We investigated the combined effect of dietary supplementation with l-arginine, which is the precursor of NO, and pharmacological treatment with atorvastatin, which is a 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitor, on the development of atherosclerosis in homozygous Watanabe heritable hyperlipidaemic rabbits. Rabbits were fed either standard rabbit chow (group C; n 9) as control, a 1·5 % l-arginine diet (group A; n 9), standard rabbit chow plus atorvastatin (2·5 mg/kg per d) in drinking water (group S; n 8), or standard rabbit chow plus a 1·5 % l-arginine diet with atorvastatin (group SA; n 8). Blood was sampled at 2-week intervals. After 8 weeks (T8), the aorta was harvested for topographic and histological analysis. Only the SA group showed decreases in total area of lesions (21 %) and the area of abdominal lesions (44 %) compared with the control group (P = 0·019). Furthermore, plaques in the SA group were smaller and less thick than those observed in the S group. Unexpectedly, plasma nitrite + nitrate levels were not modified under either the l-arginine diet alone or under l-arginine plus atorvastatin. The present study is the first to demonstrate that diet supplementation with l-arginine associated with a statin (atorvastatin) is more efficient in reducing lesion size than treatment with l-arginine or a statin alone. This is a relatively novel therapeutic approach associating a macronutrient and a drug.

Malnutrition is associated with an increased incidence of perioperative morbidity and mortality. To evaluate the effect of malnutrition on the metabolic and inflammatory response to surgery in patients with oesophageal cancer, we studied the effects of oesophagectomy in six patients with major (13·9 (se 1·3) %) weight loss and five patients with minor (0·7 (se 0·6) %) weight loss in the 6 months before to surgery. Rates of appearance (Ra) of glucose, glycerol, leucine and urea were determined by stable isotopically labelled tracer infusion before and after surgery. C-reactive protein was measured as an inflammation marker. BMI was lower in the patients with major weight loss than those with minor weight loss (20·3 (se 0·7) and 24·9 (se 1·5) kg/m2, P = 0·02). With the exception of greater glucose Ra in the major weight loss than minor weight loss subjects (11·1 (se 0·3) v. 9·5 (se 0·3) μmol/kg per min, P = 0·01), there were no differences in substrate kinetics before surgery between groups. Surgery increased glucose Ra, leucine Ra and urea Ra by 41, 24 and 58 %, respectively, in the total group. Changes in substrate kinetics in response to surgery were not different between patients with major and minor weight loss. Surgery increased C-reactive protein concentrations to a comparable extent in both groups. In conclusion, major upper gastrointestinal tract surgery in patients with oesophageal cancer elicits a catabolic response, characterized by increased inflammation, glucose production and protein breakdown. However, this catabolic response does not seem to be influenced by pre-operative nutritional status.

The present study investigated the association between protein intake and lean mass according to obesity status over a 12-year period. Data on 4412 participants aged 40–69 years were obtained from the Korean Genome and Epidemiology Study. The usual dietary protein intake of these participants was assessed at baseline using a semi-quantitative FFQ. Body composition was measured using bioelectrical impedance analysis at baseline and after a 12-year follow-up. Linear mixed-effects models were used to examine the associations between lean mass after a 12-year follow-up and protein intake at baseline. After adjusting for covariates and lean mass at baseline, comparisons between the highest and lowest tertiles revealed that dietary protein intake was positively associated with lean mass in both men (β = 0·79, P = 0·001) and women (β = 0·28, P = 0·082) after the 12-year period; however, those differences were attenuated after additional adjustment for fat mass at baseline and were stronger in the normal-weight group (men, β = 0·85, P = 0·002; women, β = 0·97, P < 0·001) but were not detected in the obese group. In the obese group, age (men, β = 4·08, P < 0·001; women, β = 2·61, P < 0·001) and regular physical activity (men, β = 0·88, P = 0·054; women, β = 0·76, P < 0·001) were significantly associated with lean mass after 12 years of follow-up. The results of the present study showed that protein intake may contribute to the prevention of ageing-related lean mass loss; however, the impact of this intake may vary depending on obesity status. Therefore, the maintenance of a healthy body weight during ageing through enhanced protein intake is likely to confer health benefits.

