Human milk oligosaccharides (HMO) are considered to influence the composition of the gut microflora in breastfed infants. We investigated direct effects of milk HMO fractions or individual oligosaccharides on proliferation, differentiation and apoptosis in transformed human intestinal cells (HT-29 and Caco-2) and non-transformed small intestinal epithelial crypt cells of fetal origin (human intestinal epithelial cells; HIEC). We observed growth inhibition induced by neutral and acidic HMO fractions in HT-29, Caco-2 and HIEC cells in a dose dependent manner. However, the effects varied between cell lines, i.e. HT-29 and Caco-2 cells were more sensitive than HIEC cells. In HT-29, all 16 individual neutral and acidic oligosaccharides except from the two fucosyllactoses had an inhibitory effect on cell growth. Regarding the induction of differentiation in HT-29 and HIEC cells a threshold concentration was observed at 7·5 mg/ml for neutral and acidic HMO fractions. Among individual oligosaccharides, only sialyllactoses induced differentiation in HT-29 and HIEC cells; no effect neither of fractions nor of individual oligosaccharides was found in Caco-2 cells. A strong induction of apoptosis was only detected in HT-29 and HIEC cells for neutral oligosaccharide but not for acidic fractions. HMO were shown to induce growth inhibition in intestinal cells through two different mechanisms, by suppressing cell cycle progression through induction of differentiation and/or by influencing apoptosis. As the development and maturation of digestive and absorptive processes depend on differentiation our experiments show that oligosaccharides are effective at influencing various stages in gastrointestinal development in vitro.

Genomic effects of phytochemicals and their implication in the maintenance of health.

Diet–gene interactions have become the focus of much research in recent years. However, little is known about UK dietitians' involvement, confidence and knowledge in genetics and diet–gene interactions. A validated postal questionnaire sent to a randomly selected sample of 600 dietitians in the UK resulted in 390 responses (65 %). Most dietitians had no involvement in eleven activities relating to genetics and diet–gene interactions and lacked confidence in undertaking such activities. However, a significant positive association was found between involvement and confidence for all activities tested (P < 0·0001). A mean knowledge score of 41 % (sd 19) indicated generally low levels of knowledge in genetics and diet–gene interactions. Knowledge scores were higher for those who reported discussing the genetic basis of disease or discussing how diet–gene interactions affected risk (P < 0·05). For the majority of activities, dietitians who reported higher confidence had higher knowledge scores. Given the importance of interactions between genetics and nutrition in preventing and managing disease, this study identifies a need to increase the involvement, confidence and knowledge in genetics and diet–gene interactions of dietitians in the UK.

Proliferation of intimal vascular smooth muscle cells is an important component in the development of atherosclerosis. Ellagic acid is a phenolic compound present in fruits (raspberries, blueberries, strawberries) and walnuts. The present study investigated the effect of ellagic acid on the oxidised LDL (ox-LDL)-induced proliferation of rat aortic smooth muscle cells (RASMC). The study found that ellagic acid significantly inhibited ox-LDL-induced proliferation of RASMC and phosphorylation of extracellular signal-regulated kinase (ERK) 1/2.Furthermore, ellagic acid also blocked the ox-LDL-induced (inducible) cell-cycle progression and down regulation of the expression of proliferating cell nuclear antigen (PCNA) in RASMC. Therefore, ellagic acid reduced the amount of ox-LDL-induced proliferation of RASMC via inactivation of the ERK pathway and suppression of PCNA expression. These results may significantly advance the understanding of the role that antioxidants play in the prevention of atherosclerosis.

The present study introduced a molecular biological approach to demonstrate changes in the composition of intestinal microbiota in neonatal mice. Female BALB/c mice were fed either a control diet or a diet supplemented with fructo-oligosaccharide (FOS) at 50 g/kg diet, and then mated to male mice. A cultivation-independent approach, denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified 16S rRNA gene, was performed to characterise changes in intestinal microbial populations in pups at 0, 7, 14 and 21 d old and their dams. Comparisons of DGGE profiles were performed using the Dice similarity coefficient and the unweighted pair group method with arithmetic mean (UPGMA) cluster analysis based on numbers, positions and intensities of bands. DGGE profiles differed between dams fed control and FOS-supplemented diets. Although profiles in pups on the day of birth showed a high similarity with dams, profiles in 7-d-old pups differed from dams and showed high similarity to littermates. In 14- and 21-d-old pups, profiles again showed high similarity with dams. DGGE profiles in pups were divided into two large clusters of control and FOS-supplemented diet groups in the range of 0- to 21-d-old, suggesting modulation of intestinal microbiota in infants by manipulation of microbiota in dams. The present study shows a useful technique for demonstrating changes in intestinal microbiota and provides a mouse model for modulation of intestinal microbiota in neonatal life.

