The mink is a seasonal breeder with a propensity for seasonal fatness, and it is very responsive to changes in energy supply. The objectives of the present study were first, to validate a multi-species leptin assay for mink (Mustela vison) plasma, and second, to evaluate how plasma leptin and insulin concentrations responded to energy restriction and body-weight loss and refeeding with restoration of body reserves. The study was performed with six very fat yearling females (initial mean body weight 1451 (SD 119) g, i.e. approximately 300 g more than for a female in normal body condition). The animals were fed in restricted amounts (about 35 % metabolizable energy requirement for maintenance) in order to reach a very lean body condition. The target weight of 800 g was reached after about 1 month of restriction. The animals were then refed ad libitum until almost complete weight recovery. Blood samples were taken on days 1, 24, 34 (end of restriction), 44, 55 and 71 (end of experiment) and analysed for plasma concentrations of leptin and insulin. Three females were mated on day 44. Leptin and insulin concentrations mirrored each other and clearly reflected changes in body weight. Significant (P<0·001) Pearson correlation coefficients of 0·75 (leptin–insulin), 0·72 (leptin–body weight) and 0·59 (insulin–body weight) were found. Two of the three females that were mated gave birth to normal litters. It was concluded that the leptin assay yielded acceptable results for animals with body weight:fat content within the range investigated here, and that plasma leptin reflected body fat mass.

We previously described the effects of intake of dairy products on plasmid dissemination in the digestive tract of gnotobiotic mice associated with human faecal flora (HFF) and found that yoghurt, heat-treated yoghurt (HTY) and milk reduced population levels of transconjugants compared with findings in mice fed a standard mouse diet. In the case of lactose intake, transconjugants were not detected. The aim of the present study was to assess the possible interrelationships between these observations and other variables (bacterial ecology, pH, moisture, enzyme activities, short-chain fatty acid (SCFA) contents, lactic acid contents). Much of the interest of the present comparison lies in the fact that the animals were homogeneous in terms of age, gender, food and intestinal microflora, owing to the gnotobiotic mouse model maintained in sterile isolators. We observed no variation in SCFA and lactic acid contents or in the population levels of strictly anaerobic strains of Bacteroides and Bifidobacterium, and of the facultative anaerobic recipient Escherichia coli PG1 strain. The main modifications were the reduction of population levels of transconjugants in mice receiving yoghurt, HTY and milk, and concomitantly an increase of β-galactosidase and a decrease of β-glucosidase activities, compared with control mice fed a standard diet. Total inhibition of plasmid transfer was observed in HFF mice consuming lactose, and concomitantly the two enzyme activities (β-glucosidase and β-galactosidase) were increased, compared with the findings in control mice fed a standard diet. In axenic mice consuming lactose, plasmid transfer occurred, β-galactosidase was not detected and β-glucosidase was decreased. It is therefore proposed that these two enzyme activities influence plasmid transfer and persistence of transconjugants in the digestive tract of HFF associated mice. When both activities were increased there was a total inhibition of plasmid transfer (case of lactose intake). When β-galactosidase increased and β-glucosidase decreased (case of yoghurt, HTY and milk), plasmid transfer occurred at a lower efficiency than in the control group, resulting in lower population levels of transconjugants.

Niacin (nicotinic acid) in its non-physiological dose level is known to be an effective lipid-lowering agent; its potential risk as a therapeutic agent, however, has not been critically considered. Since niacin is excreted predominantly as methylated pyridones, requiring methionine as a methyl donor, the present study was undertaken to examine whether metabolism of the amino acid is altered in the presence of large doses of niacin. Male Sprague–Dawley rats were given a nutritionally adequate, semi-synthetic diet containing niacin at a level of either 400 or 1000 mg/kg diet (compared to 30 mg/kg in the control diet) for up to 3 months. Supplementation with niacin (1000 mg/kg diet) for 3 months resulted in a significant increase in plasma and urinary total homocysteine levels; this increase was further accentuated in the presence of a high methionine diet. The hyperhomocysteineaemia was accompanied by a significant decrease in plasma concentrations of vitamins B6 and B12, which are cofactors for the metabolism of homocysteine. The homocysteine-raising action of niacin, in particular, has an important toxicological implication, as hyperhomocysteineaemia is considered to be an independent risk factor for arterial occlusive disease. The niacin-associated change in homocysteine status may be an important limiting factor in the use of this vitamin as a lipid-lowering agent.

