi P, Costantini M, Lesti M, Ganeva M, Gherardi G, Ene-Iordache B, Gotti E, Donati D, Salvadori M, Sandrini S, Segoloni G, Federico S, Rigotti P, Sparacino V, Ruggenenti P. Mycophenolate mofetil versus azathioprine for prevention of chronic allograft dysfunction in renal transplantation: the MYSS follow-up randomized, controlled clinical trial. J Am Soc Nephrol, 2007 Jun; 18 (6): 1973-85. Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G; Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med, 2004 Nov 4; 351 (19): 1941-51. Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S, Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P. Mycophenolate mofetil versus azathioprine for prevention of acute rejection in renal transplantation (MYSS): a randomised trial. Lancet, 2004 Aug 7-13; 364 (9433): 503-12. Ruggenenti P, Perna A, Gherardi G, Benini R, Remuzzi G. Chronic proteinuric nephropathies: outcomes and response to treatment in a prospective cohort of 352 patients with different patterns of renal injury. Am J Kidney Dis, 2000 Jun; 35 (6): 1155-65. Ruggenenti P, Perna A, Gherardi G, Garini G, Zoccali C, Salvadori M, Scolari F, Schena FP, Remuzzi G. Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet, 1999 Jul 31; 354 (9176): 359-64. Ruggenenti P, Perna A, Gherardi G, Gaspari F, Benini R, Remuzzi G. Renal function and requirement for dialysis in chronic nephropathy patients on long-term ramipril: REIN follow-up trial. Gruppo Italiano di Studi Epidemiologici in Nefrologia (GISEN). Ramipril Efficacy in Nephropathy. Lancet, 1998 Oct 17; 352 (9136): 1252-6.

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Norberto Perico got his Medicine degree in 1983 at the University of Milano, Italy. He got his specialization in Pharmacological Research in 1986 at IRFMN, Bergamo and in Clinical Nephrology in 1989 at the University of Verona, Italy. Educational training: in 1982 Fellow, Department of Pharmacology, New York Medical College, Valhalla, New York, USA; in 1984-1988 Post Doctoral Fellow, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; in 1988-1989 Researcher in the same laboratory. Areas of interest: pathophysiology and pharmacology of cyclosporine nephrotoxicity; new immunosuppressive strategies to prevent renal graft rejection; innovative approach to induce tolerance to organ transplantation; mechanism(s) and management of progression of chronic renal diseases. Employment: in 1990-1994 Head, Renal Physiology Unit, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; in 1990-2000 Assistant Professor, Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Italy; in 1994 1999 Head, Laboratory of Transplant Immunology, IRFMN, Bergamo, Italy; from January 2000 Head, Laboratory of Drug Development, Department of Renal Medicine, IRFMN, Bergamo, Italy; from September 2000 Health Director, Dacc Center, IRFMN, Bergamo, Italy. From October 2002 hes Member, ISN-COMGAN Research Committee of the International Society of Nephrology.
Selected publications: Dahlke MH, Hoogduijn M, Eggenhofer E, Popp FC, Renner P, Slowik P, Rosenauer A, Piso P, Geissler EK, Lange C, Chabannes D, Mazzanti B, Bigenzahn S, Bertolino P, Kunter U, Introna M, Rambaldi A, Capelli C, Perico N, Casiraghi F, Noris M, Gotti E, Seifert M, Saccardi R, Verspaget HW, van Hoek B, Bartholomew A, Wekerle T, Volk HD, Remuzzi G, Deans R, Lazarus H, Schlitt HJ, Baan CC; MISOT Study Group. Toward MSC in solid organ transplantation: 2008 position paper of the MISOT study group. Transplantation, 2009 Sep 15; 88 (5): 614-9. Cattaneo D, Cortinovis M, Baldelli S, Gotti E, Remuzzi G, Perico N. Limited sampling strategies for the estimation of sirolimus daily exposure in kidney transplant recipients on a calcineurin inhibitor-free regimen. J Clin Pharmacol, 2009 Jul; 49 (7): 773-81. Cattaneo D, Ruggenenti P, Baldelli S, Motterlini N, Gotti E, Sandrini S, Salvadori M, Segoloni G, Rigotti P, Donati D, Perico N, Remuzzi G; Mycophenolate Steroids Sparing (MYSS) Genetics Study Group. ABCB1 genotypes predict cyclosporinerelated adverse events and kidney allograft outcome. J Am Soc Nephrol, 2009 Jun; 20 (6): 1404-15. Perico N, Bravo RF, De Leon FR, Remuzzi G. Screening for chronic kidney disease in emerging countries: feasibility and hurdles. Nephrol Dial Transplant, 2009 May; 24 (5): 1355-8. Perico N, Benigni A, Remuzzi G. Present and future drug treatments for chronic kidney diseases: evolving targets in renoprotection. Nat Rev Drug Discov, 2008 Nov; 7 (11): 936-53. Ruggenenti P, Perico N, Gotti E, Cravedi P, D'Agati V, Gagliardini E, Abbate M, Gaspari F, Cattaneo D, Noris M, Casiraghi F, Todeschini M, Cugini D, Conti S, Remuzzi G. Sirolimus versus cyclosporine therapy increases circulating regulatory T cells, but does not protect renal transplant patients given alemtuzumab induction from chronic allograft injury. Transplantation, 2007 Oct 27; 84 (8): 956-64. Baldelli S, Merlini S, Perico N, Nicastri A, Cortinovis M, Gotti E, Remuzzi G, Cattaneo D. C-440T/T-331C polymorphisms in the UGT1A9 gene affect the pharmacokinetics of mycophenolic acid in kidney transplantation. Pharmacogenomics, 2007 Sep; 8 (9): 1127-41.

Annalisa Perna got her Statistical Sciences degree in 1984 at the University of Bologna, Italy. Educational training: She completed her research training at IRFMN, Bergamo Labs. and at the Dacc Center. Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology, statistical methods for calculating sample size and for meta-analytic techniques. She is also involved in performing systematic reviews for the Cochrane Collaboration Renal Review Group. Employment: she is Head of the Laboratory of Biostatistics - Department of Renal Medicine at Dacc Center, Ranica (Bergamo).
Selected publications: Ruggenenti P, Iliev I, Filipponi M, Tadini S, Perna A, Ganeva M, Ene-Iordache B, Cravedi P, Trevisan R, Bossi, and Remuzzi G. Effect of Trandolapril on Regression of Retinopathy in Hypertensive Patients with Type 2 Diabetes: A PreSpecified Analysis of the Benedict Trial, Journal of Ophthalmology, 2010; 2010:106384. Epub 2010 Jun 10. Ruggenenti P, Cattaneo D, Rota S, Iliev I, Parvanova A, Diadei O, Ene-Iordache B, Ferrari S, Bossi AC, Trevisan R, Belviso A, Remuzzi G; for the Ezetimibe and Simvastatin in Dyslipidemia of Diabetes (ESD) Study Group. Effects of combined ezetimibe and simvastatin therapy as compared with simvastatin alone in patients with type 2 diabetes: a prospective randomized double-blind clinical trial. Diabetes Care. Sep;33(9):1954-6, 2010. Ruggenenti, P, Perna, A, Remuzzi, G. Effects of combined ezetimibe and simvastatin therapy as compared to simvastatin alone in patients with type 2 diabetes: a prospective, randomized, double-blind, clinical trial. Diabetes Care: e133; 2010. Ruggenenti P, Perna A, Tonelli M, Loriga G, Motterlini N, Rubis N, Ledda F, Rota S Jr, Satta A, Granata A, Battaglia G, Cambareri F, David S, Gaspari F, Stucchi N, Carminati S, Ene-Iordache B, Cravedi P, Remuzzi G; for the ESPLANADE

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Study Group. Effects of Add-on Fluvastatin Therapy in Patients with Chronic Proteinuric Nephropathy on Dual ReninAngiotensin System Blockade: The ESPLANADE Trial. Clin J Am Soc Nephrol. Nov;5(11):1928-38, 2010. Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Bravo RF, Carminati S, De Leon FR, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A, Remuzzi G. Sirolimus therapy to halt the progression of ADPKD. J Am Soc Nephrol. Jun;21(6):1031-40, 2010. Ruggenenti P, Fassi A, Parvanova Ilieva A, Petrov Iliev I, Chiurchiu C, Rubis R, Gherardi G, Ene-Iordache B, Gaspari G, Perna A, Cravedi P, Bossi A, Trevisan R, Motterlini N, Remuzzi G, for the BENEDICT-B Study Investigators. Effects of verapamil added-on trandolapril therapy in hypertensive type 2 diabetes patients with microalbuminuria: the BENEDICT-B randomized trial. J Hypert. In press.

Aneliya Parvanova Ilieva got her Medical Doctor degree at the Faculty of Medicine, Thracian University (former Higher Medical Institute), Stara Zagora, Bulgaria in 1988, and the specialization in Pharmacology in Department of Pharmacology, University of Medicine, Sofia in 1992. Her medical degree is recognized in Italy in 2009. Educational training: in 1989-1998 teaching of 3rd, 4th and 5th-year medical students and 2nd and 3rd-year clinical nurses in a general pharmacology and clinical pharmacology, Thracian University, Stara Zagora, Bulgaria; examiner of these students in theoretical and practical, oral and written exams and tests and State examination. In 1993 Course on investigation of isolated organs Bulgarian Academy of Sciences, Sofia. In 1998 visiting scientist, IRFMN, Ranica, Bergamo, Italy. In 1998 proficiency in the methods for insulin sensitivity evaluation (hyperinsulinemic euglicaemic clamp technique), in renal hemodynamic measurements - glomerular filtration rate (plasma clearance of iohexol and inulin), in renal plasma flow (plasma clearance of para-aminohippuric acid), glomerular size selectivity (plasma clearance of neutral dextrans) and in twenty four-hour blood pressure monitoring. Areas of interest: primary and secondary prevention of the chronic microvascular diabetic complications (diabetic nephropathy, diabetic retinopathy and diabetic neuropathy); role of insulin resistance, arterial hypertension, dyslipidemia and hyperhomocysteinemia in micro- and macrovascular diabetic complications; possibilities for pharmacological treatment of these pathological conditions. Employment: she participates as investigator in several clinical studies. She is Head of The Unit of Early Clinical Evaluation of Drugs at IRFMN since 2000. She is a member of the Union of Bulgarian Doctors (since 1989), of the Union of Pharmacologists in Bulgaria (since 1990), of the Union of Scientists in Bulgaria (since 1991), and member of the Union of Medical Doctors and Dentists, Bergamo, Italy (since 05.03.2009).
Selected publications: Ruggenenti P, Cattaneo D, Rota S, Iliev I, Parvanova A, Diadei O, Ene-Iordache B, Ferrari S, Bossi AC, Trevisan R, Belviso A, Remuzzi G. Effects of combined ezetimibe and simvastatin therapy as compared with simvastatin alone in patients with type 2 diabetes: a prospective randomized double-blind clinical trial. Diabetes Care 2010 Sep; 33(9): 1954-6. Epub 2010 Jun 21. Vogt L, Chiurchiu C, Chadha-Boreham H, Danaietash P, Dingemanse J, Hadjadj S, Krum H, Navis G, Neuhart E, Parvanova AI, Ruggenenti P, Woittiez AJ, Zimlichman R, Remuzzi G, de Zeeuw; for the PROLONG (PROteinuria Lowering with urOteNsin receptor antaGonists) Study Group. Effect of the Urotension Receptor Antagonist Palosuran in Hypertensive patients with type 2 diabetic nephropathy. Hypertension 2010 May; 55(5): 1206-9. Epub 2010 Mar 15. Ruggenenti P, Iliev I, Costa GM, Parvanova A, Perna A, Giuliano GA, Motterlini N, Ene-Iordache B, Remuzzi G. Preventing left ventricular hypertrophy by ACE inhibition in hypertensive patients with type 2 diabetes: a perspecified analysis of the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). Diabetes Care, 2008 Aug; 31 (8): 1629-34. Parvanova A, Trevisan R, Iliev I, Dimitrov BD, Vedovato M, Tiengo A, Remuzzi G, Ruggenenti P. Insulin resistance and microalbuminuria, A Cross-sectional, case-control study of 158 patients with type 2 diabetes and different degrees of urinary albumin excretion. Diabetes, 2006; 55: 1456-1462. Parvanova A, Chiurchiu C, Ruggenenti P, Remuzzi G. Inhibition of the renin-angiotensin system and cardio-renal protection: focus on losartan and angiotensin receptor blockade. Expert Opinion on Pharmacotherapy, 2005 Sep; 6 (11):1931-1942. Ruggenenti P, Fassi A, Parvanova A, Bruno S, Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, EneIordache, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini A, Remuzzi G. Preventing Microalbuminuria in Type 2 Diabetes. NEJM, 2004;351 (19): 1941-51. Parvanova A, Iliev I, Filipponi M, Dimitrov BD, Vedovato M, Tiengo A, Trevisan R, Remuzzi G, Ruggenenti P. Insulin resistance and proliferative retinopathy: a cross-sectional, case-control study in 115 patients with type 2 diabetes. J Clin Endocrinol Metab, 2004 Sep; 89 (9): 4371-6. The BENEDICT Group. The Bergamo Nephrologic DIabetes Complications Trial (BENEDICT): design and baseline characteristics. Controlled Clinical Trials, 2003; 24: 442-461. Parvanova A, Iliev I, Dimitrov BD, Arnoldi F, Zaletel J, Remuzzi G, Ruggenenti P. Hyperhomocysteinemia and increased risk of retinopathy: a cross-sectional, case-control study in patients with type 2 diabetes. Diabetes Care, 2002; 25 (12): 2361.

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Nadia Rubis got her degree in licensed practical nurse in 1995 at the Nursing School of Azienda Ospedaliera Ospedali Riuniti di Bergamo. Education training: she completed her research training at IRFMN - Centro di Ricerche Cliniche per le Malattie Rare Aldo e Cele Dacc Bergamo, Clinical Research Nurse Course (1998). Areas of interest: coordination of clinical studies, monitoring activities and data management, pharmacovigilance. Employment: in 1998-2003 involved as co-promoter and tutor for the Clinical Research Course for Nurse at IRFMN - Centro Dacc. In 1998-2008 clinical monitor, Drug Monitoring Unit of Centro Dacc. Since September 2008 Head of the Drug Monitoring Unit.
Selected publications:

Piero Ruggenenti, Anna Fassi, Aneliya Parvanova Ilieva, Ilian Petrov Iliev, Carlos Chiurchiu, Nadia Rubis, Giulia Gherardi, Bogdan Ene-Iordache, Flavio Gaspari, Annalisa Perna, Paolo Cravedi, Antonio Bossi, Roberto Trevisan, Nicola Motterlini, Giuseppe Remuzzi, for the BENEDICT-B Study Investigators. Effects of verapamil added-on trandolapril therapy in hypertensive type 2 diabetes patients with microalbuminuria: the BENEDICT-B randomized trial. J Hypertens 29:207-216, 2011. In Press. Piero Ruggenenti, Annalisa Perna, Marcello Tonelli, Giacomina Loriga, Nicola Motterlini, Nadia Rubis, Franca Ledda, Stefano Rota Jr., Andrea Satta, Antonio Granata, Giovanni Battaglia, Francesco Cambareri, Salvatore David, Flavio Gaspari, Nadia Stucchi, Sergio Carminati, Bogdan Ene-Iordache, Paola Cravedi and Giuseppe Remuzzi for the ESPLANADE study group. Effects of Add-on Fluvastatin Therapy in Patients with Chronic Proteinuric Nephropathy on Dual Renin-Angiotensin System Blockade: The ESPLANADE Trial. Clin J Am Nephrol 5: 1928-1938, 2010. Ene-Iordache B, Carminati S, Antiga L, Rubis N, Ruggenenti P, Remuzzi G and Remuzzi A. Developing regulatorycompliant electronic case report forms for clinical trials: experience with the demand trial. J Am Med Inform Assoc, 2009 May-Jun;16 (3): 404-8. Fassi A, Rubis N, Parvanova A, Iliev I, Zamora J, Giuliano GA, Ene-Iordache B, Perna A, Anabaya A, Motterlini N, Ruggenenti P, Remuzzi G for the BENEDICT Study Investigators. Randomized and non-randomized patients in the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). Abstract. 29th Annual Meeting of the Society for Clinical Trials St Louis, Missouri, May 18-21, 2008. Ruggenenti P, Fassi A, Parvanova Ilieva A, Bruno S, Petro Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi A, Ganeva M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G, for the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med, 2004; 351: 1941-51. Ruggenenti P, Perna A, Gherardi G, Benini R, Rubis N, Gritti D, Ciocca I, Stucchi N and Remuzzi G for the GISEN Group. In chronic renal disease, hypertension and type 2 diabetes predict fast progression. Proteinuria < 2 g/24 hours, type 2 diabetes and polycystic kidneys are associated with poor response to ACE inhibition. ASN, November 5-8, 1999.

INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES

The Department of Renal Medicine was established on 1999 at the Clinical Research Center for Rare Diseases Aldo e Cele Dacc Villa Camozzi, Ranica to coordinate the activities of 6 Laboratories and 2 Units. The activities of the Department are mainly focused on the study of the mechanisms of progression of chronic nephropathies, of new prevention and intervention strategies for diabetic nephropathy, non diabetic chronic nephropathies, chronic allograft dysfunction, of cardiovascular complications of diabetes, chronic renal disease, dialysis and transplantation and of thrombotic microangiopathies. The main aims of these activities are: 1. To identify screening and intervention strategies aimed to prevent the onset of nephropathy and of other chronic complications of diabetes and/or hypertension. 2. To define intervention strategies to prevent or slow the progression of chronic nephropathies and eventually obtain remission/regression of renal dysfunction. 3. To optimize immunosuppressive protocols in kidney transplantation and to define new donor selection criteria in order to expand the pool of available organs. These aims will be pursued through the following modalities:

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1. Pilot pathophysiology and clinical pharmacology studies fully finalized at the Clinical Research Center to test new pathogenetic hypotheses and new treatment modalities. 2. National and international networks and multicenter trials aimed to verify the efficacy of treatments of potential interest identified as described at point 1. 3. Meta-analyses and probabilistic models to test new risk factors and treatments in large samples of patients and to transfer this information at individual level. Many of these activities rest on the possibility of a tight cooperation with the Department of Molecolar Medicine, the Department of Bioengineering and the Public-Private Department of Specialist and Transplant Medicine. This cooperation allows to plan the research activities of the Department on the basis of new information derived from basic research and of problems of major clinical relevance emerging from routine clinical activities.

FINDINGS/MAIN RESULTS
Definition and validation of specific treatments aimed to prevent the development and progression of nephropathy and related micro and macrovascular complications in subjects with type 2 diabetes. Definition and validation of new integrated treatment protocols aimed to slow the progression and/or to achieve remission/regression of diabetic and non-diabetic chronic nephropathies. Institution of a standardized protocol on line (The Remission Clinics) finalized to achieve regression/remission of chronic nephropathies and limit overall renal and radiovascular risk in hospital practice in the setting of a multicenter Network. Characterization of the antiproteinuric, nephroprotective and cardioprotective effect of maximized and polypharmacologic renin-angiotensin system inhibition, intensified blood pressure and lipid control and identification of novel treatments to reduce the blood pressure and ameliorate insulin sensitivity in subjects at increased cardiovascular risk. Identification of acquired or congenital risk factors for chronic complications of diabetes and cardiovascular morbidity and mortality Identification and validation of early markers of acute kidney failure and of methods for direct and indirect measurements of kidney function and GFR decline. Identification of safety and efficacy profile of new treatments for the Autosomal Polycistic Kidney Disease (APKD). Definition and validation of new, specific treatments for idiopathic membranous nephropathy and for HUS forms associated with genetic defect of complement factors including the standardization of combined liver and kidney transplantation to prevent post transplant recurrence of genetic associated HUS. Definition and validation of new laboratory procedures and predictive models to help monitoring and optimizing immunosuppressive therapy in clinical transplantation with particular focus on pharmacokynetic markers of drug exposure and genetic predictors of drug tolerability and efficacy. Definition and validation of selection and allocation criteria of kidneys from marginal and oldvery old donors to increase the donor pool and the transplant activity.

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Finalization and activation of multicenter clinical trials aimed to prevent onset and progression of diabetic and non-diabetic chronic nephropathies, to achieve remission of the nephrotic syndrome in primary glomerular diseases, minimize maintenance immunosuppression in kidney transplantation and prevent cardiovascular morbidity and mortality in chronic hemodialysis. Computerization of data acquisition and monitoring procedures for the conduction of controlled clinical trials.

NATIONAL COLLABORATIONS
AO Bolognini Seriate, Ospedale Bolognini, Seriate (BG) AO Ospedali Riuniti, Bergamo AO Treviglio, Ospedale di Treviglio, Treviglio (BG) AO Treviglio, Ospedale SS. Trinit, Romano di Lombardia (BG) AO Treviglio, Poliambulatorio extra-ospedaliero, Ponte San Pietro (BG) ASL Bergamo, Bergamo Istituto Humanitas Gavazzeni, Bergamo AO Spedali Civili di Brescia, Presidio Ospedaliero di Montichiari, Montichiari (BS) AO Spedali Civili, Spedali Civili, Brescia AO Ospedale SantAnna, Presidio Ospedaliero SantAnna, Como Azienda Ospedaliera Istituti Ospedalieri di Cremona, Cremona AO Ospedale San Carlo Borromeo, Milano AO San Gerardo, Ospedale Bassini, Cinisello Balsamo (MI) AO San Gerardo, Ospedale San Gerardo, Monza (MI) AO San Paolo Polo Universitario, Milano ASL Provincia di Milano 2, Ospedale A. Uboldo, Cernusco sul Naviglio (MI) Fondazione IRCCS Ca Granda, Ospedale Maggiore Policlinico, Milano Fondazione IRCCS Istituto Neurologico Carlo Besta, Milano IRCCS Fondazione Centro San Raffaele del Monte Tabor, Milano IRCCS Istituto Clinico Humanitas, Rozzano (MI) IRCCS Multimedica di Sesto San Giovanni, Sesto San Giovanni (MI) AO Pavia, Ospedale Civile di Voghera, Voghera (PV) AO Valtellina e Valchiavenna, Ospedale di Sondrio, Sondrio AO Universitaria, Ospedale di Circolo e Fondazione Macchi, Varese AO Ospedale Infantile Regina Margherita-SantAnna di Torino, Ospedale Infantile Regina Margherita, Torino AO Ordine Mauriziano, Ospedale Mauriziano Umberto I, Torino ASL TO2, Ospedale San Giovanni Bosco, Torino AULSS 17 Este, Ospedale di Monselice, Monselice (PD) AULSS 9 di Treviso, Ospedale Santa Maria di Ca' Foncello, Treviso AO Universistaria degli Ospedali Riuniti di Trieste, Ospedale di Cattinara, Trieste IRCCS Materno-Infantile Burlo Garofalo, Trieste AO Universitaria di Bologna, Policlinico SantOrsola-Malpighi, Bologna AUSL Forl, Ospedale G. B. Morgagni - L. Pierantoni, Forl AO Universitaria di Parma, Ospedale di Parma, Parma AUSL Ravenna, Ospedale Santa Maria delle Croci, Ravenna AO Reggio Emilia, Arcispedale Santa Maria Nuova, Reggio Emilia AUSL Rimini, Ospedale Infermi, Rimini AO Universitaria Careggi, Firenze

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Universit degli Studi, Firenze AUSL 2 Lucca, Ospedale Campo di Marte, Lucca AO Universitaria Pisana, Ospedale Santa Chiara, Pisa ASUR Zona Territoriale 13, Ospedale Mazzoni, Ascoli Piceno IRCCS Pediatrico Bambino Ges, Roma AUSL Latina, Presidio di Formia, Latina AUSL Rieti, Rieti AO Ospedale San Giuseppe Moscati, Avellino AO Antonio Cardarelli, Napoli AO Pediatrica Santobono-Pausilipon, Ospedale Santobono, Napoli Universit Azienda Ospedaliera Universitaria Federico II, Napoli Universit Azienda Ospedaliera Universitaria Federico II, Policlinico Nuovo, Napoli ASL Salerno, Ospedale Maria SS. Addolorata, Eboli (SA) ASL Teramo, Presidio Ospedaliero Giuseppe Mazzini, Teramo Centro Nazionale Ricerche, Reggio Calabria AS 3 Rossano, Ospedale Civile Nicola Giannettasio, Rossano (CS) AO Ospedale Cannizzaro, Catania AO Universitaria Policlinico-Vittorio Emanuele, Presidio Ospedaliero Vittorio Emanuele, Catania AUSL 3 Catania, Presidio Ospedaliero di Acireale, Acireale (CT) ASP 5 Messina, Ospedale di Milazzo, Milazzo (ME) AO Civico-Di Cristina-Benfratelli, Presidio Ospedaliero Civico e Benfratelli, Palermo AO Universitaria Policlinico Paolo Giaccone, Universit degli Studi, Palermo Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione (IsMeTT), Palermo Fondazione Istituto San Raffaele G. Giglio di Cefal, Cefal (PA) Azienda Ospedaliera, Ospedale Umberto I, Siracusa IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo (FG) AUSL Le/1, Ospedale Vito Fazzi, Lecce ASL Taranto 1, Presidio Ospedaliero Valle DItria di Martina Franca, Martina Franca (TA) AO Brotzu, Ospedale San Michele, Cagliari ASL Sanluri, Presidio Ospedaliero Nostra Signora di Bonaria, San Gavino Monreale (VS) ASL Olbia, Ospedale San Giovanni di Dio, Olbia (OT) ASL Sassari, Azienda Ospedaliero-Universitaria di Sassari, Sassari

INTERNATIONAL COLLABORATION
- University Medical Center, Ljubljana (Slovenia) - Service de Pharmacologie Clinique, Facult de Mdecine, Lyon (France) - Hospital Universitario de Canarias, La Laguna, Tenerife (Spain) Department of Primary Health Care , University of Oxford, Oxford (UK) Department of Clinical Sciences/Diabetes & Endocrinology Lund University, Skne University Hospital, Malm (Sweden) University Medical Center Groningen, Groningen, (The Netherlands) Department of Clinical Pharmacology, Groningen (The Netherlands) Leiden University Medical Center, Leiden (The Netherlands)

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University Medical Center, Groningen (The Netherlands) Mariinskaya Hospital, Saint-Petersburg (Russia) Moscow State University of Medicine and Dentistry, Mosca (Russia) Chiinu Hospital, Chiinu (Moldova) Damanhour Medical National Institute, Damanhour, Beheira (Egypt) Health Sciences University, Ulaanbaator (Mongolia) BP Kerala Institute of Health, Dharan (Nepal) Division of Cardiology, Brigham and Women's Hospital, Boston, MA (USA) National Kidney Foundation, New York, NY (USA) New England Medical Center, Boston, MA (USA) Complejo Hosp Metropolitano de la Caja de Seguro Social, Panama City (Panama) Hospital Juan XXIII, La Paz (Bolivia) Hospital Maciel, Montevideo (Uruguay) Cochrane Collaboration, Cochrane Renal Group, Centre for Kidney Research, NHMRC Centre for Clinical Research Excellence in Renal Medicine, The Children's Hospital at Westmead, Westmead, (Australia).

EDITORIAL BOARD MEMBERSHIP

Current Diabetes Reviews (Piero Ruggenenti) Clinical Journal of the American Society of Nephrology (Piero Ruggenenti) Journal of Nephrology (Piero Ruggenenti) Nephron (Norberto Perico) The Open Hypertension Journal (Paolo Cravedi)

PEER REVIEW ACTIVITIES

Acta Diabetologica Acta Pharmacologica Sinica American Journal of Hypertension American Journal of Kidney Diseases American Journal of Pathology American Journal of Transplantation

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Archives of Medical Science Bentham Science Blood Purification British Medical Journal (BMJ) Circulation American Hearth Association (AHA) Clinical Journal of the American Society of Nephrology (CJASN) Clinical Nephrology EMBO Molecular Medicine Expert Opinion on Pharmacotherapy Expert review of Clinical Immunology Heart Failure Reviews Indian Journal of Nephrology Internal Urology and Nephrology International Journal of Clinical Practice Islets Journal of the American Society of Nephrology (JASN) Journal of Hypertension Journal of Nephrology Kidney International Mediterranean Journal of Hematology And Infection Diseases Nature Communications Nature Reviews Nephrology Nephrology Dialysis Transplantation Nephron New England Journal of Medicine The International Journal of Artificial Organs The Lancet Translational research Transplant International Transplantation

PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS INVOLVED

Studi clinici controllati. Presentation at the Specialization Course in Nephrology at the University of Firenze. Firenze (Italy). March 2nd, 2010. Surrogate markers for micro- and macro-vascular hard endpoints for innovative diabetes tools. Malmo (Sweden)., March 10th-11th, 2010. Kidney Master Class. Bergamo (Italy). March 17th, 2010. Il continuum cardiorenale ed i farmaci del sistema RAAS. In the Corso Diabete e Malattie Cardiovascolari. Perugia (Italy). March 19th-21st, 2010. Albuminuria nella nefropatia diabetica. In the Seminario Angelini. Londra (UK). March 24th, 2010.

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Nuove evidenze nel trattamento e prevenzione della CKD-MBD. Firenze (Italy). April 2010.

8th

ARCADIA Study: secondo investigator meeting. Ranica, Bergamo (Italia). April 16th, 2010. ADPKD in Europe. Bruxelles (Belgium), May 27th, 2010. ADPKD: nuove strategie terapeutiche. In the Giornate Cefaludesi di Nefrologia 2010. Cefal (Italy). May 29th, 2010. ATHENA Study: secondo investigator meeting.Bergamo (Italy), July 12th, 2010. New ways in treating membranous glomerulonephritis. Goteborg (Sweden). September 22nd, 2010. Congresso di Oncologia. Verona (Italy). September 24th, 2010. Il futuro gi tra noi? Il progetto Remission Clinic un progetto ambizioso, una sfida per il futuro. In the 2 Congresso Percorso Integrato Territoriale in Diabetologia. Palermo (Italy). October 1st-2nd, 2010. The third WHO meeting on a prioritized research agenda for prevention and control of noncommunicable diseases. Ginevra (Switzerland). October 20th-21st, 2010. Outcome trials and differential drug responses in CKD and diabetes. In the Conference How can medical innovations reduce the cardiovascular disease burden?. Bruxelles (Belgium). November 4th-5th, 2010. Early detection and prevention of CKD in resource poor regione. In the 43rd Annual Meeting of the American Society of Nephrology. Denver (USA). November 20th, 2010. ACEi+ARBs (kidney outcome). In the 2nd International Symposium on Albuminuria. Amsterdam (Netherlands). December 12th-14th, 2010. La Remission clinic nella pratica clinica: nuove prospettive di prevenzione e trattamento per le nefropatie croniche progressive. Ranica, Bergamo (Italy). December 18th, 2010. Limportanza di far sapere cosa si fa e per chi. Informare sui risultati. Come si presenta al CEI la domanda per la sperimentazione. Gli stati di avanzamento e la chiusura delle sperimentazioni cliniche.. In the Fare bene. Presentare e gestire con competenza una sperimentazione clinica. Seriate, Bergamo (Italy). December 18th, 2010.

