Mutagenesis vol. 33 no. 4 pp. e1–e7
1093/mutage/gey022
Abstracts of the UK Molecular Epidemiology Group Spring Meeting on Nutrition and
Human Health: From Molecules to Man. Aberdeen Exhibition & Conference Centre,
Aberdeen, UK. 24th March 2017
Dampening inflammation and oxidative stress in
colorectal cancer patients
Rune Blomhoff
Department of Nutrition, University of Oslo and Division
of Cancer Medicine, Oslo University Hospital, Oslo,
Norway
While it is well established that an unhealthy diet increases
risk of colorectal cancer (CRC), few studies have focused
on the effect of diet in CRC survivors. There are two major
molecular pathways leading to CRC, both of which involve
inflammation and oxidative stress as major driving forces.
In various models systems (transgenic reporter mice, cell
culture, clinical trials, cohorts), we have identified a num-
ber of plant foods (e.g. berries, nuts, spices, coffee and
specific fruits and vegetables) with the potential of damp-
ening inflammation and oxidative stress [e.g. 1–3]. In the
CRC-NORDIET study, patients (N=500) diagnosed with
primary invasive CRC (Stage I-III) are recruited to this
randomized controlled trial after curative surgery [4]. The
intervention group receives an intensive dietary interven-
tion for 12  months and a subsequent maintenance inter-
vention for 14  years. Primary outcomes are disease-free
survival and overall survival. Secondary outcomes are time
to recurrence, compliance to the dietary recommendations
and the effects of the intervention on new comorbidities,
intermediate biomarkers, nutrition status, physical activity,
physical function and quality of life.
References
1. Carlsen MH et al (2010) The total antioxidant content of more than 3100
foods, beverages, spices, herbs and supplements used worldwide. Nutr
J., 9:3.
2. Paur I  et  al (2010) Extract of oregano, coffee, thyme, clove, and walnuts
inhibits NF-kappaB in monocytes and in transgenic reporter mice. Cancer
Prev Res. 3(5):653–63
3. Bohn SK et al. (2010) Blood cell gene expression associated with cellular
stress defense is modulated by antioxidant-rich food in a randomised con-
trolled clinical trial of male smokers. BMC Med. 8:54.
4. Henriksen HB et al. (2017) The Norwegian dietary guidelines and colorectal
cancer survival (CRC-NORDIET) study: a food-based multicentre rand-
omized controlled trial. BMC Cancer 17(1):83.
Folate and HPV-linked cancers: from the epidemiology to
the molecular
Hilary J. Powers1, Vanessa Hearnden1, Craig Murdoch2
1
Department of Oncology and Metabolism, University of
Sheffield, UK; 2School of Clinical Dentistry, University of
Sheffield, UK
Epidemiological studies have suggested that poor folate
status may increase the risk of cancers at some sites. There
is a mechanistic basis for a role for folate status in car-
cinogenesis through effects on DNA synthesis and repair,
and the methylation of cancer-relevant genes, with subse-
quent impact on their expression. The experimental evi-
dence to support a role for low folate in carcinogenesis
© Published by Oxford University Press on behalf of The UK Environmental Mutagen Society 2018
doi:10.
currently outstrips the strength of the epidemiological evi-
dence. Some studies have shown that a high folate status
may drive the progression of pre-cancerous lesions to inva-
sive cancer.
There is current interest in the contribution that folate
Downloaded from https://academic.oup.com/mutage/article-abstract/33/4/e1/5126847 by Adam Ellsworth, Adam Ellsworth on 13 October 2018
status may play in determining the risk of cancers linked
with infection of human papillomaviruses (HPVs), which
include anogenital and head and neck cancers. Infection
with pathogenic (high-risk, HR) HPV is common and tran-
sient; persistence of HR-HPV infection is strongly causally
associated with progression to cancer.
We have explored the role of folate and other methyl
donors in determining the phenotype and gene expression
in head and neck squamous cell carcinoma (HNSCC) cells.
A model of methyl donor deficiency (low folate, choline and
methionine) was established using HPV-positive UD-SCC2
cells. Methyl donor deficiency led to significantly increased
cell doubling time, reduced cell proliferation, impaired cell
migration and increased apoptosis. Methyl donor deficient
cells showed a significantly increased expression of the
pro-apoptotic genes DAPK1 and PUMA, and of DNMT3a
and TET1, both important for the regulation of DNA
methylation.
Methyl donor depletion of HNSCC cells established a bene-
ficial phenotype, supporting evidence that high intakes of
methyl donors may have adverse effects in pre-neoplastic
lesions of established cancers.
Reference
Hearnden, V., Powers, H.J., Elmogassabi, A., Lowe, R., Murdoch, C. (2017)
Methyl donor depletion of head and neck cancer cells in vitro established
a less aggressive tumour cell phenotype. Eur J Nutr Mar 1. doi: 10.1007/
s00394-017-1411-5.