Vitamin C (ascorbic acid) is an essential nutrient that is involved in a number of cellular processes. However, unlike most mammals, man is unable to synthesize vitamin C and it must therefore be acquired from the diet. Absorption of vitamin C is achieved by two transporters, SVCT1 and SVCT2, recently cloned from rat and human kidney. SVCT1 is thought to be the predominant transporter in the intestine. Vitamin C supplements are increasingly common, thus contributing to an increased dietary load, and therefore the aim of the present study was to investigate the effect of high doses of ascorbic acid on SVCT1 expression. Using the Caco-2 TC7 cell model of small intestinal enterocytes, we measured the effects of ascorbic acid (4·5 mg/ml culture medium) on L-[14C]ascorbic acid uptake and SVCT1 expression (determined by reverse transcription-polymerase chain reaction). Ascorbic acid uptake was decreased significantly in Caco-2 TC7 cells exposed to ascorbate for 24 h (-50 %, P<0·0005). Expression of SVCT1 was also significantly reduced by exposure to elevated levels of ascorbate for 24 h (-77 %, P<0·005). Taken together these results suggest that high-dose supplements might not be the most efficient way of increasing the body pool of vitamin C.

Recent studies underline the importance of the immunoinflammatory processes in the pathology of Mg deficiency. Neutrophils possess a superoxide anion-generating NADPH oxidase and its inappropriate activation may result in tissue damage. The aim of the present study was to assess the effect of experimental Mg deficiency in the rat on polymorphonuclear leucocytes (PMN) activity and the role of increasing extracellular Mg. Weaning male Wistar rats were fed either a Mg-deficient or a control diet for 8 d. In Mg-deficient rats, the characteristic inflammatory response was accompanied by a marked increase in the number of PMN. Higher plasma interleukin 6 and NO concentrations and increased lipid peroxidation in the heart were found in Mg-deficient rats as compared with control rats. As shown by chemiluminescence studies, basal neutrophil activity from Mg-deficient rats was significantly elevated when compared with neutrophils from control rats. Moreover, the chemiluminescence of PMN from Mg-deficient rats was significantly higher than that of control rats following phorbol myristate acetate or opsonized zymosan activation. PMN from Mg-deficient rats also showed an increased activity of phagocytosis in comparison with neutrophils from control animals. Increasing extracellular Mg concentration in the incubating medium of PMN (0·8 v. 8·0 mM) decreased the chemiluminescence activity of PMN from control rats following opsonized zymosan activation. Chemiluminescence activities of PMN from Mg-deficient rats following phorbol myristate acetate or opsonized zymosan challenge were also decreased by high extracellular Mg concentration. From this work, it appears that PMN activation is an early consequence of Mg deficiency and that high extracellular Mg concentration inhibits free radicals generation.

The metabolizable energy content of low-digestible carbohydrates does not correspond with their true energy value. The aim of the present study was to determine the tolerance and effects of two polyols on digestion and energy expenditure in healthy men, as well as their digestible, metabolizable and net energy values. Nine healthy men were fed for 32 d periods a maintenance diet supplemented either with dextrose, Lycasin®HBC (Roquette Frères, Lestrem, France), or the hydrogenated polysaccharide fraction of Lycasin®HBC, at a level of 100 g DM/d in six equal doses per d according to a 3×3 Latin square design with three repetitions. After a 20 d progressive adaptation period, food intake was determined for 12 d using the duplicate meal method and faeces and urine were collected for 10 d for further analyses. Subjects spent 36 h in one of two open-circuit whole-body calorimeters with measurements during the last 24 h. Ingestion of the polyols did not cause severe digestive disorders, except excessive gas emission, and flatulence and gurgling in some subjects. The polyols induced significant increases in wet (+45 and +66 % respectively, P<0·01) and dry (+53 and +75 % respectively, P<0·002) stool weight, resulting in a 2 % decrease in dietary energy digestibility (P<0·001). They resulted also in significant increases in sleeping (+4·1 %, P<0·03) and daily energy expenditure (+2·7 and +2·9 % respectively, P<0·02) compared with dextrose ingestion. The apparent energy digestibility of the two polyols was 0·82 and 0·79 respectively, their metabolizable energy value averaged 14·1 kJ/g DM, and their net energy value averaged 10·8 kJ/g DM, that is, 35 % less than those of sucrose and starch.

