Preterm neonates show enhanced sensitivity to nutrient maldigestion and bacteria-mediated gut inflammatory disorders, such as necrotising enterocolitis (NEC). We hypothesised that preterm birth increases the sensitivity of intestinal nutrient absorption to endotoxins and that feeding after birth reduces this response. Hence, we investigated the postnatal development of nutrient digestive and absorptive capacity in the preterm and term pig intestine, and its responsiveness to endotoxins. Pigs were delivered by caesarean section at preterm (n 20) or term (n 17) gestation, and the small intestine was collected at birth or after 2 d of colostrum feeding, followed by ex vivo stimulation with lipopolysaccharide endotoxins and mixed gut contents collected from pigs with NEC. Brush border enzyme activities were reduced in newborn preterm v. term pigs (39–45 % reduction, P < 0·05), but normalised after 2 d of feeding. Ex vivo leucine and glucose uptake increased with prenatal age. Bacterial stimulation reduced the nutrient uptake similarly at birth and after 2 d in preterm and term pigs (23–41 % reduction, P < 0·05), whereas IL-6 and TNF-α expression was stimulated only at birth. Toll-like receptor-4 expression increased markedly at day 2 for preterm and term pigs (22–33-fold, P < 0·05) but with much lower expression levels in newborn preterm pigs (approximately 95 %, P < 0·01). In conclusion, digestive and absorptive functions mature in the prenatal period, but are similarly affected by postnatal feeding and bacterial exposure in both preterm and term pigs. Nutrient maldigestion may contribute to NEC development, while a prematurity-related hyper-responsiveness to endotoxins could be less important, at least in pigs.

As part of its work on setting standards and establishing guidelines for nutrigenomics research, the European Nutrigenomics Organisation (NuGO) is developing bioethical guidelines for those engaged in human nutrigenomics studies. A NuGO working group developed a set of draft guidelines addressing four areas: (1) information and consenting prior to a nutrigenomicsstudy; (2) the generation and use of genotype information; (3) the establishment and maintenance of biobanks; (4) the exchange of samples and data. NuGO convened a workshop with a panel of invited external experts to assess the draft guidelines. The panel of experts confirmedthat these areas are important and that the development of specific bioethical guidelines for nutrigenomics research would therefore enhance the application of established international guidelines in this field of biomedical research.

1. The influence of three diets ((1) high-fibre–low-fat, (2) low-fibre–high-fat, (3) commercial breeding diet) on the concentration of short chain fatty acids (SCFA) has been investigated in male and female rats up to 14 weeks of age.

2. Hydrogen was detected in respired gas in all rats with no significant differences between diets or sex. Methane was detected only in control-fed rats at 12 weeks of age and thereafter. Caecal contents contained higher concentrations of SCFA than were found in faeces. There were no significant differences in the total faecal SCFA excreted by rats on the three diets. The proportions of SCFA in the caecum and faeces were influenced by diet.

3. The production and excretion of hydrogen, methane and SCFA in the rat appeared to reflect different aspects of bacterial metabolism. Comparison of caecal and faecal SCFA may indicate differential absorption from the colon or differential metabolism by bacteria or colonic mucosa which may be influenced by diet.

Effects of cassava (Manihot esculenta Crantz)-borne organic cyanide and inorganic cyanide in the form of sodium cyanide on bone and muscle development were investigated in eighteen dogs of Nigerian breed. After 16 weeks of stabilization in the laboratory from the time of purchase when the dogs were fed on the same diet, they were randomly assigned to three experimental groups of six dogs each. The control group was fed on rice while the other two groups were fed on either cassava (gari) or rice plus cyanide. The three diets were made isoenergetic and isonitrogenous by varying the quantity of meat incorporated into them. The results obtained after 14 weeks of feeding the respective diets indicated that there was retardation of muscle development in the gari-fed dogs. This may have resulted from gluconeogenesis from muscle protein associated with suppression of production of insulin by the pancreas in this group. The results indicated also that the effects of inorganic dietary cyanides on muscle development were different. Both forms of dietary cyanides, however, had no adverse effect on bone development

