There is a consensus that interventions to prevent and treat childhood obesity should involve the family; however, the extent of the child's involvement has received little attention. The goal of the present study was to evaluate the relative efficacy of treating childhood obesity via a family-based health-centred intervention, targeting parents alone v. parents and obese children together. Thirty-two families with obese children of 6–11 years of age were randomised into groups, in which participants were provided for 6 months a comprehensive educational and behavioural programme for a healthy lifestyle. These groups differed in their main agent of change: parents-only v. the parents and the obese child. In both groups, parents were encouraged to foster authoritative parenting styles (parents are both firm and supportive; assume a leadership role in the environmental change with appropriate granting of child's autonomy). Only the intervention aimed at parents-only resulted in a significant reduction in the percentage overweight at the end of the programme (P=0·02) as well as at the 1-year follow-up meeting.The differences between groups at both times were significant (P<0·05). A greater reduction in food stimuli in the home (P<0·05) was noted in the parents-only group. In both groups, the parents' weight status did not change. Regression analysis shows that the level of attendance in sessions explained 28% of the variability in the children's weight status change, the treatment group explained another 10%, and the improvement in the obesogenic load explained 11% of the variability. These results suggest that omittingthe obese child from active participation in the health-centred programme may be beneficial for weight loss and for the promotion of a healthy lifestyle among obese children.

Probiotic bacteria may provide protection against intestinal damage induced by pathogens, but the underlying mechanisms are still largely unknown. We investigated whether Bifidobacterium animalis MB5 and Lactobacillus rhamnosus GG (LGG) protected intestinal Caco-2 cells from the inflammation-associated response induced by enterotoxigenic Escherichia coli (ETEC) K88, by inhibiting pathogen attachment to the cells, which is the first step of ETEC pathogenicity, and regulating neutrophil recruitment, a crucial component of inflammation. A partial reduction of ETEC adhesion was exerted by probiotics and their culture supernatant fractions either undigested or digested with proteases. ETEC viability was unaffected by the presence of B. animalis, LGG or their supernatant fractions in the culture medium, indicating an absence of probiotic bactericidal activity. Probiotics and their supernatant fractions, either undigested or digested with proteases, strongly inhibited the neutrophil transmigration caused by ETEC. Both B. animalis and LGG counteracted the pathogen-induced up regulation of IL-8, growth-related oncogene-α and epithelial neutrophil-activating peptide-78 gene expression, which are chemokines essential for neutrophil migration. Moreover, the probiotics prevented the ETEC-induced increased expression of IL-1β and TNF-α and decrease of transforming growth factor-α, which are regulators ofchemokine expression. These results indicate that B. animalisMB5 and LGG protect intestinal cells from the inflammation-associated response caused by ETEC K88 by partly reducing pathogen adhesion and by counteracting neutrophil migration, probably through the regulation of chemokine and cytokine expression.

There is a relationship between the fatty acid profile in skeletal muscle phospholipids and peripheral resistance to insulin in adults, but similar data have not been reported in infancy and childhood. The objective of this study was to investigate the fatty acid composition of skeletal muscle and adipose tissue across the paediatric age range. The fatty acid profile of skeletal muscle phospholipids and adipose tissue triacylglycerols was analysed in ninety-three healthy Spanish infants and children distributed into four groups: group 1 (0 to <2 years, n 10); group 2 (2 to <5 years, n 41); group 3 (5 to <10 years, n 24); group 4 (10 to 15 years, n 18). In skeletal muscle phospholipids, oleic acid (18: 1n-9cis) content decreased significantly whereas that of linoleic (18: 2n-6) acid increased significantly with age (P for trend <0·01). In adipose tissue, the contents of triacylglycerol and linoleic acid increased significantly across the paediatric age range (P for trend <0·01), whereas dihomo-γ-linolenic (20: 3n-6) and arachidonic (20: 4n-6) showed significant differences between groups. The variations in fatty acid composition observed with age indicated an imbalance in dietary n-3/n-6 long-chain PUFA.