Diet and the human gut metabolome
Wendy Russell
The Rowett Institute, University of Aberdeen, Aberdeen, UK
The gut microbiota plays an important role in the devel-
opment of many human dietary related disorders such as
type II diabetes, cardiovascular disease and cancer. The
link between microbial diversity and metabolic func-
tionality is far from clear and although the products of
metabolism are considered to have an important impact
on immune function and inflammation, the metabolites
and the bacteria responsible for their formation are mostly
unknown. Tremendous advances have been made towards
determining the genetic identity of the many commensal
organisms that colonise the human gut and work at the
Rowett Institute has generated a unique collection of
human colonic bacteria, many genome-sequenced through
international projects. Research from our laboratory has
identified key molecular transformations performed by
these bacterial isolates and this is allowing us achieve our
overall aim; to elucidate the complex interplay between
diet, the intestinal microbiota and health.1 Through tightly
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Abstracts
controlled human dietary interventions we have shown
that macronutrient composition can impact on both gut
and systemic health. In particular, high-protein low-carbo-
hydrate diets were found to be detrimental if used in the
longer-term,2 and that replacement of animal protein with
an energy equivalent amount of plant protein ameliorated
this risk. As we look towards the use of plant products to
support global protein production in the human diet, an
understanding of how these products are metabolised by
the gut microbiota becomes essential.
References
1. Russell W. R., Duncan S. H., Scobbie L., Duncan G., Cantlay L., Calder
A. G., Anderson S. E. and Flint H.J. (2013). Major phenylpropanoid-de-
rived metabolites in the human gut can arise from microbial fermentation
of protein. Molecular Nutrition and Food Research, 57(3), 523–35.
2. Russell W.  R. Gratz S.  W., Duncan S.  H., Holtrop G., Ince J., Scobbie
L.  Duncan G.  Johnstone A.  M., Lobley G.  E., Wallace R.  J., Duthie
G.  G.  and Flint H.  J. (2011). High protein, reduced carbohydrate weight
loss diets promote metabolite profiles likely to be detrimental to colonic
health. American Journal of Clinical Nutrition, 93 (5), 1062–1072.
Dietary agents - and their microbial metabolites - as
epigenetic modulators in human health and disease
Clarissa Gerhauser
German Cancer Research Center (DKFZ) Heidelberg,
Germany
Dietary agents from various sources target mechanisms
involved in epigenetic gene regulation, including DNA
methylation, histone modifications and noncoding RNAs
[1–3]. Recent research has indicated that gut microbial
metabolites might be important mediators of these diet-
epigenome interaction [4]. Dietary polyphenols undergo
biotransformation by Phase II and gut microbial metab-
olism. Extensive metabolism of catechol groups alters
the availability of methyl groups for DNA and histone
methylation reactions. Berries and nuts are rich in ella-
gitannins, which are hydrolysed to ellagic acid and fur-
ther microbially metabolized to urolithins. Whereas
ellagitannin regulated the expression of small non-coding
microRNAs, urolithins inhibited the activities or expres-
sion of histone deacetylases (HDACs) and histone acetyl
transferases (HATs) in cultured cells. Glucosinolates from
cruciferous vegetables are enzymatically cleaved to iso-
thiocyanates (ITCs) and other reactive compounds. When
the plant-derived enzymes are inactivated by cooking, this
reaction is dependent on gut bacterial thioglycosidases.
The diversity of the gut microbiome therefore affects the
bioavailability of ITCs, which can be further metabolized
to HDAC inhibitors. Fermentation of dietary fiber leads
to formation of short chain fatty acids including butyrate.
Butyrate has a dose-dependent dual effect on HATs and
HDACs and both promotes proliferation and induces cell
cycle arrest and differentiation [5]. Long-chain omega-
3-fatty acids (LC ω-3-FA) regulate histone methylation
and miRNA expression in vitro and in vivo. The avail-
ability and efficacy of LC ω-3 FA might be affected by
gut microbial metabolism to conjugated linolenic acids
(CLA), which were shown to modify methylation pat-
terns of genes and expression of miRNAs involved in
energy balance and lipid metabolism [6–7].
e2
Overall, these examples indicate that the gut microbiota
interacts with the epigenome in various ways. Future stud-
ies need to integrate information on dietary intake, micro-
biome, and metabolome with genome-wide epigenetic
profiling results to draw conclusions on how to influence
this complex system in the direction of improved human
health.
References
1. Huang, J., Plass, C., Gerhauser, C. (2011) Cancer chemoprevention by tar-
Downloaded from https://academic.oup.com/mutage/article-abstract/33/4/e1/5126847 by Adam Ellsworth, Adam Ellsworth on 13 October 2018
geting the epigenome. Curr Drug. Targets 12, 1925–1956.
2. Gerhauser, C. (2012) Cancer cell metabolism, epigenetics and the poten-
tial influence of dietary components - A perspective. Biomed Res-India 23,
69–89.