Several studies have failed to demonstrate an association between Fe status and intake of dietary Fe. However, in the long term, it Seems logical to presume that body Fe reserves are, fundamentally, dependent on the intake of bioavailpble dietary Fe. This discrepancy may depend on several factors(1)interindividual variation in biological availability of dietary Fe (differences in intestinal absorption),(2) interactions between dietary Fe and absorption enhancers and inbibitors, (3) variatiom in physiological (menstruation, childbirth) or unphysiological (blood donation) Fe losses, (4) the failure to adjust dietary intake data for Fe supplements, (5) uncertain food composition data (discrepancies between calculated and chemically measured Fe content inthe diet), and (6) diet reporting error (reported intake of dietary Fe may deviate considerably from the true intake). The present study examined associations between dietary intake of Fe (assessed by diet history interview) and Fe status (assessed by femtin status) among 167 Danish women aged 35–65 years, who were not blood donors, by taking into account diet reporting error (assessed from p-amino benzoic acid-validated urinary N), physiological blood losses (menstruation, Childbirth, abortions), and Fe supplementation. Our results indicate that the lack of a general association between Fe status and dietary Fe intake may, in part, be caused by selective dietreporting error.

We aimed to investigate the relationship between the neutrophil to lymphocyte ratio (NLR) and nutritional parameters in chronic kidney disease (CKD) patients. In this cross-sectional study, 187 non-dialysis CKD patients were enrolled. Daily dietary energy intake (DEI) and daily dietary protein intake (DPI) were assessed by 3-d dietary records. Protein-energy wasting (PEW) was defined as Subjective Global Assessment (SGA) class B and C. Spearman correlation analysis, logistic regression analysis and receiver operating characteristic (ROC) curve analysis were performed. The median NLR was 2·51 (1·83, 3·83). Patients with CKD stage 5 had the highest NLR level. A total of 19·3 % (n 36) of patients suffered from PEW. The NLR was positively correlated with SGA and serum P, and the NLR was negatively correlated with BMI, waist and hip circumference, triceps skinfold thickness, mid-arm muscle circumference, DPI and Hb. Multivariate logistic regression analysis adjusted for DPI, DEI, serum creatinine, blood urea nitrogen, uric acid and Hb showed that a high NLR was an independent risk factor for PEW (OR = 1·393, 95 % CI 1·078, 1·800, P = 0·011). ROC analysis showed that an NLR ≥ 2·62 had the ability to identify PEW among CKD patients, with a sensitivity of 77·8 %, a specificity of 62·3 % and an AUC of 0·71 (95 % CI 0·63, 0·81, P < 0·001). The NLR was closely associated with nutritional status. NLR may be an indicator of PEW in CKD patients.