GRANTS AND CONTRACTS

AIFA (Agenzia Italiana del Farmaco) Innovative Medicine Initiative Joint Undertaking (IMI JU) International Society of Nephrology (ISN) Ministero della Salute

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Abbott Srl Baxter SpA Boheringer Ingelheim Italia SpA Genzyme Europe BV Otsuka Pharmaceutical Europe Ltd Sanofi-Aventis SpA Sigma-Tau Industrie Farmaceutiche Riunite SpA Solvay Pharmaceuticals

SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2010

Cravedi P, Perico N, Remuzzi G. Non-immune interventions to protect kidney allografts in the long term. Kidney Int. 2010 Dec; 78 Suppl 119: S71-5. Perico N, Remuzzi G. Do mTOR inhibitors still have a future in ADPKD? Nat Rev Nephrol. 2010 Dec; 6 (12): 696-8. Cravedi P, Ruggenenti P, Remuzzi G. Which antihypertensive drugs are the most nephroprotective and why? Expert Opin Pharmacother. 2010 Nov; 11 (16): 2651-63. Sharma SK, Zou H, Togtokh A, Ene-Iordache B, Carminati S, Remuzzi A, Wiebe N, Ayyalasomayajula B, Perico N, Remuzzi G, Tonelli M. Burden of CKD, proteinuria, and cardiovascular risk among Chinese, Mongolian, and Nepalese participants in the International Society of Nephrology screening programs. Am J Kidney Dis. 2010 Nov; 56 (5): 915-27. Cravedi P, Ruggenenti P, Remuzzi G. Old Donors for Kidney Transplantation: How Old? Gerontology. 2010 Oct 26. Perico N, Casiraghi F, Introna M, Gotti E, Todeschini M, Cavinato RA, Capelli C, Rambaldi A, Cassis P, Rizzo P, Cortinovis M, Maras M, Golay J, Noris M, Remuzzi G. Autologous Mesenchymal Stromal Cells and Kidney Transplantation: A Pilot Study of Safety and Clinical Feasibility. Clin J Am Soc Nephrol. 2010 Oct 7. Cravedi P, Ruggenenti P, Remuzzi G. Sirolimus for calcineurin inhibitors in organ transplantation: contra. Kidney Int. 2010 Dec; 78 (11): 1068-74. Cravedi P, Maggiore U, Mannon RB. Low-density array PCR analysis of reperfusion biopsies: an adjunct to histological analysis. Nephrol Dial Transplant. 2010 Dec; 25 (12): 4077-86. Ruggenenti P, Perna A, Tonelli M, Loriga G, Motterlini N, Rubis N, Ledda F, Rota S Jr, Satta A, Granata A, Battaglia G, Cambareri F, David S, Gaspari F, Stucchi N, Carminati S, EneIordache B, Cravedi P, Remuzzi G; ESPLANADE Study Group. Effects of add-on fluvastatin therapy in patients with chronic proteinuric nephropathy on dual renin-angiotensin system blockade: the ESPLANADE trial. Clin J Am Soc Nephrol. 2010 Nov; 5 (11): 1928-38. Ruggenenti P, Iliev I, Filipponi M, Tadini S, Perna A, Ganeva M, Ene-Iordache B, Cravedi P, Trevisan R, Bossi A, Remuzzi G. Effect of trandolapril on regression of retinopathy in hypertensive patients with type 2 diabetes: a prespecified analysis of the benedict trial. J Ophthalmol. 2010; 2010: 106384.

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Bell KJ, Hayen A, Macaskill P, Craig JC, Neal BC, Fox KM, Remme WJ, Asselbergs FW, van Gilst WH, Macmahon S, Remuzzi G, Ruggenenti P, Teo KK, Irwig L. Monitoring initial response to Angiotensin-converting enzyme inhibitor-based regimens: an individual patient data meta-analysis from randomized, placebo-controlled trials. Hypertension. 2010 Sep; 56 (3): 5339. Herz M, Gaspari F, Perico N, Viberti G, Urbanowska T, Rabbia M, Kirk DW. Effects of high dose aleglitazar on renal function in patients with type 2 diabetes. Int J Cardiol. 2010 Sep 11. Ruggenenti P, Cattaneo D, Rota S, Iliev I, Parvanova A, Diadei O, Ene-Iordache B, Ferrari S, Bossi AC, Trevisan R, Belviso A, Remuzzi G; Ezetimibe and Simvastatin in Dyslipidemia of Diabetes (ESD) Study Group. Effects of combined ezetimibe and simvastatin therapy as compared with simvastatin alone in patients with type 2 diabetes: a prospective randomized double-blind clinical trial. Diabetes Care. 2010 Sep; 33 (9): 1954-6. De Santo RM, Bilancio G, Santoro D, Vecchi ML, Perna A, De Santo NG, Cirillo M. A longitudinal study of sleep disorders in early-stage chronic kidney disease. J Ren Nutr. 2010 Sep; 20 (5 Suppl): S59-63. Hoogduijn MJ, Popp FC, Grohnert A, Crop MJ, van Rhijn M, Rowshani AT, Eggenhofer E, Renner P, Reinders ME, Rabelink TJ, van der Laan LJ, Dor FJ, Ijzermans JN, Genever PG, Lange C, Durrbach A, Houtgraaf JH, Christ B, Seifert M, Shagidulin M, Donckier V, Deans R, Ringden O, Perico N, Remuzzi G, Bartholomew A, Schlitt HJ, Weimar W, Baan CC, Dahlke MH; MISOT Study Group. Advancement of mesenchymal stem cell therapy in solid organ transplantation (MISOT). Transplantation. 2010 Jul 27; 90 (2): 124-6. Perico N, Antiga L, Caroli A, Ruggenenti P, Fasolini G, Cafaro M, Ondei P, Rubis N, Diadei O, Gherardi G, Prandini S, Panozo A, Bravo RF, Carminati S, De Leon FR, Gaspari F, Cortinovis M, Motterlini N, Ene-Iordache B, Remuzzi A, Remuzzi G. Sirolimus therapy to halt the progression of ADPKD. J Am Soc Nephrol. 2010 Jun; 21 (6): 1031-40. Ruggenenti P, Cravedi P, Remuzzi G; Medscape. The RAAS in the pathogenesis and treatment of diabetic nephropathy. Nat Rev Nephrol. 2010 Jun; 6 (6): 319-30. Ruggenenti P, Van der Meer IM, Remuzzi G. Oral acetyl-L-carnitine therapy and insulin resistance. Hypertension. 2010 Jun; 55 (6): e26. Gaspari F, Cravedi P, Mandal M, Perico N, de Leon FR, Stucchi N, Ferrari S, Labianca R, Remuzzi G, Ruggenenti P. Predicting Cisplatin-induced acute kidney injury by urinary neutrophil gelatinase-associated lipocalin excretion: a pilot prospective case-control study. Nephron Clin Pract. 2010; 115 (2): c154-60. Vogt L, Chiurchiu C, Chadha-Boreham H, Danaietash P, Dingemanse J, Hadjadj S, Krum H, Navis G, Neuhart E, Parvanova AI, Ruggenenti P, Woittiez AJ, Zimlichman R, Remuzzi G, de Zeeuw D; PROLONG (PROteinuria Lowering with urOteNsin receptor antaGonists) Study Group. Effect of the urotensin receptor antagonist palosuran in hypertensive patients with type 2 diabetic nephropathy. Hypertension. 2010 May; 55 (5): 1206-9. van der Meer IM, Ruggenenti P, Remuzzi G. The diabetic CKD patient-a major cardiovascular challenge. J Ren Care. 2010 May; 36 Suppl 1: 34-46.

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van der Meer IM, Cravedi P, Remuzzi G. The role of renin angiotensin system inhibition in kidney repair. Fibrogenesis Tissue Repair. 2010 May 4; 3: 7. Cravedi P, van der Meer IM, Cattaneo S, Ruggenenti P, Remuzzi G. Successes and disappointments with clinical islet transplantation. Adv Exp Med Biol. 2010; 654: 749-69. Cravedi P, Ruggenenti P, Remuzzi G. Low-dose rituximab for posttransplant recurrent membranous nephropathy. Am J Transplant. 2010 May; 10 (5): 1336. Caroli A, Antiga L, Cafaro M, Fasolini G, Remuzzi A, Remuzzi G, Ruggenenti P. Reducing polycystic liver volume in ADPKD: effects of somatostatin analogue octreotide. Clin J Am Soc Nephrol. 2010 May; 5 (5): 783-9. Persy VP, Remuzzi G, Perico N, Benghanem Gharbi M, Adu D, Jha V, Rizvi A, Ben Ammar M, Fongoro S, De Broe ME. Prevention and transplantation in chronic kidney disease: what is achievable in emerging countries? Meeting report: Bamako meeting december 4-6, 2008. Nephron Clin Pract. 2010; 115 (2): c122-32. Mignani R, Feriozzi S, Schaefer RM, Breunig F, Oliveira JP, Ruggenenti P, Sunder-Plassmann G. Dialysis and transplantation in Fabry disease: indications for enzyme replacement therapy. Clin J Am Soc Nephrol. 2010 Feb; 5 (2): 379-85. Cortinovis M, Gotti E, Remuzzi G, Perico N, Cattaneo D, Baldelli S. Omega-3 polyunsaturated fatty acids affect sirolimus exposure in kidney transplant recipients on calcineurin inhibitor-free regimen. Transplantation. 2010 Jan 15; 89 (1): 126-7.

RESEARCH ACTIVITIES Laboratory of Biostatistics Low-molecular-weight heparin in pregnant women with previous unexplained pregnancy complications. A multicenter, randomized, controlled trial: first interim analysis.
HAPPY is a multicenter, randomized, controlled clinical trial aimed to assess the effect of lowmolecular-weight heparin in women with previous unexplained pregnancy complications. The study was supported by a grant for independent research by the Italian agency of the drug (Agenzia Italiana del Farmaco), on behalf of the Ministry of Health (trial registration: EudraCT 2006-004205-26). An interim analysis was foreseen when 50% of the women to be enrolled were randomly allocated. In 2010 the first planned interim analysis was performed.

A prospective, randomized, double-blind, placebo-controlled trial to evaluate the effect of 6-month acetyl-L-carnitine therapy on arterial blood pressure, lipid and metabolic profile, and kidney function in hypertensive patients with type 2 diabetes on background simvastatin therapy (DIABASI STUDY): interim analysis.
DIABASI is a randomized, double-blind, placebo-controlled clinical trial aimed to asses the effect of 6-month therapy with Acetyl-L-Carnitine compared to placebo on systolic blood

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pressure in patients with type 2 diabetes, arterial hypertension and dyslipidemia on stable background antihypertensive, hypoglycemic, and lipid lowering therapy. In 2010 the first planned interim analysis was performed.

BErgamo NEphrologic DIabetes Complications (BENEDICT-B) follow up extension.

Trial

Phase

The main objective of the follow up extension of the BENEDICT-B study was to evaluate the effect of albuminuria levels (among the microalbuminuric range) on cardiovascular disease (CVD) in the long term in type 2 diabetic patients. The BENEDICT phase B core study was a randomized, double blind clinical trial that evaluated diverse therapeutic strategies to prevent macroalbuminuria in 281 patients. In 2010 the follow up extension has been started.

Pre- vs Post-Surgery Low-Dose Rabbit Thymoglobulin (RATG) Plus Basiliximab Induction in Renal Transplantation: A Prospective, Matched Cohort Study: final analysis.
In high-risk kidney transplant recipients, low-dose RATG plus basiliximab prevented acute rejection as effectively as standard RATG induction, but was safer and cheaper. In a singlecenter, prospective, matched-cohort study, we compared the outcome of 21 consecutive patients induced with a 6-day low-dose (0.5 mg/kg/day) RATG course started on average 6 h before graft reperfusion with that of 42 gender- and age- (5 years) matched reference-patients given the same RATG course started post-surgery. All subjects were managed by the same team and received the same concomitant therapy with basiliximab (20 mg on day 0 and 4 post-transplant), cyclosporine and mychophenolate mofetil or azathioprine, without steroids. Each couple of reference patients had the same observation period of corresponding patient. In 2010 the final analysis was performed.

Development of a new algorithm for predicting GFR in diabetic patients.

It remains a strong limitation in clinical practice that the available formulas do not properly reflect actual Glomerular Filtration Rate (GFR) in type 2 diabetes, particularly in patients with normal or supra-normal renal function, where preventive strategies are more useful. It was attempted to develop a new formula for estimating GFR in about 600 patients enrolled in DEMAND and BENEDICT Phase B trials, where GFR was measured (ioexhol clearance, mGFR). A linear regression model was used, where mGFR was the dependent variable, evaluating the role of different predictive covariates. The performance of the formula was assessed through the Lin concordance correlation coefficient (c). In 2010 the final analysis was performed.

Laboratory of Coordination and Conduction of Controlled Clinical Trials

The main aim of the Laboratory is to implement and coordinate all the activities needed to fulfil the trials planned by the Renal Medicine Department, according to the study protocols and the Good Clinical Practice (GCP). To Laboratory staff collaborates with all the Laboratories/Unit of the Mario Negri Institute involving in the clinical studies coordinated by the Renal Medicine Department taking care of: to guarantee the flow of the information between the Laboratories/Units of the Renal Medicine Department and to guarantee a continous updating of the trial status; to develop the case report form of studies; to develop the database of studies; to implement and update a centralized system of data management easily enjoyable by researchers of the Renal Medicine Department; to promote training activities for young investigators; to implement and update the SOPs needed for the trial protocols.

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Moreover part of the Laboratory staff is specifically devoted to the clinical study monitoring which purposes are to verify that: the rights and well-being of human subjects are protected; the reported trial data are accurate, complete, and verifiable from source documents; and the conduct of the trial is in compliance with the currently approved protocol/ amendments, with GCP, and with applicable regulatory requirements. Clinical monitors acting as the main line of communication between the sponsor and the investigator; verifying that the investigator has adequate qualifications and resources and these remain adequate throughout the trial period, and that the staff and facilities, including laboratories and equipment, are adequate to safely and properly conduct the trial and these remain adequate throughout the trial period; verifying that the investigator follows the approved protocol and all approved amendments; discussing wiht the investigator any protocol deviation; verifying that each adverse event is well-documented and timely notified to Authorities as required by GCP; verifying the subjects recruitment rate and discussing with Sponsor and investigator new strategies to implement the enrolment. For the investigational medicinal product (IMP), verifying: that storage times and conditions are acceptable, and that supplies are sufficient throughout the trial; that the IMP are supplied only to subjects who are eligible to receive it and at the protocol specified doses; that subjects are provided with necessary instruction on properly using, handling, storing, and returning the IMP; that the receipt, use, and return of the IMP at the trial sites are controlled and documented adequately; that the disposition of unused IMP the trial sites complies with applicable regulatory requirements and is in accordance with the sponsors authorized procedures. In 2010 have been coordinated and monitored 18 clinical studies perfomed in collaboration with 80 Italian Hospital where 350 monitoring visits have been done. Moreover, the training activity of Drug Monitoring Unit personnel continued throughout the year with single and group lessons and tutoring during the monitoring visit at Centers. During the last year, two clinical study monitors have been trained. In collaboration with Laboratory of Regulatory Affairs for Clinical Studies were held three Investigator Meetings for Investigators involved in A.TH.E.N.A., A.R.CA.DIA. and Remission Clinic studies.

