Total body water (TBW) was measured by deuterium oxide (D2O) dilution and predicted from bioelectrical impedance (Z) in nineteen anorexic and twenty-seven control women. The equation of Kushner et al. (1992) based on the impedance index (ZI = height2/Z) gave biases of 0.9 (sd 2.5) and 0.8 (sd 2.5) litres in controls and patients respectively (NS, ANOVA). The ZI-based equation of Deurenberg et al. (1993) gave biases of 1.5 (sd 2.4) litres (NS) and 3.0 (sd 2.1) litres (P <0.001) in controls and patients respectively. Despite the fact that weight was the most powerful predictor of TBW on the study sample (n 46, r2 0.90, P < 0.0001, se of the estimate 1.6 litres, CV 5.7%), the formulas of Segal et al. (1991) and Kushner et al. (1992) based on the association of weight and ZI gave an inaccurate prediction of TBW in both control and anorexic subjects, with a bias ranging from -3.2 (sd 2.4) to 2.9 (sd 2.1) litres (P ≤0.001). Population-specific formulas based on ZI (n 46) gave a more accurate prediction of TBW by bioelectrical impedance analysis on the study subjects, with biases of -0.1 (sd 1.8) and 0.5 (sd 1.7) litres in controls and patients respectively (NS). However, the individual bias was sometimes high. It is concluded that bioelectrical impedance analysis can be used to predict TBW in anorexic women at a population level, but the predictions are less good than those based on body weight alone.

Concern has been expressed that high dietary concentrations of non-milk extrinsic sugars (NMES) may potentially compromise nutrient intakes in population groups with low energy intakes (Department of Health, 1991). The objective of the present study was to examine data from the National Diet and Nutrition Survey of Children Aged 1.5 to 4.5 years (Gregory et al. 1995) for evidence of an inverse association between energy from NMES and micronutrient intakes, and if possible to quantify a level of NMES-energy at which micronutrient intakes may, theoretically, be compromised. Energy and nutrient intakes were compared across quintiles of NMES-energy for boys (n 848) and girls (n 827). As the concentration of NMES increased, energy intake rose (in boys only) while percentage energy from fat fell from 40 to 32 % across quintiles 1 to 5. Intakes of most micronutrients also fell, while intakes of vitamin C rose. Mean intakes of most micronutrients (Ca, thiamin, riboflavin, niacin, folate and vitamin C) were adequate in comparison with dietary reference values. However, intakes of Fe, Zn and vitamin D were low at all levels of NMES-energy and fell below the estimated average requirement for Fe and Zn for NMES concentrations exceeding 24 % of energy. Lower intakes of milk, meat, bread and vegetables, and higher intakes of fruit juice largely explain the observed trends in micronutrient intake. It is concluded that the inverse association of NMES with micronutrient intakes is of most significance for the 20 % of children with diets highest in NMES. However, further work is required to establish whether the associations observed have biological significance with regard to micronutrient status.

Exchangeable pools of Se after an intravenous injection of 74Se-enriched isotope as sodium selenite were measured in two groups (n 9) of elderly women (free-living aged 64–82 years and institutionalized aged 68–82 years), and a comparison group (n 9) of young women aged 31–40 years to evaluate the effect of age and institutionalization on Se reserves. Dietary Se intake was not different among the three groups. Plasma Se and glutathione peroxidase (EC 1.11.1.9) levels were significantly lower in the institutionalized elderly women (P < 0.05). In each of the three groups, two pools were determined from our model. The size of the first pool and the sum of the two pools were lower in the group of institutionalized elderly women than in the other two groups. The significant correlation between plasma Se level and total Se pool size (r 0.66, P < 0.01) indicated that this last variable could serve as a new marker of Se status. Finally, these data suggest that the Se status of elderly women is more related to lifestyle, in terms of institutionalization or not, than to age per se.

A model is proposed that allows study of the short-term dynamics of gas exchanges (and heat production) in large open-circuit respiration chambers. The model describes changes in [O2] and [CO2] in the respiration chamber by a series of differential equations based on animal metabolism and physical characteristics of gas exchange. The model structure was similar for O2 and CO2, although model parameters differed. A constant level of O2 consumption (and CO2 production) was assumed for resting animals which was different for fed and fasted animals. The adaptation from a fed to a fasting state was described as a first-order process. Physical activity (standing or sitting) was recorded and was included in the model as a constant. Thermic effect of feed comprised the O2 consumption and CO2 production related to several relatively rapidly occurring processes after ingestion of a meal (e.g. ingestion, digestion or absorption). In the model, these processes were pooled into a single phenomenon. Model parameters were obtained statistically by comparing model predictions (based on the numerically integrated differential equations) with the observed [O2] and [CO2]. The model was evaluated by studying gas exchanges in growing pigs that were fasted for 31 h and re-fed a single meal thereafter. The model fitted the data well over the 47 h measurement range. Traditional methods in which heat production is calculated suffer from noisy data when the interval between observations becomes too short. The proposed method circumvents this by modelling the observed concentration of gases in the respiration chamber rather than the calculated heat production.