The effects of ammoniation of wheat straw with or without supplementation of protein sources of either high (casein) or relatively low (potato protein) rumen degradability on intake and digestion were studied with four sheep in a 4 × 4 Latin square design. Rations offered were: (1) untreated wheat straw (UWS), (2) ammoniated wheat straw (AWS), (3) AWS supplemented with 3·2 g casein/kg live weight (W)0·75 per d (AWSC) and (4) AWS supplemented with 3·9 g potato protein/kg W0·75per d (AWSP). Straw was offered ad lib. and all rations were supplemented with sugarbeet pulp and a mineral mixture. NH3 treatment increased intake and digestion. Supplementation of AWS with potato protein increased total digestible organic matter intake (DOMI) compared with AWS whereas supplementation with casein did not affect total DOMI. Protein supplementation of AWS significantly reduced rumen digestion of cellulose, and when the supplementation was with casein it reduced rumen digestion of neutral-detergent fibre and hemicellulose also. This lower rumen digestion was compensated by a higher proportion of digestion occurring in the hindgut for hemicellulose (P<0·05 for AWSC, P>0·05 for AWSP), but not for cellulose. Across all rations, rumen fluid volume increased with increasing cell-wall intake. The efficiencies of microbial protein synthesis were (average of three different methods of estimation) 23·3, 26·2, 34·8 and 31·7gN/kg apparently-rumen-degraded organic matter for UWS, AWS, AWSC and AWSP respectively. The difference between UWS and AWS was not significant, but values for AWSC and AWSP were significantly higher than that for AWS. The rumen digestion of feed aimno acid-N (AA-N) was significantly higher for AWSC than for the other rations. The apparent small-intestinal digestion of AA-N and N was significantly higher for AWSP than for the other rations. The true small-intestinal digestion values were 0·86, 0·84 and 0·68 for AA-N, N and non-protein-N respectively. Heal endogenous losses of AA-N were approximately 6 mg/g duodenal non-protein dry-matter flow. Linear relationships were observed between DOMI and N balance and truly absorbed AA-N, indicating that DOMI could have been limited by small-intestinal amino acid availability. Regression of N balance v. truly absorbed AA-N resulted in an estimate of net efficiency of utilization of truly absorbed AA-N of 0·54.

The role of diet in asthma is still debated. In France, a front-of-pack (FOP) nutrition label based on a modified Food Standards Agency Nutrient Profiling System/High Council for Public Health (FSAm-NPS/HCSP) has recently been implemented to help consumers to make healthier food choices during purchase. At the individual level, the FSAm-NPS dietary index (DI) has been shown to reflect the nutritional quality of the diet. The aim of the present study was to investigate the association between the FSAm-NPS DI and the asthma symptom score. In total, 34 323 participants (25 823 women and 8500 men) from the NutriNet-Santé cohort were included. The overall nutritional quality of the diet was assessed using the FSAm-NPS DI. Increasing FSAm-NPS DI reflects decreasing overall diet quality. Asthma was defined by the asthma symptom score (sum of five questions). Negative binomial regression was used to evaluate the association between the FSA-NPS DI and the asthma symptom score. Overall, mean participant’s age was 54 ± 14 years, and about 27 % reported at least one asthma symptom. We observed a significant positive association between less healthy diet, as expressed by higher FSAm-NPS DI, and the asthma symptom score. The adjusted OR were 1·27 (95 % CI 1·17, 1·38) among women and 1·31 (95 % CI 1·13, 1·53) among men. Unhealthy food choices, as reflected by a higher FSAm-NPS DI, were associated with greater asthma symptoms. These results reinforce the relevance of public health approach to orient consumers towards healthier food choices by using a clear and easy-to-understand FOP nutrition label based on the FSAm-NPS, such as the Nutri-Score.

In the present study, we analysed the effects of SNP rs174547 (T/C) in the fatty acid desaturase 1 (FADS1) gene on long-chain PUFA levels. Four databases were searched to retrieve related literature with keywords such as fatty acid (FA), SNP, FADS1 and rs174547. A meta-analysis of the data was performed using Stata12.0 software, including summary statistics, test for heterogeneity, evaluation of publication bias, subgroup analysis and sensitivity analysis. The associations between rs174547 in FADS1 and seven types of FA, and Δ-5 (D5D) and Δ-6 fatty acid desaturase (D6D) activity were assessed based on the pooled results from eleven papers. A total of 3713 individuals (1529 TT and 2184 TC + CC) were included. The results demonstrated that minor C allele carriers of rs174547 had higher linoleic acid (LA; P < 0·001) and α-linolenic acid (P = 0·020) levels, lower γ-linolenic acid (GLA; P = 0·001) and arachidonic acid (P = 0·024) levels, and lower D5D (P = 0·005) and D6D (P = 0·004) activities than the TT genotype group. Stratification analysis showed that minor C allele carriers of rs174547 had higher LA and lower GLA levels and lower D6D activities in plasma (LA, P < 0·001; GLA, P < 0·001; D6D activity, P < 0·001) samples and in Asian populations (LA, P < 0·001; GLA, P = 0·001; D6D activity, P = 0·001) than the TT genotype group. In conclusion, minor C allele carriers of the SNP rs174547 were associated with decreased activity of D5D and D6D.

