The suppression of prolactin production with bromocriptine (BRO) in the last 3 d of lactation reduces milk yield (early weaning) and increases the transfer of leptin through the milk, causing hyperleptinaemia in pups. In adulthood, several changes occur in the offspring as a result of metabolic programming, including overweight, higher visceral fat mass, hypothyroidism, hyperglycaemia, insulin resistance, hyperleptinaemia and central leptin resistance. In the present study, we investigated whether overweight rats programmed by early weaning with maternal BRO treatment have hypothalamic alterations in adulthood. We analysed the expression of neuropeptide Y (NPY), cocaine- and amphetamine-regulated transcript (CART), pro-opiomelanocortin (POMC) and α-melanocyte-stimulating hormone (α-MSH) by immunohistochemistry in the following hypothalamic nuclei: medial and lateral arcuate nucleus (ARC); paraventricular nucleus (PVN); lateral hypothalamus (LH). Additionally, we sought to determine whether these programmed rats exhibited hypothalamic inflammation as indicated by astrogliosis. NPY immunostaining showed a denser NPY-positive fibre network in the ARC and PVN (+82 % in both nuclei) of BRO offspring. Regarding the anorexigenic neuropeptides, no difference was found for CART, POMC and α-MSH. The number of astrocytes was higher in all the nuclei of BRO rats. The fibre density of glial fibrillary acidic protein was also increased in both medial and lateral ARC (6·06-fold increase and 9·13-fold increase, respectively), PVN (5·75-fold increase) and LH (2·68-fold increase) of BRO rats. We suggest that early weaning has a long-term effect on the expression of NPY as a consequence of developmental plasticity, and the presence of astrogliosis indicates hypothalamic inflammation that is closely related to overweight and hyperleptinaemia observed in our model.

1. The effect of daily supplements of 20–30 mg inorganic iron as ferrous sulphate on the growth, activity and haematological status of preschool children was studied for 3.5, 7 and 12 months and compared to that of children who served as controls. All children were given their daily requirements of energy and protein. In addition, they received 5 μg cyanocobalamin and 200 μg folic acid.

2. Fe supplementation increased the haemoglobin, serum Fe and percentage saturation of transferrin and reduced the unsaturated Fe-binding capacity significantly compared to corresponding values for the controls.

3. Height and activity were unaffected by Fe supplements.

4. Of the children 45% had haemoglobin values below 110 g/l at the end of 7–12 months of Fe supplementation.

The effects of high dietary levels of phosphatidylethanolamine (PE) on plasma concentrations of homocysteine (tHcy) have not previously been studied. Eighteen mink (Mustela vison) studied were fed one of three diets during a 25d period in a parallel-group design. The compared diets had 0, 17 and 67% extracted lipids from natural gas-utilising bacteria (LNGB), which were rich in PE. The group with 0% LNGB was fed a diet of 100% soyabean oil (SB diet). Phospholipids are the main lipid components in LNGB and Methylococcus capsulatus is the main bacteria (90%). The fasting plasma concentration of tHcy was significantly higher when the mink consumed the diet with 67% LNGB than when they consumed the SB diet (P=0·039). A significantly lower glutathione peroxidase activity was observed in mink consuming the 17% LNGB diet or the 67% LNGB diet than was observed in mink fed the SB diet. The lack of significant differences in the level of plasma PE due to the diets indicates that most of the PE from the 67% LNGB diet was converted to phosphatidylcholine (PC) in the liver. It has previously been hypothesised that phosphatidylethanolamine N-methyltransferase is an important source of tHcy. The present results indicate that plasma tHcy is at least partly regulated by phospholipid methylation from PE to PC. This methylation reaction is a regulator of physiological importance.

1. In hay-fed, cannulated sheep the apparent degradation in and outflow from the rumen were determined for graded doses of mixtures of the amino acids lysine, threonine and methionine, administered intraruminally and using polyethylene glycol (PEG) as a liquid marker. The doses ranged between 2.5 and 15 g for each amino acid in the mixtures.