3. Gerhauser, C. (2013) Cancer chemoprevention and nutriepigenetics: state
of the art and future challenges. Top. Curr. Chem. 329, 73–132.
4. Hullar, M.A, Fu, B.C. (2014) Diet, the gut microbiome, and epigenetics.
Cancer J. 20, 170–175.
5. Donohoe, D.R., Collins, L.B., Wali, A., Bigler, R., Sun, W., Bultman, S.J.
(2012) The Warburg effect dictates the mechanism of butyrate-mediated
histone acetylation and cell proliferation. Mol. Cell. 48, 612–626.
6. Chaplin, A., Palou, A., Serra, F. (2015) Methylation analysis in fatty-acid-
related genes reveals their plasticity associated with conjugated linoleic
acid and calcium supplementation in adult mice. Eur. J. Nutr. 56, 879–891.
7. Parra, P., Serra, F., Palou, A. (2010) Expression of adipose microRNAs is
sensitive to dietary conjugated linoleic acid treatment in mice. PLoS One 5,
e13005.
Dietary habits and their influence in genomic (in)stability
– results from a Portuguese food frequency questionnaire.
Carina Ladeira1,2,3, Elisabete Carolino1, Manuel C Gomes4,
Miguel Brito2
1
Environment and Health Research Group, Escola Superior
de Tecnologia da Saúde de Lisboa, ESTeSL, Instituto
Politécnico de Lisboa, Av. D. João II, Lote 4.69.01, 1990-
096 Lisboa, Portugal; 2Grupo de Investigação em Genética
e Metabolismo, Escola Superior de Tecnologia da Saúde
de Lisboa – IPL, Portugal; 3Centro de Investigação e
Estudos em Saúde Pública, Escola Nacional de Saúde
Pública, ENSP, Universidade Nova de Lisboa; 4Faculdade
de Ciências da Universidade de Lisboa, Portugal
The past decades have witnessed an increasing number
of studies attempting to associate nutrition and disease.
Indeed, dietary habits are recognized important modifiable
environmental factors with a putative causal or preventive
role in many diseases, namely those linked with genomic
instability, such as cancer.
Epidemiological studies involving food frequency ques-
tionnaires, have been developed to gather information con-
cerning diet and cancer, yet diet is a complex composite
of various nutrients (macro and micronutrients) and non-
nutritive food constituents, making the search for specific
factors almost limitless.
The goal of this study is to investigate the role of nutrients
- assessed by a food frequency questionnaire (FFQ) - in
genomic instability, as assessed by comet assay and cyto-
kinesis blocked micronucleus assay.
The study was based upon 46 subjects (34 females and 12
males) between 20 and 61 years of age, in Portugal, and the
FFQ had information regarding 92 Portuguese food items.
Two negative correlations were found between calories and
omega-6 and DNA damage measured by comet assay. Five
positive correlations were found between micronucleus and

caffeine, calcium, magnesium, zinc and protein (p<0.05,
Spearman correlation). Some of our results are corrobo-
rated by other studies, but some are controversial, because
in contradiction with present biological plausibility know-
ledge. Increasing sample size, making it more representa-
tive could eventually lead to more elucidative results. It is
important to understand the role of diet factors as genetic
modulators, in a world where future trends point to per-
sonalized diet intakes based on requirements for individual
genetic stability.
Serum metabolites and colon cancer risk within the EPIC
cohort study
Jelena  Bešević1, Marc  Chadeau-Hyam2,
Amanda J.  Cross and Marc J.  Gunter on behalf of the
1 3
European Prospective Investigation into Cancer and
Nutrition (EPIC) Investigators
1
Department of Epidemiology and Biostatistics, School of
Public Health, Imperial College London, London, United
Kingdom; 2MRC-PHE Centre for Environment and Health,
Department of Epidemiology and Biostatistics, School of
Public Health, Imperial College London, London, UK;
3
International Agency for Research on Cancer, Nutrition
and Metabolism Section, 69372 Lyon CEDEX 08, France
Colorectal cancer (CRC) is the third most common cancer
worldwide. Colon cancer accounts for approximately two
thirds of all CRC cases. Several modifiable factors have
been positively associated with colon cancer, including
obesity and type 2 diabetes. The mechanisms underlying
the association of obesity and metabolic health with colon
cancer are not fully understood. The aim of this study
was to apply metabolomic profiling to identify novel
biochemical pathways underlying colon cancer develop-
ment. A nested case-control study was conducted within
the European Prospective Investigation into Cancer and
Nutrition cohort. A  total of 485 incident colon cancer
cases were matched to 485 controls by risk set sampling.
Metabolites were quantified in mainly fasting serum sam-
ples using the Biocrates AbsoluteIDQ p180 kit which
captures metabolites from six biochemical families: acyl-
carnitines, amino acids, biogenic amines, glycerophos-
pholipids, sphingolipids and sugars. Metabolite levels
were categorised into quartiles according to the distribu-
tion of metabolite concentrations among the controls.