TNF-α is a pro-inflammatory cytokine that is involved in type 1 diabetes (T1D) pathogenesis. The TNFa gene is subject of epigenetic regulation in which folate and homocysteine are important molecules because they participate in the methionine cycle where the most important methyl group donor (S-adenosylmethionine) is formed. We investigated whether TNFa gene promoter methylation status in T1D patients was related to blood folate, homocysteine and TNF-α in a transversal case–control study. We studied T1D patients (n 25, mean=13·7 years) and healthy control subjects (n 25, mean=31·1 years), without T1D and/or other autoimmune diseases or direct family history of these diseases. A blood sample was obtained for determination of serum folate, plasma homocysteine and TNF-α concentrations. Whole blood was used for the extraction of DNA to determine the percentage of methylation by real-time PCR and melting-curve analysis. Results are expressed as means and standard deviations for parametric variables and as median (interquartile range) for non-parametric variables. T1D patients showed a higher TNFa gene promoter methylation (39·2 (sd 19·5) %) when compared with control subjects (25·4 (sd 13·7) %) (P=0·008). TNFa gene promoter methylation was positively associated only with homocysteine levels in T1D patients (r 0·55, P=0·007), but not in control subjects (r −0·122, P=0·872). To our knowledge, this is the first work that reports the methylation status of the TNFa gene promoter and its relationship with homocysteine metabolism in Chilean T1D patients without disease complications.

Low-molecular-weight fucoidan (LMWF) has been broadly studied in recent years due to its numerous biological properties. Nevertheless, there have been no reports about the effects of LMWF on myogenic differentiation (MyoD). The objective of the present study was to demonstrate the impact of LMWF on myogenesis in C2C12 cells. The ultimate aim was to establish whether LMWF regulates myogenesis similar to heparin as a partial regulator of myogenesis. LMWF was prepared at a minimal size by ultra-filtration compared with crude fucoidan. We treated C2C12 cells with LMWF and/or heparin during MyoD. The data from the present study are the first to suggest that LMWF suppresses the expression of the myogenic regulatory factors and the myocyte enhancer factors as well as the morphological changes that occur during differentiation. Additionally, the expression of the mitogen-activated protein kinase (MAPK) family was significantly inhibited by LMWF when compared with controls. The LMWF-treated group showed significantly decreased expression of reactive oxygen species (ROS) enzymes compared with control cells. Heparin was used as a positive control because it has been reported to activate MyoD. Taken together, these results suggest that LMWF might regulate MyoD through the MAPK pathway and by regulating ROS activity in C2C12 cells.

Long-chain fatty acids are capable of inducing alterations in the homoeostasis of glucose-stimulated insulin secretion (GSIS), but the effect of medium-chain fatty acids (MCFA) is poorly elucidated. In the present study, we fed a normoenergetic MCFA diet to male rats from the age of 1 month to the age of 4 months in order to analyse the effect of MCFA on body growth, insulin sensitivity and GSIS. The 45 % MCFA substitution of whole fatty acids in the normoenergetic diet impaired whole body growth and resulted in increased body adiposity and hyperinsulinaemia, and reduced insulin-mediated glucose uptake in skeletal muscle. In addition, the isolated pancreatic islets from the MCFA-fed rats showed impaired GSIS and reduced protein kinase Bα (AKT1) protein expression and extracellular signal-related kinase isoforms 1 and 2 (ERK1/2) phosphorylation, which were accompanied by increased cellular death. Furthermore, there was a mildly increased cholinergic sensitivity to GSIS. We discuss these findings in further detail, and advocate that they might have a role in the mechanistic pathway leading to the compensatory hyperinsulinaemic status found in this animal model.