2. Relative rate of apparent degradation in the first 4 h was highest for lysine, and lowest for methionine. The apparent degradation in 24 h was highest for lysine and lowest for threonine. Conversely the fraction flowing out of the rumen in intact form in 24 h was highest for threonine and lowest for lysine. Rates of apparent degradation as well as outflow were dose-dependent.

3. The validity of the estimated outflow of amino acids from the rumen was corroborated by measurements of concentrations of the amino acids in duodenal contents and in blood plasma which were also dose-dependent.

4. It was concluded that part of the requirement for the essential amino acids threonine and methionine may be met, even when these amino acids are delivered in unprotected form, given as a feed supplement.

The contribution of binge eating (BE) behaviour to cardiometabolic risk factors has been scarcely investigated so far. Previous studies have not considered the nutritional status and lifestyle of subjects suffering from BE. The aim of this study was to evaluate the contribution of BE to the metabolic syndrome (MS), its components, high total cholesterol and high LDL in a large sample of subjects, taking into account nutritional status, dietary habits, smoking status and physical activity. For this purpose, 5175 adults seeking a weight loss or maintenance programme were recruited. Anthropometrical measurements and blood parameters were measured. BE was evaluated using the Binge Eating Scale (BES). A fourteen-item questionnaire was used to evaluate the adherence to the Mediterranean diet. Smoking status and physical activity were investigated by interview. BE prevalence was 0·16 (95 % CI 0·15, 0·17). A sex- and age-adjusted Poisson regression model showed a higher prevalence of MS in binge eaters (0·33; 95 % CI 0·28, 0·37) compared with non-binge eaters (0·27; 95 % CI 0·25, 0·28, P=0·011). However, the statistical difference was lost after inclusion of BMI and lifestyle parameters in the multiple-adjusted model. We also evaluated the association between the continuous outcomes of interest and the BES score using a multivariable median regression model. We observed a positive, but clinically irrelevant, association between BES score and HDL levels (P<0·001). In conclusion, BE does not seem to be independently related to cardiometabolic risk factors. However, the screening and treatment of BE are of clinical relevance in order to reduce the risk of developing obesity.

Nighttime eating is often associated with a negative impact on weight management and cardiometabolic health. However, data from recent acute metabolic studies have implicated a benefit of ingesting a bedtime snack for weight management. The present study compared the impact of ingesting a milk snack containing either 10 (BS10) or 30 g (BS30) protein with a non-energetic placebo (BS0) 30 min before bedtime on next morning metabolism, appetite and energy intake in mildly overweight males (age: 24·3 (sem 0·8) years; BMI: 27·4 (sem 1·1) kg/m2). Next morning measurements of RMR, appetite and energy intake were measured using indirect calorimetry, visual analogue scales and an ad libitum breakfast, respectively. Bedtime milk ingestion did not alter next morning RMR (BS0: 7822 (sem 276) kJ/d, BS10: 7482 (sem 262) kJ/d, BS30: 7851 (sem 261) kJ/d, P=0·19) or substrate utilisation as measured by RER (P=0·64). Bedtime milk ingestion reduced hunger (P=0·01) and increased fullness (P=0·04) during the evening immediately after snack ingestion, but elicited no effect the next morning. Next morning breakfast (BS0: 2187 (sem 365) kJ, BS10: 2070 (sem 336) kJ, BS30: 2582 (sem 384) kJ, P=0·21) and 24 h post-trial (P=0·95) energy intake was similar between conditions. To conclude, in mildly overweight adults, compared with a non-energetic placebo, a bedtime milk snack containing 10 or 30 g of protein does not confer changes in next morning whole-body metabolism and appetite that may favour weight management.