Multivariable conditional logistic regression models,
adjusted for body mass index, physical activity, education,
and alcohol consumption, were used to calculate the odds
ratios (OR) and 95% confidence intervals (CI). A  total
of 135 known, quantified metabolites were available for
analysis. Mean length of follow-up for the study subjects
was 6.3 years. Twenty-seven metabolites were nominally
significantly associated with colon cancer; however, fol-
lowing Bonferroni correction only the amino acid histi-
dine remained significantly associated (Q4 v.  Q1 OR=
0.47, 95% CI 0.31–0.71, p-trend<0.001). Histidine is a
major substrate for gut microbial metabolism where it is
converted to histamine. Histamine plays an important role
in the inflammatory response but its specific role in colon
tumorigenesis is thus far unknown. This analysis provides
novel insights into metabolic pathways that may play a
Abstracts
role in colon cancer and highlights a possible role for his-
tidine metabolism in the development of this malignancy.
Dietary carbohydrate specialisation in beneficial human
gut bacteria
Paul O. Sheridan, Sylvia H. Duncan, Alan W. Walker and
Harry J. Flint.
The Rowett Institute, University of Aberdeen, Aberdeen, UK
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The human gut microbiota can have profound effects on
human health. An important mechanism by which benefi-
cial effects are imparted is via the degradation of dietary
carbohydrates and subsequent production of metabolites
such as short chain fatty acids (SCFAs) including acetate,
propionate and butyrate. In particular, colonic production
of butyrate influences a myriad of intestinal functions that
promote intestinal health, including reinforcing the colonic
defence barrier and reducing intestinal inflammation. The
majority of fermentable carbohydrates reaching the colon
are in the form of complex polysaccharides, and the mech-
anisms through which gut microbes degrade these poly-
saccharides into simpler carbohydrates and utilise them to
produce SCFAs including butyrate are of great importance
in understanding how diet influences human health.
Representative strains from the dominant butyrate-pro-
ducing Roseburia genus were studied using a combination
of in silico and in vitro techniques to determine the mecha-
nisms through which these bacteria degrade a variety of
dietary polysaccharides. These studies revealed a conserved
genus-wide capacity to utilise starch, which is the most
abundant source of carbohydrate entering the colon in
western diets. Moreover, certain species within this genus
possessed specialities enabling them to utilise alternative
types of dietary polysaccharide. These specialities allow
Roseburia to maintain butyrate production in the colonic
environment where carbohydrate availability can vary.
Gaining a fundamental understanding of the mechanisms
involved in complex carbohydrate breakdown should
facilitate the rational design of nutritional interventions to
improve human health.
The effect of amino acid deprivation on the transfer of
iron through Caco-2 cell monolayers
Guenievre  Roussel, Valerie  Stevens, Sarah  Cottin,
Harry J. McArdle
Rowett Institute of Nutrition and Health, University of
Aberdeen, Aberdeen, UK
The main mechanism of regulation of iron occurs at the
gut, where uptake is limited via expression of different
regulatory genes and proteins. How these are affected
by other nutritional factors is poorly understood. Amino
acids (AA) play a central role in major biological processes,
such as protein synthesis and energy supply. However, the
influence of AA status on iron metabolism has not been
investigated. In the present study, we tested whether AA
alters iron metabolism in an intestinal cell model, the
Caco-2 cells.
Caco-2 cells differentiated on membrane to form a mono-
layer were deprived of either essential AA or non-essential
AA. We measured the impact of overnight depletion on
e3
Abstracts
radioactive 59Fe transfer through Caco-2 cells, gene expres-
sion by RT-qPCR and protein expression by ELISA.
Non-essential AA deficiency significantly increased both
Fe uptake and transfer across the cell monolayer (both
p<0.001) while essential AA deficiency only increased Fe
transfer (p<0.0001). Both essential and non-essential AA
deficiency decreased DMT1 (±IRE) exon1A mRNA expres-
sion (respectively p=0.0007 and p=0.006) and increased
expression of ferritin heavy chain (p=0.01). DMT1+IRE
(exon1A/1B) mRNA levels were decreased by essential AA
deficiency (p=0.012). The mRNA levels of total DMT1
were also decreased by essential, but not non-essential,
AA deficiency (p=0.006). Hepcidin levels were increased
significantly by non-essential amino acid deprivation
(p=0.047). AA deficiency had no effect on protein expres-
sion of ferroportin and ferritin heavy chain. Consequently,
ferroportin and/or ferritin do not affect Fe efflux though
iron content of ferritin could be increased.
Our data demonstrate, for the first time, that AA status –
essential and non-essential – affects iron transport and the
expression of genes related to iron metabolism in Caco-2
cells. We hypothesise that the effect on Fe transfer is medi-
ated through an increased movement across the cell layer,
rather than transfer across the cell membranes.