Increasing evidence suggests that a low dietary intake of the n-3 long-chain PUFA EPA and DHA may contribute not only to the risks for various physical illnesses (particularly cardiovascular and immune system disorders), but also to many disorders of mental health and performance1,2. From their recent comprehensive review of the evidence for these n-3 fatty acids in the prevention and treatment of psychiatric disorders3, the American Psychiatric Association (APA) made the following recommendations:

The present paper explores the relative importance of liking and taste preferences as correlates of fruit and vegetable (FV) intakes among schoolchildren in Europe. The paper first provides an overview of potential determinants of food choice among children and subsequently summarizes the results of two recent observational studies on determinants of FV intakes among school-aged children. It is proposed that taste preferences and liking are important for children's food choices as part of a broader spectrum of nutrition behaviour determinants. Taste preferences and liking are important for motivation to eat certain foods, but social-cultural and physical environmental factors that determine availability and accessibility of foods, as well as nutrition knowledge and abilities should also be considered.

Study 1 shows that children with a positive liking for FV have a greater likelihood to eat fruits (odds ratio (OR) = 1·97) or vegetables (OR = 1·60) every day, while ability and opportunity related factors such as knowledge, self-efficacy, parental influences and accessibility of FV were also associated with likelihood of daily intakes (ORs between 1·16 and 2·75). These results were consistent across different countries in Europe. Study 2 shows that taste prerences were the stongest mediator of gender differences in FV intakes among children; the fact that girls eat more could for a large extend be explained by there stronger taste preferences.

The essentiality of copper (Cu) in humans is demonstrated by various clinical features associated with deficiency, such as anaemia, hypercholesterolaemia and bone malformations. Despite significant effort over several decades a sensitive and specific Cu status biomarker has yet to be identified. The present article updates a comprehensive review recently published by the authors which assesses the reliability and robustness of current biomarkers and outlines the on-going search for novel indicators of status(1). The essential features of this earlier review are reiterated whilst considering whether there are other approaches, not yet tested, which may provide valuable information in the quest for an appropriate measure of copper status. Current biomarkers include a range of cuproenzymes such as the acute phase protein caeruloplasmin and Cu-Zn-superoxide dismutase all of which are influenced by a range of other dietary and environmental factors. A recent development is the identification of the Cu chaperone, CCS as a potential biomarker; although its reliability has yet to be established. This appears to be the most promising potential biomarker, responding to both Cu deficiency and excess. The potential for identifying a ‘suite’ of biomarkers using high-throughput technologies such as transcriptomics and proteomics is only now being examined. A combination of these technologies in conjunction with a range of innovative metal detection techniques is essential if the search for robust copper biomarkers is to be successful.

Minor components from dietary oils can modulate the atherogenic response of the TAG-rich lipoproteins (TRL) in which they are transported. In the present study we investigated the influence of TRL isolated from man after the intake of oleic acid-rich oils with different minor component compositions on VLDL secretion by rat primary hepatocytes. TRL were isolated from nine men after the intake of meals enriched with high-oleic sunflower oil (HOSO) or virgin olive oil (VOO) or VOO enriched with minor components (EVO). TRL were incubated with rat primary hepatocytes and the lipid accumulation was analysed in the cells and the secreted VLDL. The expression of genes for proteins related to hepatic lipid metabolism and VLDL production was also measured. Incubation of hepatocytes with TRL derived from HOSO as compared to VOO led to lower intracellular lipid accumulation and VLDL production despite higher mRNA expression for diacylglycerol-acyltransferase, microsomal TAG transfer protein, apoB and PPARα. When TRL derived from EVO were used there were no changes in VLDL secretion. These results suggest that incorporation of minor components from dietary high-oleic oils into TRL modulates the effect of these atherogenic particles on VLDL secretion.