The relationships of N input or protein status and the concentrations of serum insulin-like growth factor-1 (IGF-1), plasma fibronectin (FN) and total protein (TP) were examined in three experiments with steers and sheep nourished by intragastric infusion of nutrients. In Expt 1, three steers (340 kg live weight) were infused with three levels of volatile fatty acids (0, 300 and 600 kJ/kg metabolic weight (W0.75) per d) and six levels of casein (0, 200, 400, 650, 1500 and 2500 mg N/kg W0.75 per d). Each N treatment was imposed for 5 d. In Expts 2 and 3, five groups of sheep (about 35 kg live weight) were infused with casein at 500 mg N/kg W0.75 per d for 2 weeks followed by 1500, 500 or 50 mg N/kg W0.75 per d in Expt 2, and in Expt 3, with 100 mg N/kg W0.75 per d for 6 weeks or 10 mg N/kg W0.75 per d for 4 weeks. Non-protein energy was maintained constant at 500 kJ/kg W0.75 per d throughout. Daily N balance and total body N content at the end were measured, and protein status was defined as a percentage of cumulative N accretion or depletion in relation to the total body N content at maintenance. It was found that IGF-1 and FN responded rapidly and substantially to altered N input, and that when daily N input was maintained constantly at sub-maintenance, their continuous declines were related closely to progressive protein depletion in the sheep. Plasma TP concentration was independent of N input when N input was altered acutely in the steers, but declined significantly and gradually with severe, chronic body protein depletion in the sheep. Plasma content of TP in the sheep however reduced acutely with a reduction in N input. Plasma volume fell substantially over the first 2 weeks of protein depletion, compensating for the declines in TP content and maintaining TP concentration plateau. The possible implications of the changes in TP concentration and content (concentration x volume) to body protein loss in sheep are discussed.

Daily secretion of pancreatic juice, including postprandial responses to food, was investigated in two groups of calves: preruminant (fed with liquid food) and ruminant (fed with solid food). Male Friesian calves (1 week old and 6 weeks old) were surgically fitted with a pancreatic duct catheter, duodenal cannula and two duodenal electrodes. Continuous 24 h collections of pancreatic juice and myoelectrical recordings were performed with minimal restraint and disturbance of animals. In both groups of calves clear periodic fluctuations in pancreatic juice secretion (volume, protein output and trypsin activity) coinciding with duodenal migrating myoelectric complexes (MMC) were recorded. Secretion of juice per cycle and per day was greater in ruminant calves, but the frequency and amplitude of cycles were lower in this group. There were no differences between day and night-time preprandial pancreatic cycles and duodenal MMC in preruminant calves, whilst in ruminant calves, evening MMC were longer than morning MMC. The pancreatic cephalic phase (increase of volume flow, protein output and trypsin activity during and just after food intake) was significant only in preruminant calves following morning feeding. Postprandial pancreatic cycles did not differ from preprandial cycles, except the pancreatic cycle (juice volume and trypsin activity) in which food was offered in preruminant calves. No gastric or intestinal phase was observed in either group of calves. In conclusion, biological cycles of the gastrointestinal tract are present in both preruminant and ruminant calves, and these cycles evolve along with the change from liquid to solid food.

Following 10 weeks of fat supplementation a group of aerobically trained thoroughbred horses exhibited a significant decrease in postprandial plasma triacylglycerol concentration. This decrease was associated with a mean 50% increase in plasma total lipase activity following pentosan polysulfate administration and an increase in postprandial plasma cholesterol concentration. A significant increase in the activity of muscle citrate synthase (EC 4.1.3.7), expressed as a ratio to the total fractional area occupied by type I and type IIa muscle fibres, was also observed. No significant change in the concentration of resting muscle glycogen or triacylglycerol occurred as a result of fat supplementation. These results suggest that there was improved management of the fat load and that the triacylglycerol-clearing capacity of the horses was increased as a result of fat supplementation. It is suggested that the increase in plasma total lipase activity following pentosan polysulfate administration may have reflected an increase in muscle lipoprotein lipase (EC 3.1.1.34) activity, which would increase the capacity of muscle for free fatty acid uptake from circulating triacylglycerol-rich plasma lipoproteins. Fat supplementation may also enhance the oxidative capacity of muscle, as suggested by the significant increase in muscle citrate synthase and the trend towards an increase in β-hydroxyacyl CoA dehydrogenase (EC 1.1.1.35) following 10 weeks of fat supplementation.

A 120 d feeding study with adult rats was conducted to evaluate the influence of two protein sources (casein and gelatin), two protein levels (50 and 300g/kg diet) and two fat levels (50 and 150g/kg diet) on serum lipids (total cholesterol, HDL-cholesterol and triacylglycerols) and liver polyunsaturated fatty acid levels. In general, the concentrations of serum triacylglycerols and total cholesterol and liver phospholipid levels of arachidonic acid (AA) and docosahexaenoic acid (DHA) were higher in rats fed on casein diets compared with those fed on the gelatin diets. These effects were more pronounced in rats fed on the high-casein (300 g/kg)-high-fat (150g/kg) diet. Gelatin was hypocholesterolaemic and also suppressed the liver phospholipid levels of AA and DHA (reported for the first time). The difference in the amino acid composition between casein and gelatin may be responsible for the observed effects. Casein contains higher levels of glutamic acid, methionine, phenylalanine and tyrosine, while gelatin contains higher levels of arginine, glycine and hydroxyproline. It is suggested that a protein source which increases serum cholesterol may also increase the concentrations of AA and DHA in rat tissues.