Experiments studied the effect of yoghurt on the recovery of defence mechanisms against Streptococcus pneumoniae respiratory infection in malnourished mice. Weaned mice were malnourished with a protein-free diet (PFD) for 21d. Malnourished mice were made replete with a balanced diet (BD), yoghurt, or the BD with supplemental yoghurt (BD+Y) for 7, 14 or 21d. The normal control (NC) group was fed the BD whereas malnourished control (MC) mice consumed only the PFD. Mice were challenged with pneumococci at the end of each dietary treatment. MC mice showed increased susceptibility to pneumococcal infection. Blood leucocytes, phagocyte activity and serum and bronco-alveolar anti-pneumococcal IgG and IgA were significantly lower in the MC than in the NC group. Repletion of malnourished mice with the BD for 21d was necessary to obtain a response to infection similar to that of NC mice; however, administration of the BD+Y for 14d was enough to normalise the immune defence mechanisms. Histological examination of MC lungs showed progressive loss of alveolar architecture. Lung injuries were significantly less pronounced in NC mice. Mice treated with the BD+Y for 14d showed histological signs similar to the NC group. The present study showed that administration of yoghurt to malnourished mice induced an early recovery of the immunological parameters studied. Despite the uncertainties about the mechanisms involved and about the human relevance of the effects observed in animal models, the present study provides a strong rationale for the hypothesis that yoghurt consumption by malnourished hosts will accelerate the recovery of the immune mechanisms involved in the protection against respiratory infections

Clinical trials have shown that soya protein reduces the concentrations of some atherogenic lipids in subjects with normal renal function. The present study examined the effects of soya protein on serum lipid concentrations and lipoprotein metabolism in patients on hypercholesterolaemic haemodialysis. Twenty-six hypercholesterolaemic (total cholesterol ≥6·21mmol/l) patients on haemodialysis were studied in a randomized, double-blind, placebo-controlled clinical trial. After a 4-week run-in phase, the subjects were randomly assigned to two groups. Isolated soya protein or milk protein 30g was consumed daily as a beverage at breakfast or post-dialysis for 12 weeks. Soya protein substitution resulted in significant reductions in total cholesterol (17·2 (sd8·9)%), LDL-cholesterol (15·3 (sd12·5)), apo B (14·6 (sd12·1)%) and insulin (23·8 (sd18·7)%) concentrations. There were no significant changes in HDL-cholesterol or apo A-I. These results indicate that replacing part of the daily protein intake with soya protein has a beneficial effect on atherogenic lipids and favourably affects lipoprotein metabolism in hypercholesterolaemic patients undergoing haemodialysis.

One of the most frequently reported immunomodulatory actions of n−3 PUFA is their ability to diminish in vivo lymphocyte proliferation. The purpose of this study was to determine if n−3 PUFA intake affects the kinetics or magnitude of the antigen-driven expansion of CD8+T-lymphocytes in vivo. In this study we utilized a well-characterized model of T-cell immunity (i.e. infection with the intracellular bacterium, Listeria monocytogenes). Weanling BALB/c mice were fed one of two experimental diets that differed solely in fat source. Our control diet contained lard (180g/kg) and was devoid of long-chain n−3 PUFA. The experimental diet contained 150g/kg menhaden fish oil and 30g/kg corn oil, thus providing approximately 8% of energy from long-chain n−3 PUFA. After 4 weeks, mice were infected intravenously with 106 colony-forming units of actA-deficient L. monocytogenes. Clonal expansion of antigen-specific CD8+T-cells in the spleen was measured at 5, 7, 9 and 14d post-challenge using a class I MHC tetramer loaded with the immunodominant peptide from this pathogen (i.e. Kd:LLO91–99). We report that feeding mice a diet rich in n−3 fatty acids did not significantly impact either the kinetics or magnitude of in vivo, antigen-driven expansion of CD8+T-cells. Furthermore, contraction of this T-cell population was not affected by n−3 PUFA treatment. To our knowledge this is the first time MHC tetramers have been used to investigate the influence of n−3 PUFA on in vivo CD8+T-cell proliferation.

