Red meat consumption is associated with increased risk of colorectal cancer. We have previously shown that haemin, Hb and red meat promote carcinogen-induced preneoplastic lesions: aberrant crypt foci (ACF) and mucin-depleted foci (MDF) in rats. We have also shown that dietary Ca, antioxidant mix and olive oil inhibit haemin-induced ACF promotion, and normalize faecal lipoperoxides and cytotoxicity. Here we tested if these strategies are effective also against red meat promotion in dimethylhydrazine-induced rats. Three diets with 60 % beef meat were supplemented with calcium phosphate (31 g/kg), antioxidant agents (rutin and butylated hydroxyanisole, 0·05 % each) and olive oil (5 %). ACF, MDF, faecal water cytotoxicity, thiobarbituric acid reactive substances (TBARS) and urinary 1,4-dihydroxynonane mercapturic acid (DHN-MA) were measured. Beef meat diet increased the number of ACF (+30 %) and MDF (+100 %) (P < 0·001), which confirms our previous findings. Promotion was associated with increased faecal water TBARs ( × 4) and cytotoxicity ( × 2), and urinary DHN-MA excretion ( × 15). Ca fully inhibited beef meat-induced ACF and MDF promotion, and normalized faecal TBARS and cytotoxicity, but did not reduce urinary DHN-MA. Unexpectedly, high-calcium control diet-fed rats had more MDF and ACF in the colon than low-Ca control diet-fed rats. Antioxidant mix and olive oil did not normalize beef meat promotion nor biochemical factors. The results confirm that haem causes promotion of colon carcinogenesis by red meat. They suggest that Ca can reduce colorectal cancer risk in meat-eaters. The results support the concept that toxicity associated with the excess of a useful nutrient may be prevented by another nutrient.

Studies have shown that soya consumption has been associated with low incidence of CVD. Because the chemical structures of soya isoflavones are similar to oestrogen, the beneficial outcome may be attributed to the oestrogenicity of these compounds. In this study, effect of the soya isoflavone genistein on the mRNA expression of apoA-1 in the human hepatoma HepG2 cell was investigated. Without oestrogen receptor (ER) α transfection, soya isoflavones in the physiological range had no effect on the apoA-1 transcription. Once ERα was ectopically expressed in these cells, soya isoflavone dramatically increased the apoA-1 mRNA abundance quantified by real-time PCR. ApoA-1-reporter assays with plasmid constructed from the 5′-flanking segment upstream to the coding region revealed that the transactivation of the apoA-1 promoter was induced by the soya isoflavone in HepG2 cells expressing ERα. This induction was reduced by the anti-oestrogen ICI 182780, but not the inhibitors of protein kinase (PK) C, PKA, or mitogen-activated PK. Based on the previously identified response elements on the promoter, a series of truncated promoter reporter plasmids were then constructed. An induction profile of genistein was built and insulin response core element at − 411 to − 404 appeared to be a potential site of interaction. This study illustrated that soya isoflavones at physiological concentrations could up regulate apoA-1 mRNA expression in ERα-transfected HepG2 cells.

The inclusion of insoluble NSP (iNSP) in weaner pig diets has been reported to decrease post-weaning colibacillosis (PWC). Conversely, soluble NSP (sNSP) have been shown to exacerbate PWC. The present study investigated the effect of NSP solubility and inclusion level on the health and performance of newly weaned pigs challenged with enterotoxigenic Escherichia coli (ETEC), using NSP sources known not to affect digesta viscosity, in a 2 ×  2 ×  2 factorial combination of NSP solubility (iNSP v. sNSP), inclusion level (low (L; 50 g/kg) v. high (H; 150 g/kg)) and ETEC challenge (infected v. sham). Infection had no effect on pig health, but reduced performance to a larger extent in pigs on the L diets compared with those on the H diets. The inclusion of sNSP significantly decreased the occurrence of diarrhoea (P < 0·001) and improved gut health, as indicated by a lower caecal digesta pH (P = 0·008) and increased (P = 0·002) Lactobacillus:coliform ratio, when compared with the iNSP diet on day 14 post-weaning. There was no effect of NSP solubility on ETEC shedding, digesta viscosity or pig performance. Pigs on the H diets had fewer cases of diarrhoea and shed fewer ETEC than those on the L diets. Increasing NSP inclusion significantly increased colonic Lactobacillus:coliform ratio, volatile fatty acid concentration and caecal digesta viscosity, but decreased performance. These results suggest that sNSP per se are not detrimental to pig health and that increasing the concentration of NSP in weaner diets that do not increase digesta viscosity may have a beneficial effect on gut health and protect against PWC.