Laboratory of Pharmacokinetics and Clinical Chemistry Performance of GFR estimation equations in Autosomal Dominant Polycystic Kidney Disease (ADPKD) and type 2 diabetes patients with virtually identical renal function
The methods for precise measurement of glomerular filtration rate (GFR), the best overall index of renal function, require the infusion of specific exogenous substances such as inulin, 51CrEDTA, 99mTc-DTPA or unlabelled contrast agents such iohexol. These methods, however, are laborious and expensive. Therefore, in clinical practice formula-derived estimates of GFR have been adopted that include serum creatinine and anthropometric indexes such as gender, age, and weight to account for between-individual differences in muscle mass and the consequent differences in creatinine generation. The evaluation of GFR is of critical importance in the clinical management of ADPKD patients. This is the most common hereditary renal disease, responsible for the 8% to 10% of the cases of end-stage renal disease (ESRD) in Western Countries. The ADPKD progression is largely dependent on the development and growth of cysts and secondary disruption of normal tissue and, eventually, the renal function will decrease. Similarly, diabetic patients also necessitate a careful monitoring of renal function since diabetic nephropathy affects 25-40% of patients with diabetes, that is the leading cause of end-stage renal disease. Scant data in both ADPKD and type 2 patients on prediction formulas

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performance are available so far, and, interestingly, in literature there are no any described equation specifically developed on these populations. Therefore we aimed to explore in ADPKD and type 2 diabetic patients the performance in predicting GFR of fourteen formulas compared to the reference iohexol measured GFR, to evaluate the possible differences in accuracy of GFR estimation between these populations and to assess whether the peculiar demographic, anthropometric and biochemical characteristics of these subjects may play a relevant and different role in predicting renal function. We studied 97 ADPKD patients with normal renal function or mild to moderate renal insufficiency. Each ADPKD patient was matched with a diabetic subject of the same gender and with a measured GFR differing no more than 1 mL/min/1.73m2. GFR determined by the plasma clearance of iohexol was compared with values estimated by means of 14 prediction equations. The selection criterion segregated two cohorts of patients with mean GFR values virtually identical: 81.4026.41 and 81.6826.24 mL/min/1.73m2 in ADPKD and diabetic subjects, respectively. GFR ranged between 30 to about 140 mL/min/1.73m2. Subjects had similar serum creatinine levels (1.150.43 vs. 1.040.40 mg/dL), however diabetic patients were significantly shorter, heavier, older and had a higher body mass index than ADPKD subjects. Glucose levels as well as glycated hemoglobin, total cholesterol, triglycerides and systolic blood pressure were higher in diabetics. Each prediction equation significantly correlated with measured GFR. However, the correlation coefficient was significantly higher in ADPKD subjects. Performance of GFR equations showed that both bias and mean percent error (MPE) in GFR prediction were quite different for each equation in the two patients cohorts. Considering all the models as a whole, median values were positive (bias: 2.61, MPE: 4.17) in ADPKD and negative (bias: -1.37; MPE: -3.20) in diabetic subjects. In ADPKD patients, CKD-Epi and Jelliffe-2 equations gave the best accuracy; however only approximately 49% of estimated GFR were within an error between 10% (i.e. values virtually identical to measured GFR). In diabetic patients all the equations showed markedly lower accuracy. To further clarify the different performance of the prediction formulas in the two populations, the patients were classified in stages of kidney function according to K/DOQI guidelines. Median bias was similar in the three stages for ADPKD patients (1.78, 5.36, 1.03) and smaller than in diabetic patients for stage 1 and 3. In diabetic subjects bias was similar in stage 3 and stage 2 (5.16, 4.34), but was markedly negative in stage 1 patients (-12.67), indicating that at higher levels of renal function (GFR >90 mL/min/1.73m2) in these patients prediction equations consistently underestimate the true GFR. MPE values showed a similar trend. These findings showed a different performance of prediction formulas in ADPKD than in diabetic patients, despite they had identical measured GFR. This is true especially in subject with normal or near normal renal function, suggesting that the different biochemical and metabolic characteristics of the patients may have a relevant impact on accuracy of GFR estimation. Furthermore, our data documented that these prediction formulas must be used cautiously, especially in clinical trials, since they do not represent a valid substitute of the direct measurement of renal function: no more than 50% of GFR were estimated within 10% error.

Development of a simple and reliable method to measure GFR in the rat

Glomerular filtration rate (GFR) is the best index to evaluate renal function. While in human patients a lot of methods are available, in the small laboratory animals methodological and analytical difficulties do still exist. The common techniques for assessing GFR in animals models are cumbersome, invasive and can required the animals sacrifice, that prevent the repetition of these measurements in the same subject at different intervals. Conventional measurements necessitate also extensive surgery and a degree of diuresis, that involves bladder

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catheterization to warrant accurate urine collections. Moreover, the reference techniques rely on anaesthesia, which may influence the renal function in different experimental situation. Considering all these drawbacks, we planned to apply on rats our standardized method for GFR evaluation in humans, based on plasma clearance determination of iohexol, a non-radioactive contrast medium, as a reliable marker of GFR. The modified method required a very small sample volume (15 l) and the determination of iohexol concentrations by high performance liquid chromatographic in whole blood. Plasma concentrations were eventually calculated by correcting by the hematoctrit value. After intravenous injection of iohexol, blood samples were drawn in rats according to a 2compartment kinetic profile. A total of 11 samples per animal were obtained. Iohexol clearance was determined as Dose/AUC (area under the blood-time concentration). The AUC was determined according to 3 different approaches: by using the whole profile (2-compartment model, reference method) and determining AUC by the trapezoidal rule; by the slope-intecept method by using all the time-points belonging to the elimination phase; by the slope-intercept method by using only 4 iohexol samples taken at selected time points (simplified methods). The mean GFR according to the 2-compartment model was 1.060.29 mL/min/100g; with the slope-intercept approach was 1.180.32 mL/min/100g and by using only 4 time points was 1.200.35 mL/min/100g. We found a constant ratio (0.90) between each of the two the slopeintercept and the reference method. This value has been used as a correction factor to obtain the true clearance from the simplified 4-points slope-intercept approach. The procedure we set up to measurement of GFR in conscious rats was proven to be reliable, requires the drawing of small volume of whole blood at 4 selected time-points without the need of urine collections and catheterization of the animals. This method allows repeated GFR determinations in the same rat in long term experiments.

Laboratory of Advanced Development of Drug Renal graft function and low-dose cyclosporine affect mycophenolic acid pharmacokinetics in kidney transplantation
The pharmacokinetics of mycophenolic acid (MPA), the active compound of mycophenolate mofetil (MMF), is influenced by concomitant cyclosporine (CsA) administration and renal function. In this study we evaluated the impact of minimization of CsA dose on MPA exposure in stable kidney transplant recipients (> 6 months post-surgery) with varying degree of graft function deterioration. To address this issue, we performed a retrospective analysis on 135 complete MPA profiles collected from 56 kidney transplant recipients given MMF and lowdose CsA. In particular, dose-adjusted MPA pharmacokinetic parameters were correlated with the area under the concentration-time curve from 0 to 12 h (AUC0-12) of CsA and with renal function, measured as glomerular filtration rate (GFR) by the plasma clearance of iohexol. Moreover, the relative contribution of CsA exposure and GFR to MPA kinetics as compared to other clinical and hematochemical parameters was determined in multivariate analysis. We found that dose-adjusted MPA AUC0-12 negatively and significantly correlated with CsA AUC012. Although the correlation was quite weak, dose-adjusted MPA AUC0-12 significantly increased when CsA AUC0-12 was below 2000 ng*h/mL. An inverse and significant association was also documented between daily MPA exposure and GFR. Moreover, stratification of MPA profiles according to three stages of renal dysfunction showed that dose-adjusted MPA AUC0-12 was significantly higher in patients with severe renal insufficiency. The lowest CsA dose given in patients with severe renal insufficiency might have also contributed to this trend. In multivariate analysis GFR, albumin and hemoglobin levels and use of gastric pH modulators were identified as independent determinants of MPA AUC0-12. Altogether, these data suggest that minimization of CsA dose should be accompanied by monitoring of MPA concentrations, especially in

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patients with severe renal insufficiency, in order to maintain adequate immunosuppression while avoiding the risk of MPA over-exposure.

Pharmacogenomics of azathioprine in kidney transplantation to predict tolerability and graft outcome

In kidney transplantation azathioprine (AZA) and mycophenolate mofetil (MMF) have been shown equally effective in preventing both acute rejection and chronic allograft injury in combination with cyclosporine and short-term corticosteroid. This is of particular importance for developing countries, as AZA is 16-fold less expensive than MMF. The main enzyme responsible of AZA metabolism is thiopurine methyltransferase (TPMT). Eleven percent of individuals are heterozygous for non-functional alleles of TPMT gene and exhibit reduced enzyme activity. AZA given to TPMT-deficient patients may induce fatal myelosuppression. The risk of AZA complications is very high in patients homozygous for TPMT non-functional alleles, but considerably lower in heterozygous individuals. This suggests that factors other than TPMT genotype may affect the enzyme activity. S-adenosylmethionine (SAM) is a TPMT stabilizing cofactor whose endogenous levels are influenced by 5,10-methylenetetrahydrofolate reductase (MTHFR) activity. Two single nucleotide polymorphisms (SNPs) in the MTHFR gene, 677C>T and 1298A>C, are associated with decreased MTHFR activity, which reduces SAM availability and TPMT stability. Other enzymes, such as Inosine triphosphatase (ITPA), are involved in AZA metabolism and SNPs in their genes may affect drug tolerability by causing accumulation of AZA toxic metabolites, as documented in patients with inflammatory bowel disease. So far SNPs in MTHFR and ITPA genes have not been assessed in transplant setting. Our preliminary data confirm the presence of SNPs in TPMT, MTHFR and ITPA genes in kidney transplant recipients. Altogether, these observations suggest that individualization of AZA dosage according to TPMT, MTHFR and ITPA genotypes could limit AZA-related side effects and optimize treatment for long-term graft survival in kidney transplant recipients. Specifically, in this study we want to: 1) investigate the relative contribution of TPMT, MTHFR and ITPA allelic variants to the likelihood of developing thiopurine-related side effects in kidney transplant recipients given AZA as part of their immunosuppressive therapy; 2) assess whether TPMT, ITPA and MTHFR genotypes correlate with renal function decline and/or graft survival over 3-year follow-up and 3) develop algorithms able to bring together genetic and clinical data aimed to predict the best AZA dosage for each kidney transplant recipient to minimize side effects and optimize graft outcome.

Effects of sirolimus on disease progression in patients with autosomal dominant polycystic kidney disease (The SIRENA study)
Activation of mammalian target of rapamycin (mTOR) pathways may contribute to uncontrolled cell proliferation and secondary cyst growth in patients with autosomal dominant polycystic kidney disease (ADPKD). To assess the effects of mTOR inhibition on disease progression, we performed a randomized, crossover study (The SIRENA Study) comparing a 6month treatment with sirolimus or conventional therapy alone on the growth of kidney volume and its compartments in 21 patients with ADPKD and GFR>or=40 ml/min per 1.73 m2. In 10 of the 15 patients who completed the study, aphthous stomatitis complicated sirolimus treatment but was effectively controlled by topical therapy. Compared with pre-treatment, post-treatment mean total kidney volume increased less on sirolimus (46+/-81 ml; P=0.047) than on conventional therapy (70+/-72 ml; P=0.002), but we did not detect a difference between the two treatments (P=0.45). Cyst volume was stable on sirolimus and increased by 55+/-75 ml (P=0.013) on conventional therapy, whereas parenchymal volume increased by 26+/-30 ml (P=0.005) on sirolimus and was stable on conventional therapy. Percentage changes in cyst and parenchyma volumes were significantly different between the two treatment periods. Sirolimus had no appreciable effects on intermediate volume and GFR. Albuminuria and proteinuria

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marginally but significantly increased during sirolimus treatment. In summary, sirolimus halted cyst growth and increased parenchymal volume in patients with ADPKD. Whether these effects translate into improved long-term outcomes requires further investigation. The results of this study have been published in Journal of American Society of Nephrology 2010; 21: 1031-40.

Effects of sirolimus on disease progression in patients with autosomal dominant polycystic kidney disease and severe renal insufficiency (The SIRENA II study)
The results of the SIRENA study documented that in patients with autosomal dominant polycystic kidney disease (ADPKD) and normal kidney function or moderate renal dysfunction, a short-course treatment with sirolimus halted cyst growth and increased parenchymal volume. Moreover, a retrospective study in a small number of sirolimus-treated ADPKD transplant patients showed that the treatment significantly reduced native kidney volumes over an average of 24 month follow-up. This reduction was three times higher than that reported in a control group of ADPKD transplant recipients not given sirolimus over a 40 month period. These observations provide the rationale for testing the efficacy of sirolimus even in adults ADPKD patients with severe renal insufficiency (15-40 mL/min/1.73m2). In particular, the aim of this study is to assess the safety and the short-term efficacy of 1 year treatment with sirolimus (on the top of the best available therapy) in slowing renal function decline as compared to conventional therapy. Thereafter, an extension phase of two year period is planned to primarily evaluate the long-term impact of sirolimus treatment on renal function decline and on slowing kidney (and liver) growth rate.

Ex-vivo expanded mesenchymal stem cells to repair the kidney and improve function in cisplatin-induced acute renal failure in patients with solid organ cancer
Since its introduction into clinical trials, cisplatin has had a major impact in cancer medicine, changing the course of therapeutic management of several tumours, such as that of ovary. Unfortunately, this compound causes dose-dependent and cumulative nephrotoxicity sometimes requiring dose reduction or discontinuation of treatment. Present strategies for the treatment of acute renal failure induced by cisplatin have focused on targeting individual mechanisms thought to contribute to ischemic or toxic insults to the kidney. However, translational research efforts in patients have yielded disappointing results. An alternative possibility is to adopt a strategy aimed to regenerate the injured renal tissue. Attempts to accelerate recovery have focused on administration of growth factors. Although this strategy has been successful in experimental models, no beneficial effects have been observed in clinical trials. The ability of extrarenal cells to participate in the regenerative response following post-transplant acute renal failure may hold true for acute renal failure that develops in native kidneys after cisplatin therapy. The rationale for this approach rests on the recent demonstration in mice that MSCs infusion repairs acute tubular damage in animals given cisplatin. Similarly, consistent evidence of the beneficial effect of bone-marrow derived cell therapy has been recently reported in humans with ischemic heart disease. These observations raises the possibility that adult-derived bone marrow cells could be administered to enhance the recovery from renal injury. The aim of this study is to evaluate the efficacy of ex-vivo expanded donor mesenchymal stem cells infusion in the acceleration of tubular regeneration, and thus renal function recovery, in patients with cisplatin-induced acute renal failure, a disease that so far has not cure. Acute renal injury will be diagnosed by measuring urine concentrations of neutrophil gelatinase-associated lipocalin (NGAL).

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Effect of induction and maintenance immunosuppressive therapies on de novo anti-human leukocyte (HLA) antibodies development in kidney transplantation
The post-transplant production of anti-human leukocyte antigen (HLA) antibodies represents an important risk factor for the development of both acute rejection and chronic allograft injury in kidney transplantation. Notwithstanding, while the relevance of pre-formed anti-HLA antibodies has been extensively studied, the role of anti-HLA class I and II antibodies developed after kidney transplantation has not been fully elucidated so far. The preliminary results of a prospective multicenter study on 2278 kidney graft recipients showed that, at one year after transplant, 6.6% of patients who lost their graft had anti-HLA antibodies, as compared with 3.3% of those who were negative for antibodies. Further small studies confirmed the presence of anti-HLA antibodies as a risk factor for both graft loss and graft function decline. During the past decade, anti-HLA antibody tests have moved from complement-dependent citotoxicity to solid-phase assays, which showed increased sensitivity and accuracy to detect anti-HLA antibodies. Luminex technology is the most widely used method to detect and define low levels of these antibodies. The primary aim of this study is to use the Luminex technology to analyze de novo anti-HLA antibodies development after kidney transplantation, taking into account the immunosuppressive treatment. Specifically, in this study we want to: 1) assess the impact of two different induction therapies, namely Campath-1H or Basiliximab plus low-dose Rabbit anti-thymocyte globulin, on anti-HLA antibody development post-transplantation; 2) to evaluate the effect of maintenance immunosuppressive therapy with mycophenolate mofetil administered with sirolimus or cyclosporine on anti-HLA antibody production; 3) to evaluate whether antiHLA antibodies development post-transplantation translates into worse renal function and/or graft survival compared with that of patients without anti-HLA antibodies over 4-year followup.