The aim of this study was to explore the effect of single-dose albendazole and vitamin A intervention on the anaemic status and Fe metabolism of pre-school children. This study was a randomised, placebo-controlled and double-blinded intervention trial. All eligible anaemic pre-school children were randomly divided into three groups: group 1 received no intervention, which served as the control group, group 2 received 400 mg single-dose albendazole administration and group 3 received a 60000 μg vitamin A capsule combined with 400 mg single-dose albendazole at the beginning of the study. The follow-up period was for 6 months. Anthropometry and biochemical index about Fe metabolism were measured before and after intervention. A total of 209 pre-school anaemic children were randomly divided into three intervention groups (sixty-four, sixty-two and sixty for groups 1, 2 and 3, respectively). The mean age of the children in the study was 4·4 (sd 0·7) years and 50·5 % of the children were female (94/186). After a follow-up period of 6 months, the levels of serum retinol, ferritin, transferrin receptor-ferritin index and body total Fe content of children in group 3 were significantly higher compared with children in groups 1 and 2 (P<0·05). Moreover, the proportion of vitamin A deficiency, marginal vitamin A deficiency and Fe deficiency among children in group 3 were markedly lower compared with children in groups 1 and 2 (P<0·05). Albendazole plus vitamin A administration showed more efficacy on the improvement of serum retinol and Fe metabolic status.

The correlation of glutathione S-transferase (GST) M1/T1 genetic polymorphisms with oxidative stress-related chronic diseases was proved recently. The aim of the present study was to investigate the association of GSTM1/T1 genetic polymorphisms with antioxidant biomarkers and consumption of fruits and vegetables (F&V) in healthy subjects. In this study, for conducting a 3 d dietary survey, 190 healthy adults were recruited. After DNA extraction, a multiple PCR method was used for GSTM1/T1 genotyping. A spectrophotometer method was applied for the determination of plasma total antioxidant capacity (T-AOC), vitamin C level and erythrocyte GST enzyme activity. A general linear model was used to compare the mean values of antioxidant parameters for different GSTM1/T1 genotypes and consumption of F&V. Polymorphisms of GSTM1/T1 had no effects on plasma T-AOC and vitamin C levels. Deletion of the GSTM1 gene decreased the erythrocyte GST activity. There was correlation between plasma T-AOC and consumption of F&V in the GSTM1− or GSTT1+ subjects. A similar pattern was evident for erythrocyte GST activity in the GSTM1− subjects. No association was found among consumption of F&V and GSTM1/T1 genotypes and plasma vitamin C level. Different consumption of F&V had no impact on plasma T-AOC and vitamin C levels in the GSTM1−/GSTT1+ or GSTM1−/GSTT1− subjects. The erythrocyte GST activity was more sensitive to consumption of F&V in the individuals with the GSTM1−/GSTT1+ genotype. Association was found among GSTM1/T1 genotypes, antioxidant parameters and consumption of F&V. Large-scale and multiple ethnic studies are needed to further evaluate the relationship.

1. Pregnant rats were given control (46 mg iron/kg, 61 mg zinc/kg), low-Zn (6.9 mg Zn/kg) or low-Zn plus Fe (168 mg Fe/kg) diets from day 1 of pregnancy. The animals were allowed to give birth and parturition times recorded. Exactly 24 h after the end of parturition the pups were killed and analysed for water, fat, protein, Fe and Zn contents and the mothers' haemoglobin (Hb) and packed cell volume (PCV) were measured.

2. There were no differences in weight gain or food intakes throughout pregnancy. Parturition times were similar (mean time 123 (SE 15) min) and there were no differences in the number of pups born. Protein, water and fat contents of the pups were similar but the low-Zn Fe-supplemented group had higher pup Fe than the low-Zn unsupplemented group, and the control group had higher pup Zn than both the low-Zn groups. The low-Zn groups had a greater incidence of haemorrhaged or deformed pups, or both, than the controls.