The nutritional consequences of algae consumption in young populations consuming hypercholesterolaemic diets have hardly been investigated. This study tests the effect of algae supplementation of cholesterol-enriched balanced diets on growth, dietary efficiency ratio, mineral intake and absorption, organ weight and structure and cholesterolaemia in growing Wistar rats. Three groups of ten rats each were fed for 3 weeks with experimental diets containing 93 % casein-soyabean oil base with 2·4 % cholesterol-raising agent and 7 % supplement. The control group received cellulose (35 %), group 2 consumed Nori (33·8 % fibre) and group 3 consumed Konbu (36·1 % fibre). Food intake and body weight gain were not significantly affected. Algae groups presented significantly higher dietary efficiency ratio values than control rats. Apparent absorption of several minerals appeared significantly affected, mainly in Nori-fed rats, with a significant decrease in the ratio of Zn and Cu intakes and apparent absorption. Nori diet significantly decreased plasma cholesterol. Algae supplement did not significantly affect organ size and structure. Control and Konbu rats showed severe liver fat infiltration, while Nori rats exhibited a significantly lower degree of lipid-like hepatocyte vacuolization but light to moderate leukocyte infiltration. Light to moderate scaling off of the epithelium and moderate submucosa oedema was observed in all groups. Although long-term studies are needed to check the possible extrapolation of these data to human subjects, it can be concluded that a Nori, but not a Konbu, dietary supplement reverses the negative effect of dietary cholesterol intake and also appears to be related to mineral availability in growing subjects.

Adiponectin is discussed to regulate energy balance and insulin sensitivity. Several studies indicated an association of fasting adiponectin with parameters of the metabolic syndrome. We investigated postprandial adiponectin release and its relation to traits of the metabolic syndrome. Serum adiponectin concentration after an oral glucose tolerance test and after ingestion of a standardised mixed, fat-containing meal in 110 male non-diabetic subjects was assessed. Fasting and postprandial adiponectin and the decrease of adiponectin were correlated with anthropometric and metabolic parameters. Subjects were genotyped for adiponectin − 11 388 G/A promoter single nucleotide polymorphism. Adiponectin slightly decreased after both test meals. A significant decrease was attained 5 and 6 h after the lipid load and 2 h after the glucose load. Particularly, the mixed meal postprandial adiponectin showed stronger correlations with most traits of the metabolic syndrome than fasting adiponectin: postprandial adiponectin with HDL (r 0·30) v. fasting adiponectin with HDL (r 0·23); with postprandial insulin (area under the curve): r − 0·20 v. r − 0·16; with fasting insulin: r 0·10 v. r 0·14; with BMI: r − 0·23 v. r − 0·20; with waist: r − 0·18 v. − 0·16; with systolic blood pressure: r − 0·14 v. r − 0·12; with diastolic blood pressure: r − 0·18 v. r − 0·15. In multivariate analysis, postprandial TAG were the only independent predictor of adiponectin. There was no significant association of adiponectin, NEFA and TAG with − 11 388 G/A adiponectin promoter polymorphism. Our findings favour the interpretation that postprandial adiponectin has the strongest and independent associations to postprandial TAG metabolism.

Nutrigenomics represents a shift of nutrition research from epidemiology and physiology to molecular biology and genetics. Nutrigenomics seeks to understand nutrition influences on homeostasis, the mechanism of genetic predispositions for diseases, to identify the genes influencing risk of diet related diseases. This review presents some in vitro models applicable in nutrigenomic studies, and discuses the use of animal models, their advantages and limitations and relevance for human situation. In vitro and in vivo models are suitable for performance of DNA microarrays, proteomic and transcriptomic analyses. In vitro models (intracellular organelles and suborganellar compartments, cell cultures, or tissue samples/cultures) give insight in metabolic pathways and responses to test stimuli on cellular and molecular levels. Animal models allow evaluation of the biological significance of the effects recorded in vitro and testing of the hypothesis on how a specific factor affects specific species under specific circumstances. Therefore, the evaluation of the data in relation to human organism should be done carefully, considering the species differences. The use of in vitro and in vivo models is likely to continue as the effects of nutrition on health and disease cannot be fully explained without understanding of nutrients action at nuclear level and their role in the intra- and intercellular signal transduction. Through advances in cell and molecular biology (including genomic and proteomic), the use of these models should become more predictively accurate. However, this predictive value relies on an underpinning knowledge of the advantages and limitations of the model in nutrigenomic research as in other fields of biomedical research.