Autologous Mesenchymal Stromal Cells and Kidney Transplantation: A Pilot Study of Safety and Clinical Feasibility
Mesenchymal stromal cells (MSCs) abrogate alloimmune response in vitro, suggesting a novel cell-based approach in transplantation. Moving this concept toward clinical application in organ transplantation should be critically assessed. A safety and clinical feasibility study (ClinicalTrials.gov, NCT00752479) of autologous MSCs infusion was conducted in two recipients of kidneys from living-related donors. Patients were given T cell-depleting induction therapy and maintenance immunosuppression with cyclosporine and mycophenolate mofetil. On day 7 posttransplant, MSCs were administered intravenously. Clinical and immunomonitoring of MSC-treated patients was performed up to day 360 postsurgery. We found that serum creatinine levels increased 7 to 14 days after cell infusion in both MSC-treated patients. A graft biopsy in patient 2 excluded acute graft rejection, but showed a focal inflammatory infiltrate, mostly granulocytes. In patient 1 protocol biopsy at 1-year post-transplant showed a normal graft. Both MSC-treated patients are in good health with stable graft function. A progressive increase of the percentage of CD4(+)CD25(high)FoxP3(+)CD127(-) Treg and a marked inhibition of memory CD45RO(+)RA(-)CD8(+) T cell expansion were observed posttransplant. Patient T cells showed a profound reduction of CD8(+) T cell activity. Findings from this study in the two patients show that MSC infusion in kidney transplant recipients is feasible, allows enlargement of regulatory T in the peripheral blood, and controls memory CD8(+) T cell function. The results of this study have been published in Clinical Journal of American Society of Nephrology 2011; 6. This study is currently underway and the protocol of autologous MSCs infusion has also been applied to other patients.

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Efficacy of paricalcitol in reducing parathyroid hormone levels and ameliorating markers of bone marrow remodelling in renal transplant recipients with secondary hyperparathyroidism (APPLE study)
The risk of fracture for kidney transplant recipients is four times higher than that of the general population. The pathogenesis of post-transplant bone disease is multifactorial, but hyperparathyroidism plays a key role in the maintenance or development of bone remodelling. Vitamin D and its analogues are key components of treatment aimed to prevent or ameliorate secondary hyperparathyroidism in patients with chronic kidney disease (CKD). In hemodialysis patients, intravenous paricalcitol, a new vitamin D analogue, achieves a faster and more effective normalization of parathyroid hormone (PTH) levels than calcitriol, an effect that is associated with smaller changes in serum calcium and phosphorous levels. Preliminary evidence is also available that in pre-dialysis patients with CKD and secondary hyperparathyroidism, treatment with oral paricalcitol may also reduce urinary protein excretion, an effect that is independent of concomitant treatment with agents that block renin-angiotensin system and that in the long-term might translate into slower progression to end stage kidney disease and need for renal replacement therapy. Renal transplant patients are at high risk of hyperparathyroidism, largely because of long-lasting renal insufficiency before transplant, and of progressive deterioration of kidney function because of chronic allograft nephropathy (a disease of proteinuria and progressive decline of the glomerular filtration rate). The aims of this study are: 1) evaluate whether 6-month treatment with paricalcitol may achieve a prompt and effective reduction of PTH serum levels in stable renal transplant patients with secondary hyperparathyroidism; 2) assess whether oral paricalcitol may also achieve reduction of urinary protein excretion; 3) verify whether amelioration of hyperparathyroidism (if any) may translate into an improvement of bone remodelling. Laboratory of Clinical Pathophysiology of Renal Disease and Transplantation Main objective of the Laboratory of Clinical Pathophysiology of Renal Disease and Transplantation is the study of pathophysiological mechanisms underlying the progression of chronic kidney disease and the identification of new therapeutic strategies for diabetic kidney disease and nondiabetic proteinuric nephropathies and chronic rejection. The multidisciplinary nature of these lines of research necessarily requires a close integration between various skills of clinical physiology and clinical pharmacology, and of molecular biology. For this reason, the laboratory includes not only researchers from the Department of Renal Medicine, which touch on the laboratory, but also the Department of Public-Private Medical Specialist and Transplants. From an operational standpoint, the laboratory interacts closely with the Laboratory for the coordination and conduct of controlled clinical trials which finalizes the protocols designed for clinical research within the Department of Renal Medicine.

A prospective, sequential study to assess the efficacy of rituximab therapy in maintaining remission of NEphrotic syndrome after steroid and immunosuppressive therapy withdrawal in patients with steroiddependant or Multirelapsing minimal change disease Or focal segmental glomerulosclerosis (NEMO Study)
Patients, especially children, with steroid-dependant or multirelapsing nephrotic syndrome (NS) secondary to minimal change disease (MCD) or idiopathic focal and segmental glomerulosclerosis (FSGS) on continuous treatment with steroids and/or other immunosuppressive agents to limit or prevent recurrences are at increased risk of severe drugrelated adverse events. Case reports suggest that Rituximab, a B cell depleting monoclonal

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antibody, could be a safe and effective alternative to steroid or immunosuppressants to achieve and maintain remission in this population. The study is primarily aimed at evaluating whether Rituximab may maintain stable NS remission after tapering and withdrawal of steroid and immunosuppressive therapy in patients with MCD or FSGS and steroid-dependent or multirelapsing NS. Secondarily, the study will assess whether Rituximab allow reducing maintenance doses of steroids and other immunosuppressants (in those who relapse), thus limiting treatment related side effects and costs. This prospective, sequential, open, study will include 20 patients with histology evidence of MCD or FSGS and steroid-dependant or multirelapsing NS, who are on stable complete or partial remission since at least 1 month and, based on their previous history, are expected to invariably relapse after steroid/immunosuppression withdrawal. After baseline evaluation of clinical, laboratory and kidney function parameters [including GFR, RPF, albumin and sodium fractional clearance and the glomerular albumin permeability assay (Palb)], patients will receive one Rituximab infusion that will be repeated 1 week later if CD20 cells are not fully depleted from the circulation. Then ongoing immunosuppression will be progressively tapered up to complete withdrawal over 6 to 9 months. 24h proteinuria will be monitored monthly and spot urine will be tested daily by albustix to early detect disease relapses. Baseline evaluations will be repeated at study end (1 year). Relapses will be treated with high-dose steroids as per center practice and the last immunosuppressive therapy effective in preventing disease reactivation will be reintroduced. Rituximab is expected to prevent NS recurrence following tapering and discontinuation of steroid and other immunosuppressants. Maintaining remission without chronic immunosuppression is expected to minimize risks and costs of therapy and to remarkably improve patient outcomes.

A prospective, randomized, open label, blinded end-point (PROBE) trial to evaluate whether, at comparable blood pressure control, Ace inhibitor therapy more effectively than non Ras inhibitor therapy reduces CArdiovascular morbidity and mortality in chronic DIAlysis patients with left ventricular hypertrophy (ARCADIA Study)
Angiotensin-converting-enzyme (ACE) inhibitors have a specific cardioprotective effect and, compared to treatment not directly interfering with the renin-angiotensin-system (RAS), significantly reduce cardiovascular (CV) mortality and morbidity in subjects with normal renal function. Patients with end stage renal disease (ESRD) have a 10-20 fold excess cardiovascular risk compared to subjects with normal renal function and excess risk is even higher in those with left ventricular hypertrophy (LVH). Despite CV events are the leading cause of death in these patients, no adequately powered trial so far evaluated the specific cardioprotective effect of ACE inhibitors in this population. The study is primarily aimed at evaluating whether, at comparable blood pressure (BP) control, ACE inhibitor as compared to non-RAS inhibitor therapy significantly reduces the incidence of a composite end point of CV death (including sudden death) and non-fatal myocardial infarction or stroke in hypertensive (pre-dialysis systolic/diastolic BP >140/90 mmHg or postdialysis systolic/diastolic BP >130/80 mmHg or antihypertensive therapy) chronic dialysis patients with LVH. Secondarily, the study will compare the incidence of single components of the primary outcome, new onset paroxysmal or persistent atrial fibrillation, thrombosis of the artero-venous fistula, progression or regression of LVH, changes in components of the metabolic syndrome, the safety profile of the two treatment regimens and their cost/effectiveness. This prospective, randomized, open label, blinded end point (PROBE) trial will include 624 hypertensive ESRD patients with echocardiography evidence of LVH who are on chronic hemodialysis since >6 months. After 1 month wash-out period from previous RAS inhibitor

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therapy and stratification for diabetes YES/NO, they will have a baseline evaluation of main clinical and laboratory parameters and will be randomized to 2-year treatment with an ACE inhibitor or a BP lowering regimen not including RAS inhibitors. Treatment will be adjusted to achieve and maintain a target BP <140/90 mmHg (pre-dialysis) and a target BP <130/80 mmHg (post-dialysis) in both groups. ACE inhibitor compared to non-RAS inhibitor therapy is expected to reduce more effectively fatal and non-fatal CV events, limit progression or induce regression of LVH, improve some components of the metabolic syndrome, and reduce treatment costs for cardiovascular complications. These findings might help achieving more effective cardioprotection in people on chronic dialysis at lower costs.

A prospective, randomized, open label blinded end point (PROBE), CrossOver study to compare the effects of Telmisartan ANd losartan on metabolic profile of renal Transplant patients (COSTANT Study)
In renal transplant recipients, residual renal insufficiency combined to the effects of immunosuppressive therapy with steroids or calcineurin inhibitors may reduce insulin activity and may contribute to several of the abnormalities associated with the metabolic syndrome, such as hypertension, glucose intolerance and hyperlipidemia1,2. In turn, insulin resistance, hypertension, hyperglycemia and dyslipidemia may importantly contribute to the excess cardiovascular risk of renal transplant patients (an excess comparable to that of diabetes subjects with over diabetic nephropathy) and may also accelerate progressive renal function deterioration and promote graft loss. Recently, drugs such as peroxisome proliferators-activated receptor-g (PPARg) activators, that ameliorate insulin sensitivity and metabolic syndrome, have become available. Recent studies showed that telmisartan, an angiotensin II (AII) type 1 receptor antagonist, in addition to block the AII receptor - a key surface receptor involved in the regulation of blood pressure - may also activate PPARg, thus improving some of the features of the metabolic syndrome, such as hyperglycemia and dyslipidemia in people with hypertension and/or diabetes. Thus, in addition to control high blood pressure and to limit some of the adverse effects of angiotensin II, including target organ damage, graft fibrosis and CsA nephrotoxicity, telmisartan may also substantially reduce the overall cardiovascular and renal risk of renal transplant recipients by ameliorating some of the modifiable components of the metabolic syndrome, such as hypertension, glucose intolerance and hyperlipidemia. On the other hand, telmisartan is devoided of the adverse effects of PPARg activators such as fluid retention, and has therefore a remarkably better risk/benefit profile. Primary aim of the present study is to compare the short-term effects of telmisartan and losartan on insulin sensitivity in kidney transplant recipients with stable renal function and concomitant treatment with steroids and/or calcineurin inhibitors. In this Prospective, Randomized, Open label Blinded End point (PROBE), crossover study10, after four weeks wash-out period from previous ACE inhibitors and AII receptor antagonist, eligible patients will be randomized to two treatment arms with telmisartan or losartan. At the end of the first treatment period with telmisartan or losartan, each patient will crossover to the other treatment. After the second treatment period, there will be a four-week recovery period.

A randomized, prospective, multicenter trial to compare THe Effect on chronic allograft Nephropathy prevention of mycophenolate mofetil versus Azathioprine as the sole immunosuppressive therapy for kidney transplant recipients (ATHENA Study)
Steroid-free, low-dose cyclosporine (CsA) and mycophenolate mofetil (MMF) therapy

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combined to induction with basiliximab and low-dose Rabbit Anti-human Thymocyte Globulin (RATG) effectively prevented acute rejection in kidney transplant recipients. Steroid-free therapy with CsA and MMF has been also associated with a reduced risk of chronic allograft nephropathy (CAN), a frequent cause of late graft loss. Whether basiliximab/low-dose RATG induction plus MMF monotherapy may prevents CAN even in a CsA-free regimen, without increasing the risk of acute rejection, is worth investigating. The MYSS trial found that MMF and AZA are equally effective in preventing acute rejection after steroid withdrawal. Whether AZA might also be as effective as MMF in preventing CAN in a CsA-free immunosuppressive regimen is unknown. The study will primarily compare the incidence of biopsy-proven CAN three years posttransplant in kidney recipients randomly allocated to MMF or AZA, after induction therapy with basiliximab and low-dose RATG, and sequential steroid and CsA withdrawal. Secondarily, the study will compare acute rejections after CsA withdrawal, long-term patient and graft survival, and graft function and prevalence/severity of CAN at study end. Two-hundred-twenty-four kidney transplant recipients from deceased donors given induction therapy with two 20 mg basiliximab injections 4 days apart and a seven-day course of RATG (0.5 mg/kg/day), will be randomly allocated on a 1:1 basis to 3-year treatment with low-dose MMF or AZA, added-on CsA maintenance therapy. At 1 year, rejection-free patients with no evidence of tubulitis at kidney biopsy will withdraw CsA and will have a kidney biopsy 3 year posttransplant for evaluating the presence and severity of CAN. Should the cumulative incidence of acute rejection exceed 15% during CsA withdrawal the study will be stopped. Should the incidence differ by >30% between the two treatment arms, all patients will be given the most effective treatment and the follow up will be continued. A final biopsy will be repeated 4 years posttransplant. Most patients are expected to be effectively maintained on single drug immunosuppression, which implies less steroid- and CsA- related complications and treatment costs. MMF is expected to prevent CAN more effectively than AZA. However, should AZA be more or as effective compared to MMF, at study end all patients could be shifted to AZA, that is 15-fold less expensive than MMF.

Laboratory of Regulatory Affairs for Clinical Studies

Since 1999, the Department of Renal Medicine, in collaboration with other Departments of the Institute and with the Hospital of Bergamo, within the framework of activities of the PublicPrivate Department of Specialist and Transplant Medicine, designs, promotes and coordinates projects of clinical research. These are mainly independent research projects; some of them are carried out exclusively in the Dacc Centre, but most of them are multi-centre trials, in collaboration with Hospitals, Research Centres and Universities, both Italian and foreign. The Laboratory of Regulatory Affairs for Clinical Studies has been created in relation to this activity, to verify that the studies are conducted in compliance with patients safety and rights and according to the legislation in force. In the execution of its activities, the Laboratory, interacts with several interlocutors, among them the Ministry of Health, the Italian Agency for Drugs (AIFA), the Local Ethic Committees, the ASL and the Pharmaceutical companies. The Laboratory, supported by a secretariat, participates in all the phases of planning, organizing, conducting and managing the clinical studies. During the preparatory phase, preceding the onset of the study, the Laboratory collaborates with the researchers in order to establish the protocol and the studys documents in accordance with the principles of Good Clinical Practice. In addition it verifies the adherence of these documents to regulatory and ethical aspects. Once the protocol has been established, the Laboratory

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proceeds to the input of the trial in the National Monitoring Centre on Clinical Research with Medicines (OsSC) and subsequently submits all the documents to the competent authorities, in order to obtain the ethical and administrative approvals and manages all the bureaucratic procedures. During the course of the study, the Laboratory is responsible for complying with the legal obligations to keep AIFA and the Ethic committees constantly updated on the progress of the experimentation. Moreover, it occupies with the collection and maintenance of all essential documents for the conduction of clinical studies. Since 2005, according to the rules set by the International Committee of Medical Journal Editors, the Laboratory ensures that all the clinical studies promoted by the Centre Dacc are registered to the NIH international registry, clinicaltrials.gov. During 2010, the Laboratory has carried out the activities described above for all the ongoing studies at the Department of Renal Medicine and in collaboration with the Hospital of Bergamo and other centers, both Italian and foreign, participating in our research projects. The studies performed in 2010 are 35 and involve about 80 centers. The studies for which approval was requested in the year 2010 are 10. In collaboration with the Monitoring Unit were also organized the Investigator Meetings of studies ARCADIA, ATHENA and Remission Clinic. In respect to the recent sanitary accreditation of Dacc Centre, the Laboratory has maintained relations with ASL and other competent authorities, for everything concerning the maintenance of requirements and other legal purposes.

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RARE DISEASES DOCUMENTATION AND RESEARCH

STAFF

Head

Erica DAINA, M.D.

Genetics for Clinical Research

Head Elena BRESIN, M.D.