Identification and characterisation of phytochemicals
activating endogenous cytoprotective mechanisms
Jennifer A. Harbottle and Andreas F. Kolb
Rowett Institute of Nutrition and Health, University of
Aberdeen, UK
Improved health span and lifespan extension in a wide
phylogenetic range of species is associated with upregula-
tion of the environmental stress response (ESR)1. The ESR,
driven by the transcription factor Nrf2, provides endogen-
ous antioxidant defence to maintain intracellular redox
balance. Phytochemicals which stimulate the ESR thus
upregulate cytoprotective mechanisms and ultimately con-
tribute towards delaying the onset of age-related degenera-
tive diseases.
Two bioluminescent reporter cell lines were constructed to
identify and characterise inducers of the ESR, specifically
induction of Nrf2 target genes NQO1 and HMOX1. The
first cell line was generated using random integration of an
exogenous luciferase gene under control of an NQO1 gene
promoter, and allowed rapid and sensitive high throughput
screening of natural chemical libraries. The second assay
system employed a more targeted strategy using an adeno-
associated virus (AAV) vector to mediate the site-specific
integration of a luciferase reporter into the locus of the
endogenous ESR gene HMOX1, thus allowing predict-
ive indexing of the gene’s transcription. The two cell lines
were validated with potent ESR inducers sulforaphane
and curcumin. A compound identified in the screening of
a chemical library from the Marine Biodiscovery Centre
(University of Aberdeen, UK) was further characterised in
a microarray experiment and compared to known phyto-
chemicals. In combination, these in vitro cell-based assay
systems will allow a better understanding of the activity-
structure relationship of ESR inducers, and assess the bio-
activity of phytochemical metabolites from human serum
samples following intervention trials.
e4
Reference
Lewis KN, Wason E, Edrey YH, Kristan DM, Nevo E, Buffenstein R. (2010)
Nrf2, a Guardian of Healthspan and Gatekeeper of Species Longevity.
Integr Comp Biol 50 (5): 829–843.
Dorsal Raphe 5-HT neurons respond to dietary nutrients
Teodora Georgescu, David J Lyons and Lora K Heisler
Rowett Institute of Nutrition and Health, University of
Aberdeen, Aberdeen, Scotland
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Genetic and pharmacological research has identified the
5-hydroxytryptamine (5-HT; serotonin) system as a funda-
mental regulator of energy balance and body weight. We
hypothesize that circulating nutrients act as endogenous
regulators of the 5-HT appetitive brain circuit. Using c-fos
immunohistochemistry as a marker of neuronal activation,
here we demonstrate that a subset of 5-HT neurons in the
dorsal raphe nucleus (DRN) of mice increase their activ-
ity in response to the ingestion of food. Next, we used a
gavage technique to remove variables associated with the
sight, smell and taste of food in addition to mastication
and gastric distension and found that nutrients increased
the activity of a subset of DRN 5-HT neurons compared
to a gavage of water. Using ex vivo slice electrophysiologi-
cal techniques in a line of mice with fluorescently labelled
5-HT neurons, we also reveal that DRN 5-HT neurons pos-
sess KATP channels, which suggests that they are capable of
directly responding to nutritional cues. Finally, we deter-
mined that individual nutrients directly affect the firing rate
of a subset of DRN 5-HT neurons, ex vivo. These findings
suggest that 5-HT neurons are directly activated by dietary
nutrients and this may be an essential mechanism through
which 5-HT regulates appetite and body weight.
Dietary exposure and intestinal metabolism of the
mycotoxin deoxynivalenol
Silvia W.  Gratz1, Valerie  Stevensen1, Reshma  Dinesh1,
Susan McDonald2
1
Rowett Institute, University of Aberdeen, UK; 2Fera
Science Ltd, York, UK
Mycotoxins are toxic fungal metabolites that commonly
contaminate cereal grains. Fusarium moulds are the most
prevalent moulds in temperate climates and produce a host
of different mycotoxins. Deoxynivalenol (DON) is the
most common mycotoxin found in small grain cereals and
has been shown to be a potent ribotoxin disrupting intes-
tinal barrier function and immune function in vivo. A tol-
erable daily intake (TDI) has been set at 1 µg/kg BW and
maximum permitted levels in food have been set by EFSA.
In addition to parent mycotoxins plant-derived masked
metabolites commonly co-contaminate cereals and their
contribution to toxicity is unknown. Our work assesses
the metabolism and transport of the common masked
mycotoxin DON3glucoside in the human gut in vitro.
DON-3-glucoside was stable under conditions prevailing
in the upper GI tract and was not absorbed intact through
epithelial monolayers. Unabsorbed masked mycotoxins
are therefore likely to be delivered to the colon where we
found human gut microbiota to hydrolyse DON3Glc effi-
ciently and release DON. This free DON is likely to be

directly toxic to colonic epithelium and to be absorbed
into systemic circulation.
To assess exposure, urinary DON excretion is used as a
validated biomarker. A  survey in adult volunteers (n=15)
shows high prevalence of exposure (100% of samples
tested DON-positive) at low levels (0% exceeded TDI), but
significantly higher levels can be detected during years of
high Fusarium prevalence (13% exceeded TDI). Children
are at risk to exceed TDI levels due to high cereal intakes
relative to body weight, and we found that over 50% of
children (n=12) exceeded TDI.