An experiment was conducted to determine whether adding oat hulls to weaner pig diets based on extruded rice or unprocessed wheat influenced post-weaning diarrhoea (PWD) and protein fermentation in the large intestine. Ninety-six male piglets (5·16 (sem 0·08) kg) were allocated to (i) extruded rice plus animal proteins (RAP); (ii) RAP with added oat hulls (20 g/kg); (iii) wheat plus animal proteins (WAP); (iv) WAP with added oat hulls (20 g/kg). Blood and faecal samples were collected on days 7 and 14 after weaning at about age 21 d. Pigs fed RAP had more PWD than pigs fed WAP (P < 0·05). Oat hull supplementation to diet RAP decreased the incidence of PWD (P < 0·05). The total-tract digestibility of DM, starch and energy was higher in rice-based diets than in wheat-based diets (P < 0·001); however, oat hulls decreased digestibility of DM and gross energy (P < 0·001). Pigs fed RAP had higher plasma creatinine concentrations (P < 0·01), which were positively correlated to cumulative β-haemolytic Escherichia coli scores after weaning (R2 0·928; P = 0·015). Addition of oat hulls decreased plasma urea concentrations only in pigs fed RAP (interaction; P < 0·05). Pigs fed RAP had lower faecal total biogenic amine concentrations than pigs fed WAP (P < 0·001). Oat hull supplementation tended to decrease total biogenic amine concentrations (P = 0·103). These data indirectly suggest that a mostly insoluble dietary fibre source such as oat hulls can decrease PWD in dietary situations where there may be a misbalance of carbohydrate to protein entering the hindgut.

In the elderly, nutritional deficiencies, such as low energy and protein intake, are suggested to increase the risk of osteoporotic fractures. Modulation of the amount and quality of protein intake under energy deficient conditions represents an interesting strategy to prevent aged-related bone loss. We investigated the effect of a 5-month dietary restriction on bone status in 16-month-old male rats. Rats were randomised into six groups (n 10 per group). Control animals were fed a normal diet containing either casein (N-C) or whey protein (N-WP). The other groups received a 40 % protein and energy-restricted diet with casein or whey protein (PER-C and PER-WP) or a normal protein and energy-restricted diet (ER-C and ER-WP). Both restrictions (PER and ER) induced a decrease in femoral bone mineral density (BMD), consistent with impaired biomechanical properties and a reduced cortical area at the diaphysis. Plasma osteocalcin and urinary deoxypyridinoline levels suggested a decrease in bone turnover in the PER and ER groups. Interestingly, circulating insulin-like growth factor 1 (IGF-1) levels were also lowered. Overall, normal protein intake did not elicit any bone sparing effect in energy-deficient rats. Regarding protein quality, neither casein nor WP appeared to significantly prevent the BMD decrease. This study confirms that nutritional deficiencies may contribute to osteopenia through decreased IGF-1 levels. Moreover, it seems that impaired bone status could not be significantly prevented by modulating the amount and quality of dietary proteins.

Nutrition research has slowly started to adopt the proteomics techniques to measure changes in the protein complement of a biological system. This enables modelling of biological processes in response to dietary interventions, as well as the elucidation of novel biomarkers for health or disease that are sensitive to such interventions. There are limited studies on the effect of micronutrients on the proteome, so this review concentrates rather more on dietary intervention studies that have used proteomics (mainly classical 2D gel electrophoresis combined with mass spectrometry) to elucidate changes in pathways that relate to glucose and fatty acid metabolism, oxidative stress, anti-oxidant defence mechanisms and redox status. The ability to measure regulation of more low abundant proteins, such as those involved in inflammatory pathways, as well as the evaluation and validation of newly discovered candidate biomarkers in human biofluids, may depend on the introduction of more quantitative and sensitive methods like multiple reaction monitoring (MRM) and multiplexed immunoassays in nutrition research.