Modulating the gut microbiota via dietary interventions is a common strategy to enhance the natural defence mechanisms of the host. Several in vitro studies have highlighted the probiotic potential of Bifidobacterium thermophilum RBL67 (RBL67) selected for its anti-Salmonella effects. The present study aimed to investigate the impact of RBL67 alone and combined with fructo-oligosaccharides (FOS) on the gut microbiota of Göttingen minipigs. Minipigs were fed a basal diet supplemented with 8 g/d probiotic powder (1×109 CFU/g in skim milk matrix) (probiotic diet (PRO)), 8 g/d probiotic powder plus 8 g/d FOS (synbiotic diet (SYN)) or 8 g/d skim milk powder (control), following a cross-sectional study design. Faecal and caecal microbiota compositions were analysed with pyrosequencing of 16S rRNA genes and quantitative PCR. Metabolic activity in the caecum and colon was measured by HPLC. 16S rRNA gene amplicon sequencing revealed that minipig faeces show close similarity to pig microbiota. During the treatments and at the time of killing of animals, RBL67 was consistently detected in faeces, caecum and colon at numbers of 105–106 16S rRNA copies/g content after feeding PRO and SYN diets. At the time of killing of animals, significantly higher Bifidobacterium numbers in the caecum and colon of SYN-fed minipigs were measured compared with PRO. Our data indicate that the Göttingen minipig may be a suitable model for gut microbiota research in pigs. Data from this first in vivo study of RBL67 colonisation suggest that the combination with FOS may represent a valuable symbiotic strategy to increase probiotic bacteria levels and survival in gastrointestinal tracts for feed and food applications.

European Union Regulation 258/97 defines novel foods as food products and food ingredients that have not been consumed to a significant degree in the European Union before May 1997. However, there are new foods that for some reason are not considered as novel foods, though we think that safety of these products is not always a priori established. We defined a ‘grey area’ which consists of such foods, and the present paper intends to raise awareness of this ‘grey area’ of unidentified novel foods. The grey area of novel foods is divided into two categories: (1) food products or ingredients for which the current Regulation leaves too much space for different interpretations and (2) food products or ingredients that are not novel according to the current Regulation, because the current Regulation contains gaps. These categories are illustrated by means of products already on the market in The Netherlands. We found about two dozen examples of products that had not been identified as novel foods according the current Regulation, yet could be considered to be classified as novel foods and hence for which a safety evaluation (toxicological and/or nutritional) would be indicated.

In epidemiological studies, chronic dairy milk consumption is associated with improved vascular health and reduced age-related increases in blood pressure. Although milk protein supplementation augments conduit artery flow-mediated dilation, whether or not acute dairy milk intake may improve microvascular function remains unclear. We hypothesised that dairy milk would increase direct measurement of endothelial nitric oxide (NO)-dependent cutaneous vasodilation in response to local skin heating. Eleven men and women (61 (sem 2) years) ingested two or four servings (473 and 946 ml) of 1 % dairy milk or a rice beverage on each of 4 separate study days. In a subset of five subjects, an additional protocol was completed after 473 ml of water ingestion. Once a stable blood flow occurred, 15 mm-NG-nitro-l-arginine methyl ester was perfused (intradermal microdialysis) to quantify NO-dependent vasodilation. Red-blood-cell flux (RBF) was measured by laser-Doppler flowmetry, and cutaneous vascular conductance (CVC=RBF/mean arterial pressure) was calculated and normalised to maximum (%CVCmax; 28 mm-sodium nitroprusside). Full expression of cutaneous vasodilation was not different among dairy milk, rice beverage and water, and there was no effect of serving size on the total vasodilatory response. Contrary to our hypothesis, NO-dependent vasodilation was lower for dairy milk than rice beverage (D: 49 (sem 5), R: 55 (sem 5) %CVCmax; P<0·01). Acute dairy milk ingestion does not augment NO-dependent vasodilation in the cutaneous microcirculation compared with a rice beverage control.