In summary, DON exposure occurs frequently and exceeds
the safe level in children. Colonic hydrolysis of masked
DON is likely to contribute to exposure, but the direct tox-
icity remains to be confirmed.
This study received funding from the Scottish Government
and Food Standards Agency.
References
1. Gratz SW et  al. (2017): Mol Nutr Food Res. DOI: 10.1002/
MNFR.201600680.
2. Gratz SW et al. (2014): Food Additiv Contamin A, 31(9): 1579–85.
Nutritional protection against obesity
Andreas F.  Kolb, Linda  Petrie, Diana Moreno  Sanchez,
Alina Zitskaja
Rowett Institute, University of Aberdeen, Aberdeen, UK
Obesity is a pressing public health problem caused by an
interaction of several factors including genetic susceptibil-
ity and lifestyle changes (e.g. consumption of energy-dense,
high fat/sugar foods and, possibly, increases in sedentary
behaviour). Public health messages have had limited success
in combating obesity. Energy dense foods often have high
palatability which makes them attractive to consumers. Food
additives which improve the metabolic health profile of these
foods without changing their sensory qualities would provide
an alternative public health intervention to combat obesity.
We have investigated the effects of blueberry (BB) extract sup-
plementation as an anti-obesogenic intervention. The supple-
mentation completely prevented the effects of a high fat diet in
a C57B/6 mouse obesity model. Many physiological param-
eters including body weight, body composition, organ weight,
serum lipids, adipose tissue inflammation, food intake, and
glucose tolerance were all normalised by the supplementation.
Liver lipid metabolism was significantly changed in response
to BB supplementation leading to a significant reduction in
liver MUFA storage. BB supplementation resulted in a signifi-
cant increase in energy excretion. In addition, energy balance
calculations suggest that energy expenditure is also signifi-
cantly increased in response to BB supplementation.
The results suggest that BB extracts can mitigate against
the obesogenic effects of a high fat diet by reducing energy
uptake and increasing energy expenditure.
Effect of myricetin nano and bulk forms on lymphocytes
from blood cancer and pre-cancerous patients compared
to those from healthy individuals.
Shabana Akhtar, Rajendran C. Gopalan, Mojgan Najafzadeh,
Mohammad Isreb and Diana Anderson
University of Bradford, Bradford, UK
Abstracts
Cancer is one of the leading causes of death universally
which needs appropriate prevention and cost-effective
treatment. Several recent studies have suggested that die-
tary intake of various flavonoids have a protective effect
against different types of cancers and cardiovascular dis-
eases. This present study is conducted to investigate the
effects of myricetin nano and bulk forms on the lympho-
cyte from blood cancer and pre-cancerous patients com-
pared to those from healthy individuals. Lymphocytes
from the patients showed higher levels of DNA damage
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compared to the healthy individuals which was observed
throughout the in-vitro treatment. Myricetin in both forms
has not induced any significant DNA damage; in fact the
results demonstrated a reduction in damage upon treat-
ing with the myricetin nano form in lymphocytes from
the patient group. DNA repair capacity of myricetin bulk
and nano was determined by co-treating the drug with
hydrogen peroxide (H2O2). Both forms of myricetin signifi-
cantly reduced the oxidative stress caused by H2O2, where
myricetin nano form seemed to be more effective in both
healthy individuals and patients using the Comet assay
(p<0.001). This could be because NPs have a larger surface
area which could improve their reactivity and the anti-can-
cer properties of this compound. Overall myricetin showed
protective and anti-genotoxic effects by demonstrating the
reduction of the DNA damage caused by over-production
of ROS and oxidative stress. Results are consistent in both
the Comet and MN assay.
Association between vitamin D and colorectal cancer: a
2-sample Mendelian randomization approach
Kawthar  Al-Dabhani1, Kostas  Tsilidis1,2, Marc  Gunter3,
Ioanna Tzoulaki1
1
Imperial College London, London, UK; 2University of
Ioannina, Ioannina, Greece; 3International Agency for
Research on Cancer, Lyon, France
Colorectal cancer is the third most common cancer in
the world with approximately 70–80% of the variation
in risk potentially explained by environmental factors.
Observational studies have reported that higher levels of
serum vitamin D are associated with reduced risk of colo-
rectal cancer. Mendelian randomization analysis can be
used to provide more robust evidence for causality.
Summary-level data was obtained from a previously pub-
lished study to ascertain the association estimates of the
genetic variants (rs12785878, rs1993116, rs2282679, and
rs17217119) associated with circulating concentrations of
25-hydroxyvitamin D.  Individual-level data was obtained
from 126,947 UK Biobank participants with available
genetic data, in order to estimate the association between the
aforementioned genetic variants with colorectal cancer risk.