Psyllium, a dietary fibre rich in soluble components, has both cholesterol- and TAG-lowering effects. Many studies have verified these actions using liver samples, whereas little information is available on the effects of psyllium treatment on other organs. The purpose of the present study was to evaluate the possible beneficial effects of psyllium. We investigated the gene expression profiles of both liver and skeletal muscle using DNA microarrays. C57BL/6J mice were fed a low-fat diet (LFD; 7 % fat), a high-fat diet (HFD; 40 % fat) or a HFD with psyllium (40 % fat+5 % psyllium; HFD+Psy) for 10 weeks. Body weights and food intake were measured weekly. After 10 weeks, the mice were killed and tissues were collected. Adipose tissues were weighed, and plasma total cholesterol and TAG blood glucose levels were measured. The expression levels of genes involved in glycolysis, gluconeogenesis, glucose transport and fatty acid metabolism were measured by DNA microarray in the liver and skeletal muscle. In the HFD+Psy group, plasma total cholesterol, TAG and blood glucose levels significantly decreased. There was a significant reduction in the relative weight of the epididymal and retroperitoneal fat tissue depots in mice fed the HFD+Psy. The expression levels of genes involved in fatty acid oxidation and lipid transport were significantly up-regulated in the skeletal muscle of the HFD+Psy group. This result suggests that psyllium stimulates lipid transport and fatty acid oxidation in the muscle. In conclusion, the present study demonstrates that psyllium can promote lipid consumption in the skeletal muscle; and this effect would create a slightly insufficient glucose state in the liver.

Fruit intake is associated with lower risk of cardiometabolic diseases. However, effects of dried fruits on cardiometabolic health are not well researched. We investigated the effect of daily dried fruit consumption compared with a carbohydrate-rich snack on cardiometabolic disease risk factors in adults with increased cardiometabolic risk. A two-period randomised crossover trial was conducted in adults (n 55) with elevated BMI and at least one additional risk factor for cardiometabolic disease to compare the effects of consuming 3/4 cup/d mixed dried fruits (plums, figs, dates and raisins) or an energy- and carbohydrate-matched control snack for 4 weeks. The primary outcome was LDL-cholesterol; secondary outcomes included other lipids and lipoproteins, glucose and insulin, C-reactive protein, blood pressure and vascular stiffness. Linear mixed models were used for data analysis. Lipid and lipoprotein concentrations did not differ between conditions; however, dried fruit increased LDL-cholesterol (0·10 mmol/l, 95 % CI 0·01, 0·20) compared with baseline. Compared with the control, dried fruit increased mean fasting glucose (0·08 mmol/l, 95 % CI 0·005, 0·16; P = 0·038). Vascular outcomes, fasting insulin and C-reactive protein did not differ between conditions. Mean weight changes did not differ (P = 0·55) but tended to increase after both conditions (dried fruit 0·3 kg, 95 % CI –0·09, 0·65; control 0·4 kg, 95 % CI 0·01, 0·75). Thus, short-term daily consumption of a large portion of mixed dried plums, figs, dates and raisins, without structured dietary guidance, did not improve cardiometabolic risk factors, compared with carbohydrate-rich snacks, in adults with increased baseline cardiometabolic risk.

1. The responses of germ-free (GF) and conventional (CV) chicks to suboptimal amounts of dietary calcium pantothenate (CaPa) were compared. At the lower levels of supplementation the signs of deficiency were more severe in the CV chicks.

2. Calculations based on mean body-weibts of chicksgiven graded dietary supplements of calcium pantothenate (CaPa) in both environments showed that, to achieve the same body-weight and freedom from signs of deficiency, the GF birds required only approximately two-thirds the amount of CaPa as was needed by corresponding CV birds.

3. Increasing the dietary supplement of CaPa resulted in increased contents of PaA in the livers. GF birds had more PaA per liver than their CV counterparts. In both instances the effect was due to the larger liver size. Concentration of PaA in the liver was not al€ected by dietary supplement nor by the environment.

4. CV birds were deemed to be under greater stress than GF birds because their adrenal glands were heavier and contained less cholesterol.

5. CV chicks deliberately stressed by injections of adrenocorticotrophic hormone had heavier adrenals with lower cholesterol contents than unstressed controls, but the condition was not altered by increasing the dietary supply of CaPa.

High dietary energy density (ED) has been associated with weight gain. However, little is known about the long-term effects of ED on weight changes among free-living subjects, particularly in Japanese and other Asian populations. In this study, we assessed dietary habits and weight changes in participants (5778 males and 7440 females, 35–69 years old) of the Takayama study. ED was estimated using a validated FFQ at baseline only. Information on body weight (BW) was obtained by self-administered questionnaires at baseline and follow-up. Mean BW difference in 9·8 years was 17 (se 4221) g for men and −210 (se 3889) g for women. In men, ED was positively associated with BW at follow-up after controlling for age, BW, height, physical activity score, alcohol consumption, energy intake, years of education at the baseline and change of smoking status during the follow-up. On average, men in the highest quartile of ED (>5·322 kJ/g (>1·272 kcal/g)) gained 138 (se 111) g, whereas men in the lowest ED (<1·057) lost 22 (se 111) g (Pfor trend=0·01). The association between ED and BW gain was stronger in men with normal weight. In women, the association between ED and weight change was not statistically significant. In conclusion, contrary to some studies that report an association between ED and weight gain in the overweight only, our data suggest that high-ED diets may be associated with weight gain in the lean population as well, at least in male subjects.