It is believed that frequent Fe doses decrease the efficiency of absorption as a consequence of the loading of intestinal mucosal cells with Fe from the previous supplemental dose. We examined this premise in thirty anaemic Sprague-Dawley rats given Fe supplements as FeSO4 in 1 g preparations of a 50:50 (w/w mixture of low-Fe diet and sucrose under one of the following regimens: one 3 mg Fe dose daily for 3d, four 0.75mg doses daily at 6h intervals for 3d, and one 9mg dose on day 1 followed by two placebo (low-Fe diet) doses on days 2 and 3. All groups were fed on two low-Fe meals daily (8.3 mg Fe/kg diet). After an overnight fast rats were dosed with 1 ml of an 59Fe-labelled ferric nitrilotriacetic acid solution (37 kBq 59Fe, 50 μg Fe) orally and killed 10 h later. Absorption of 59Fe was measured as the percentage of the 59Fe retained by the carcass without the gastrointestinal tract 10 h after dosing relative to the initial 59Fe dose. Haemoglobin-Fe gain, liver non-haem-Fe, and mucosal duodenal ferritin were determined after the 3 d supplementation period. Absorption of the test dose in rats supplemented once 3 d before assessment of Fe absorption was 2.6-fold greater than those supplemented with daily single doses and 1.9-fold greater than those supplemented with daily multiple doses. Our data indicate that both mucosal ferritin and liver Fe levels account for the higher absorption efficiency found in rats supplemented once to simulate intermittent regimens.

Cholesterol oxidation products (COP) have been reported to influence vital cellular processes such as cell growth, cell proliferation, membrane function and de novo sterol biosynthesis. The objectives of the present study were: (1) to develop an in vitro model using newborn rat kidney (NRK) cells to investigate the actions of COP; (2) to investigate the effect of COP on cell viability, endogenous antioxidant enzymes activities, i.e. superoxide dismutase (EC 1.15.1.1; SOD) and catalase (EC 1.11.1.6; CAT), and the extent of lipid peroxidation in this model; (3) to determine whether the addition of 100–1000 nm-α-tocopherol, β-carotene or butylated hydroxytoluene (BHT) could protect against COP-induced cytotoxicity. NRK cells were cultured in the presence of various concentrations (5–50 μM) of cholesterol or cholestan-3β,5α,6β-triol (cholestantriol) for a period of 24 h. Cholesterol over the range 5–50 μM did not induce cytotoxicity as indicated by the neutral-red-uptake assay or the lactate dehydrogenase (EC 1.1.1.27)-release assay. However, cell viability was compromised by the addition of > 10 μM-cholestantriol (P < 0.05). The addition of β-carotene (100–1000 nM) did not increase cell viability significantly in cholestantriol-supplemented cells. However, the addition of α-tocopherol (1000 nM) and BHT (1000 nM) significantly increased percentage cell viability above that of the cholestantriol-supplemented cells but not back to control levels. SOD and CAT activities in NRK cells significantly decreased (P < 0.05) following incubation with cholestantriol. The addition of > 750 nM-α-tocopherol, β-carotene or BHT returned SOD and CAT activities to that of the control. Lipid peroxidation was significantly induced (P < 0.05) in the presence of cholestantriol. Supplementation of the cells with α-tocopherol (250, 500 or 1000 nM) or BHT (750 or 1000 nM) resulted in a reduction in the extent of lipid peroxidation (P < 0.05). The addition of β-carotene over the concentration range of 250–1000 nM did not reduce lipid peroxidation significantly compared with cells exposed to cholestantriol alone. These findings suggest that addition of exogenous antioxidants may be beneficial in the prevention of COP-induced toxicity in vitro.

Since experimental Se deficiency results in a significant increase in plasma cholesterol concentration the present investigation was undertaken to assess further the influence of this deficiency on the expression of proteins involved in hepatic lipid metabolism. Se deficiency was induced by feeding weanling male Wistar rats on a deficient diet for 6 weeks. Hypercholesterolaemia associated with Se deficiency was related to increased 3-hydroxy-3-methylglutaryl-coA (HMG-CoA) reductase (EC 1.1.1.34) activity in liver microsomes as compared with control animals. Hepatic lipoprotein receptor levels (LDL-receptor and HDL-binding proteins, HB1 and HB2) were not significantly affected by Se deficiency, as assessed by immunoblotting. Plasma triacylglycerol concentrations tended to decrease in Se-deficient rats in concert with their reduced post-Triton secretion. There was no significant effect of Se deficiency on the hepatic synthesis of apolipoproteins. These results point to the need for further investigations into the mechanism related to the increased activity of HMG-CoA reductase and the enhanced cholesterogenesis in the liver of Se-deficient rats likely to result from this.

Selenium: Cholesterol: Triacylglycerol: HMG-CoA reductase

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.