Two different approaches of Mendelian randomization for
summary data were used in this study were: the inverse-vari-
ance weighted approach and the likelihood-based approach.
These methods were applied to estimate causal inference of
the association between the allele risk score, the metabolism
allele score (DHCR7 and CYP2R1)), and the synthesis allele
(CYP24A1 and GC)) with colorectal cancer risk.
Among the 126,947 UK Biobank participants, there were
712 prevalent and incident cases of colorectal cancer. No
association was found for any of the four genetic variants
e5
Abstracts
of 25-hydroxyvitamin D and genetic risk scores with colo-
rectal cancer risk in the UK Biobank population. There was
no evidence of a causal association between the genetic
variants of 25-hydroxyvitamin D and the risk of colorectal
cancer (OR: 1.01, 95%CI: 0.97–1.04).
Our results do not provide evidence for a causal associ-
ation between 25-hydroxyvitamin D concentration and
the risk of colorectal cancer. This may have been due to the
low power in this study as a result of the small number of
colorectal cancer cases in the UK Biobank and the limita-
tions of the Mendelian randomisation assumptions.
Protective effect of both nano and bulk forms of aspirin
against tertiary butyl hydroperoxide (TBHP) and
bleomycin-induced oxidative stress in lymphocytes from
prostate cancer patients and healthy individuals
Azeza Guma1, Adolf Baumgartner2, Mojgan Najafzadeh1,
Mohammad Isreb1 and Diana Anderson1
1
University of Bradford, Bradford, UK; 2 York St John
University, York, UK
Prostate cancer is one of the most frequently diagnosed can-
cers and accounts for nearly a quarter of all new male cancer
diagnoses in the United Kingdom. Reactive oxygen species
(ROS) are elements with great oxidative activity. Intracellular
ROS oxidizes DNA, which leads to DNA damage promot-
ing the initiation of carcinogenesis and malignant transfor-
mation of cells. The ROS production rate is generally higher
in the cancer cells than in the normal cells. A growing body
of evidence suggests that aspirin is a potent antioxidant
which has an ability to protect the cells against oxidative
stress. Nanoscience and nanotechnology have been grow-
ing and expanding at a fast pace in the past two decades,
driving the development of nanomaterials with huge poten-
tial. It has been shown that there are a number of proper-
ties of nano-sized components which do not exist in their
bulk forms which make them more versatile and diffusible
into biological systems. The present study was undertaken
to investigate the cytotoxicity, genotoxicity and protective
effects of aspirin bulk and nano forms, against the oxidative
damage induced by TBHP and bleomycin in lymphocytes
from healthy individuals and prostate cancer patients. The
levels of reactive oxygen species and DNA strands break
were measured as biomarkers of cellular oxidative stress.
Cell cytotoxicity and genotoxicity were measured using the
3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bro-
mide (MTT) and the Comet assay, respectively. These results
have clearly shown that aspirin has protected the lympho-
cytes against oxidative damage by suppressing ROS produc-
tion and reducing DNA damage. The protective effect of
both forms of aspirin on lymphocytes from prostate cancer
patients could be due to their antioxidant properties.
Soyasaponin bio-availability in humans after the
consumption of high plant protein diets
Madalina  Neacsu, Yara  Benavidez-Paz, Nicholas
J. Vaughan, Sylvia H. Duncan, Gary Duncan, James Christie,
Alexandra Johnstone and Wendy R. Russell
Rowett Institute, University of Aberdeen, Aberdeen, UK.
High protein plants such as legumes are rich sources of
saponins, which are considered to have anti-nutritional
e6
properties [1]. However, epidemiological studies sug-
gest that saponins may benefit human health by lowering
cholesterol and play a role in protection from cancer [2].
Therefore, understanding their bioavailability in humans
is crucial. Volunteers (n=10; BMI > 27  kg/m2) consumed
soya-rich meals with high content of soyasaponin I (from
26.47 ± 3.73 up to 146.25 ± 10.55 mg/100 g food prod-
uct) for two weeks. Methods of extraction and LC-MS/
MS analysis of soyasaponin I  and soyasapogenol B from
diets and biological samples were developed. The con-
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centrations of soyasaponin I  and soyasapogenol B in the
gut were up to 1.2 and 17.5  mg/100  g fresh faecal sam-
ple respectively. Soyasaponin I  gut metabolism (in vitro)
was also confirmed by microbial incubation of soyasapo-
nin I (n=4 donors; 48 hours). From 1 mol of soyasaponin
I was recovered 0.98 mol soyasapogenol B. Soyasapogenol
B was the major metabolite in the plasma, with an average
concentration of 73.3  ±  12.1  ng/mL, and found only in
conjugated form. Soyasaponin I was absent in plasma sam-
ples from fasted volunteers and soyasapogenol B present
at unchanged levels from baseline for up to five hours fol-
lowing consumption of soya-rich meals. This demonstrates
that soyasapogenol was absorbed from the colon as a gut
metabolite of soyasaponin and could be partly responsible
for the decrease in cholesterol, observed following con-
sumption of soya diet [3].