Few studies have been interested in the relationship between dietary patterns and activity limitation in older adults yet. We analysed the association between dietary patterns and the risk of self-reported activity limitation – that is mobility restriction, limitation in instrumental activities in daily living (IADL) and in activities in daily living (ADL) – in community-dwellers aged 67+ years initially free of activity limitation in 2001–2002 and re-examined at least once over 10 years – that is 583 participants for mobility restriction, 1114 for IADL limitation and 1267 for ADL limitation. At baseline, five sex-specific dietary clusters were derived by hybrid clustering method from weekly frequency of intake of twenty food and beverage items. Self-reported mobility restriction, limitations in IADL and in ADL were assessed using the Rosow–Breslau, the Lawton–Brody and the Katz scales, respectively. Associations between dietary clusters and the risk of each activity limitation were assessed using Cox proportional hazard models. In models adjusted for socio-demographic and health-related covariates, compared with the ‘Healthy’ cluster the ‘Biscuits and snacking’ cluster was associated with a higher risk of mobility restriction (hazard ratio (HR)=3·0; 95 % CI 1·6, 5·8) and limitation in IADL (HR=2·1; 95 % CI 1·1, 4·2) in men and limitation in ADL in women (HR=2·3; 95 % CI 1·3, 4·0). In this French cohort of community-dwellers aged 67+ years, some unhealthy dietary patterns may increase the risk of activity limitation all along the disablement process in older adults.

1. An experiment is reported in which the effects of giving copper sulphate-supplemented diets and control unsupplemented pair-fed diets to laying hens were compared.

2. The level of food intake significantly adversely affected mean body-weight, egg number, egg weight, liver, kidney, oviduct and ovary weights. Gizzard weight/kg body-weight was significantly increased both with decreasing levels of food intake and increased CuSO4 supplementation.

3. There was evidence of a depressing effect of CuSO4per se on egg production and possibly on oviduct and ovary weight.

4. Liver lipid concentration was significantly decreased with decreasing levels of food intake and the results also suggest a depressing effect of CuSO4.

5. The Cu concentrations and total contents in liver and kidneys were significantly increased by dietary added CuSO4. Liver and kidney Zn and Fe concentrations were increased with decreasing levels of food intake rather than by CuSO4. addition.

The bacterial single-cell protein BioProtein (BP; Norferm Danmark, Odense, Denmark), produced by fermentation of natural gas with methanotrophic bacteria, is a potential protein source for man and animals. For human consumption, removal of the nucleic acid is necessary. Preliminary studies have shown that ingested BP induces a specific immune response. The objective of the present study was to characterize the type of response, its development over time and product-related causative factors. Mice were fed with diets containing 60 g nucleic acid-reduced BP/kg, 240 g nucleic acid-reduced BP/kg, 240 g untreated BP (basic BP)/kg or 240 g casein/kg (control). In another study, mice were fed 240 g basic BP/kg, whole cell-free BP-culture homogenate or control diet. The immune response was monitored using an ELISA for BP-specific immunoglobulin in blood and BP-specific immunoglobulin A in blood and saliva. Ingested BP induced a steady specific mucosal and systemic immune response, characterized by a dose-dependent production of immunoglobulin and immunoglobulin A in blood and immunoglobulin A in saliva. Basic BP and nucleic acid-reduced BP induced identical responses. However, feeding mice BP-culture homogenate induced immunoglobulin A in saliva but there was no systemic response. The antibodies from BP-fed mice cross-reacted with BP-culture homogenate revealing the presence of the same antigenic components in the two products despite the different oral immunogenicity. Thus, ingestion of BP induces a persistent mucosal and systemic immune response of which the systemic response can be avoided by ingesting a BP preparation free of whole cells. This indicates the importance of the non-particulate constitution of single-cell protein products intended for human or animal consumption.

1. Two experiments involving 106 lambs were conducted to study the functioning of the oesophageal groove reflex in lambs sucking from either individual feeders or from a bulk dispenser. The effects on the performance of lambs given either a high-protein or a low-protein creep feed in conjunction with either a ewe's milk substitute, Ewelac, or a homogenate of calcium caseinate, Casilan, were studied. Two milk substitutes, one with a long and one with a short clotting time were compared in terms of their effects on the incidence of diarrhoea.