3. Pregnant rats were given diets of adequate Zn level (40 mg/kg) but with varying Fe:Zn (0.8, 1.7, 2.9, 3.7). Zn retention from the diet was measured using 65Zn as an extrinsic label on days 3, 10 and 17 of pregnancy with a whole-body gamma-counter. A group of non-pregnant rats was also included as controls. The 65Zn content of mothers and pups was measured 24–48 h after birth and at 14, 21 and 24 d of age.

4. In all groups Zn retention was highest from the first meal, fell in the second meal and then rose in the third meal of the pregnant but not the non-pregnant rats. There were no differences between the groups given diets of varying Fe:Zn level. Approximately 25% of the 65Zn was transferred from the mothers to the pups by the time they were 48 h old, and a further 17% during the first 14 d of lactation. The pup 65Zn content did not significantly increase after the first 20 d of lactation but the maternal 65Zn level continued to fall gradually.

Dietary polychlorinated biphenyls (PCBs) caused hypercholesterolaemia in rats. The concentration and output of biliary cholesterol was significantly lower than that of the control group. Biliary output of total bile acids was significantly decreased in rats given the PCB-supplemented diet. Faecal excretion of total steroids (sum of neutral steroids and acidic steroids) was not significantly changed in rats given the PCB-supplemented diet. The present results indicate that dietary PCBs cause hypercholesterolaemia without modifying the faecal total steroids excretion. These results suggest that PCBs produce hyper-cholesterolaemia accompanied by changes in biliary or faecal excretion of bile acids and neutral steroids in addition to an increase in hepatic cholesterol synthesis.

1. Excreta were collected for four consecutive days from 4- to 18-week-old cockerels following subcutaneous injection of Nτ-[14CH3]methylhistidine.

2. The recoveries of radioactivity in excreta were incomplete and progressively decreased with increasing age.

3. Most of the radioactivity not recovered in excreta after 4 d was found in skeletal muscle where > 55% of the radioactivity present was in the Nτ-methylhistidine-containing dipeptide, balenine.

4. This peptide appeared to be relatively stable so that most of the labelled Nτ-methylhistidine incorporated was not released during the period of the recovery measurements.

5. The total pool of non-protein bound Nτ-methylhistidine (free Nτ-methylhistidine+balenine) in pectoral and mixed thigh muscles increased with age and relative to the daily excretion of Nτ-methylhistidine. At 18 weeks the pool was 3.3 times the daily excretion of Nτ-methylhistidine.

6. These observations account for the decreasing recoveries of radioactivity in excreta described previously, due to progressive dilution of labelled Nτ-methylhistidine in an expanding pool of non-protein-bound Nτ-methylhistidine, part of which was relatively stable.

7. It is concluded that excretion of Nτ-methylhistidine by 4- to 18-week-old cockerels cannot be used as a reliable index of muscle protein breakdown in vivo.

1. The contribution of school milk to the nutrition of 396 Kent primary schoolchildren aged 8–11 years was assessed using information collected in a survey which included a weighed-diet record, a socio-economic questionnaire and a medical examination.

2. Over half the children (59% for boys, 54% for girls) drank school milk every school day of the diet-record week.

3. Children who drank school milk every school day, when compared with those not drinking it, had a significantly higher mean daily intake of liquid milk, a higher total daily intake of several important nutrients including animal protein, calcium, thiamin and riboflavin, and a diet richer in Ca and riboflavin (boys) and animal protein and Ca (girls). They were also less likely to have intakes of Ca and riboflavin below the recommended daily intakes for these nutrients (Department of Health and Social Security, 1969).

4. The increased nutritional intake associated with school milk consumption was not related to any differences in height, weight, arm circumference or skinfold thickness.

