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.

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.

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.

Altered fatty acid (FA) composition is related to insulin resistance and CVD. One possible mediator may be inflammation, but longitudinal data relating FA composition to inflammation taking insulin resistance into account are limited. We investigated the long-term association between FA composition and C-reactive protein (CRP) concentrations in a large population-based cohort study in 767 men followed for 20 years. The association between FA composition in serum cholesteryl esters at age 50 and CRP concentrations at age 70 was investigated using linear regression. In addition, desaturase activities (stearoyl-CoA desaturase-1 (SCD-1), Δ5- and Δ6-desaturase) were estimated using FA product-to-precursor ratios. Insulin resistance was measured directly at follow-up by euglycaemic clamp. After adjusting for confounders (smoking, physical activity, alcohol intake, obesity and erythrocyte sedimentation rate) CRP concentrations were inversely associated with the proportion of 18 : 2n-6 (P = 0·002) and positively associated with 16 : 1n-7 (P = 0·008), 18 : 1n-9 (P = 0·0003), 20 : 5n-3 (P = 0·04) and estimated SCD-1 (P = 0·005) and Δ6-desaturase (P = 0·02) activities. After adding insulin resistance to the model, 18 : 1n-9, 18 : 2n-6 and SCD-1 remained significant predictors of CRP. A FA composition indicating low intake of 18 : 2n-6, high intake of SFA and high SCD-1 activity is, in a Swedish population of middle-aged men, associated with CRP concentrations 20 years later, even independently of obesity and insulin resistance.

Numerous mechanisms have been proposed to explain the anti-carcinogenic effects of Se, among them altered carcinogen metabolism. We investigated the effect of Se supplementation on activities of glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione S-transferase (GST) in different blood compartments, and expression of selected phase 1 and phase 2 genes in leucocytes (GPX1, γ-glutamylcysteine ligase catalytic subunit (GCLC), AP-1 transcription factor Fos-related antigen 1 (Fra1), NAD(P)H:quinone oxidoreductase (NQO1), and aryl hydrocarbon receptor repressor (AhRR)). Healthy elderly Danes (n 105; age 71·3 (sd 4·26) years; 36 % reporting use of multivitamin/mineral supplements) participated and were supplemented daily for 5 years with placebo, 100 μg, 200 μg or 300 μg Se as Se-enriched yeast (SelenoPrecise®). Blood samples were collected after 5 years of intervention. When all four groups were compared we found no effect of Se supplementation on plasma GPX or GR, on erythrocyte GPX, GR or GST, or on thrombocyte GR or GST. We found increased thrombocyte GPX activity at the two highest dosage levels in women only, but not in men. No effects on GPX1, NQO1 or AhRR gene expression were found. When all Se-supplemented groups were pooled we found significant down regulation of the expression of some phase 2 genes (GCLC, Fra1). A significant increase in AhRR gene expression with smoking was found but was independent of Se supplementation. Down regulation of phase 2 genes could increase the risk of cancer. However, further studies are needed to establish whether the observed effect in leucocytes reflects a similar expression pattern in target tissues.

Oxidation of LDL is hypothesised as an early and critical event in atherogenesis. Oxidised LDL (oxLDL) favour the transformation of macrophages into foam cells, an important cell involved in atherosclerosis. Furthermore, oxLDL cause multiple changes in macrophage functions. Thus, oxLDL induces certain genes, suppresses others and alters cell lipid metabolism. Consumption of a Mediterranean diet is associated with a low incidence of atherosclerotic disease, but data about the specific dietary constituents involved and mechanisms conferring cardioprotection are still sparse. The aim of the present study was to determine the effect of representative minor components of wine and olive oil on reactive oxygen species and eicosanoid synthesis induced by oxLDL-stimulated macrophages. We observed that exposure to non-toxic oxLDL concentrations leads to the production of H2O2 by RAW 264.7 macrophages and this effect was reverted by apocynin, a NADPH oxidase inhibitor. Moreover, oxLDL induced arachidonic acid (AA) release, cyclo-oxygenase-2 overexpression and subsequent PGE2 release. We observed that resveratrol and tyrosol revert H2O2 production induced by oxLDL as well as AA release and PGE2 synthesis and that these effects were not as a consequence of these compounds interfering with the oxLDL binding to their receptors. Interestingly, β-sitosterol presence enhances these polyphenol actions. Thus, we found a synergistic action of polyphenols of olive oil and wine and β-sitosterol of olive oil led to the modulation of the effects of oxLDL on oxidative stress and PGE2 synthesis.