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium verticillioides, a fungus that commonly contaminates maize. Sex-related effects of FB1 have been observed with respect to carcinogenicity in rodents, to performances in pigs and immunosuppression in mice. In the present study the sex-related effect of FB1 on the pig immune response was determined. Female and castrated male piglets received for 28d either control feed or feed contaminated with 8mg FB1/kgfeed in the form of F. verticillioidesculture material. At day 7 and day 21, animals were immunised subcutaneously with a Mycoplasma agalactiae vaccine. Ingestion of FB1-contaminated feed significantly decreased weight gain in males but had no effect in females. No sex-related difference was observed in biochemical parameters, but a higher level of creatinine was noted in toxin-treated animals. FB1 also altered the pig immune response in a sex-specific manner. In males, ingestion of FB1-contaminated feed significantly decreased specific antibody levels after vaccination as well as the mRNA expression level of IL-10. In females, the toxin has no effect on specific antibodies or on cytokine mRNA levels. The results of the present study indicatethat FB1 is immunosuppressive in pigs. The magnitude of this FB1-induced immunosuppression is highly dependent on sex, with males being more susceptible than females.

The present study reports the activities of quercetin and its main circulating conjugates in man (quercetin-3′-sulphate (Q3′S) and quercetin-3-glucuronide (Q3G)) on in vivo angiogenesis induced by vascular endothelial growth factor (VEGF) and examines the effects of these molecules on cultured endothelial cells. We found opposing effects of quercetin and its metabolites on angiogenesis. While quercetin and Q3G inhibited VEGF-induced endothelial cell functions and angiogenesis, Q3′S per se promoted endothelial cell proliferation and angiogenesis. The inhibitory effect elicited by Q3G was linked to inhibition of extracellular signal-regulated kinases 1/2 (ERK1/2) phosphorylation elicited by VEGF. The activation of endothelial cells by Q3′S was associated to stimulation of VEGF receptor-2 and to downstream signalling activation (phosphatidylinositol-3 kinase/Akt and nitric oxide synthase pathways), ultimately responsible for ERK1/2 phosphorylation. These data indicate that the effects of circulating quercetin conjugates on angiogenesis are different depending on the nature of the conjugate. Q3G andQ3′S are the two major conjugates in plasma, but their ratio is dependenton several factors, so thatinhibition or activation of angiogenesis could be subtly shifted as a result of metabolismin vivo.

Consumption of some dietary fibres may benefit bowel health; however, the effect of Australian sweet lupin (Lupinus angustifolius) kernel fibre (LKFibre) is unknown. The present study examined the effect of a high-fibre diet containing LKFibre on bowel function and faecal putative risk factors for colon cancer compared to a control diet without LKFibre. Thirty-eight free-living, healthy men consumed an LKFibre and a control diet for 1 month each in a single-blind, randomized, crossover study. Depending on subject energy intake, the LKFibre diet was designed to provide 17–30g/d fibre (in experimental foods) above that of the control diet. Bowel function self-perception, frequency of defecation, transit time, faecal output, pH and moisture, faecal levels of SCFA and ammonia, and faecal bacterial β-glucuronidase activity were assessed. In comparison to the control diet, the LKFibre diet increased frequency of defecation by 0·13 events/d (P=0·047), increased faecal output by 21% (P=0·020) and increased faecal moisture content by 1·6% units (P=0·027), whilst decreasing transit time by 17% (P=0·012) and decreasing faecal pH by 0·26 units (P<0·001). Faecal butyrate concentration was increased by 16% (P=0·006), butyrate output was increased by 40% (P=0·002)and β-glucuronidase activity was lowered by 1·4μmol/h per g wet faeces compared to the control diet (P<0·001). Addition of LKFibre to the diet incorporated into food products improved some markers of healthy bowel function and colon cancer risk in men.