Epidemiological studies suggest that the incidence of CVD and postmenopausal osteoporosis is low in the Mediterranean area, where herbs and nuts, among others, play an important role in nutrition. In the present study, we sought a role of walnuts (Juglans regia L.) in endothelial and bone-cell function. As the endothelial cell expression of adhesion molecules has been recognised as an early step in inflammation and atherogenesis, we examined the effect of walnut methanolic extract and ellagic acid, one of its major polyphenolic components (as shown by HPLC analysis), on the expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in human aortic endothelial cells. After incubating the cells with TNF-α (1 ng/ml) in the absence and in the presence of walnut extract (10–200 μg/ml) or ellagic acid (10− 7–10− 5m), the VCAM-1 and ICAM-1 expression was quantified by cell-ELISA. We further evaluated the effect of walnut extract (10–50 μg/ml), in comparison with ellagic acid (10− 9–10− 6m), on nodule formation in the osteoblastic cell line KS483.Walnut extract and ellagic acid decreased significantly the TNF-α-induced endothelial expression of both VCAM-1 and ICAM-1 (P < 0·01; P < 0·001). Both walnut extract (at 10–25 μg/ml) and ellagic acid (at 10− 9–10− 8m) induced nodule formation in KS483 osteoblasts. The present results suggest that the walnut extract has a high anti-atherogenic potential and a remarkable osteoblastic activity, an effect mediated, at least in part, by its major component ellagic acid. Such findings implicate the beneficial effect of a walnut-enriched diet on cardioprotection and bone loss.

The role of zinc deficiency as an important cause of morbidity and impaired linear growth has prompted the need to identify indicators of population zinc status. Three indicators have been recommended – prevalence of zinc intakes below the estimated average requirement (EAR), percentage with low serum zinc concentrations, and percentage of children aged < 5 years who are stunted. This review outlines steps to estimate the prevalence of inadequate intakes, and confirm their validity based on the EARs set by International Zinc Nutrition Collaborative Group. Next, the appropriateness of serum zinc as a biochemical marker for population zinc status is confirmed by a summary of: (a) the response of serum zinc concentrations to zinc intakes; (b) usefulness of serum zinc concentrations to predict functional responses to zinc interventions; (c) relationship between initial serum zinc and change in serum zinc in response to interventions. Height- or length-for-age was chosen as the best functional outcome after considering the responses of growth, infectious diseases (diarrhoea, pneumonia), and developmental outcomes in zinc supplementation trials and correlation studies. The potential of other zinc biomarkers such as zinc concentrations in hair, cells, zinc-metalloenzymes, and zinc-binding proteins, such as metallothionein, is also discussed. Molecular techniques employing reverse transcriptase (RT)-polymerase chain reaction to measure mRNA in metallothionein and ZIP1 transporter hold promise, as do kinetic markers such as exchangeable zinc pools (EZP) and plasma zinc turnover rates. More research is needed to establish the validity, specificity, sensitivity, and feasibility of these new biomarkers, especially in community-settings.

Inulin, a linear β fructan, is present in a variety of plants including chicory root and wheat. It exhibits prebiotic properties and has been shown to enhance mineral absorption and increase beneficial bacteria in the colon. The aim of the present study was to assess the effect of dietary inulin on the gene expression of selected intestinal Fe transporters and binding proteins. Anaemic piglets at age 5 weeks were allocated to a standard maize–soya diet (control) or the same diet supplemented with inulin at a level of 4 %. After 6 weeks, the animals were killed and caecum contents and sections of the duodenum and colon were removed. Segments of the genes encoding for the pig divalent metal transporter 1 (DMT1) and duodenal cytochrome-b reductase (Dcytb) were isolated and sequenced. Semi-quantitative RT-PCR analyses were performed to evaluate the expression of DMT1, Dcytb, ferroportin, ferritin, transferrin receptor (TfR) and mucin genes. DMT1, Dcytb, ferroportin, ferritin and TfR mRNA levels in duodenal samples were significantly higher in the inulin group (P ≤ 0·05) compared with the control. In colon, DMT1, TfR and ferritin mRNA levels significantly increased in the inulin group. Additionally, the caecal content microflora was examined using 16S rDNA targeted probes from bacterial DNA. The Lactobacillus and Bifidobacterium populations were significantly increased in the inulin group (P ≤ 0·05) compared with the control group. These results indicate that dietary inulin might trigger an up regulation of genes encoding for Fe transporters in the enterocyte. The specific mechanism for this effect remains to be elucidated.