References
1. Campos-Vega, R., Oomah, B. D.(2010) Minor components of pulses and
their potential impact on human health. Food Res. Int., 461–482.
2. Rochfort, S., Panozzo, J.(2007) Phytochemicals for health, the role of puls-
es. J. Agric. Food Chem., 55(20), 7981–7994.
3. Neacsu, M., Fyfe, C.,Horgan, G.,Johnstone, A.(2014) Appetite control and
biomarkers of satiety with vegetarian(soy) and meat-based high-protein di-
ets for weight-loss in obese men: a randomized-crossover trial. Am J. Clin.
Nutr., 100(2), 548–558.
Nucleus of the solitary tract neuropeptide Y cells Are
glucose-sensing
Minos  Kritikos1, Claudia  Cristiano1, Guenievre  Roussel1,
Mark L. Evans2, and Lora K. Heisler1
1
Rowett Institute of Nutrition and Health, University
of Aberdeen, Aberdeen, UK; 2Department of Medicine
and Institute of Metabolic Science, University of
Cambridge, Wellcome Trust/Medical Research Council,
Cambridge, UK
Glucose homeostasis is critical for survival. As such, spe-
cific neuronal populations within the brain can be activated
and respond to blood glucose fluctuations. Some of these
neurons expressing neuropeptide Y (NPY) located in the
hypothalamic arcuate nucleus and the lateral hypothalamic
area (LHA) were previously shown to be glucose inhibited.
Another brain structure, the nucleus of the solitary tract
(NTS), is known to contain glucose-sensitive neurons and
receiving direct input from the periphery. NPY-expressing
neurons have also been identified within the NTS. We
first characterized the neurochemical phenotype of NTS
NPY cells using a transgenic NPY-green fluorescent pro-
tein (GFP) mouse line. We found that a subpopulation of
NTS NPY-GFP co-localized with nesfatin (72%), Phox2b
(57%), tyrosine hydroxylase (24%) and GAD67 (22%).
Next, we characterized the neurochemical properties of

these NTS NPY-expressing neurons in response to glu-
cose deprivation. Hypoglycemia was induced in NPY-GFP
mice using insulin or 36h-fasting and characterization of
NPY-expressing neurons was performed using immuno-
histochemistry. A  subpopulation of NTS NPY neurons
was activated in vivo both by insulin-induced and fasting-
induced hypoglycemia. The present findings indicate that a
NTS NPY-expressing neuronal subpopulation responds to
glucose fluctuations and this thereby adds a new player to
the central regulation of glucose homeostasis.
Environment, DNA methylation and risk of childhood
acute lymphoblastic leukaemia: a novel Mendelian
randomization study
Jessica A.  Timms1, Caroline L.  Relton2, Judith  Rankin1,
Gordon Strathdee3, Jill A. McKay1
1
Institute of Health and Society, Newcastle University, U.K.,
2
MRC Integrative Epidemiology Unit, School of Social and
Community Medicine, University of Bristol, 3Northern
Institute for Cancer Research, Newcastle University, UK
Previous literature has identified multiple environmen-
tal exposures to be associated with an increased risk of
acute lymphoblastic leukaemia (ALL). DNA methylation is
modifiable by the environment and is therefore a plausible
mediating mechanism of environmental exposures on ALL
risk. An altered methylation has also been found in ALL
patients. Therefore two sample Mendelian randomization
(MR) will be used to strengthen evidence about the causal
pathway for childhood ALL, and determine whether DNA
Abstracts
methylation acts as a mediator between early life environ-
mental exposures and ALL development.
MR is a relatively new approach to causal inference.
It addresses limitations of observational epidemiology
study designs (i.e. bias, confounding and reverse
causation) in assessing the casual role of modifiable
exposures/risk factors, and can be used to examine
causality when randomized controlled trials (RCTs)
are not feasible. The approach uses genetic variation
(such as single nucleotide polymorphisms (SNP)) as
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an instrumental variable (proxy) for an “exposure” of
interest; the alleles of this exposure–associated genetic
variant are randomly distributed amongst the study
population (thus analogous to a RCT). The advantage
of using the genetic proxy is that the direction of caus-
ation is always from the genetic polymorphism to the
outcome. The underlying principle of the approach is
that, if a genetic variant alters or emulates the bio-
logical effect of an environmentally-modifiable expos-
ure then, if that exposure is truly casually-related to
the disease, the genetic variant should also be related
to disease risk.
An epigenome-wide association study (EWAS) previ-
ously carried out identified altered methylation at individ-
ual CpG sites associated with an ALL-risk exposure (e.g.
maternal smoking, alcohol consumption, and folic acid
supplementation). SNPs which correspond to these CpG
sites will be selected for the two-step Mendelian random-
ization analysis and investigated within Genome-wide
association (GWAS) data for 2487 childhood ALL cases
and 1014 cancer-free control children.
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