Network Development for Rare Diseases

Head Sara GAMBA, Research Nurse

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CURRICULA
Erica Daina got her degree in Medicine at the University of Milan in 1987 and the specialisation in Medical Nephrology in 1990 at the same University. She performed her training at the II Medical Division - San Raffaele Hospital - Milan, and at the Division of Nephrology and Dialysis - Riuniti Hospital - Bergamo. In March 1988 she started her collaboration with the Mario Negri Institute and since June 1993 she works as full-time clinical researcher at the Clinical Research Center for Rare Diseases Aldo e Cele Dacc. 1996 2009: Chief, Information Center for Rare Diseases June 2009: Chief, Laboratory of Rare Diseases Documentation and Research Areas of interest: Rare diseases, Takayasus Arteritis, Hemolytic Uremic Syndrome/Thrombotic Thrombocytopenic Purpura, genetic kidney diseases.
Selected publications
1. 2. 3. 4. Warnock DG, Daina E, Remuzzi G, West M. Enzyme Replacement Therapy and Fabry Nephropathy. Clin J Am Soc Nephrol. Epub 2009 Dec. 10. 2010 feb; 5(2):371-8 Bresin E, Gastoldi S, Daina E, Belotti D, Pogliani E, Perseghin P, Scalzulli PR, Paolini R, Marcen R, Remuzzi G, Galbusera M. Rituximab as pre-emptive treatment in patients with thrombotic thrombocytopenic purpura and evidence of anti-ADAMTS13 autoantibodies. Thromb Haemost 2009; 101(2):233-8 Schieppati A, Henter JI, Daina E, Aperia A. Why rare diseases are an important medical and social issue. Lancet. 2008 Jun 14;371(9629):2039-41. Bresin E, Daina E, Noris M, Castelletti F, Stefanov R, Hill P, Goodship T H J, Remuzzi G, International Registry Recurrent Familial HUS/TTP. Outcome of renal transplantation in patients with non-Shiga toxin-associated Hemolytic Uremic Syndrome: Prognostic significance of genetic background. Clinical Journal American Society Nephrology 2006; 1: 88-99 Daina E, Schieppati A, Remuzzi G. Mycophenolate mofetil for the treatment of Takayasu arteritis: report of three cases. Ann Intern Med. 1999 Mar 2;130(5):422-6.

5.

Elena Bresin got her degree in Medicine at the University of Padua in 1994 and the specialisation in Medical Genetics at the University of Verona in 2000. She performed her training at the Department of Pediatrics of the University Policlinic of Padua, then at the Department of Biology and Genetics of the University Policlinic of Verona and since 2000 at t the Clinical Research Center for Rare Diseases Aldo e Cele Dacc. Since January 2001 she works as full-time clinical researcher at the Clinical Research Center for Rare Diseases Aldo e Cele Dacc and since June 2009 she is Unit Head of Genetics for Clinical Research. Areas of interest: thrombotic microangiopathies, membranoproliferative glomerulonephritis, familial focal segmental glomerulosclerosis.
Selected publications
1. Noris M, Caprioli J, Bresin E, Mossali C, Pianetti G, Gamba S, Daina E, Fenili C, Castelletti F, Sorosina A, Piras R, Donadelli R, Maranta R, van der Meer I, Conway EM, Zipfel PF, Goodship TH, Remuzzi G Relative Role of Genetic Complement Abnormalities in Sporadic and Familial aHUS and Their Impact on Clinical Phenotype. Clin J Am Soc Nephrol, 2010; 5(10):1844-59 Bresin E, Gastoldi S, Daina E, Belotti D, Pogliani E, Perseghin P, Scalzulli PR, Paolini R, Marcen R, Remuzzi G, Galbusera M. Rituximab as pre-emptive treatment in patients with thrombotic thrombocytopenic purpura and evidence of anti-ADAMTS13 autoantibodies. Thromb Haemost 2009; 101(2):233-8 Castelletti F, Donadelli R, Banterla F, Hildebrandt F, Zipfel PF, Bresin E, Otto E, Skerka C, Renieri A, Todeschini M, Caprioli J, Caruso RM, Artuso R, Remuzzi G, Noris M. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Proc Natl Acad Sci U S A 2008 Feb 19;105(7):2538-43 Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P, Mele C, Bresin E, Cassis L, Gamba S, Porrati F, Bucchioni S, Monteferrante G, Fang CJ, Liszewski MK, Kavanagh D, Atkinson JP, Remuzzi G. Genetics of HUS: the impact of MCP, CFH and IF mutations on clinical presentation, response to treatment, and outcome. Blood 2006;108(4):1267-79 Bresin E, Daina E, Noris M, Castelletti F, Stefanov R, Hill P, Goodship T H J, Remuzzi G, International Registry Recurrent Familial HUS/TTP. Outcome of renal transplantation in patients with non-Shiga toxin-associated Hemolytic Uremic Syndrome: Prognostic significance of genetic background. Clinical Journal American Society Nephrology 2006; 1: 88-99

2. 3. 4.

5.

Sara Gamba got her Nurse Diploma on 1994 at Bolognini Hospital Nurses School, Seriate (Bergamo). Educational training: Clinical Research Nurse Diploma on 1996 at IRFMN Dacc Center. First Level Master on Clinical Research at Milan University on 2008.

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Areas of interest: rare diseases studied by the Laboratory staff. She is involved with the Italian rare diseases patients Associations and she coordinates the documentation service addressed to the patient with rare conditions, their relatives and the health care professionals. Employment: In 1997-2003 she was involved as co-organizing, speaker, co-speaker and tutor for the Clinical Research Course for Nurses at IRFMN Dacc Center. Since 2009 she is Chief of the Unit Network Development for Rare Diseases.
Selected publications
1. Noris M, Caprioli J, Bresin E, Mossali C, Pianetti G, Gamba S, Daina E, Fenili C, Castelletti F, Sorosina A, Piras R, Donadelli R, Maranta R, van der Meer I, Conway EM, Zipfel PF, Goodship TH, Remuzzi G Relative Role of Genetic Complement Abnormalities in Sporadic and Familial aHUS and Their Impact on Clinical Phenotype. Clin J Am Soc Nephrol, 2010; 5(10):1844-59 Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P, Mele C, Bresin E, Cassis L, Gamba S, Porrati F, Bucchioni S, Monteferrante G, Fang CJ, Liszewski MK, Kavanagh D, Atkinson JP, Remuzzi G. Genetics of HUS: the impact of MCP, CFH and IF mutations on clinical presentation, response to treatment, and outcome. Blood 2006;108(4):1267-79 Krulichova I, Gamba S, Ricci E, Garattini L on behalf of the Italian Takayasu Arteritis Study Group. Direct medical costs of monitoring and treating patients with Takayasu arteritis in Italy. Eur J Health Econ Vol. 49, August 2004 Noris M, Brioschi S, Caprioli J, Todeschini M, Porrati F, Gamba S, Remuzzi G for the International Registry of Recurrent and Familial HUS/TTP. Familial haemolytic uraemic syndrome and an MCP mutation. The Lancet, Vol. 362, November 8, 2003 Gamba S, Cicci L, Stefanov R. Qualit della vita in pazienti con malattie rare:lesperienza degli infermieri del Centro di Ricerche Cliniche per le Malattie Rare Aldo e Cele Dacc. Assistenza Infermieristica e Ricerca, Vol. 22, N. 3, JulySeptember 2003.

2.

3. 4. 5.

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ACTIVITIES
Rare Diseases (RD) represent about ten percent of all human medical illnesses and infirmities. It is difficult to define what exactly is intended as a RD. The US Congress in the Orphan Drug Act has given the first definition in 1983. Under this law it is considered rare a disease that affects less than 200 000 Americans (prevalence 0.75 per 1 000). The European Parliament adopted a more strict definition; they consider rare a condition that affects not more than five individuals per 10 000 in the European Community (prevalence 0.5 per 1 000). According to the WHO, there are 5-7 000 rare diseases and most of them (about 4 000) are of genetic origin. Rarity often brings a difficult and/or late diagnosis, and represents a difficulty in implementing experimental and clinical research studies. RD comprehend heterogeneous groups of diseases and often require a multidisciplinary approach. The greatest barrier to prevention, diagnosis and treatment of RD is inadequate knowledge. Once a diagnosis of RD is made, a major complaint of patients and of those involved in their care is the difficulty to obtain pertinent information about causes, symptoms and either established or experimental treatments. Often, patients with RD are willing to participate in clinical studies, but they do not know where and how, and physicians or health authorities are seldom able to help them. RD is not a very attracting field for basic and clinical investigators for several reasons: it is difficult to find adequate animal models for many rare disorders; clinical trials may require more patients than available; financial support is insufficient. Few countries have a central body or system to disseminate information on RD. Accurate information on the incidence and prevalence of RD is extremely important for both basic and clinical investigators. Invaluable help to research advances in RD would come from the availability of registries and databases containing diagnostic, clinical and biological data of patients with rare disorders. The Laboratory has been established whit the aim to collect different skills to support patients with rare diseases. Since the beginning of the activities, a Database for rare diseases was created with updated information for patients, their families and professionals. Specific research projects has been developed for some rare conditions such as familial and recurrent forms of Hemolytic Uremic Sindrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP), Fabry disease, Alport Syndrome, Takayasu arteritis, hereditary nephrotic syndromes. During the years the Laboratory role has changed, because of the major attention on rare diseases developed at the Institutional level. In 2001 it has been nominated Coordinating Centre of the Regional Network for Rare Diseases in the Lombardy Region, an area of 9 million people in Northern Italy. As Coordinating Centre, it is also working with the National Centre of Rare Diseases at Istituto Superiore di Sanit in Rome. All the information regarding the activities of the Coordinating Centre are available at the web site: http://malattierare.marionegri.it In 2009 the Genetics for Clinical Research Unit was established with the aim to support research projects on hereditary rare diseases ongoing at the Dacc Centre, through a close collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation and the Laboratory of Cell Biology and Xenotransplantation Negri Bergamo. The Network Development for Rare Diseases Unit (also established in 2009) represents a suitable tool for the enhancement of collaborations already activated in the past years and for the development of new network on rare diseases. In addition, very important are the collaborations with Italian patients organisations and whit national (UNIAMO-FIMR) and

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international (NORD, EURORDIS) Federations for rare diseases. Moreover, particular attention is dedicated to keep up-to-date documentation and scientific bibliography on rare diseases. An help-line service to the public it is also maintained.

MAIN FINDINGS
The database of the Information Centre for Rare Diseases contains data about 11292 patients affected by 911 different rare disorders. In the Bank of biological materials, samples from 1793 patients with rare conditions and their families have been collected. The Centre has established contacts with 349 Italian Associations for rare diseases. It was even possible that patients with 92 different rare diseases - for which no Associations have been established in Italy yet - to meet among themselves. In 2000, the European Commission recognized the Laboratory as a site for "Postgraduate training on rare diseases" (contract No. QLK4-1999-50547). In December 2001 (Delibera della Giunta Lombarda N. 7328), the Centre was identified as "Coordinating Centre of the Regional Network for Rare Diseases". The Laboratory coordinates the International Registry of Recurrent and Familial Hemolytic Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP), since 1996. The research projects developed in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation and with the Laboratory of Cell Biology and Xenotransplantation, have allowed to better comprehend the pathogenesis of these diseases.

NATIONAL COLLABORATIONS
Italian National Institute of Health Centro di Ricerche di Immunopatologia e Documentazione su Malattie Rare - Struttura Complessa a Direzione Universitaria di Immunologia Clinica, Torino Coordinamento Interregionale delle Malattie Rare del Piemonte e della Valle dAosta Riuniti Hospital, Bergamo Italian Registry of Membranoproliferative Glomerulonephritis Registry of Steroid-Resistant Nephrotic Syndrome Biotechnology Laboratory, IRCCS Policlinico San Matteo, Pavia Assessorato alla Sanit, Lombardia Region University of Turin, School of Clinical Pathology, Faculty of Medicine and Surgery Italian Network for Promotion of Folic Acid to Prevent Birth Defects University of Turin, Department of Experimental Medicine and Oncology, 2nd Level Master in Rare Diseases Italian Society of Neonatology (Lombardy section), Rare Congenital Respiratory Diseases Study Group University of Milan, 1st Level Master in Clinical Research BergamoScienza Association

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INTERNATIONAL COLLABORATIONS
International Registry of Recurrent and Familial Hemolytic Uremic Syndrome and Thrombotic Thrombocytopenic Purpura Information Centre for Rare Diseases and Orphan Drugs ICRDOD, Bulgaria Podonet: Consortium for Clinical, Genetic and Experimental Research into Hereditary Diseases of the Podocyte Coordinator: Heidelberg University Hospital, Germany International Network and Registry for Thrombotic Microangiopathy (TMA) Coordinator: Schneider Childrens Hospital, Albert Einstein College of Medicine, USA

EDITORIAL COMMITTEE MEMBERSHIP

Quaderni di Farmacoeconomia

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Network for Rare Diseases Lombardy Region (Delibera Regione Lombardia N7328, 11/12/2001) Working group Classification and coding of rare diseases coordinated by Italian National Institute of Health Working group National Registry of Rare Diseases coordinated by Italian National Institute of Health

EVENT ORGANIZATION
Open Day: informazione, ricerca e cure Un percorso dedicato alle malattie rare Ranica, (Bergamo) February 26, 2010 Pharma Research & Innovation- Educational Workshop Ranica, (Bergamo) June 17-18, 2010 Riunione Tecnica dei referenti delle ASL del Gruppo di Lavoro Rete Regionale Malattie Rare Ranica (Bergamo) October, 29, 2010

CONFERENCE AND WORKSHOP CONTRIBUTIONS

Malattie Rare, Quatto Incontri I incontro: Le sfide della transizione dallet pediatrica allet adulta Milano, January 15, 2010 13 Convegno Patologia Immune e Malattie Orfane Torino, January 21-23, 2010 Rare Diseases and Orhpan Drugs Roma, Jebruary 22-25, 2010 Terza giornata Internazionale delle malattie Rare

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Le Malattie Rare in Lombardia: Rete, Governo Clinico, Partecipazione e ricerca Milano, March 04, 2010 I Meeting Congiunto Gruppo Italiano Pazienti Fabry (G.I.P.F. Onlus) e Associazione Italiana Pazienti Anderson Fabry (A.I.P.A.F. Onlus) Bologna, March 13, 2010 Progetto di formazione Conoscere per assistere - UNIAMO FIMR Milano, March 2, 2010 Corso Farmaci Orfani e accessibilit al trattamento delle malattie rare Roma, April 15-17, 2010 Le Malattie Rare. La ricerca scientifica, i problemi sociali, le iniziative politiche Napoli, May 7, 2010 Regional Network for Rare Diseases Coordinating Centre meet Dutch Delegation Ranica (Bergamo), June 08, 2010 2nd International Conference HUS-MPGN-PNH. Current diagnosis and therapy of thrombotic microangiopathies Innsbruck, June 13-15, 2010 Progetto di formazione Conoscere per assistere UNIAMO FIMR Genova, September 18, 2010 Renal Master Class Bergamo, October 7-9, 2010 Teleangectasia emorragica ereditaria: 10 riunione nazionale pazienti Crema, October 09, 2010 2 Seminario del Progetto Il Codice di Atlantide UNIAMO FIMR e Telethon Milano, October 9, 2010 XII Convegno Nazionale Societ Italiana di Genetica Umana (SIGU) Firenze October 14-17, 2010 Malattie Rare: valutazione e gestione dei bisogni socio-sanitari Milano, November 1, 2010 Conferenza nazionale EUROPLAN Firenze, November 12-13, 2010 Malattie Rare, Quatto Incontri II incontro: Le sfide della transizione condividere i problemi e le soluzioni Milano, November 26, 2010 Le malattie metaboliche curabili in et pediatrica Brescia, November 27, 2010

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GRANTS AND CONTRACTS

"Fondazione Aiuti per la Ricerca sulle Malattie Rare", Bergamo Lombardy Region Fondazione Telethon

SCIENTIFIC PUBLICATIONS (2010)

Schieppati A, Daina E Rare autoimmune diseases. Adv Exp Med Biol, 2010:365-74. Noris M, Caprioli J, Bresin E, Mossali C, Pianetti G, Gamba S, Daina E, Fenili C, Castelletti F, Sorosina A, Piras R, Donadelli R, Maranta R, van der Meer I, Conway EM, Zipfel PF, Goodship TH, Remuzzi G Relative Role of Genetic Complement Abnormalities in Sporadic and Familial aHUS and Their Impact on Clinical Phenotype. Clin J Am Soc Nephrol, 2010; 5(10):1844-59 Warnock DG, Daina E, Remuzzi G, West M Enzyme replacement therapy and Fabry nephropathy. Clin J Am Soc Nephrol, 2010 Feb;5(2):371-8

RESEARCH ACTIVITIES Bank of biological samples and description of hereditary nephropathies

The aim of this project is to collect clinical data and biological samples from patients and their families with rare genetic conditions. A database with clinical data and a Bank for biological samples collection and preservation has been created. The availability of clinical data and biological samples is useful to perform new biochemical and genetic tests within specific research projects aimed to better reveal the mechanisms of the diseases, their manifestation and therapeutic opportunities. In particular, the attention is focused on rare genetic disorders of the kidney. A thorough clinical evaluation, including clinical data collection, medical physical examination, renal ultrasonography, laboratory tests of blood and urine is offered to patients, affected by hereditary nephropathies (Alport syndrome, Fabry disease, Familial Focal Glomerulosclerosis, Glomerulopathy with Fibronectin deposits, Membranoproliferative glomerulonephritis, Medullary Cystic Kidney disease, Cystinuria), who are addressing our Centre. After obtaining a written informed consent, biological samples from patients and their relatives are collected, labelled with specific codes to assure the anonymity and conserved in the Bank for biological samples. In case the responsible gene for a hereditary nephropathy is known (Alport syndrome, Fabry disease, Medullary Cystic Kidney disease, Cystinuria), the blood samples are redirected to the relevant Laboratory of reference. For other nephropathies, where the identification of the gene mutation is unknown or still in course, the blood samples are conserved with the aim to be used in specific future research projects.