Iodine deficiency is an important clinical and public health problem. Its prevention begins with an adequate intake of iodine during pregnancy. International agencies recommend at least 200 μg iodine per d for pregnant women. We assessed whether iodine concentrations in the amniotic fluid of healthy pregnant women are independent of iodine intake. This cross-sectional, non-interventional study included 365 consecutive women who underwent amniocentesis to determine the fetal karyotype. The amniocentesis was performed with abdominal antisepsis using chlorhexidine. The iodine concentration was measured in urine and amniotic fluid. The study variables were the intake of iodized salt and multivitamin supplements or the prescription of a KI supplement. The mean level of urinary iodine was 139·0 (sd 94·5) μg/l and of amniotic fluid 15·81 (sd 7·09) μg/l. The women who consumed iodized salt and those who took a KI supplement had significantly higher levels of urinary iodine than those who did not (P = 0·01 and P = 0·004, respectively). The urinary iodine levels were not significantly different in the women who took a multivitamin supplement compared with those who did not take this supplement, independently of iodine concentration or multivitamin supplement. The concentrations of iodine in the amniotic fluid were similar, independent of the dietary iodine intake. Urine and amniotic fluid iodine concentrations were weakly correlated, although the amniotic fluid values were no higher in those women taking a KI supplement. KI prescription at recommended doses increases the iodine levels in the mother without influencing the iodine levels in the amniotic fluid.

To assess the effect of lupin protein on concentrations of lipids in plasma lipoproteins and liver and hepatic mRNA concentrations of genes involved in lipid metabolism, adult rats were fed egg albumin-based diets containing either lupin protein from Lupinus albus or casein (50 g/kg) supplemented (hypercholesterolaemic) or not (normolipaemic) with a cholesterol–cholate mixture for 20 d. Lupin protein compared with casein lowered the concentrations of TAG in liver (P < 0·01) and circulating VLDL + chylomicrons (P < 0·05) of hypercholesterolaemic rats, but not of normolipaemic rats. Hepatic mRNA concentrations of genes involved in fatty acid synthesis such as sterol regulatory element-binding protein-1c, glucose-6-phosphate dehydrogenase, fatty acid synthase, stearoyl-CoA desaturase-1 and acyl-CoA:glycerol-3-phosphate acyltransferase were lower and mRNA concentrations of lipoprotein lipase, hepatic lipase and apoA5 involved in TAG hydrolysis were higher in rats fed lupin protein than in rats fed casein. These effects were stronger in hypercholesterolaemic rats than in normolipaemic rats. Hypercholesterolaemic rats fed the lupin protein had higher liver cholesterol concentrations (P < 0·01) and lower levels of LDL-cholesterol (P < 0·05) than rats fed casein. No effect of lupin protein was observed on cholesterol concentration in VLDL + chylomicrons and HDL and hepatic mRNA concentrations of genes involved in cholesterol and bile acid metabolism. In conclusion, the present study shows that lupin protein has hypotriacylglycerolaemic action possibly via down regulation of fatty acid synthesis genes and up regulation of genes involved in TAG hydrolysis. Alterations in cholesterol metabolism could not be explained on the basis of mRNA data.

Micronutrient deficiencies are frequently associated with tuberculosis (TB) worldwide. We tested the effect of exogenous vitamin E on proliferation and cytokine production of peripheral blood mononuclear cells (PBMC) from TB patients and healthy purified protein derivative (PPD)+ volunteers. Proliferation was stimulated with mycobacterial antigen (PPD) and evaluated by the incorporation of tritiated thymidine in PBMC cultured with or without 50 μm-vitamin E for 6 d. Cytokine production (IL-2 and interferon (IFN)-γ) was determined by intracellular cytokine staining and by ELISA in the supernatant of PBMC stimulated for 24 h with phytohaemagglutinin or PPD. Our results show that culture with vitamin E increased (P ≤ 0·05) the antigen-induced proliferation of PBMC in TB patients but not in healthy PPD+ volunteers. No significant changes in the number of cytokine-producing cells or in the production of IFN-γ were observed with vitamin E treatment. These results indicate that vitamin E may enhance the antigen-specific in vitro response of PBMC from TB patients.

Omnivores have the advantage of a variety of food options but face a challenge in identifying foods that are safe to eat. Not surprisingly, therefore, children show a relative aversion to new foods (neophobia) and a relative preference for familiar, bland, sweet foods. While this may in the past have promoted survival, in the modern food environment it could have an adverse effect on dietary quality. This review examines the evidence for genetic and environmental factors underlying individual differences in children's food preferences and neophobia. Twin studies indicate that neophobia is a strongly heritable characteristic, while specific food preferences show some genetic influence and are also influenced by the family environment. The advantage of the malleability of human food preferences is that dislike of a food can be reduced or even reversed by a combination of modelling and taste exposure. The need for effective guidance for parents who may be seeking to improve the range or nutritional value of foods accepted by their children is highlighted.