2. The location of the liquid feeds in the stomach was studied radiologically.

3. In both experiments the sucking behaviour of the lambs was closely related to thefunctioning of the oesophageal groove reflex and performance of the lambs. In the first experi-ment the milk substitute consistently entered the abomasum in seven of the ten individually fed lambs and in twelve of the twenty-two group-fed lambs. The corresponding figures for the second experiment were twenty-two of the twenty-four individually fed and thirty-four of the fifty group-fed lambs.

4. When the milk substitute entered the abomasum it always did so throughout the fattening period even in those instances where the milk had been withheld for 1 month.

5. Poor sucking, low milk consumption and the entry of the milk into the rumen resulted in very poor performance by the lambs. Such lambs demonstrated a non-specific unthriftiness similar to the ‘tail enders’ sometimes observed with lambs reared with ewes.

6. Continued feeding with either the milk substitute or casein homogenate with a low-protein creep feed did not improve performance.

7. The clotting time of the milk substitute did not appear to influence the incidence of diarrhoea among the lambs.

Basolateral and brush-border membranes were prepared from the intestines and kidneys of spontaneously hypertensive (SHR) and normotensive (WKY) rats fed on a calcium-adequate diet and assayed for their enzyme activities. In intestinal basolateral membranes the activities of Na+ K+-ATPase (EC 3.6.1.37) Ca2+-ATPase (EC 3.6.1.38) and alkaline phosphatase (EC 3.1.3.1) were lower in SHR rats when compared with WKY rats, whilst 5'-nucleotidase (EC3.1.3.5) (a marker for basolateral membranes) was unaffected. In kidney basolateral membranes all enzymes were similar in activity in SHR and WKY rats. In intestinal brush-border membranes the activities of Ca2+-ATPase and alkaline phosphatase were lower in SHR rats when compared with WKY rats, whilst microvillus aminopeptidase (EC 3.4.11.2) (a marker for brush-border membranes) was unaffected. In kidney brush-border membranes all enzymes were similar in activity in SHR and WKY rats. The blood pressures of the SHR rats were considerably higher than those of the WKY rats. When SHR rats were fed on a Ca-deficient diet the activities of Na+K+-ATPase, Ca2+-ATPase and alkaline phosphatase in basolateral membranes and Ca2+-ATPase and alkaline phosphatase in brush-border membranes were all increased in the intestine when compared with SHR rats fed on a Ca-adequate diet. The equivalent enzymes in the kidneys of SHR rats, and the intestines and kidneys of WKY rats, were not affected by altering the Ca in the diet. The blood pressures of SHR rats fed on a Ca-deficient diet were higher than in those fed on a Ca-adequate diet. Blood pressures of WKY rats were not affected by altering the diet in this way. The results indicate that the absorption of Ca by active mechanisms may be reduced in SHR rats compared with WKY rats. Changing the level of Ca in the diet modified both blood pressure and the activities of enzymes which catalyse active Ca transport. The implications of these results to the aetiology, and possible nutritional treatment, of essential hypertension are discussed.

1. Lipid extracted from breast and leg muscle of 10-week-old turkeys was fractionated by preparative thin-layer chromatography and five individual ‘neutral’ and eight individual phospholipid fractions, representing 95% by weight of the extractable lipid, were recovered from the plates for analysis.

The ‘neutral’ lipids from breast and leg muscle consisted mainly of triglyceride (202–497 and 1644–2333 mg/100 g), together with cholesterol (74 and 103 mg/100 g), free fatty acid (27 and 123 mg/100 g), diglyceride (17 and 66 mg/100 g) and cholesterol ester (9 and 12 mg/100 g).

The phospholipids contained phosphatidylcholine (367 and 500 mg/100 g), phosphatidylethanolamine (157 and 279 mg/100 g), phosphatidylinositol (60 and 109 mg/100 g), sphingomyelin (43 and 62 mg/100 g), phosphatidylserine (31 and 61 mg/100 g), ‘cardiolipin’ (23 and 35 mg/100 g), lysophosphatidylcholine (9 and 11 mg/100 g) and ‘origin fraction’ (6 and 7 mg/100 g), accounting together (700 mg and 1070 mg/100 g for breast and leg muscle respectively) for 98% of the lipid phosphorus extracted.

Partial replacement of carbohydrate in the cereal-based diet by beef fat (2·5%) or anchovy oil (2·5 or 5·0%) had no effect on the amount of any of the lipid fractions, except for triglyceride, which varied considerably and was lowest in tissue from groups receiving 2·5% anchovy oil.