International Registry of Recurrent and Familial Hemolytic Uremic Syndrome and Thrombotic Thrombocytopenic Purpura
Hemolytic Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP) are rare diseases of microangiopathic haemolytic anemia and thrombocytopenia with signs of renal (most prevalent in HUS) and cerebral (most prevalent in TTP) damage. There are extremely rare forms of HUS and TTP, often occurring in families, in which the patients relapse even after complete recovery of the first episode with permanent renal and neurological sequelae. In the

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familial and recurrent forms of HUS and TTP, the attention is concentrated mainly on the genetic predisposition to the disease. The Laboratory coordinates the International Registry of Recurrent and Familial HUS/TTP, since 1996. The Registry has collected more than 700 cases of HUS/TTP referred from 100 Italian and 80 European and extra-European Centres. Clinical and laboratory data of all patients referred to the Registry are collected by a uniform data extraction form. The family history and also the personal data of the unaffected relatives are collected, when possible. Biological samples are collected from all patients and available relatives, for the biochemical and genetic analyses. Many research projects in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation and the Laboratory of Cellular Biology and Xenotransplantation Negri Bergamo are developed and have still allowed to identify some genetically determined forms of HUS and TTP. The maintenance of a centralised bank of biological samples ensure the availability of clinical material for new investigative approaches as they will be developed.

Rituximab in relapsing and chronic thrombotic thrombocytopenic purpura

In most cases, TTP is related to an acquired deficiency of ADAMTS13 activity, due to the presence of autoantibodies anti-ADAMTS13. The absence of ADAMTS13 activity leads to the accumulation of von Willebrand Factor ultralarge multimers (normally cleaved to smaller molecular forms by ADAMTS13 enzyme), which probably induces platelets aggregation and the ensuing process of thrombotic microangiopathy. The purpose of the project is to evaluate the use of a monoclonal anti-CD20 antibody (Rituximab) in patients with relapsing and chronic thrombotic thrombocytopenic purpura, related to the presence of anti-ADAMTS13 antibodies. Rituximab is a humanized monoclonal antibody that targets the CD20 molecule on B cells (white cells producing antibodies). Administration of Rituximab leads to a selective B cell depletion that usually lasts 6 to 9 months. Inhibition of B cell activation or growth by rituximab treatment limit the uncontrolled synthesis of autoantibodies, that may predispose to chronic relapsing TTP. After Rituximab treatment, the patients are periodically controlled with medical examination, blood and urine exams, and with ADAMTS13 plasma activity assay and research of antiADAMTS13 autoantibodies, in collaboration with the Laboratory of Cellular Biology and Xenotransplantation Negri Bergamo. Observational period lasts 12 months.

MA.RES.CO.: Congenital Respiratory Malformations database

The Project originated from the collaboration between the Italian Society of Neonatology (Lombardy section) and the Regional Coordinating Centre for Rare Diseases. The MA.RES.CO (Congenital Respiratory Malformations) Database is conceived to facilitate the diagnostic and assistance procedures for pediatricians facing with respiratory malformations. For each syndrome a description is available with indications about Reference Centres. A link with the OMIM database is also provided. The Database is available at the web site of the Regional Coordinating Center for Rare Diseases: http://malattierare.marionegri.it/

Identification of new genes associated to the Autosomal Dominant Familial form of Focal Segmental Glomerulosclerosis
Focal segmental glomerulosclerosis (FSGS) is a pathological entity, and is a significant cause of end-stage renal disease (ESRD). Glomerular disease is the third leading cause of ESRD, and FSGS comprises a significant proportion of this subgroup: up to 5% of adults and 20% of children with ESRD. The diagnosis of FSGS is based on renal pathology. The clinical hallmarks include proteinuria, nephrotic syndrome, occasional hematuria and frequent progression to ESRD. Hypertension is also a common finding. At present, there are no consistently reliable treatments for FSGS and

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response rates to available treatments have been estimated at <30-50%. Because FSGS has become an important cause of ESRD, it is essential to understand the molecular basis and pathogenesis of this disease process. There are various subtypes of FSGS, which include primary (idiopathic or sporadic), secondary, familial and FSGS associated with congenital syndromes. Among familial forms of FSGS, both autosomal recessive and dominant inheritance patterns have been reported. The study, in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation, is aimed at identifying the genetic causes of the autosomal dominant familial forms of FSGS, in order to help in the clinical management. Patients with an autosomal dominant familial form of FSGS (at least two affected subjects in two different generations within a family) and their available relatives, will be enrolled. Clinical data and blood samples will be collected and used to perform routine laboratory exams and to obtain DNA for the genetic analyses. Genetic counselling will be provided to all patients and to their relatives.

The Registry of Rare Diseases of Lombardy

The Registry of Rare Diseases of Lombardy has the objectives of providing information for health planning, monitoring and studying the epidemiology of Rare Diseases. The Registry collect demographic, public assistance and clinical information. Data are collected by specialists of the Reference Centres of the Rare Diseases Network; at present the implementation of the Registry is web-based using a specific software called Sistema Malattie Rare. The Coordinating Center is in charge of Registry management which consists in analyzing data and sending minimum dataset (data required by the National Registry of Rare Diseases) to Istituto Superiore di Sanit. On 31 December 2010, the Registry contains 11512 records of diagnosis of rare disease. Most reported Rare Diseases are those in the category of the endocrine glands, nutrition, metabolism and immune disorders. Twenty-five percent of the patients received one or more medications for the treatment of their Rare Diseases. Fifteen percent of reported patients live in another Italian region. The Registry of Rare Diseases of Lombardy represent a remarkable resource for the study of epidemiology and health planning for rare diseases. Periodic Review Reports has been implemented and are available at the Coordinating Centre web site (http://malattierare.marionegri.it).

Complement abnormalities in primary Membranoproliferative glomerulonephritis

Primary membranoproliferative glomerulonephritis (MPGN) is an uncommon cause of chronic renal disease that occurs principally in children and young adults, and that often has an unfavourable prognosis. Data on incidence and prevalence of MPGN are scanty, thus a first aim of this project is to collect through a Registry clinical data and biological samples from well characterized patients with primary MPGN. Since 2006 the Laboratory coordinates the Italian Registry of primary form of MPGN and till now 93 cases (25 of type II MPGN, the rarest form) have been collected. The Registry contains homogeneous clinical data for all recruited patients. The family history is also recorded and biological samples are collected from the patients to perform biochemical and genetic analyses. Genetic counselling is provided to all patients and relatives. The pathogenesis of primary MPGN is ill defined. The available data indicate that excessive activation of complement due to autoimmune and genetic abnormalities plays a role in inducing damage to the kidney and other organs. The complement is a complex system that mediates the immune response against bacteria and viruses. In normal conditions human cells are protected from complement attack by several inhibitors that restrict complement activation to the surface of pathogens. Such regulatory system is defective in MPGN. Thus a major goal of this project, in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ Transplantation, will be the identification of the genetic and acquired defects underlying

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complement activation in MPGN and to establish whether specific complement abnormalities are associated with disease progression, response to therapy and recurrence on a transplanted kidney. As yet, a genetic predisposing factor of complement system has been identified in 10% of cases (50% are patients with MPGN type II). Otherwise, a positive activity of C3Nef (an antibody binding C3-convertase) has been in 30% of cases.

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INTERNATIONAL RELATIONS OFFICE OF RARE DISEASES

STAFF

Head

Arrigo SCHIEPPATI, M.D.

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CURRICULUM
Arrigo Schieppati got his degree in Medicine at the University of Milan in 1978 and the specialisation in Medical Nephrology in 1984 at the same University. He performed his training at the Mario Negri Bergamo Laboratories with Dr. Remuzzi, and completed it with stages at the laboratories of prof. Patrono (Catholic University in Rome), prof. John Gordon (Cambridge, GB), and at the Division of Renal Diseases - University of Colorado Medical School, directed by Dr. Schrier (Denver, USA). Since 1982 he works at the Division of Nephrology and Dialysis Riuniti Hospital Bergamo, where he is in charge of Outpatients Clinic and Day Hospital. 1991-1995: Chief, Information Center for Rare Diseases 1996- 2008: Chief, Laboratory for Coordination of Information and Diagnosis of Rare Diseases 2009 to date: Chief, International Relations Office of Rare Diseases. Areas of interest: diagnosis and therapy of chronic renal diseases, hypertension and rare kidney diseases. Affiliations: ethical committee Riuniti Hospital - Bergamo; member of the working group of the regional network for rare diseases in Lombardy; scientific committee Bolognini Hospital Seriate (BG); member of the Task Force on Rare Diseases (DG Health and Consumer Protection); International Society of Nephrology; American Society of Nephrology; Editorial Board Journal of Nephrology.
Principali pubblicazioni 1. Ramaswami U, Najafian B, Schieppati A, Mauer M, Bichet DG. Assessment of renal pathology and dysfunction in children with Fabry disease. Clin J Am Soc Nephrol. 2010 Feb;5(2):365-70. 2. Schieppati A, Henter J I, Daina E, Aperia A. Why rare diseases are an important medical and social issue. Lancet. 2008 June 14; 371:2039-55. 3. Schieppati A, Remuzzi G. Chronic renal diseases as a public health problem: epidemiology, social, and economic implications. Kidney Int Suppl. 2005 Sep;(98):S7-S10. 4. Remuzzi G, Schieppati A, Boissel JP, Garattini S, Horton R. Independent clinical research in Europe. Lancet. 2004 Nov 612;364(9446):1723-6. 5. Schieppati A, Perna A, Zamora J, Giuliano GA, Braun N, Remuzzi G. Immunosuppressive treatment for idiopathic membranous nephropathy in adults with nephrotic syndrome. Cochrane Database Syst Rev. 2004 Oct 18;(4):CD004293. 6. Remuzzi G, Schieppati A, Ruggenenti P. Clinical practice. Nephropathy in patients with type 2 diabetes. N Engl J Med. 2002 Apr 11;346(15):1145-51. 7. Schieppati A, Remuzzi G, Garattini S. Modulating the profit motive to meet needs of the less-developed world. Lancet. 2001 Nov 10;358(9293):1638-41.

INTRODUCTION TO THE LABORATORYS ACTIVITIES

The International Relations Office of Rare Diseases has been established in 2009. It represents the evolution of the previous Laboratory mainly dedicated to information about rare diseases. The Laboratory represents also the evolution of the Mario Negri Institute activities in the field of rare diseases, with particular attention to the European collaborations. The International Relations Office of Rare Diseases is involved in the EUROPLAN project for rare diseases national plans development.

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NATIONAL COLLABORATIONS
Italian National Institute of Health Assessorato alla Sanit, Lombardia Region UNIAMO - Rare Diseases Italian Federation Riuniti Hospital, Bergamo BergamoScienza Association

INTERNATIONAL COLLABORATIONS
ICORD Society - International Conference on Rare Diseases and Orphan Drugs EURORDIS Rare Diseases Europe, non-governmental patient-driven alliance ECRIN - European Clinical Research Infrastructures Network ICRDOD - Information Centre for Rare Diseases and Orphan Drugs, Bulgaria

EDITORIAL COMMITTEE MEMBERSHIP

Journal of Nephrology

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Task Force on Rare Diseases (established by DG Health and Consumer Protection on 21 January 2004). External Advisory Board E-Rare (ERA-Net for research programs on rare diseases). Network for Rare Diseases Lombardy Region (Delibera Regione Lombardia N7328, 11/12/2001) Scientific Committee A.O. Bolognini di Seriate Ethical Committe, A.O. Ospedali Riuniti di Bergamo

EVENT ORGANIZATION
Regional Network for Rare Diseases Coordinating Centre meet Dutch Delegation Ranica (Bergamo), 08 giugno 2010

PARTICIPATION IN EVENTS IN WHICH THE LABORATORY WAS INVOLVED

Meeting EUROPLAN Roma, January 18-19, 2010 13 Convegno Patologia Immune e Malattie Orfane Torino, January 21-23, 2010 Terza giornata Internazionale delle malattie Rare Le Malattie Rare in Lombardia: Rete, Governo Clinico, Partecipazione e ricerca Milano, march 04, 2010 5th European Conference on Rare Diseases

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Cracovia, may 12-16, 2010 Pharma Research & Innovation- Educational Workshop Ranica, (Bergamo), June 17-18, 2010 Renal Master Class Bergamo, October 7-9, 2010 Prima conferenza nazionale sulla Ricerca Sanitaria (NHRC) Cernobbio (CO), November 09, 2010 Conferenza nazionale EUROPLAN Firenze, November 12-13, 2010

GRANTS AND CONTRACTS

European Commission (DG SANCO)

SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2010

Ramaswami U, Najafian B, Schieppati A, Mauer M, Bichet DG. Assessment of renal pathology and dysfunction in children with Fabry disease. Clin J Am Soc Nephrol. 2010 Feb;5(2):365-70. Schieppati A, Daina E. Rare autoimmune diseases. Adv Exp Med Biol. 2010;686:365-74. Whitfield K, Huemer KH, Winter D, Thirstrup S, Libersa C, Barraud B, Kubiak C, Stankovski L, Grhlert X, Dreier G, Geismann S, Kuchinke W, Strenge-Hesse A, Temesvari Z, Blasko G, Kardos G, O'Brien T, Cooney M, Gaynor S, Schieppati A, Serrano M, de Andres F, Sanz N, Hernndez R, Kreis G, Asker-Hagelberg C, Johansson H, Asghar A, Husson JM, Demotes J, Gluud C. Compassionate use of interventions: results of a European Clinical Research Infrastructures Network (ECRIN) survey of ten European countries. Trials. 2010 Nov 12;11:104.

