Children with epilepsy, especially those facing intractable seizures, experience a great impact on the quality of their lives. Effective treatment is essential, and although new anti-epileptic drugs have shown an improved profile of action, still a substantial number of children look for more efficacious ways of treatment that are far away from potential toxicity and ineffectiveness. The ketogenic diet is a dietary therapy for epileptic children based on manipulation of metabolism principles and brain energetics. This regimen aims to produce a controlled ketonaemia through excessive dietary fat intake, little carbohydrates and adequate (for growth) protein. The present paper is a review of previous and current papers regarding the proposed mechanisms of the ketogenic diet’s action, and the efficacy of the regimen on epileptic children. Unfortunately, a few small studies in sample size and duration tried to evaluate the potential risks of this regimen and their results were rather inconclusive. Further research needs to be done in order for the exact mechanism of the ketogenic diet to be clarified which will help to improve the diet’s application, especially for vulnerable epileptic children as far as their growth is concerned.

Dietary flavonoids can improve endothelial function, but the response varies between individuals. Wide variability is also seen in flavonoid O-methylation, a major pathway of flavonoid metabolism. The O-methylation of flavonoids could alter activity, and thus influence any effect on endothelial function. The objective of the current analysis was to investigate whether variability in the endothelial function response to ingestion of tea, a rich source of flavonoids, is related to the degree of O-methylation of flavonoids. This relationship was investigated in two studies in which endothelium-dependent flow-mediated dilatation (FMD) of the brachial artery was assessed and urinary 4-O-methylgallic acid (4OMGA) excretion was used as a marker of the O-methylation of tea-derived flavonoids. In the first study, amongst participants consuming five cups of tea per day for 4 weeks, the degree of increase in 4OMGA excretion was inversely associated with the change in FMD responses (r −078, p=0·008). In the second study, there was a significant difference in the FMD responses to acute ingestion of three cups of tea between individuals with a low (<median) and high (>median) 4OMGA response (1·94 (sem 0·79) % and −0·25 (sem 0·53) %, respectively; p=0·03). That is, any improvement in FMD following ingestion of tea may be enhanced in individuals who O-methylate less of the absorbed flavonoids. The present results are consistent with the suggestion that differences in flavonoid metabolism may influence their effect on endothelial function. Thus, differences in flavonoid metabolism could be related to the level of benefit of dietary flavonoids on the risk of CVD.

The chemopreventive effects of dietary n-3 PUFA in various pathologies has so far remained controversial, and we were interested in studying their potential influence on cell redox status. DHA (22:6n-3), a typical highly unsaturated n-3 PUFA, was used at 30µmol/l in a model of human fibroblast cell culture. A dose–response effect, roughly linear, was checked for DHA between 0 and 60µmol/l, and was accompanied by a large increase in intracellular GSH content. A time course study of this effect shows that, after a short fall, as soon as 4h after the beginning of the experiment, the large increase in the GSH content was associated with elevated catalytic activities of γ-glutamyl-cysteinyl ligase, glutathione reductase and glutathione S-transferase. This coordinated response is characteristic of an antioxidant response and was confirmed by the induction of expression of mRNA for γ-glutamyl-cysteinyl ligase, glutathione reductase and haem-oxygenase. This large increase in the GSH content contributes to decreasing the reactive oxygen species level, as assessed by the decreased accumulation of dichlorofluorescein inside cells. To our knowledge, this is the first report on a specific and potent effect of DHA for decreasing the oxidative stress of human fibroblasts.

Cattle fed grass silage diets have been reported to have high carcass fat:protein ratios. The effect of grass silage and dried grass diets, fed at different levels of intake to ensure a range of equivalent metabolisable energy intakes (MEI) from 1·1×metabolisable energy requirement for maintenance to ad libitum, on fat and protein metabolism in twenty-four Hereford×Friesian steers was investigated. After about 84d of dietary treatment rates of whole-body fat and protein metabolism were measured, as were rates of lipogenesis in omental, perirenal and subcutaneous adipose tissue. Carcass composition was determined. Animals fed silage had greater (P<0·001) carcass fat:protein ratios than animals fed dried grass at equivalent levels of MEI. Animals fed silage had lower (P<0·001) rates of protein gain. Rates of leucine entry and oxidation were lower (P<0·001) in animals fed silage, but there was no dietary difference in the rate of whole-body protein synthesis. There was no dietary difference in the rate of carcass fat gain, but rates of lipogenesis in perirenal adipose tissue were significantly (P=0·007) higher in animals fed silage. There was no dietary difference in the rate of palmitate and glycerol entry or palmitate oxidation. There were no interactions between MEI and diet, indicating that increments of energy were utilised with the same efficiency from both diets. It was concluded that the high carcass fat:protein ratios of young growing steers was due to limited rates of protein accretion and not to elevated rates of carcass fat accretion.

Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reaches the colon. However, little is known about effects on the intestinal microflora. During two 4-week periods in a double-blind, placebo-controlled, cross-over design, nineteen healthy volunteers consumed a controlled basal diet enriched with either 30g isomalt or 30g sucrose daily. Stools were collected at the end of each test phase and various microbiological and luminal markers were analysed. Fermentation characteristics of isomalt were also investigated in vitro. Microbiological analyses of faecal samples indicated a shift of the gut flora towards an increase of bifidobacteria following consumption of the isomalt diet compared with the sucrose diet (P<0·05). During the isomalt phase, the activity of bacterial β-glucosidase decreased (P<0·05) whereas β-glucuronidase, sulfatase, nitroreductase and urease remained unchanged. Faecal polyamines were not different between test periods with the exception of cadaverine, which showed a trend towards a lower concentration following isomalt (P=0·055). Faecal SCFA, lactate, bile acids, neutral sterols, N, NH3, phenol and p-cresol were not affected by isomalt consumption. In vitro, isomalt was metabolized in several bifidobacteria strains and yielded high butyrate concentrations. Isomalt, which is used widely as a low-glycaemic and low-energy sweetener, has to be considered a prebiotic carbohydrate that might contribute to a healthy luminal environment of the colonic mucosa.

Recent reports have demonstrated various cardiovascular and neurological benefits associated with the consumption of foods rich in anthocyanidins. However, information regarding absorption, metabolism, and especially, tissue distribution are only beginning to accumulate. In the present study, we investigated the occurrence and the kinetics of various circulating pelargonidin metabolites, and we aimed at providing initial information with regard to tissue distribution. Based on HPLC and LC-MS analyses we demonstrate that pelargonidin is absorbed and present in plasma following oral gavage to rats. In addition, the main structurally related pelargonidin metabolite identified in plasma and urine was pelargonidin glucuronide. Furthermore, p-hydroxybenzoic acid, a ring fission product of pelargonidin, was detected in plasma and urine samples obtained at 2 and 18h after ingestion. At 2h post-gavage, pelargonidin glucuronide was the major metabolite detected in kidney and liver, with levels reaching 0·5 and 0·15nmol pelargonidin equivalents/g tissue, respectively. Brain and lung tissues contained detectable levels of the aglycone, with the glucuronide also present in the lungs. Other tissues, including spleen and heart, did not contain detectable levels of pelargonidin or ensuing metabolites. At 18h post-gavage, tissue analyses did not reveal detectable levels of the aglycone nor of pelargonidin glucuronides. Taken together, our results demonstrate that the overall uptake of the administered pelargonidin was 18% after 2h, with the majority of the detected levels located in the stomach. However, the amounts recovered dropped to 1·2% only 18h post-gavage, with the urine and faecal content constituting almost 90% of the total recovered pelargonidin.

Previous studies have found Kenyan endurance runners to be in negative energy balance during training and prior to competition. The aim of the present study was to assess energy balance in nine elite Kenyan endurance runners during heavy training. Energy intake and expenditure were determined over 7d using weighed dietary intake and doubly labelled water, respectively. Athletes were on average in negative energy balance (mean energy intake 13 241 (SD 1330) kJ/d v. mean energy expenditure 14 611 (SD 1043) kJ/d; P=0·046), although there was no loss in body mass (mean 56·0 (SD 3·4) kg v. 55·7 (SD 3·6) kg; P=0·285). The calculation of underreporting was 13; (range −24 to +9%) and almost entirely accounted for by undereating (9% (range −55 to +39%)) as opposed to a lack of significant underrecording (i.e. total water intake was no different from water loss (mean 4·2 (SD 0·6) l/d v. 4·5 (SD 0·8) l/d; P=0·496)). Fluid intake was modest and consisted mainly of water (0·9 (SD 0·5) l/d) and milky tea (0·9 (SD 0·3) l/d). The diet was high in carbohydrate (67·3 (SD 7·8) %) and sufficient in protein (15·3 (SD 4·0) %) and fat (17·4 (SD 3·9) %). These results confirm previous observations that Kenyan runners are in negative energy balance during periods of intense training. A negative energy balance would result in a reduction in body mass, which, when combined with a high carbohydrate diet, would have the potential in the short term to enhance endurance running performance by reducing the energy cost of running.