As in man, canine obesity is associated with insulin resistance, dyslipidaemia and other chronic diseases. This study was designed to examine the effects of a nutritional supplement (green tea) on insulin sensitivity and plasma lipid concentrations in an obese insulin-resistant dog model. We also determined mRNA expression of two transcription factors, PPARγ and PPARα, and some of their target genes, including GLUT4, lipoprotein lipase (LPL) and adiponectin. Obese dogs were divided into two groups: a green tea group (n 6); a control group (n 4). Dogs in the green tea group were given green tea extract (80 mg/kg per d) orally, just before their single daily meal, for 12 weeks. Insulin sensitivity (using a euglycaemic–hyperinsulinaemic clamp) and concentrations of plasma TAG, total cholesterol and NEFA were assessed in each group. Gene expression was measured in visceral and subcutaneous adipose tissues and in liver and skeletal muscle, by real-time PCR. At 12 weeks in the green tea group, mean insulin sensitivity index was 60 (sem 11) % higher (P < 0·05) and TAG concentration 50 (sem 10) % lower (P < 0·001), than baseline. PPARγ, GLUT4, LPL and adiponectin expression were significantly higher in both adipose tissues, whilst PPARα and LPL expression were significantly higher in skeletal muscle, compared with baseline. These findings show that nutritional doses of green tea extract may improve insulin sensitivity and lipid profile and alter the expression of genes involved in glucose and lipid homeostasis.

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.

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.

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.

Dietary fatty acids have been shown to influence allergic sensitisation. Both n-3 and n-6 PUFA are involved in targeted mediation of inflammatory responses during allergic sensitisation and manifestation of atopic diseases. In the present experiments we investigated whether supplementation of DHA-enriched fish oil partly substituting dietary sunflower-seed oil, in comparison with sunflower-seed oil, supplemented to mice influences fatty acid composition of serum lipid classes. The effects of the two different diets were also investigated depending on allergic sensitisation. Supplementation of DHA and EPA in doses of 2 and 0·12 % (w/w) to non-sensitised and sensitised mice resulted in significantly increased percentile contributions of DHA to all lipid classes. In contrast, serum values of the n-6 PUFA arachidonic acid (AA) were significantly lower, both in non-sensitised and sensitised mice fed the DHA-enriched diet. The fatty acid composition of serum lipids also reflected allergic sensitisation: the EPA:AA ratio in TAG, cholesteryl esters and phospholipids in non-supplemented animals fell to 23, 29 and 29 % respectively of the original value after allergic sensitisation, whereas it decreased to 70, 80 and 76 % respectively only in the animals supplemented with DHA. In summary, allergic sensitisation alone decreased significantly the EPA:AA ratios in serum TAG, while concomitant supplementation of DHA-enriched fish oil ameliorated this decrease. We postulate from the present results that the amelioration of the severity of allergic sensitisation after DHA supplementation may be linked to altered ratios of the eicosanoid precursors EPA and AA as well as DHA needed for further metabolic activation to pro- or anti-inflammatory bioactive lipids.

Tocotrienols have been shown to inhibit proliferation and induce apoptosis in cancer cells. However, the molecular mechanisms involved in tocotrienol-induced apoptosis are still unclear. In the present study, γ-tocotrienol induced apoptosis in human gastric adenocarcinoma SGC-7901 cell line through down regulation of the extracellular signal-regulated kinase (ERK) signalling pathway. Furthermore, γ-tocotrienol-induced apoptosis was accompanied by down regulation of Bcl-2, up regulation of Bax, activation of caspase-3, and subsequent poly (ADP-ribose) polymerase cleavage. These results indicated that up or down regulation of Bcl-2 family proteins play a major role in the initiation of γ-tocotrienol-induced apoptosis as an activator of caspase-3.γ-Tocotrienol also down regulated the activation of the Raf-ERK signalling pathway, and down regulated c-Myc by decreasing the expressions of Raf-1 and p-ERK1/2 proteins. The results suggest that key regulators in tocotrienol-induced apoptosis may be Bcl-2 families and caspase-3 in SGC-7901 cells through down regulation of the Raf-ERK signalling pathway.

