Documente Academic
Documente Profesional
Documente Cultură
Diagnostic Medicine
Section of General Pathology
Center of Excellence MIUR for the
Study of Inflammation (ICSI)
University of Ferrara, Ferrara, Italy
F. Di Virgilio
Particulate air pollution increases chronic obstructive lung diseases,
congestive heart failure, myocardial infarction, and in
general hospital admissions for cardiorespiratory symptoms
• Hourly peaks in air pollution more than double stroke risk (American Heart Association, British Medical Journal)
• People with diabetes, cardiovascular diseases, COPD, rheumatoid arthritis have an increased
death risk in polluted environments (American Thoracic Society)
• Children with obesity and asthma are more affected by air pollution than other kids
(American Thoracic Society)
Emphysema
COPD/BPCO
Variazione percentuale
della mortalità aggiustata
per l’età in U.S.A.
Proporzione della frequenza del 1965
3.0
Coronaro- Infarto Altre BPCO Tutte le
2.5 patie Malattie altre cause
CV
2.0
1.5
1.0
0.5
1965 - 1998 1965 - 1998 1965 - 1998 1965 - 1998 1965 - 1998
But also in Cardiovascular diseases:
Copyright ©2005 American Society for Biochemistry and Molecular Biology Cinti, S. et al. J. Lipid Res. 2005;46:2347-2355
Inflammation as a concurrent/causative factor in age-related
deterioration (microcirculation insufficiency, muscle waste,
cognitive impairment), Alzheimer
“inflammageing”
“Notae vero inflammationis sunt quatuor: rubor et tumor cum calore et dolore”
Cornelio Celso
TISSUE INJURY
Trauma
Ischemia
Neoplasms
Infectious agents
Foreign particles
?
VASOACTIVE MEDIATORS CHEMOTACTIC FACTORS
Histamine C5a
Serotonin PRODUCTION OF
Lipoxygenase products
Bradykinin INFLAMMATORY Formylated peptides
Anaphylotoxins MEDIATORS Chemokines
Leukotriene/prostaglandins
PAF
Nitric oxide
RECRUITMENT AND
STIMULATION OF INFLAMMATORY CELLS
VASODILATATION
INCREASED VASCULAR PERMEABILITY
HOW?
Cytokine and
Chemokine release
Interleukin-1β
Interleukin-6
Interleukin-8
TNFα Local and distal effects
The Monty Python Flying Circus: and now for something completely different/and now more of the same
How do NSPs trigger inflammation?
Complement activation
ROS/RNS
PMN recruitment
Pro-coagulant activity
?????????????
HOW?
Particulate matter
“Sensor”
“Sensor”
Leucin Rich Repeat (LRR): -L-X-X-L-X-L- Modified from Roy and Zamboni, 2006, Nat Immunol 7:549-551
NLR make the inflammasome sensitive to danger signals
• Thus, it would be helpful for human health to understand what goes wrong
However
Cytokine and
Chemokine release
Interleukin-1β
Interleukin-6 Local and distal effects
Interleukin-8
TNFα
Inflammation is becoming an increasing concern for physicians (and patients!)
Why?
Cytoplasmic sensors of danger
or
cellular trasnducers of noxious signals
Effetti cellulari e molecolari
indotti da particelle ultrafini
Annamaria Colacci
Eccellenza Cancerogenesi Ambientale
ARPA - ER
Attivazione AM
Accumulo
Particelle
Infiammazione Acuta
Infiammazione Fibrosi
Crronica
Meccanismi Meccanismi
Genetici Epigenetici
CANCRO
Proliferazione
Metaplasia
Cellule Epiteliali
Qual è il ruolo delle particelle
ultrafini a livello cellulare e
d’organo?
Componente Genetica
Alimento
Componente Genetica
Soggetto
Tecnologie Biomiche:
Genomica Funzionale
Le tecnologie biomiche:
Metodologie di studio
dell’era postgenomica
o Genomica
o Trascrittomica
o Proteomica
o Metabonimica
La tecnologia microarray
si basa sull’abilità di ogni
acido nucleico a singolo
filamento di accoppiarsi
alla sequenza
complementare
controllo esposto
m-RNA m-RNA
ibridazione
lavaggi
scanner
Bioinformatica e Data Mining
Feature Extractor
Primo screening dei dati provenienti dallo scanner. Il software ha il
compito di segnalare quegli spot troppo intensi, non uniformi
Normalizzazione
Rende confrontabili i campioni etichettati con Cy3 e Cy5 ed ibridati
congiuntamente su un singolo microarray.