The present study was to investigate the effects of l-arginine (l-Arg) supplementation on cardiac oxidative stress and the inflammatory response in rats following acute exhaustive exercise on a treadmill. Rats were randomly divided into four groups: sedentary control (SC); SC with l-Arg treatment (SC+Arg); exhaustive exercise (E); exhaustive exercise with l-Arg treatment (E+Arg). Rats in groups SC+Arg and E+Arg received a 2% l-Arg diet. Rats in groups E and E+Arg performed an exhaustive running test on a treadmill at a final speed of 30m/min, 10% grade, at approximately 70–75% VO2max. The results showed a significant increase in cardiac xanthine oxidase (XO) and myeloperoxidase activities and membrane lipid peroxidation endproduct (malondialdehyde; MDA) levels of exercised rats compared with SC rats. The increased cardiac XO activity and MDA levels in exercised rats were significantly decreased in exercised rats supplemented with l-Arg. Myocardial GSSG content increased whereas the GSH:GSSG ratio was depressed in exercised rats compared with SC rats. Cardiac GSSG levels significantly decreased, whereas total glutathione, GSH and the GSH:GSSG ratio increased in exercised rats supplemented with l-Arg compared with exercised rats. The activities of creatinine kinase (CK) and lactate dehydrogenase (LDH), and lactate, uric acid, and nitrite and nitrate levels in the plasma significantly increased in exercised rats compared with SC rats. The activities of plasma CK and LDH were significantly decreased in l-Arg-supplemented plus exercised rats compared with exercised rats. These findings suggest that l-Arg supplementation reduces the oxidative damage and inflammatory response on the myocardium caused by exhaustive exercise in rats.

Rainbow trout has a limited ability to utilize digestible carbohydrates efficiently. Trout feeds generally contain high levels of DHA, a fatty acid known to inhibit a number of glycolytic and lipogenic enzymes in animals. A study was conducted to determine whether carbohydrate utilization by rainbow trout might be affected by dietary DHA level. Two low-carbohydrate (<4% digestible carbohydrate) basal diets were formulated to contain 1 (adequate) or 4 (excess)g/100g DHA diet respectively. The two basal diets were diluted with increasing levels of digestible starch (0%, 10%, 20% and 30%, respectively) to produce eight diets. These diets were fed to fish for 12 weeks at 15°C according to a pair-fed protocol that consisted of feeding the same amount of basal diet but different amounts of starch. Live weight, N and lipid gains, hepatic glycogen and plasma glucose values significantly increased, whereas feed efficiency (gain:feed) significantly decreased, with increasing starch intake (P<0·05). The retention efficiency of N (N gain/digestible N intake) improved with starch supplementation but was not affected by DHA level (P>0·05). Starch increased the activity of glucokinase, pyruvate kinase, glucose 6-phosphate dehydrogenase and fatty acid synthase (P<0·05) but did not affect hexokinase and malic enzyme activity. DHA had no effect on growth but increased plasma glucose and reduced carcass lipid and liver glycogen contents (P<0·05). Glycolytic and lipogenic enzymes were not affected by DHA level, except for pyruvate kinase, which was reduced by increasing DHA level. These results suggest only a marginal effect of dietary DHA on the ability of fish to utilize carbohydrate.

Four cattle, sheep, ponies and donkeys were fed dehydrated lucerne, early-cut hay, later-cut hay or barley straw in a Latin square-based design for four periods of 35d. In the first sub-period animals were fed the diets ad libitum (1–21d) and in the second sub-period they were fed the same diet restricted to 0·75 of ad libitum intake (days 22–35). Measurements of forage intake, apparent digestibilities and gastrointestinal mean retention times (MRT) were made in the last 7d of each sub-period. Differences between species in voluntary DM intake (VDMI; g/kg live weight LW)0·75 and g/LW) were greatest on the lucerne and least on barley straw. Cattle VDMI (g/kg LW0·75) compared with intake of the other species was > ponies > sheep > donkeys on lucerne. On barley straw VDMI (g/kg LW0·75) of cattle compared with intake of the other species was = donkey = ponies > sheep. VDMI of hays were intermediate between the lucerne and straw forages. Apparent digestibilities of DM, organic matter (OM), neutral-detergent fibre (NDF) and acid-detergent fibre (ADF) of the lucerne and hays were higher in the ruminants than in the equids. Effect of feeding level was not significant. Gastrointestinal MRT was shorter in the equids than in the ruminants. On straw diets donkeys showed similar apparent digestibilities of feed components to those of the cattle, whilst apparent digestibility of the straw diet by the ponies was lowest. Results are discussed in relation to evolutionary differences in feeding and digestion strategy associated with fore- or hind-gut fermentation in ruminants and equids.