Human health is affected by many factors. Diet and inherited genes play an important role. Food constituents, including secondary metabolites of fruits and vegetables, may interact directly with DNA via methylation and changes in expression profiles (mRNA, proteins) which results in metabolite content changes. Many studies have shown that food constituents may affect human health and the exact knowledge of genotypes and food constituent interactions with both genes and proteins may delay or prevent the onset of diseases. Many high throughput methods have been employed to get some insight into the whole process and several examples of successful research, namely in the field of genomics and transcriptomics, exist. Studies on epigenetics and RNome significance have been launched. Proteomics and metabolomics need to encompass large numbers of experiments and linked data. Due to the nature of the proteins, as well as due to the properties of various metabolites, experimental approaches require the use of comprehensive high throughput methods and a sufficiency of analysed tissue or body fluids. In this contribution, we describe the basic tools currently used in nutrigenomics studies and indicate the general requirements for future technology methodological routings.

The study aimed to investigate the in vivo digestion of Phaseolus vulgaris phaseolin types differing in their subunit pattern composition. Diets contained either casein as the sole source of protein or a mixture (1:1) of casein and pure Sanilac (S), Tendergreen (T) or Inca (I) phaseolin either unheated or heated. Rats were fed for 11 d with the experimental diets. Their ileal content and mucosa were collected and prepared for electrophoresis, Western blotting, densitometry and MS. Differences in digestion among native phaseolin types were observed for intact phaseolin at molecular weights (MW) of 47–50·5 kDa and for an undigested fragment at MW of 19–21·5 kDa in ileal digesta. In both cases, the concentration of these protein bands was lower for I phaseolin than for S or T phaseolin (P < 0·05). In the mucosa, the concentration of a protein band at MW of 20·5–21·5 kDa was lower for S phaseolin as compared to T or I phaseolin (P < 0·001). The presence of phaseolin subunits and their fragments was confirmed by Western blotting. MS analysis revealed the presence of undigested α and β subunit fragments from phaseolin and endogenous proteins (anionic trypsin I and pancreatic α-amylase) in ileal digesta. Thermal treatment improved digestion (P < 0·01), acting on both dietary and endogenous protein components. In conclusion, this study provides evidence for differences in intestinal digestion among phaseolin types, S phaseolin being more resistant and I phaseolin more susceptible. These differences were affected by the origin of the phaseolin subunit precursor. Heat treatment enhanced phaseolin digestion.

Probiotics are considered to have beneficial effects on the immune system. An association between the composition of microflora and allergies has been demonstrated and modulation of microflora of infants by probiotics might reduce the risk of allergies. To investigate immune effects of probiotics administered early after birth two animal models were used: a mouse model for respiratory allergy; a rat model for experimental autoimmune encephalomyelitis (EAE). Administration of the probiotic Lactobacillus casei Shirota (LcS) started during lactation and allergy or autoimmunity were induced at an adult age. Results were compared with similar studies in rats and mice that were exposed from an adult age. Early administration of LcS significantly increased lymphocytes in the lungs of female mice and eosinophils in the lungs of male mice. LcS had no effects on ovalbumin-specific serum IgE levels and on ovalbumin-specific cytokine production by spleen cells. In adult mice, LcS enhanced ovalbumin-specific cytokine production by the spleen, whereas other parameters were not affected. Early administration of LcS to rats significantly increased the duration of clinical symptoms of EAE. This was also demonstrated previously in adult rats exposed to LcS. Timing of administration of LcS induced divergent effects on respiratory allergy and only early administration of LcS exacerbated lung inflammation. In the EAE model, LcS stimulated autoimmunity independent of the timing of administration. Our data show that immune effects of probiotics do not necessarily induce beneficial effects. It is therefore important that, in the evaluation of probiotics, efficacy and safety should be demonstrated.