In the present study, the effect of flax hulls with or without flax oil bypassing the rumen on the expression of lipogenic genes in the mammary tissue of dairy cows was investigated. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four periods of 21 d each and four treatments: control diet with no flax hulls (CONT); diet with 9·88 % flax hulls in the DM (HULL); control diet with 500 g flax oil/d infused in the abomasum (COFO); diet with 9·88 % flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). A higher mRNA abundance of sterol regulatory element binding transcription factor, fatty acid (FA) synthase, lipoprotein lipase (LPL), PPARγ1, stearoyl-CoA desaturase (SCD) and acetyl-coenzyme A carboxylase-α was observed in cows fed HULL than in those fed CONT, and HUFO had the opposite effect. Compared with CONT, COFO and HUFO lowered the mRNA abundance of SCD, which may explain the lower proportions of MUFA in milk fat with flax oil infusion. The mRNA abundance of LPL in mammary tissue and proportions of long-chain FA in milk fat were higher in cows fed COFO than in those fed CONT. The highest proportions of trans FA were observed when cows were fed HULL. The present study demonstrates that flax hulls with or without flax oil infusion in the abomasum can affect the expression of lipogenic genes in the mammary tissue of dairy cows, which may contribute to the improvement of milk FA profile.

1. Riboflavine given orally to sheep, calves, goats and ruminating kids gave rise, in milk and urine, to extra riboflavine accompanied by hydroxyethylflavine, by formylmethylflavine and by other as yet unidentified metabolites, but subcutaneous injection of riboflavine caused prolonged excretion of riboflavine in urine and milk unaccompanied by metabolites.

2. Placement of riboflavine by means of fistulas into the rumen, the abomasum or the caecum and abstraction of samples from the rumen or caecum showed that the rumen was the site of origin of the metabolites, thus accounting for the lapse of time of several hours between the ingestion of riboflavine and the appearance of hydroxyethylflavine in the urine and the absence of metabolites after subcutaneous injection of riboflavine.

3. Kids in which rumination had not yet become established could not produce metabolites from ingested riboflavine although, when fed milk containing the metabolites, they readily excreted them in their urine.

5. Incubation in vitro of kidney or liver tissue from cattle or goats with riboflavine did not give rise to metabolites, but these tissues could both reduce formylmethylflavine to hydroxyethylflavine in vitro.

6. Post-mortem examination of animals to which riboflavine had been fed showed the presence of metabolites only in gut contents, in the urinary bladder and in the bile. When injected, riboflavine reached the bile but no metabolites were produced.

7. Metabolites were found to occur naturally in the urine of ruminants in small amounts. Hydroxyethylflavine was readily demonstrable in the urine of the rabbit, but in the urine of the dog metabolites could not be found.

8. From these and earlier and later experiments the conclusion is drawn that riboflavine metabolites arise in the ruminant from the degradation of riboflavine by rumen bacteria.

1. In order to examine the interaction of dietary fat and carbohydrate in the regulation of lipid metabolism, we have studied hepatic and extrahepatic lipogenesis, and adipose tissue lipoprotein lipase (EC 3.1.1.34) in rats fed on one of the following diets: a fructose-based diet containing 0 (F0) or 150 g maize oil (F15)/kg, or a glucose based diet containing 0 (G0) or 150 g maize oil (G15)/kg.

2. The rats were meal-fed on the diets for 2 weeks after which the activities of a number of hepatic ‘lipogenic’ enzymes were measured and the activity of epididymal-fat-pad lipoprotein lipase. The activities of the lipogenic enzymes were: F0 > G0 > G15 > F15. Lipoprotein lipase activity was F0 = G0 = F15 = G15. The percentage of total body fatty acid synthesis which occurred in the liver was F0 > G0 > F15 > G15.

3. We conclude that fructose-induced hypertriglyceridaemia is primarily a result of the increased hepatic synthesis rather than decreased adipose-tissue lipoprotein lipase activity.