Biotin is required by rumen microbes for efficient fermentation. To evaluate the role of protozoa in ruminal biotin metabolism, five diets composed of grass hay or of grass hay/cereal grain mixtures were supplied to faunated or defaunated RUSITEC fermenters. In the mixed diets, hay was replaced to 33:67 or 67:33 w/w on an air-dried basis by either wheat or maize grain in order to simulate different cellulolytic and amylolytic fermentation conditions. Defaunation increased SCFA production, whereas NH4 concentration and the release of CH4 were reduced. Biotin input declined when cereal grain was used to replace the hay. With the exception of the high-wheat treatment, defaunated fermenters yielded higher biotin outputs than faunated fermenters. The biotin balance, calculated as the difference between the total biotin output (biotin in the solid residue contained in the nylon bags after fermentation plus the biotin in the effluent) and the biotin input with the feed, was negative for all the dietary treatments apart from fermenters supplied with the high-maize diet. It was less negative or, in the case of the high-maize diets, more positive for defaunated compared with faunated fermenters. It was concluded that, under normal faunated conditions, protozoa directly utilise or indirectly affect the bacterial synthesis and/or utilisation of biotin. With diets of a high fermentation potential, as realised with the high-wheat diet, protozoa prevent the development of a bacterial population that would utilise high or synthesise low amounts of biotin.

Enteral exposure of suckling rats to phytohaemagglutinin (PHA) has been shown to induce growth and precocious functional maturation of the gastrointestinal tract. The aim of the present study was to explore the mechanism of this action. Suckling rats, 14d old, were fed a single dose of PHA (0·05mg/g body weight) or saline. The binding of PHA to the gut epithelium and its effect on the morphology and functional properties of the gut and pancreas were studied up to 3d after treatment. Initially, at 1–24h, the PHA bound along the gut mucosal lining, resulting in disturbed gut morphology with villi shortening and rapid decreases in disaccharidase activities and macromolecular absorption capacity. During a later phase, between 1 and 3d, the PHA binding had declined, and an uptake by enterocytes was observed. An increase in crypt cell proliferation and gut growth became evident during this period, together with a functional maturation, as indicated by increases in disaccharidase (maltase and sucrase) activities and the low macromolecular absorption capacity. Pancreas growth also increased, as did its content of digestive enzymes. We conclude that enteral exposure to PHA in suckling rats temporarily causes mucosal disarrangement and functional impediment of the gut, which may be explained by binding to and disruption of the gut mucosa and a two-fold increase in the plasma corticosterone concentration. These findings may lead to a better understanding of the role of diet in gastrointestinal maturation and may constitute a basis for the treatment of mammals having an immature gut.

Phaseolus vulgaris phaseolin has been shown to stimulate faecal losses of endogenous N in rats. Experiments with purified phaseolin were carried out in rats to test the hypothesis that these losses reflect intestinal disorders. Phaseolin composition varies depending on its constitutive subunits. Therefore, three phaseolin types (S, T, I) were tested. Phaseolin T was incorporated in varying levels (0, 33, 67 or 100% of the dietary protein) as raw material in experiment 1. In experiment 2, the three phaseolin types were incorporated at 50%, with or without previous thermic treatment. Raw casein was the basal protein source and was also heated in experiment 2. Faecal digestibility of phaseolin and gut integrity were evaluated in both experiments. The incorporation level or type of phaseolin had little effect on gross anatomy of gut segments but these factors influenced the weight and pH of fresh contents of the stomach and caecum (P<0·05). Raw phaseolin T incorporated at various levels led to an enlargement of duodenal villi together with a tendency for increased crypt depth in the jejunum (P=0·06). Activities of both alkaline phosphatase in the duodenum and aminopeptidase N in the ileum decreased (P<0·05) after thermal treatment of casein while they increased (P<0·05) for heat-treated phaseolin S and T, respectively. In conclusion, raw phaseolin had no effect on the tissue weight of gut segments and induced limited alterations in the small intestine. Differences due to phaseolin level or type were limited too.