We studied dietary risk factors by analysing a questionnaire administered at birth, 1 year and 2½ years of age, as well as the level of glutamic acid decarboxylase autoantibodies (GADA) and tyrosine phosphatase autoantibodies (IA-2A), in 7208 2½-year-old children from the All Babies in Southeast Sweden cohort, using the 95th percentile cut-off for autoantibodies to identify children at risk of type 1 diabetes. A total of 657 children had either IA-2A (n 360) or GADA (n 335), and thirty-eight children had both GADA and IA-2A. In univariate analysis, male gender and maternal coeliac disease implied a risk of possessing IA-2A. Maternal type 2 diabetes, a high consumption of fresh cows milk at the age of 1 year and a late introduction of gluten were associated with a risk of GADA. Early cessation of breast-feeding (≤2 months of age) was associated with a risk of the simultaneous occurrence of both IA-2A and GADA. In logistic regression analysis, a high consumption of milk at the age of 1 year (odds ratio 2·6) represented a risk for GADA, and maternal coeliac disease (odds ratio 2·9) represented a risk for IA-2A. The combination of an early introduction of cows milk formula and a late introduction of gluten-containing food gave an odds ratio of 6·0 for positivity for at least one autoantibody at 1 and 2½ years of age. The induction of autoantibodies by the age of 2½ years has a male preponderance and is more common in children with maternal type 2 diabetes or maternal coeliac disease. Dietary risk factors for the induction of β-cell autoantibodies in 2½-year-old children are a short duration of breast-feeding, an early introduction of cows milk formula and a late introduction of gluten, as well as a high consumption of milk at the age of 1 year.

A community-based, randomized, placebo-controlled, double-blind trial was conducted from March 2001 to March 2002 in Kilosa, a rural district of Morogoro Region in Tanzania. One hundred and fifty-eight infants were selected randomly from lists of local Maternal and Child Health Care Centres and received either processed complementary food (PCF) or unprocessed complementary food (UPCF) from age 6 to 12 months. Processing increased Zn solubility and energy density of the porridge prepared from the complementary food (CF) as determined in vitro. Phytate:Zn molar ratio of the PCF and UPCF was 25·8 and 47·5, respectively. Under the study conditions, the processing of CF did not improve Zn status as measured by hair analysis. No significant correlations were found between hair Zn values and anthropometric measurements. Our findings suggest that processing alone of cereal-based CF may be insufficient to ensure an adequate supply of Zn to improve growth and Zn status of infants. Dietary modification to tackle Zn deficiencies in similar target groups may therefore only be successful when other Zn-rich foods such as meat and fish are included.

Data on folate absorption from food from validated human studies using physiological folate doses are still needed to estimate dietary requirements and to formulate recommendations. The aim of the present work was to study the effects from fortified and processed foods on folate absorption in ileostomy volunteers (n 9) using the area under the plasma concentration curve (AUC) and kinetic modelling. Using a standardized single-dose protocol, dairy products fortified with a candidate fortificant (6S)-5-methyltetrahydrofolate ((6S)-5-CH3-H4folate), folic acid-fortified bread and a dessert crème containing natural yeast folate polyglutamates were compared with folate supplements. Absorbed folate was estimated by AUC and a kinetic model, and non-absorbed folate by ileostomal folate excretion. Median apparent absorption from test foods ranged from 55 to 86%. Added folate-binding proteins (FBP) significantly reduced folate absorption from dairy products, as in the absence of FBP, AUC–dose-corrected ratios were increased and ileal folate excretion decreased. After in vivo gastrointestinal passage of dairy products containing FBP, up to 43% of the ingested FBP was found in ileostomal effluent. Folate absorption was similar for (6S)-5-CH3-H4folate fortificant from fermented milk and for folic acid from fortified bread. Folic acid, ingested as food fortificant in bread, was significantly less absorbed compared with an isolated supplement. We conclude that all tested foods were suitable matrices for folate fortification. However, dairy products, fortified with the new candidate fortificant (6S)-5-CH3-H4folate, are suitable if no active FBP is present.

Infectious diarrhoea is a major problem in both children and piglets. Infection of enterotoxigenic Escherichia coli (ETEC) results in fluid secretion and electrolyte losses in the small intestine. In the present study the effect of processed and fermented soyabean products on net absorption during ETEC infection was investigated. Soyabean was processed into an autoclaved, a cooked and a mould-fermented (tempeh) product. The soyabean products were pre-digested and the effect of the products on net absorption in the small intestineof piglets was studied. Pairs of small-intestinal segments, one non-infected and the other ETEC-infected, were perfused simultaneously with the different products during 8h. Net absorption of fluid, DM, Na, chloride, K and total solutes was determined. Net fluid absorption washighest for cooked soyabean followed by autoclaved soyabean and tempeh as a result of the osmolality of these products. In ETEC-infected segments, cooked soyabean and tempeh showed minor fluid losses (27 (se 23) and 43 (se 20) μl/cm2, respectively) compared with the saline control (260 (se 23) μl/cm2). Tempeh resulted in a high uptake of solutes. Processed soyabean products, particularly cooked soyabean and tempeh, are beneficial in maintaining fluid balance during ETEC infection. Additionally, tempeh showed high DM and total solute absorption. Therefore, particularly, tempeh may bebeneficial in the case of post-weaning diarrhoeain piglets and possibly in children as well.