Dietary energetic impact on oxidative stress is incompletely understood. Therefore, effects of diets on oxidative stress were studied using a crossover block design. In Expt 1, intake of metabolizable energy (ME) was restricted or ad libitum. In Expt 2, isoenergetic and isonitrogenic diets were fed, replacing carbohydrate energy by energy of fatty acids. Circulatory lipohydroperoxides (LOOH), markers of acute oxidative stress, were expressed absolutely and in terms of cholesterol or TAG levels. In Expt 1, plasma (jugularis vein) LOOH was assayed in combination with whole-body oxidative metabolism using gas exchange and heart rate (HR) during feeding periods and at rest. In Expt 2, LOOH was assayed in plasma from portal and a large udder vein and a mesenteric artery. In Expt 1, intake increased VO2, HR and LOOH following overnight fast with higher values (P < 0·05) when feeding ME ad libitum. Intake of ME ad libitum (3 weeks) increased cardiac protein of cytochrome oxidase and endothelial-type nitric oxide synthase (P < 0·05), indicating adaptation of the heart to higher activity. Transient HR responses evoked by an antidiabetic drug (levcromakalim) revealed a linear positive correlation with relative LOOH (r2 0·79), supporting the relationship between oxidative metabolic rate and lipoperoxidation. Evidence for exogenous lipids as LOOH source provided the vessel-specific rise in LOOH through replacing carbohydrate ME by lipid ME (Expt 2). Thus, dietary energy level and energetic source are important for circulatory LOOH with a role of vascular activity in production of oxidant.

Self-report recall questionnaires used to measure physical activity and dietary intake in children can be labour intensive and monotonous and tend to focus on either dietary intake or physical activity. The web-based software, Synchronised Nutrition and Activity ProgramTM (SNAPTM), was developed to produce a novel, simple, quick and engaging method of assessing energy balance-related behaviours at a population level, combining principles from new and existing 24 h recall methodologies, set within a user-friendly interface. Dietary intake was measured using counts for twenty-one food groups and physical activity levels were measured in min of moderate to vigorous physical activity (MVPA). A combination of the mean difference between methods, type II regression and non-parametric limits of agreement techniques were used to examine the accuracy and precision of SNAPTM. Method comparison analyses demonstrated a good agreement for both dietary intake and physical activity behaviours. For dietary variables, accuracy of SNAPTM (mean difference) was within ± 1 count for the majority of food groups. The proportion of the sample with between-method agreement within ± 1 count ranged from 0·40 to 0·99. For min of MVPA, there was no substantial fixed or proportional bias, and a mean difference between methods (SNAPTM – accelerometry) of − 9 min. SNAPTM provides a quick, accurate, low-burden, cost-effective and engaging method of assessing energy balance behaviours at a population level. Tools such as SNAPTM, which exploit the popularity, privacy and engagement of the computer interface, and linkages with other datasets, could make a substantial contribution to future public health monitoring and research.

Adolescents aged 11–14 years (n 326), belonging to organized sports federations in the Federal District, Brazil were interviewed. Subjects (n 107) provided four non-consecutive days of food consumption and 219 subjects provided two non-consecutive days of intake. The objective was to assess their nutrient and water intake according to dietary reference intake values and their energy and macronutrient intake by sex and sports groups they were engaged in: endurance, strength-skill or mixed, according to the guidelines established by the American College of Sport Medicine (ACSM). Dietary data were corrected for intra-individual variation. Total energy expenditure was higher among endurance athletes (P < 0·001) following their higher training time (P < 0·001) when compared to adolescents engaged in strength-skill or mixed sports. Total energy intake was only significantly higher among endurance-engaged females (P = 0·05). Protein intake of males was above the guidelines established by the ACSM for all sports groups. All male sport groups fulfilled the intake levels of carbohydrate per kg body weight but only females engaged in endurance sports fulfilled carbohydrate guidelines. Intakes of micronutrients with low prevalence of adequate intake were: vitamins B1, E and folate, magnesium and phosphorus. Few adolescents ( < 5 %) presented adequate intake for calcium, fibre, drinking water and beverages. For micronutrients, prevalence of adequacies were lower for females than males, except for liquids and water. Nutrition guidance is needed to help adolescents fulfil specific guidelines of macronutrient intake for their sports and to improve their intake of micronutrients and water. Special attention should be given to female adolescent athletes.