Results of population studies suggest that black tea can reduce cardiovascular risk. Effects of black-tea polyphenols to reduce platelet aggregability may help to explain any benefits. Given that black tea is often consumed with and after meals, and man spends much of his life in the postprandial state, the objective of the present study was to investigate the acute effects of ingestion of black tea on postprandial platelet aggregation ex vivo. Twenty healthy participants had platelet aggregation and blood lipids assessed before and 4 h after the ingestion of 50 g dairy fat on two occasions in random order, corresponding to black tea or hot water. Black tea or hot water (one cup) was consumed immediately following the dairy fat, then after 1·5 and 3·0 h. Platelet aggregation ex vivo was assessed in platelet-rich plasma in response to three concentrations of collagen (0·2, 0·6, 2·0 μg/ml) and ADP (2, 4, 8 μM). Urinary concentrations of 4-O-methylgallic acid were used as an indicator that tea polyphenols were absorbed. Serum total cholesterol and triacylglycerol concentrations increased significantly 4 h after ingesting the dairy fat, but there was no significant difference between black tea and hot-water treatments on the cholesterol or triacylglycerol responses. Urinary 4-O-methylgallic acid concentrations were significantly increased following ingestion of black tea (P=0·0001) but not water. Black tea in comparison to hot water did not inhibit collagen or ADP-induced postprandial platelet aggregation. The results of this study do not support the suggestion that reduced postprandial platelet aggregability contributes to any benefits of black tea on cardiovascular risk.

Pancreatic islets were isolated from rats that had been nursed by dams fed with a control or an 8·7 % protein diet during the first 12 d of the lactation period. Glucose-induced insulin secretion from islets in the 8·7 % protein group was reduced 50 %. The islet insulin and DNA content were similar, whereas the pancreatic insulin content was reduced by 30 % in the rats fed 8·7 % protein. In order to elucidate the mechanism responsible for the attenuation of insulin secretion, measurements were performed of the activity of several islet enzymes that had previously been supposed to be involved in the coupling of glucose stimulation to insulin secretion. Islet glucose oxidation was unaffected, but glucose-stimulated hydrolysis of phosphatidylinositol was reduced by one-third in the islets of rats fed 8·7 % protein. The activity of mitochondrial glycerophosphate dehydrogenase was similar in islets of rats fed the 8·7 % protein diet and those fed the control diet. The activity of Ca-independent phospholipase A2 was increased fourfold in the islets of rats fed 8·7 % protein. It is concluded that impairment of glucose-induced insulin secretion in rats fed a low-protein diet may be caused by attenuation of islet phosphatidylinositol hydrolysis, and it is tentatively suggested that the increased activity of Ca-independent phospholipase A2 in islets of rats fed a low-protein diet may participate in the stimulation of apoptosis.