Despite adequate scientific evidence of the potential benefits of probiotics to human health or disease prevention, their contribution to the growth of adipose tissue remains to be established. Four-week-old male Sprague-Dawley rats were fed a diet containing skim milk (control diet) or skim milk fermented by Lactobacillus gasseri SBT2055 (LGSP diet) for 4 weeks. Their body weight gain, adipose tissue weight, adipocyte size distribution profile, blood and hepatic lipids, and serum leptin, glucose and adiponectin levels were determined. There was a significant reduction in average adipocyte size in mesenteric white adipose tissue (P = 0·004). Moreover, the rats fed the LGSP diet displayed greater numbers of small adipocytes from mesenteric and retroperitoneal adipose tissues than did those on the control diet. Whereas adiponectin concentrations did not differ between the groups, serum leptin concentrations were decreased to 32 % in the LGSP diet group compared with the control group. Concentrations of serum glucose and lipids, and liver lipids, except for the liver TAG level, were similar in the two groups. These results indicate a possible role for a fermented milk product in the regulation of adipose tissue growth.

Though common in older adults, anaemia is unexplained in about one-third of cases. As a rare cause of anaemia and neutropenia, Cu deficiency could account for some cases of unexplained anaemia. We examined the relationship between serum Cu and unexplained anaemia among 11 240 participants in the Second National Health and Nutrition Examination Survey (NHANES II): 638 (5·7 % of all adults) were anaemic; 421 (3·7 %) were not explained by deficiencies of vitamin B12, folate or Fe, chronic illness or renal disease. Spline regression showed a U-shaped relationship between serum Cu levels and unexplained anaemia, indicating that both high and low serum Cu levels are associated with unexplained anaemia in adults. Chronic inflammation and mild Fe deficiency could account for the association between unexplained anaemia and elevated Cu levels. On the other hand, the finding of hypocupraemia in a subset of adults with unexplained anaemia suggests that Cu deficiency may be a common reversible cause of anaemia in adults.

Soya protein has well-documented beneficial effects on serum lipid levels in adults, the potential beneficial effect of a prolonged soya protein-substituted diet in children and adolescents with familial (FH) and polygenic hypercholesterolaemia (PH) being unknown. To assess the effect of 3 months’ treatment of children and adolescents with FH and PH with a soya-substituted diet on serum lipids and lipoproteins, twenty-three children and adolescents were initially assigned to a standard phase 1 diet for 3 months, after which they were instructed to include soya protein (0·25–0·5 g/kg body weight) into their diet for 3 months. Sixteen patients (ten males and six females, thirteen with FH (eight males and five females), three with PH (two males and one female); mean age 8·8 (sd 4·2) years (range 4–18 years); mean BMI 16·7 (sd 2·6) kg/m2)) completed both phases. The phase 1 diet resulted in a significant reduction of total cholesterol (TC), LDL-cholesterol and apo B by 12·3, 11·8 and 10·6 %, respectively, HDL-cholesterol, TAG, apo A1 and lipoprotein(a) not being different. Dietary intake of soya protein during phase 2 resulted in a significant decrease of TC, LDL-cholesterol and apo B by 7·7, 6·4, and 12·6 %, respectively. TAG, HDL-cholesterol, apo A1, and lipoprotein(a) did not change significantly. Substitution of soya protein for animal protein in a low-fat, fat-modified diet is of additional benefit in many, but not all, children and adolescents with FH and PH when aiming at lowering serum TC, LDL and apo B. It seems to be a feasible long-term dietary lifestyle intervention and may grant additive benefit in the prevention of early vascular disease.

Micronutrients are involved in specific biochemical pathways and have dedicated functions in the body, but they are also interconnected in complex metabolic networks, such as oxidative-reductive and inflammatory pathways and hormonal regulation, in which the overarching function is to optimise health. Post-genomic technologies, in particular metabolomics and proteomics, both of which are appropriate for plasma samples, provide a new opportunity to study the metabolic effects of micronutrients in relation to optimal health. The study of micronutrient-related health status requires a combination of data on markers of dietary exposure, markers of target function and biological response, health status metabolites, and disease parameters. When these nutrient-centred and physiology/health-centred parameters are combined and studied using a systems biology approach with bioinformatics and multivariate statistical tools, it should be possible to generate a micronutrient phenotype database. From this we can explore external factors that define the phenotype, such as lifestage and lifestyle, and the impact of genotype, and the results can also be used to define micronutrient requirements and provide dietary advice. New mechanistic insights have already been developed using biological network models, for example genes and protein-protein interactions in the aetiology of type 2 diabetes mellitus. It is hoped that the challenge of applying this approach to micronutrients will, in time, result in a change from micronutrient oriented to a health oriented views and provide a more holistic understanding of the role played by multiple micronutrients in the maintenance of homeostasis and prevention of chronic disease, for example through their involvement in oxidation and inflammation.