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.

A reduced protein diet (RPD) is known to increase the level of intramuscular lipid in pig meat with a smaller effect on the amount of subcutaneous adipose tissue. This might be due to tissue-specific activation of the expression of lipogenic enzymes by the RPD. The present study investigated the effect of a RPD, containing palm kernel oil, soyabean oil or palm oil on the activity and expression of one of the major lipogenic enzymes, stearoyl-CoA desaturase (SCD) and on the level of total lipids and the fatty acid composition of muscle and subcutaneous adipose tissue in pigs. The RPD significantly increased SCD protein expression and activity in muscle but not in subcutaneous adipose tissue. The level of MUFA and total fatty acids in muscle was also elevated when the RPD was fed, with only small changes in subcutaneous adipose tissue. A positive significant correlation between SCD protein expression and total fatty acids in muscle was found. The results suggest that an increase in intramuscular but not subcutaneous adipose tissue fatty acids under the influence of a RPD is related to tissue-specific activation of SCD expression. It is suggested that the SCD isoform spectra in pig subcutaneous adipose tissue and muscle might be different.

Observational studies have shown that children in developing countries consuming diets containing high amounts of bioavailable nutrients, such as those found in animal-source foods, grow better. The present study investigated which specific nutrients from the diet of Kenyan school children predicted their growth. The children (n544, median age 7 years) participated in a 2-year long food supplementation study with animal-source foods. Height gain during the intervention period was positively predicted by average daily intakes of energy from animal-source foods, haem Fe, preformed vitamin A, Ca and vitamin B12. Weight gain was positively predicted by average daily intakes of energy from animal-source foods, haem Fe, preformed vitamin A, Ca and vitamin B12. Gain in mid-upper-arm muscle area was positively predicted by average daily intakes of energy from animal-source foods and vitamin B12. Gain in mid-upper-arm fat area was positively predicted by average daily intakes of energy from animal-source foods. Gain in subscapular skinfold thickness was not predicted by any of the nutrient intakes. Negative predictors of growth were total energy and nutrients that are contained in high amounts in plant foods. The study shows that growth was positively predicted by energy and nutrients that are provided in high amounts and in a bioavailable form in meat and milk, and their inclusion into the diets of children in developing countries should be part of all food-based programmes in order to improve micronutrient status and growth.

High-fat feeding reduces the expression of GLUT-2 and the glycolytic enzyme glucokinase (GK). The transcription factor, pancreatic duodenal homeobox-1 (Pdx-1), is important for β-cell maintenance. The aim of the present study was to determine, in weanling Wistar rats, the effect of a maternal high-fat diet (HFD) during defined periods of gestation and lactation, on body weight, circulating glucose and insulin concentrations, and the expression of GLUT-2, GK and Pdx-1. At postnatal day 21, weights were recorded and glucose and insulin concentrations were measured. The expression levels for mRNA were quantified by LightCycler PCR. Pancreatic sections, immunostained for GLUT-2, GK or Pdx-1, were assessed by image analysis. Weanlings from dams fed an HFD throughout gestation were lighter, with heavier weanlings produced from dams fed an HFD throughout gestation and lactation. Both these groups of weanlings were normoglycaemic, all the others being hyperglycaemic. Hypoinsulinaemia was evident in weanlings from dams fed an HFD throughout gestation only and also for either the first week of lactation or throughout lactation. GLUT-2 mRNA expression was reduced and GLUT-2 immunoreactivity was increased in most of the weanlings. GK mRNA expression and immunoreactivity was reduced in most of the offspring. Pdx-1 mRNA expression was increased in weanlings from dams fed an HFD throughout both gestation and lactation and reduced in those from dams only fed a lactational HFD. Normal Pdx-1 immunoreactivity was found in all of the weanlings. A maternal HFD induces hyperglycaemia in weanlings concomitant with reduced GK expression which may compromise β-cell function.