The purpose of the present study was to test whether hyperlipidaemia and insulin resistance in type 2 diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats can be improved by dietary supplementation with purified eicosapentaenoic acid (EPA) or oleic acid (OA). Male OLETF rats were fed powdered chow (510 g fat/kg) alone (n 8) or chow supplemented with 1·0 g EPA- (n 8) or OA- (n 8) rich oil/kg per d from 5 weeks until 30 weeks of age. An oral glucose tolerance test and hyperinsulinaemic euglycaemic clamp was performed at 25 and 30 weeks of age. EPA supplementation resulted in significantly (P<0.05) reduced plasma lipids, hepatic triacylglycerols, and abdominal fat deposits, and more efficient in vivo glucose disposal compared with OA supplementation and no supplementation. OA supplementation was associated with significantly increased insulin response to oral glucose compared with EPA supplementation and no supplementation. Inverse correlation was noted between glucose uptake and plasma triacylglycerol levels (r -0·86, P<0·001) and abdominal fat volume (r -0·80, P<0·001). The result of oral glucose tolerance test study showed that the rats fed EPA tended to improve glucose intolerance, although this was not statistically significant. Levels of plasma insulin at 60 min after glucose was significantly increased in rats fed OA compared with the other two groups. The results indicate that long-term feeding of EPA might be effective in preventing insulin resistance in diabetes-prone rats, at least in part, due to improving hypertriacylglycerolaemia.

The present study examined the consistency of bioelectrical impedance analysis (BIA) and anthropometric measurements in body composition analysis in a field study of prepubertal children using a representative group of 2286 5–7-year-old children from Kiel, north-west Germany. Body composition was assessed using anthropometric measures (A; four skinfolds) and BIA. Various published algorithms (according to and for A, , and for BIA and for a combined approach) were used to estimate body composition. Using A resulted in a sum of four skinfolds varying between age-dependent median values of 24·0 and 28·2 mm in boys and 30·5 and 33·3 mm in girls. When fat mass (FM) was calculated from A, age- and algorithm-dependent differences in median values were observed, with values varying between 8·5 and 14·6 % for boys and 11·1 and 14·9 % for girls. Using different algorithms ( v. ) only minor inconsistencies were observed. BIA-derived resistance index (height2/resistance) varied between 18·8 and 24·4 cm2/Ω for boys and 17·1 and 19·0 cm2/Ω for girls. Using four different algorithms to estimate FM from BIA data resulted in high intra-individual variances in percentage FM (from 13·8 to 33·4) as well as in the prevalence of overweight (from 14·7 to 98·4 % for boys and from 42·3 to 98·5 % for girls). Data obtained using the different BIA algorithms showed some, or even marked, inconsistencies as well as systematic deviations (an overestimation of FM at low percentage FM, v. ). When comparing BIA with A, BIA systematically overestimated FM. The differences between the results were influenced by BMI, gender and height. Considerable inconsistencies were observed at low BMI (<10th percentile) for girls and for small children. Although the within-observer as well as between-observer CV for both techniques are acceptable, we recommend caution in relation to the algorithms used for data analysis. The use of an interchange table of percentage FM derived from different algorithms for different percentile groups of skinfold thicknesses is recommended.

In clinical practice and epidemiological surveys, anthropometric measurements represent an important component of nutritional assessment in the elderly. The anthropometric standards derived from adult populations may not be appropriate for the elderly because of body composition changes occurring during ageing. Specific anthropometric reference data for the elderly are necessary. In the present study we investigated anthropometric characteristics and their relationship to gender and age in a cross-sectional sample of 3356 subjects, randomly selected from an elderly Italian population. In both sexes, weight and height significantly decreased with age while knee height did not. The BMI was significantly higher in women than in men (27·6 SD 5·7 V. 26·4 sd 3·7; P<0·001) and it was lower in the oldest than in the youngest subjects (P<0·05) of both genders. The 75th year of age was a turning point for BMI as for other anthropometric measurements. According to BMI values, the prevalence of malnutrition was lower than 5 % in both genders, whereas obesity was shown to have a higher prevalence in women than in men (28 % v. 16 %; P<0·001). Waist circumference and waist : hip ratio values were higher for the youngest men than for the oldest men (P<0·05), whereas in women the waist : hip ratio values were higher in the oldest women, suggesting that visceral redistribution in old age predominantly affects females. In conclusion, in the elderly the oldest subjects showed a thinner body frame than the youngest of both genders, and there was a more marked fat redistribution in women.