Isothiocyanates have been implicated in the cancer-protective effects of brassica vegetables. When cabbage is consumed, sinigrin is hydrolysed by plant or microbial myrosinase partly to allyl isothiocyanate (AITC), which is mainly excreted as N-acetylcysteine conjugates (NAC) of AITC in urine. The effect of cooking cabbage on the excretion of NAC of AITC, and glutathione-S-transferase (GST) and uridine 5′-diphospho-glucuronosyl transferase (UGT) activity in rat liver and colon was investigated. Germ-free (GF) and human faecal microbiota-associated (HFM) rats were fed a control diet containing 20 % raw, lightly cooked, or fully cooked cabbage for 14 d. When plant myrosinase was present, excretion of NAC of AITC/24 h was increased by 1·4 and 2·5 times by the additional presence of microbial myrosinase after consumption of raw and lightly cooked cabbage respectively. When plant myrosinase was absent, as after consumption of fully cooked cabbage, excretion of the AITC conjugate was almost zero in GF and HFM rats. None of the cabbage diets modified hepatic GST activity. When microbiota was absent, colonic GST was 1·3-fold higher after fully cooked cabbage, and hepatic UGT was increased by 1·4–1·8-fold after all cabbage diets, compared with the control feed. There were no differences in GST or UGT following cabbage consumption when microbiota was present. It is possible that other constituents of cabbage, rather than metabolites of glucosinolates per se, may be responsible for changes in phase 2 enzyme activity. The main effect of cooking cabbage and altering colonic microbiota was on excretion of NAC of AITC.

Dietary long-chain n-3 PUFA (n-3 LCPUFA) are thought to have immune-modulating effects, but the specific effects and mechanisms are not fully elucidated. The aim of this study was to determine whether dietary n-3 LCPUFA could affect ex vivo oxidative burst in healthy young men. The study had a randomised 2 × 2-factorial design in which subjects were randomly assigned to 8-week supplementation with capsules containing fish oil (about 2·9 g n-3 LCPUFA/d) or olive oil (control). Subjects were also randomly assigned to household use of oils and fat spreads with a high or a low 18 : 2n-6 content. At baseline and at the end of the intervention, the fatty acid composition of peripheral blood mononuclear cells (PBMC) was analysed by GLC and oxidative burst was studied in whole blood stimulated with zymosan using luminol-enhanced chemiluminescence. The PBMC content of n-3 LCPUFA was markedly increased by the fish oil-supplementation (P < 0·001, compared to the olive oil groups). No effect of the intervention was observed on neutrophil count, but one measure of the zymosan-induced oxidative burst was higher in the fish oil groups (P = 0·03) compared to the olive oil groups. The fat intervention did not in itself affect oxidative burst neither did it change the effect of the fish-oil intervention. The measures of oxidative burst at the end of the intervention period were found to be associated with the DHA content of PBMC (r 0·44, P = 0·016), suggesting a dose–response relationship. These results indicate that n-3 LCPUFA may have immuno-stimulating effects.

Arnould, S. ES78

Phytochemicals may protect cellular DNA by direct antioxidant effect or modulation of the DNA repair activity. We investigated the repair activity towards oxidised DNA in human mononuclear blood cells (MNBC) in two placebo-controlled antioxidant intervention studies as follows: (1) well-nourished subjects who ingested 600 g fruits and vegetables, or tablets containing the equivalent amount of vitamins and minerals, for 24 d; (2) poorly nourished male smokers who ingested 500 mg vitamin C/d as slow- or plain-release formulations together with 182 mg vitamin E/d for 4 weeks. The mean baseline levels of DNA repair incisions were 65·2 (95 % CI 60·4, 70·0) and 86·1 (95 % CI 76·2, 99·9) among the male smokers and well-nourished subjects, respectively. The male smokers also had high baseline levels of oxidised guanines in MNBC. After supplementation, only the male smokers supplemented with slow-release vitamin C tablets had increased DNA repair activity (27 (95 % CI 12, 41) % higher incision activity). These subjects also benefited from the supplementation by reduced levels of oxidised guanines in MNBC. In conclusion, nutritional status, DNA repair activity and DNA damage are linked, and beneficial effects of antioxidants might only be observed among poorly nourished subjects with high levels of oxidised DNA damage and low repair activity.