5. There was no evidence that school milk consumption was associated with obesity (as assessed clinically).

Several clinical reports have shown changes in plasma Se concentration with corticosteroid treatments, but the results have been inconsistent. Few experimental studies have been done on this subject. In the present study the effect of dexamethasone (DEX) treatment on Se concentrations and activities of Se-dependent glutathione peroxidase (EC 1.11.1.9; SeGPx) were examined in adult male ICR mice. In the first experiment, DEX was given via drinking water containing 5 or 50 mg DEX/I. At 1 or 3 weeks of DEX treatment, mice were dissected and the Se concentrations as well as SeGPx activities in various tissues, including plasma, were determined. At 1 week the DEX-treated groups had significantly lower hepatic Se concentrations and significantly higher plasma and cerebral concentrations than the control group. The DEX-treated groups showed lower SeGPx activities in the hepatic cytosol and higher SeGPx activities in the plasma than the saline (9 g NaCl/l)-treated group, in parallel with the changes in Se concentrations. At 3 weeks, neither hepatic nor plasma Se concentrations showed a significant change. In the second experiment, mice were injected subcutaneously with DEX and, thereafter, mice were food-deprived. The DEX-injected groups had higher plasma Se concentrations. A similar finding was obtained also when the DEX- or saline-injected mice were not food-deprived. Thus, the difference between the DEX-treated and control groups was possibly caused by redistribution of tissue Se. These results suggested that the effects of DEX on Se concentrations were tissue dependent and that the higher plasma Se observed in DEX-treated groups might be explained by the release of tissue Se into plasma as plasma SeGPx.

Stroke-prone spontaneously hypertensive (SHRSP) rats are considered a suitable model for studying the effects of dietary and other environmental factors on human essential hypertension and haemorrhagic stroke. To investigate the suitability of a control diet for this strain of rats, we studied the effects of supplementing casein and soya protein isolate (SPI) with two sulphur amino acids (methionine and cystine) on the growth and lifespan of SHRSP rats. The source of dietary protein and the type of supplemental sulphur amino acid had significant (P<0·05) effects on food intake and weight gain measured after 31d of the feeding study, while only the type of supplemental sulphur amino acid had significant effects on mean survival times and the survival rates. On average, the casein groups had higher food intake and weight gain compared with the SPI groups. The methionine-supplemented groups had lower food intake but higher weight gain than the cystine-supplemented groups. Similarly, the methionine-supplemented groups had higher mean survival times and survival rates compared with the cystine-supplemented groups. The data would suggest that a control diet based on cystine-supplemented casein (as recommended for normal healthy rats by the American Institute of Nutrition), may not meet the sulphur amino acid requirements for SHRSP rats, and that the methionine-supplemented casein would be an appropriate control diet for this animal model.

Excessive Ca intakes have been proposed to associate with vascular calcification and a higher risk of prostate cancer. We investigated the associations of supplemental and dietary Ca intake with mortality using data from 497 828 UK Biobank participants. The average follow-up was 4·2 years and 14 255 participants died, 8297 from cancer, 2959 from CVD and 572 from respiratory disease. The use of Ca supplements and milk consumption were associated with differences in mortality in younger (≤65 years) but not in older participants (>65 years, Pinteraction ≤ 0·04 for all comparisons). Among participants <65 years, there was an inverse association between Ca supplementation (OR 0·91, 95 % CI 0·83, 0·99) and milk consumption (OR 0·93, 95 % CI 0·86, 1·00) with respect to all-cause mortality. In the same age group, milk drinkers had lower odds of cancer mortality (OR 0·89, 95 % CI 0·80, 0·98) but Ca supplement use was associated with increased odds of respiratory mortality (OR 1·69, 95 % CI 1·16, 2·74). All associations in participants aged ≥65 years were null after full adjustment. In sensitivity analyses stratified by hormone replacement therapy, Ca supplement use was associated with decreased odds of cancer mortality in users but increased risk in other women (OR 0·81, 95 % CI 0·69, 0·94 v. OR 1·17, 95 % CI 1·01, 1·35, respectively). To conclude, we saw little evidence for harm with dietary or supplemental Ca. Further studies are required to confirm the proposed interaction with hormone replacement therapy and to exclude reverse causation as a determinant in the association between Ca supplements and increased risk of respiratory diseases.