Magnesium deficiency may be induced by a diet impoverished in magnesium. This nutritional deficit promotes chronic inflammatory and oxidative stresses, hyperexcitability and, in mice, susceptibility to audiogenic seizures. Potentiation by low-magnesium concentrations of the opening of N-methyl-d-aspartate (NMDA) receptor/calcium channel in in vitro and ex vivo studies, and responsiveness to magnesium of in vivo brain injury states are now well established. By contrast, little or no specific attention has been, however, paid to the in vivo NMDA receptor function/excitability in magnesium deficiency. The present work reports for the first time that, in mice undergoing chronic nutritional deprivation in magnesium (35 v. 930 parts per million for 27 d in OF1 mice), NMDA-induced seizure threshold is significantly decreased (38 % of normal values). The attenuation in the drop of NMDA seizure threshold (percentage of reversal) was 58 and 20 % upon acute intraperitoneal administrations of magnesium chloride hexahydrate (28 mg magnesium/kg) and the antioxidant ebselen (20 mg/kg), respectively. In nutritionally magnesium-deprived animals, audiogenic seizures are completely prevented by these compound doses. Taken as a whole, our data emphasise that chronic magnesium deprivation in mice is a nutritional in vivo model for a lowered NMDA receptor activation threshold. This nutritional model responds remarkably to acute magnesium supply and moderately to acute antioxidant administration.

Body composition was measured in nine healthy, normal-weight, weight-stable subjects in three different research centres. In each centre the usual procedures for the measurements were followed. It revealed that the measurement procedures in the three centres were comparable. Body composition was measured in each centre between 09.00 and 13.00 hours after a light breakfast by densitometry (underwater weighing) and bio-electrical impedance. A single, total-body-water determination by D2O dilution was used as a reference value. Body fat determined by densitometry was significantly lower in one centre, which, however, could be completely explained by a lower body weight, probably due to water loss (the subjects refrained for a longer time from food and drinks before the measurements in that centre) and, thus, by violation of the assumptions of Siri's (1961) formula. Also, body impedance was slightly higher in that centre, indicating a lower amount of body water. Mean body fat from densitometry was also slightly lower in that centre compared with body fat determined by D2O dilution. Individual differences between body fat from densitometry and from total body water were relatively large, up to 7% body fat. The relationship between fat-free mass from densitometry and bio-electrical impedance was not different between the centres. It is concluded that differences in the relationship between body composition and bio-electrical impedance, as reported in the literature, may be due to differences in standardization procedures and/or differences in reference population.

The purpose of the present study was to test the hypothesis that the already well-established mitochondria1 lesion in fatty acid oxidation in riboflavin-deficient experimental animals, might be accompanied by an alteration in vivo in the kinetics of oxidation of labelled adipic acid. This dicarboxylic acid was chosen for testing as a metabolic probe because a block in its oxidation was already apparent from urine analysis of riboflavin-deficient animals, whereas the oxidation of medium- or long-chain monocarboxylic acids seemed to be little affected by deficiency in vivo. Female adult Norwegian hooded rats fed on purified diets containing either 15 mg riboflavin/kg diet (controls) or about 0.4 mg/kg (riboflavin-deficient) received an intragastric dose of either [l,6-I4C] adipic acid or [l-'4C] octanoic acid. Expired carbon dioxide was then collected in an alkaline trap over 3 h, for determination of radioactivity.This test was repeated at intervals for up to 2 weeks following riboflavin repletion of the deficient animals, and in riboflavin-dosed controls. Whereas the rate and extent of ['4C]octanoic acid oxidation was not significantly affected by the deficiency or repletion, the extent of [I4C]adipic acid oxidation was markedly and significantly increased during repletion of the deficient animals. The time-course indicated a temporary overshoot, followed by a slow return to the control values over 1–2 weeks. Adipate oxidation was also much less affected by a preceding period of overnight starvation, than was octanoate oxidation. Thus, adipic acid (or a related metabolic probe) may have appropriate properties for the design of a functional test of fatty acid oxidation efficiency, during riboflavin deficiency or allied metabolic conditions in human subjects.

1. The effects on body composition, measured by direct techniques, of a controlled 25% body-surfacearea thermal injury have been studied in two groups of forty male Wistar rats.

2. The extent of weight loss in the animals was directly related to their energy deficit resulting from a combination of injury, food intake and rate of wound healing.

3. Body fat proved the most labile source of tissue energy, decreasing to a minimum of approximately 30 g/kg body-weight.

4. Relationships between water and fat, and water and protein seen in control animals were not significantly different in the traumatized group.