Elimina errori sistematici dovuti a:
o diversa quantità di mRNA associata ad ogni marcatore
o diversa efficienza di incorporazione dei due marcatori
o diversa intensità di radiazione emessa dai due marcatori differentemente
misurata dal tubo fotomoltiplicatore
o differenti intensità misurate in diverse parti del microarray a causa del
posizionamento non perfettamente orizzontale del microarray rispetto al
laser dello scanner
o differenze tra gruppi di spot creati da differenti penne
Controllo qualità della slide
Analisi statistica dei dati
Maanova di R
LIMMA
Interpretazione biologica:
GoMINER High-Throughput
o Integra i risultati di molteplici microarray in una singola analisi
o Organizza la lista di geni modulati nel contesto di Gene Ontology un database
che contiene una classificazione dei geni in categorie gerarchiche basate su
processo biologico, funzione molecolare e compartimento cellulare
Tossicogenomica:
la validità dell’approccio
1 sola dose
Profilo Tossicologico
della miscela
complessa
3 dosi 3 dosi
Relazioni dose-risposta
Predizione
del rischio cancerogeno
ESPOSTI CONTROLLI NON ESPOSTI
MICROARRAY
Exposure to
137Cs radiation
source exposed not exposed
not exposed cells
Size <20μm
Polluted food
Pulmonary Mesothelioma
Asbestos fiber
Lung
Ni Group :
nodules observed on both sides
(particles + bulk material) in all
animals 6 months after
implantation
Co Group :
Nodules observed on the left
side (nanoparticles IM) in all
cases 8 months after
implantation
– nothing on the right side (bulk
material implanted SC)
Nickel and Cobalt nanoparticles
induced rabdomiosarcoma after
6-month implantation in rats,
the bulk samples only fibrotic
capsules or granulomas.
RF 4
Nanoparticles of Gold
in a liver granuloma.
The patient was
treated with colloidal
gold particles for knee
arthrosis
10 μm
Bladder Carcinoma in a mine-sweeper
Gulf-War kidney
20 μm
Hg
Barium-sulphate
nanoparticles found
inside a thrombus
10 μm
10 μm
BLOOD
Soldier’s wife
affected by
Burning Semen
disease
Red cells
292
99 GW
10 μm
Bladder cancer with Cobalt & Tungsten
2 μm
Colon cancer
with clusters of Silver nanoparticles
Industrial Hamburger
Hay
Gas emission of a diesel car
CrFeNi
Ag
Blood clot in vivo
Ag
Cancerogenic liver tissue with nanoparticles
of Titanium inside a cell nucleous
Liver cancer
Fe
Section of a lymph node
in a civilian of Sarajevo
affected by Hodgkin
Disease (FEG-ESEM)
5 μm
Particle of Antimony- Cobalt found in a soldier’s
sperm
ESEM image of sperm with a foreign body
spermatozoon
Lead nanoparticle
20 μm
Malformed lamb born inside a
groundfire in Sardinia, 2003
Internal organs of the 291CA
malformaed foetus
Liver
Brain
Testicle
Malformed lamb fromSardinia
Pb
Kidney
Pb
Sb
Neu-Laxova syndrome
Neu-Laxova syndrome is a rare congenital
abnormality characterised by intrauterine
growth restriction, microcephaly, facial
dysmorphy, short neck, edema, scaly skin
and perinatal death. Additional features such
as spina bifida, cryptorchidism and shallow
orbital cavities have been reported.
Chromosomal analysis in reported cases has
revealed a normal karyotype and an
autosomal recessive inheritance has been
postulated.
Liver 111 MA
Polvere cittadina con Cerio Palladio Zirconio
platinoplatino
Particelle su
filtro trovate in
area urbana
Polmone
___10μ____
Ce Nd La
Cigarette from
Sarajevo
Cigarette from
Bagdhad
Anchovy’s liver from the Adriatic sea
Co Cr W Nb Nd
Cl Sb
Vongola
_____10μ____
Al Fe
Uncontaminated
bread
Pane Italia (1)
PHYSICAL
Foreign body
Size CHEMICAL
Composition BIOLOGICAL
Shape
Surface area Corrodibility Organ (cell) involved
Speciation Health condition
Concentration
Individual variability
Intake velocity
Radioactivity
What is the cell reaction to a discrete,
non continous stimulus?
Discrete stimulus
Scheme of the
cell membrane
Balb Cells with
Co nanoparticles
by
Sabbioni JRC-
Ispra
Hematite nanoparticles inside a 3T3 cell nucleus
during the mitotic phase
DIPNA
Development of an Integrated Platform for Nanoparticle
Analysis to verify their possible toxicity and the eco-toxicity
Project objectives
- In-vitro tests of interaction of engineered
nanoparticles (NP) with cells,
- Identification of the modes of NP-cell interaction
- Application of the laboratory-developed cellular
Specific challenges:
models on the field investigations -Development of :
-1 in vitro nanotoxicological tests,
and single cell-to-NP interaction
-2 new sensors to detect specific NP-
induced biological reactivities ,
STREP - 3 evaluation of NP pollution in
industrial settings
Expected impact:
- Better understanding of the possible risks related to nanopollution and
criteria to assess the risk case by case.
-Assessment of the health risk for nanotechnological operators, citizens
and end-users and identification of safety procedures
- Criteria for prevention and contribution to Standards for policy
makers
Acronym:
STREP
DIPNA
Coordinator Partners
-University of Salzburg, A
Dr. Antonietta M. Gatti -Fraunhofer Institute of Biomedical
Consorzio Nazionale Interuniversitario Engineering, DE
Sviluppo Materiali - University of -Consiglio Nazionale delle Ricerche, I
Modena and Reggio Emilia Italy -Università della Magna Graecia, I
-Grimm Aerosol, DE
-VITO n.v. NL
EU funding: 2 793 235 Euro -CSEM SA, CH
-Institut Català de Nanotecnologia, ES
Duration: 36 months -Commission of the European
Communities - Directorate General -
Start date: x Joint Research Centre Ispra –
European Center Validation
End date: x
Alternative Methods, I
Project’s website:
in progress
Memento quia pulvis es et in pulverem reverteris