RESEARCH ACTIVITIES EUROPLAN - European Project for Rare Diseases National Plans Development
EUROPLAN is a three-year project of the Programme of Community action in the field of Public Health (2003 - 2008), which began in April 2008. The main goal is to provide National Health Authorities with a supporting tools for the development and implementation of National Plans and Strategies for rare diseases (RDs) following the recently agreed European Council Recommendation on an action in the field of RDs (2009/C 151/02). This supporting tools will be composed of three documents focused on defined priority areas: the Guidance document on recommendations for the definition and implementation of National Plans and Strategies for rare diseases; the report on current practices and relevant cases in the field of rare diseases; and the document on the recommended set of indicators for monitoring and evaluating the implementation of national initiatives.

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The National Centre for Rare Diseases (Italian Institute of Health - Istituto Superiore di Sanit, Italy) is the leading partner that organizes the contributions from 31 countries and Eurordis (the European Organisation for rare diseases) ensuring a broad representation of different EU contexts and experiences and patients point of view. The Mario Negri Institute collaborates in the EUROPLAN Project as scientific counsellor. In addition, the project ensures an inclusive and wide engagement of stakeholders - Ministries, regional and local authorities, health care planners, programme managers, health care professionals, researchers and patients. EUROPLAN Projects is divided in eight work-packages. Till now have been completed some of the activities requested by the work-package (i.e. information about initiatives for rare diseases already implemented by the Member States, development of monitoring tools of national plans and creation of contents for EUROPLAN recommendations). EURORDIS, one of the EUROPLAN associated partners, has in charge the organization of 15 National Conferences envisaged from May to December 2010.

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The Transplant Research Center Chiara Cucchi De Alessandri e Gilberto Crespi

The Transplant Research Center (CRT) was set up in 2002 to support and promote the work of outstanding research scientists throughout the world and to carry out major organ transplant research programs. The Center is housed in the Villa Camozzi, at Ranica, under the same roof as the Mario Negri Institute in Bergamo and is managed in collaboration with the Institute. The Centers staff is mainly made up of senior and junior researchers that were trained in the laboratories of the Mario Negri Institute in Bergamo, focusing on transplant immunology, research for less toxic immunosuppressant drugs, and new gene therapy techniques to prevent acute rejection of transplanted organs. Information on the Centers activities can be found in the sections addressed to the Department of Molecular Medicine (Laboratory of Immunology and Genetics of Organ Transplantation and Rare Diseases) and the Department of Renal Medicine (Laboratory of Pharmacokinetics and Clinical Chemistry).

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EDUCATIONAL ACTIVITIES
Dean, Educational Activities Dr. Mario Salmona

The Mario Negri Institute holds a well established expertise in educational training of young post-degree students in biomedicine, that, since 1963, when the Institute started its activities, amount to more than 6000. Excellence of the educational courses is confirmed by the fact that Mario Negri Institute diplomas are widely considered a guarantee of an excellent theoretical and practical training, and students who earn their specialization title at the Mario Negri can easily find positions in academic and industrial research laboratories both in Italy and abroad. The Pharmacological Research Specialists. Recognized by Lombardy Region, was the first educational program of the Institute. In 2009 the Lombardy Region started reviewing its occupational training courses and established an ad hoc register for Regional Occupational Standards (Quadro Regionale degli Standard Professionali - QRSP) , This lists all the training courses and the standards reached by pupils. The Lombardy Region Decree No. 14355 dated 22/12/2009 approved the new occupational profile known as Biomedical Research Specialists (formerly Pharmacological Research Specialists) presented by the Mario Negri Institute. The Biomedical Research Specialists will receive diplomas issued officially by the Lombardy Region and the Mario Negri Institute for Pharmacological Research. These have legal value throughout Italy, and are recognised in competitions for public posts, where they are worth a certain number of points In the last ten years new post-degree courses have been introduced. In particular, in 1996 the International Graduate Program has been introduced in the three Institute locations (Milano, Bergamo, Ranica (BG)), organized in collaboration with the Open University, UK (Milton Keynes, UK) . More recently, in the Bergamo campuses, a collaboration with the university of Groningen and Maastricht (The Netherlands) have been started. These courses confer a PhD title recognized worldwide.. The Research Degree Coordinator acts as the local contact person and is responsible for all communications with the foreign universities, coordinates and supervises the teaching, training, financial and administrative activities of the School. According consolidated European procedures, the viva defense of the thesis is done in English language, with external examinators non involved in the student projects. These procedures further confer excellence to the Institute educational activities. The Open University PhD degree earned at the Institute has legal value throughout Europe and in the USA. Since January 2009 the Institute started a two-year Advanced School in Applied Pharmacology (SAFA). The course provides advanced teaching and practical experience with experimental work in the laboratories. The SAFA course is aimed at preparing young researchers and enabling them to specialize and work for the pharmaceutical industry, for other research institutes and for public institutions In January 2009, the Institute started running courses for Pharmacological Research Doctorates,

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recognised by the Ministry for the University and Research with Ministerial Decree dated 11 November 2008. The main feature of these courses is that students receive their training "on site". They work full-time in research programs of a high scientific standard, using advanced equipment and learning the latest methods, in regular contact with colleagues in different countries. Besides its scientific value, this approach provides an excellent preparation on the human and personal scale. Students are usually assigned to one of the Institutes laboratories, where they gradually gain specialized skills by working on specific research projects. They are expected to attend lessons, seminars, courses and congresses and learn to make full use of the Institutes well-stocked library. Students all have access to the internet and to biomedical databases and can print out articles they need to consult, from major international journals. Should the opportunity arise, students are expected to be available for trips abroad, to participate in conferences or courses. Students enrolled in formal courses are assigned study grants. Between 1963 and 2009 the Mario Negri Institute awarded 7000 grants, 732 of them to foreign researchers who came to the Institute for special training. Everything possible is done to help students find work once they finish the course. At the moment the following courses are available: Three-year course for graduates, in Milan or Bergamo, leading to a diploma as Biomedical Research Specialist. Three-year course for diploma-holders, in Milan or Bergamo, leading to a diploma as Biochemical Research Technician. Research doctorates (PhD), run under an agreement with the Open University (UK) and the Universities of Maastricht and Groningen (NL). Two-year Advanced School in Applied Pharmacology (SAFA) for graduate students, in Milan and Bergamo. Three-year course for Pharmacological Research Doctorates, in Milan or Bergamo, recognised by the Ministry for the University and Research. First Level Master in Clinical Research, in collaboration with the University of Milan. Second Level Master Course in Rare Diseases, in collaboration with the University of Turin.

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Other training opportunities PREPARING A DEGREE THESIS Students can prepare their thesis in scientific subjects at the Institute, with the approval of their university faculty. These students must work at the Institute for at least two years. SUMMER STUDENTS

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In June and July each year the Institute accepts a certain number of students in their last two years at high school, to give them experience as part of school/work programs. Since 2003 the Institutes training schemes have been certified according to UNI EN ISO 9001:2008 requirements for the Design and provision of specialized training courses in biological and medical fields

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Executive Offices Services and Offices

STAFF

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Prof. Silvio Garattini

Silvio Garattini was born in Bergamo (Italy) on 12 November 1928. He earned a diploma in Chemistry, then a degree in Medicine and was appointed lecturer in Chemotherapy and Pharmacology. He held the post of Assistant then Deputy Professor at the Milan University Institute of Pharmacology, until 1962. A founder of the Mario Negri Institute for Pharmacological Research, when it opened in 1963 he was its director. The Institute now has four locations (Milan, Bergamo, Ranica (Bg), S. Maria Imbaro (Ch)) and more than 950 people work there. Professor Garattini is a member of the Gruppo 2003 (a group of the most cited Italian scientists in international scientific literature) and has hundreds of publications in Italian and English in international scientific journals, and texts on pharmacology. He was a founder of the European Organisation for Research and Treatment of Cancer (EORTC). In the last ten years Professor Garattini has acted in various organizations, including the Italian National Research Council (CNR) - Committee on Biology and Medicine; the National Health Council, the Committee for Italian Research Policy, set up by the Presidency of the Council of Ministers; the Ministry of Health Commissione Unica del Farmaco (CUF). He has held the following posts: President of the UICC Committee on Antitumoral Chemotherapy, President of the European Organisation for Research and Treatment of Cancer (EORTC), Consultant to the World Health Organisation; President of the European Society of Biochemical Pharmacology; member of the Committee for Proprietary Medicinal Products (CPMP) of the European Agency for the Evaluation of Medicinal Products (EMEA); member of the Committee of Experts of Research Policy CEPR - at the Ministry for University and Scientific and Technological Research; member of the Scientific Committee of the Lega Italiana per la Lotta Contro i Tumori; member of the Board of the Istituto Superiore di Sanit; Chairman of the Committee for Research of the Italian Agency for Drugs (AIFA). Other appointments include: Vice-president of the Consiglio Superiore di Sanit; President of the Research and Development Commission of the Agenzia Italiana del Farmaco (AIFA); President of the Angelo and Angela Valenti Foundation and the "Via di Natale" Foundation; President of the Technical Commission for Pharmaceutical Assistance of the Regione Autonoma of Sardinia. He is a member of the Strategic Committee for Welfare, Regione Lombardia, the Consiglio Superiore di Sanit and the Comitato Nazionale di Bioetica, the International Scientific Committee, Centro di Riferimento Oncologico, Aviano, and the Scientific Committee of AISLA, Member Comitato Scientifico, Dipartimento Politiche Antidroga Presidenza del Consiglio dei Ministri, Member, Advisory Board ADAMO Onlus, Milan, Member of Committee of the Recommendation for the Call, Wemos Foundation, Amsterdam, , Member, Development Advisory Board of the International Center for Biomedical Sciences (ICBMS), Luxu Town, Wujiang City, China, Honorary Member AREAS-CCI Onlus. Silvio Garattini is a Fellow of the New York Academy of Sciences, the American Association for the Advancement of Science, Honorary Fellow of the Royal College of Physicians (Pharmaceutical Medicine), London, Honorary Fellow of the Italian Society of Pharmacology and a member of numerous other Italian and international scientific societies. He has received many awards for his work, including the French Legion d'Honneur for scientific merit, and the Grand Ufficiale della Repubblica Italiana, and holds honorary degrees from the Universities of Bialystok in Poland, and Barcelona in Spain. Recent awards include the Ippocrate Prize, 2003; Mens Sana in Corpore Sano; Nuova Spoleto, 2003; Angelo dellanno; Alkmeon International Prize; International Prize SantAgostino Citt di Bergamo; Il Campione della Scienza; Natta Medal; Coppola Prize, Scienza e Societ in the framework of the Premio Citt di Firenze and Premio Rana dOro, Casalbeltrame (Novara), Premio Barocco Citt di Lecce, Premio Nazionale Tv LAltra Italia, Caserta; Premio Chirone 2009, Centro Ricerche e Studi Direzionali (Cerisdi), Palermo. In its 47 years, the Mario Negri Institute for Pharmacological Research, under Professor Garattini's leadership, has published more than 11,105 scientific papers and more than 250 books, on topics ranging from cancer and its treatment to tumour immunology, neuropsychopharmacology, and cardiovascular and renal pharmacology. More than 6100 young Italian and 670 foreign graduates and technicians have obtained specialist qualifications at the Institute.

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Prof. Giuseppe Remuzzi

Giuseppe Remuzzi was born in Bergamo, Italy in 1949. Upon completion of his medical training at the University of Pavia in 1974, he received specialty training in Hematology and Nephrology at the University of Milan. Since 1975, he has pursued his academic career at the Ospedali Riuniti of Bergamo, where he was appointed Professor of Nephrology and Director of the Department of Medicine and Transplantation in 1996. Since 1999, he is Director of the Department of Nephrology and Dialysis of the same hospital. The Negri Bergamo Laboratories, which he has directed since 1984, is a group of basic scientists, physiologists, pharmacologists, molecular and cellular biologists, pathologists and clinicians devoted to the study of renal disease. As Research Coordinator of the Negri Bergamo Laboratories and the affiliated Clinical Research Center for Rare Diseases "Aldo e Cele Dacco", both divisions of the Mario Negri Institute for Pharmacological Research, he conduct a diverse team of researchers studying human renal diseases and their corresponding experimental models from the perspective of pathophysiology and therapeutic intervention. He touched major advances in many areas of nephrology, with particular focus on platelet endothelial interactions, vascular prostaglandin biology, coagulation and renal disease, progression of renal disease, experimental models of glomerular damage, and transplant immunology and tolerance. Particularly his contributions to the understanding of the pathophysiology of hemolytic uremic syndrome, prostaglandin metabolism in pregnancy, renal vascular biology in uremia, the role of protein trafficking in renal disease progression, the induction of graft tolerance by intrathymic injection of donor antigens, the role of the co-stimulatory CD28-B67 pathway in transplant rejection and the prevention of renal and cardiovascular damage in diabetes. Prof. Remuzzi serves on editorial boards of numerous journals including the prestigious New England Journal of Medicine and is member of the International Advisory Board of The Lancet. In recognition of his achievements, he has been awarded in 1998 honorary memberships of the Association of American Physicians and the British Royal College of Physicians. In 2001 he was nominated Chairman of the Research Committee of the COMGAN (Commission on Global Advancement of Nephrology) of the International Society of Nephrology (ISN). He received an Honorary Professorship at the University of Maastricht in 2003. In 2005 during the World Congress of Nephrology in Singapore he received the ISN Jean Hamburger Award and in November 2007 he received during the annual congress of the American Society of Nephrology the precious John P. Peters Award. In August 2008, he was appointed "Honoris Causa Professor" by the Catholic University of Cordoba (Argentina). In april 2011 he received the ISN AMGEN Award (World Congress of Nephrology, Vancouver) where he was also appointed as President- elect of the ISN for the period 2013-2015. Prof. Remuzzi has authored and co-authored more than 1065 scientific articles, reviews and monographs.

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Mario Negri Institute Milan

Executive Office Director Prof. Silvio GARATTINI, M.D. Administrative Office Maria Grazia PEZZONI, Chief Technical Office Fabio BRIGHENTI, D. Arch. General Maintenance Emanuele SINELLI Studies Office Armanda JORI, Pharm.D. Press Office Sergio VICARIO, Econ. D. Public Relations Office Claudio PANTAROTTO, Comm. Prevention and Safety Office Emilio BENFENATI, Chem.D. Annamaria SEGALINI, Phys.D. English Style Editor Judi BAGGOTT Photography and Audio-Visual Service Felice DE CEGLIE Purchasing Office Eufrasia COVIELLO Directors Office Elvira CARCANO, Head of the Secretariat Office

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Negri Bergamo Laboratories

Executive Offices Director Research Coordinator Scientific Secretariat

Prof. Silvio GARATTINI, M.D. Giuseppe REMUZZI, M.D. Ariela BENIGNI, Biol.Sci.D., Ph.D.

Administration of Research Projects/Admn. Assistance Daniela MELACINI, Biol.Sci.D. Press and Communication Office Francesca Di Fronzo, Mod. Lit. D. Audio-Visual Service Antonella PICCINELLI, Biol.Sci.D. Prevention and Safety Office Chief Annamaria SEGALINI, Phys.D. Library Chief

Valeria MIGLIOLI

General Maintenance Giancarlo GASPARI Directors Office Antoinette van ENGELEN, Chief

ANNUAL REPORT

391

2010

IRFMN

ANNUAL REPORT

392

2010

IRFMN

Centro Aldo e Cele Dacc Ranica (Bg)

Executive Offices Director Research Coordinator Scientific Secretariat Health Director

Prof. Silvio GARATTINI, M.D. Giuseppe REMUZZI, M.D. Ariela BENIGNI, Biol.Sci.D., Ph.D. Norberto PERICO,M.D.

Press and Communication Office Francesca Di Fronzo, Mod. Lit. D. Prevention and Protection Office Chief Annamaria SEGALINI, Phys.D. Paola BOCCARDO, Biol.Sci.D. Library Mansueto Astori Chief Valeria MIGLIOLI Monica MINALI Directors Office Barbara REMONTI, Oriental Languages D. Clinical Trials Office Paola BOCCARDO, Biol.Sci.D. Custodian/general maintenace Giampiero CUGUSI

ANNUAL REPORT

393

2010