Blood pressure (BP) shows a continuous relationship with the risk of CVD. There is substantial evidence that dietary potassium exerts an anti-pressor effect. Most clinical trials have used KCl. However, the chloride ion may have a pressor effect and in foods potassium is associated with organic anions. In a double-blind randomized placebo-controlled trial we explored the effect on BP of two salts of potassium, KCl and potassium citrate (K-cit), in predominantly young healthy normotensive volunteers. The primary outcome was the change in mean arterial pressure as measured in a clinic setting. After 6 weeks of supplementation, compared with the placebo group (n 31), 30 mmol K-cit/d (n 28) changed mean arterial pressure by − 5·22 mmHg (95 % CI − 8·85, − 4·53) which did not differ significantly from that induced by KCl (n 26), − 4·70 mmHg ( − 6·56, − 2·84). The changes in systolic and diastolic BP were − 6·69 (95 % CI − 8·85, − 4·43) and − 4·26 (95 % CI − 6·31, − 2·21) mmHg with K-cit and − 5·24 (95 % CI − 7·43, − 3·06) and − 4·30 (95 % CI − 6·39, − 2·20) mmHg with KCl, and did not differ significantly between the two treatments. Changes in BP were not related to baseline urinary electrolytes. A greater treatment-related effect was observed in those with higher systolic BP. Increasing dietary potassium could therefore have a significant impact on the progressive rise in BP in the entire population.

Dietary carotenoids are associated with a reduced risk of chronic diseases. Raw food diets are predominantly plant-based diets that are practised with the intention of preventing chronic diseases by virtue of their high content of beneficial nutritive substances such as carotenoids. However, the benefit of a long-term adherence to these diets is controversial since little is known about their adequacy. Therefore, we investigated vitamin A and carotenoid status and related food sources in raw food diet adherents in Germany. Dietary vitamin A, carotenoid intake, plasma retinol and plasma carotenoids were determined in 198 (ninety-two male and 106 female) strict raw food diet adherents in a cross-sectional study. Raw food diet adherents consumed on average 95 weight% of their total food intake as raw food (approximately 1800 g/d), mainly fruits. Raw food diet adherents had an intake of 1301 retinol activity equivalents/d and 16·7 mg/d carotenoids. Plasma vitamin A status was normal in 82 % of the subjects ( ≥ 1·05 μmol/l) and 63 % had β-carotene concentrations associated with chronic disease prevention ( ≥ 0·88 μmol/l). In 77 % of subjects the lycopene status was below the reference values for average healthy populations ( < 0·45 μmol/l). Fat contained in fruits, vegetables and nuts and oil consumption was a significant dietary determinant of plasma carotenoid concentrations (β-carotene r 0·284; P < 0·05; lycopene r 0·168; P = 0·024). Long-term raw food diet adherents showed normal vitamin A status and achieve favourable plasma β-carotene concentrations as recommended for chronic disease prevention, but showed low plasma lycopene levels. Plasma carotenoids in raw food adherents are predicted mainly by fat intake.

Lignans, derived from flaxseed, are phyto-oestrogens being increasingly studied for their health benefits. An 8-week, randomised, double-blind, placebo-controlled study was conducted in fifty-five hypercholesterolaemic subjects, using treatments of 0 (placebo), 300 or 600 mg/d of dietary secoisolariciresinol diglucoside (SDG) from flaxseed extract to determine the effect on plasma lipids and fasting glucose levels. Significant treatment effects were achieved (P < 0·05 to < 0·001) for the decrease of total cholesterol (TC), LDL-cholesterol (LDL-C) and glucose concentrations, as well as their percentage decrease from baseline. At weeks 6 and 8 in the 600 mg SDG group, the decreases of TC and LDL-C concentrations were in the range from 22·0 to 24·38 % respectively (all P < 0·005 compared with placebo). For the 300 mg SDG group, only significant differences from baseline were observed for decreases of TC and LDL-C. A substantial effect on lowering concentrations of fasting plasma glucose was also noted in the 600 mg SDG group at weeks 6 and 8, especially in the subjects with baseline glucose concentrations ≥ 5·83 mmol/l (lowered 25·56 and 24·96 %; P = 0·015 and P = 0·012 compared with placebo, respectively). Plasma concentrations of secoisolariciresinol (SECO), enterodiol (ED) and enterolactone were all significantly raised in the groups supplemented with flaxseed lignan. The observed cholesterol-lowering values were correlated with the concentrations of plasma SECO and ED (r 0·128–0·302; P < 0·05 to < 0·001). In conclusion, dietary flaxseed lignan extract decreased plasma cholesterol and glucose concentrations in a dose-dependent manner.