Fish-bone peptides (FBP) with a high affinity to Ca were isolated using hydroxyapatite affinity chromatography, and FBP II with a high ratio of phosphopeptide was fractionated in the range of molecular weight 5·0–1·0kDa by ultramembrane filtration. In vitro study elucidated that FBP II could inhibit the formation of insoluble Ca salts in neutral pH. In vivo effects of FBP II on Ca bioavailability were further examined in the ovariectomised rat. During the experimental period, Ca retention was increased and loss of bone mineral was decreased by FBP II supplementation in ovariectomised rats. After the low-Ca diet, the FBP II diet, including both normal level of Ca and vitamin D, significantly decreased Ca loss in faeces and increased Ca retention compared with the control diet. The levels of femoral total Ca, bone mineral density, and strength were also significantly increased by the FBP II diet to levels similar to those of the casein phosphopeptide diet group (no difference; P>0·05). In the present study, the results proved the beneficial effects of fish-meal in preventing Ca deficiency due to increased Ca bioavailability by FBP intake

The present study was designed to examine the effects of habitual consumption of Taiwanese vegetarian diets on hormonal secretion, and on lipid and glycaemic control. Of the ninety-eight healthy female adults recruited from Hualien, Taiwan (aged 31–45 years), forty-nine were Buddhist lactovegetarians and forty-nine were omnivores. Dietary intakes were measured, and blood levels of nutrients and hormones were analysed. Vegetarians consumed less energy, fat and protein, but more fibre than the omnivores. Compared with the omnivores, the vegetarians had, on average, lower BMI and smaller waist circumference. Except for slightly lower levels of thyroxine (T4) in vegetarians, vegetarians and omnivores both showed similar levels of triiodothyronine (T3), free T4, thyroid-stimulating hormone, T3:T4 ratio and cortisol. Compared with the omnivores, the vegetarians had significantly lower levels of fasting insulin (median: 35·3 v. 50·6pmol/l) and plasma glucose (mean: 4·7 (se 0·05) v. 4·9 (se 0·05) mmol/l). Insulin resistance, as calculated by the homeostasis model assessment method, was significantly lower in the vegetarians than in the omnivores (median: 1·10 v. 1·56), while β-cell function was not different between the two groups. BMI and diet were both independent predictors for insulin resistance, and contributed 18 and 15% of the variation in insulin resistance, respectively. In conclusion, Taiwanese vegetarians had lower glucose and insulin levels and higher insulin sensitivity than did the omnivores. Diet and lower BMI were partially responsible for the high insulin sensitivity observed in young Taiwanese vegetarians.

The present study is the first to examine the hypothesis that dietary supplementation with β-casein A1 promotes an increased risk relative to supplementation with β-casein A2 in patients traditionally at high risk of developing CVD. The study was conducted in fifteen asymptomatic participants (six male; nine female) at high risk of developing CVD. A double-blind cross-over study design was used with a total duration of 24 weeks. Dietary intervention was a daily supplementation (25g) of either casein A1 or A2 (for 12 weeks each). Surrogate measures of cardioprotection studied included the examination of vascular (endothelium and arterial) function, resting blood pressure, plasma lipids and biochemical markers of inflammation. Total plasma cholesterol levels were significantly lower following 12 weeks of both casein A1 and A2 interventions but the decrease was not different between intervention. Plasma insulin, homocysteine, C-reactive protein, fibrinogen, protein C and S and von Willebrand factor levels were not different between the two casein supplements. Endothelium function, measured as a vascular response using venous occlusion plethysmography to intra-aterial infusions of the endothelium-dependent agonist acetylcholine, were not different between the two casein interventions. Similarly, neither blood pressure nor measures of large artery stiffness were affected by differing the casein variant. We therefore conclude that there is no evidence from the present study that supplementation with casein A1 has any cardiovascular health disadvantage over consumption of casein A2.