Bowel cancer risk is strongly influenced by lifestyle factors including diet and physical activity. Several studies have investigated the effects of adherence to the World Cancer Research Fund (WCRF)/American Institute for Cancer Research (AICR) cancer prevention recommendations on outcomes such as all-cause and cancer-specific mortality, but the relationships with molecular mechanisms that underlie the effects on bowel cancer risk are unknown. This study aimed to investigate the relationships between adherence to the WCRF/AICR cancer prevention recommendations and wingless/integrated (WNT)-pathway-related markers of bowel cancer risk, including the expression of WNT pathway genes and regulatory microRNA (miRNA), secreted frizzled-related protein 1 (SFRP1) methylation and colonic crypt proliferative state in colorectal mucosal biopsies. Dietary and lifestyle data from seventy-five healthy participants recruited as part of the DISC Study were used. A scoring system was devised including seven of the cancer prevention recommendations and smoking status. The effects of total adherence score and scores for individual recommendations on the measured outcomes were assessed using Spearman’s rank correlation analysis and unpaired t tests, respectively. Total adherence score correlated negatively with expression of Myc proto-oncogene (c-MYC) (P=0·039) and WNT11 (P=0·025), and high adherers had significantly reduced expression of cyclin D1 (CCND1) (P=0·042), WNT11 (P=0·012) and c-MYC (P=0·048). Expression of axis inhibition protein 2 (AXIN2), glycogen synthase kinase (GSK3β), catenin β1 (CTNNB1) and WNT11 and of the oncogenic miRNA miR-17 and colonic crypt kinetics correlated significantly with scores for individual recommendations, including body fatness, red meat intake, plant food intake and smoking status. The findings from this study provide evidence for positive effects of adherence to the WCRF/AICR cancer prevention recommendations on WNT-pathway-related markers of bowel cancer risk.

Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3·770 and 7·256 MBq/g, respectively. Male Sprague–Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex® Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0·04, 0·03 and 0·01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1·99 and 1·54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8·53 and 9·06 % of the kudzu dose, respectively, and 3·60 and 5·64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation.

Dietary peptides are absorbed along the intestine through peptide transporter 1 (PepT-1) which is highly responsive to dietary protein level. PepT-1 is also involved in gut homeostasis, both initiating and resolving inflammation. Low-birth-weight (LBW) neonates are routinely fed a high-protein (HP) formula to enhance growth. However, the influence of this nutritional practice on PepT-1 activity is unknown. Intestinal PepT-1 activity was compared in normal-birth-weight (NBW) and LBW piglets. The effect of HP v. normal-protein (NP) formula feeding on PepT-1 activity and gut homeostasis in LBW piglets was evaluated, during the neonatal period and in adulthood. Flux of cephalexin (CFX) across the tissue mounted in Ussing chambers was used as an indicator of PepT-1 activity. CFX flux was greater in the ileum, but not jejunum or colon, of LBW than NBW piglets during the neonatal period. When LBW piglets were formula-fed, the HP formula increased colonic CFX during the 1st week of life. Later in life, intestinal CFX fluxes and barrier function were similar whether LBW pigs had been fed NP or HP formula. However, colonic permeability of HP- but not NP-fed pigs increased when luminal pH was brought to 6·0. The formyl peptide N-formyl methionyl-leucyl-phenylalanine conferred colonic barrier protection in HP-fed piglets. Heat shock protein 27 levels in the colonic mucosa of HP-fed LBW pigs correlated with the magnitude of response to the acidic challenge. In conclusion, feeding a HP formula enhanced colonic PepT-1 activity in LBW pig neonates and increased sensitivity of the colon to luminal stress in adulthood.