A large variety of phytochemicals are found in vegetables, fruits, spices and staple plant foods(1,2).

Effects of n-acetyl cysteine (NAC), s-ethyl cysteine (SEC), s-propyl cysteine (SPC) and cysteine on enzymes participating in biosynthesis of TAG and cholesterol, and antioxidant protection in liver from mice consuming a high-saturated fat diet was examined. The high-fat diet provided 70 % fat energy, in which saturated fat was 55 % of total fat. NAC, SEC, SPC or cysteine, each agent at 1 g/l, was directly added into the drinking water as a supplement for 4 weeks. Results showed high saturated fat significantly increased hepatic TAG and total cholesterol contents (P < 0·05) via enhancing the activity and mRNA expression of malic enzyme, fatty acid synthase and 3-hydroxy-3-methylglutaryl coenzyme A reductase (P < 0·05). The intake of NAC, SEC or SPC significantly decreased TAG and total cholesterol levels (P < 0·05) via lowering the activity and mRNA expression of these three lipogenic-related enzymes (P < 0·05). NAC, SEC or SPC treatment also significantly suppressed high saturated fat-induced hepatic mRNA expression of sterol regulatory element-binding protein (SREBP)-1c and SREBP-2 (P < 0·05). High saturated fat decreased hepatic content of glutathione, and the activity of catalase and glutathione peroxidase (P < 0·05). The intake of NAC, SEC or SPC significantly increased hepatic glutathione content (P < 0·05), restored the activity and mRNA expression of glutathione peroxidase, and alleviated the high saturated fat-induced oxidative stress (P < 0·05). These results support that NAC, SEC and SPC are potent agents for affecting hepatic biosynthesis of TAG and cholesterol, and protecting liver against high saturated fat-associated oxidative damage.

The present paper is a review of available data on effects of parental feeding attitudes and styles on child nutritional behaviour. Food preferences develop from genetically determined predispositions to like sweet and salty flavours and to dislike bitter and sour tastes. There is evidence for existence of some innate, automatic mechanism that regulate appetite. However, from birth genetic predispositions are modified by experience. There are mechanisms of taste development: mere exposure, medicine effect, flavour learning, flavour nutrient learning. Parents play a pivotal role in the development of their child's food preferences and energy intake, with research indicating that certain child feeding practices, such as exerting excessive control over what and how much children eat, may contribute to childhood overweight. Mothers are of particular interest on children's eating behaviour, as they have been shown to spend significantly more time than fathers in direct interactions with their children across several familial situations.

A recent paper describes two primary aspects of control: restriction, which involves restricting children's access to junk foods and restricting the total amount of food, and pressure, which involves pressuring children to eat healthy foods (usually fruits and vegetables) and pressuring to eat more in general.

The results showed significant correlations between parent and child for reported nutritional behaviour like food intake, eating motivations, and body dis- and satisfaction. Parents create environments for children that may foster the development of healthy eating behaviours and weight, or that may promote overweight and aspects of disordered eating. In conclusion positive parental role model may be a better method for improving a child's diet than attempts at dietary control.

Green tea catechins are known to have hypocholesterolaemic effects in animals and human subjects. In the present study, we investigated the effects of green tea catechins on the mRNA level and promoter activity of hepatic cholesterol 7α-hydroxylase (CYP7A1), the rate-limiting enzyme in the conversion of cholesterol to bile acids, in human hepatoma cells. Real-time PCR assays showed that different catechins, ( − )-epicatechin gallate (ECG), ( − )-epigallocatechin-3-gallate (EGCG), ( − )-epigallocatechin (EGC) and ( − )-epicatechin (EC), up regulated the CYP7A1 mRNA level by 5·5-, 4·2-, 2·9- and 1·9-fold, respectively, compared with the control. The − 1312/+358 bp of the CYP7A1 promoter was subcloned into the pGL3 basic vector that includes luciferase as a reporter gene. ECG or EGCG significantly increased CYP7A1 promoter activity by 6·0- or 4·0-fold, respectively, compared with the control. Also, EGCG stimulated CYP7A1 at both mRNA level and promoter activity in a dose-dependent manner. These results suggest that the expression of the CYP7A1 gene may be directly regulated by green tea catechins at the transcriptional level.