The benefit of the introduction of mandatory folic acid fortification of all flour products in the USA in 1998 has been amply demonstrated in a reduction of neural tube defect births. Doubt has been cast on the actual level of fortification and recent calculations have shown that the level of folic acid fortification is likely to have been over twice the amount mandated. The implication of this is that a greater proportion of the population are likely to have consumed folic acid at >1mg/d, the Food and Drug Administration safe upper level of intake. Using the criteria of appearance of synthetic folic acid in serum, the objective of this pilot study was to investigate the consequences of consumption of baked bread preparations containing 1mg folic acid. Four healthy adult volunteers undertook each dosing schedule 2 weeks apart. This consisted of a single dose of 1000μg, two doses of 500μg, three doses of 333μg, five doses of 200μg and, finally, ten doses of 100μg. Serum was collected pre- and postprandially and analysed for synthetic folic acid by a combined HPLC–microbiological assay for folic acid. Folic acid appeared in all subjects at all test doses, with the effect more pronounced as the standard dose was administered in smaller amounts over the test period. Approaches to optimise folic acid intake in target populations as part of a universal fortification strategy should take into consideration the potential hazard of over-exposure in groups consuming high amounts of flour-based products.

Lipid, oxidative and inflammatory parameters are frequently altered in dialysis patients and may be worsened by intravenous lipid emulsions (ILE). We assessed the efficacy and tolerance of olive as compared with standard soybean oil-based ILE during intradialytic parenteral nutrition (IDPN). IDPN mixtures containing amino acids, glucose, and either olive oil (OO group, n 17) or soybean oil-based ILE (SO group, n 18) were administered in a 5-week randomized, double-blind study. On days 0 and 35, patients' nutritional status was assessed by BMI, normalized protein catabolic rate, predialytic creatinine, serum albumin and transthyretin; lipid metabolism by plasma LDL- and HDL-cholesterol, triacylglycerols, phospholipids, apo A-I, A-II, B, C-II, C-III, E and lipoprotein (a); oxidative status by α-tocopherol, retinol, selenium, glutathione peroxidase, malondialdehyde and advanced oxidized protein products; inflammatory status by serum C-reactive protein, orosomucoid, IL-2 and IL-6. No serious adverse event was observed. Significant changes were observed from day 0 to day 35 (P<0·05): nutritional criteria improved (albumin in OO; albumin, transthyretin and creatinine in SO); LDL-cholesterol, apo B, C-II, C-III and apo A-I/A-II ratio increased in both groups. HDL-cholesterol decreased in OO; apo E increased and lipoprotein (a) decreased in SO; α-tocopherol/cholesterol ratio increased in OO; malondialdehyde decreased in both groups; IL-2 increased in both groups. The between-group comparison only showed the following differences: α-tocopherol/cholesterol increased in OO; lipoprotein (a) decreased in SO. From these data, it was concluded that OO- and SO-based IDPNs similarly improved nutritional status and influenced plasma lipid, oxidative, inflammatory and immune parameters.

Glucagon-like peptide 1 (GLP-1) is a peptide hormone that is released in response to nutrient ingestion. Postprandial GLP-1 release has been reported to be attenuated in obese subjects, but reports on the effect of weight loss on GLP-1 are conflicting. The aim of the present study was to clarify the effect of a weight-loss period and a consecutive weight-maintenance period on nutrient-stimulated GLP-1 release in obese subjects. Nutrient-stimulated (standard breakfast; 1·9MJ) GLP-1 release was investigated in thirty-two obese subjects on three occasions: before weight loss (T1) (BMI 30·0 (sd 2·5) kg/m2); after a 6-week very-low-energy diet (VLED) (T2) (BMI 27·6 (sd 2·3) kg/m2); after a 3-month weight-maintenance period (T3) (BMI 27·9 (sd 2·3) kg/m2). At each occasion, following a fasting blood sample the test meal was fed and blood was drawn every 30min for 2h relative to ingestion in order to determine plasma GLP-1, insulin, glucose and NEFA concentrations. Subjects lost 7 (sd 3·4) kg during the VLED (P<0·0001) and regained 1 (sd 3·2) kg during the weight-maintenance period (NS). The area under the curve for nutrient-stimulated plasma GLP-1 (pmol/l×h) was significantly decreased (P=0·01) at T2 (6·8 (sd 1)) compared with T1 (12·8 (sd 2·9)) and T3 (11·1 (sd 1·5)). Since we found a rebound of concentrations after a weight-maintenance period, decrease after weight loss seems to be transient and possibly due to a negative energy balance.