Severe malnutrition is widely distributed throughout the world, showing a high prevalence in developing countries. Experimental animal models have been useful to study the effects of malnutrition at different levels and ages. Apoptosis is a well recognised process of cell death occurring under several physiological and pathological conditions. It represents the principal mechanism involved in cell selection in the thymus. Thymocyte apoptosis induction by dexamethasone is one of the best characterised experimental models of programmed cell death. The aim of the present study was to determine whether severe malnutrition increased spontaneous and/or dexamethasone-induced apoptosis in vivo in thymocytes of experimentally malnourished rats during lactation. Thymocytes were obtained from malnourished rats at weaning (21d of age). Apoptosis frequency was estimated by the terminal transferase-mediated dUTP nick end labelling assay. Spontaneous apoptosis was 1·9 (SD 1·0) % IN WELL NOURISHED RATS IN CONTRAST TO 13·3 (sd 3·8) % in malnourished animals; this is seven times greater (P<0·001). Interestingly, the frequency of dexamethasone-induced apoptosis was similar in both groups of animals (47·9 (sd 10·1) % in well nourished rats and 53·8 (sd 8·0) % in malnourished rats). The results obtained in the present study indicate that malnutrition is associated with a significant increase of spontaneously apoptotic cells. In addition, the data showed that the fraction of thymocytes susceptible to dexamethasone-induced apoptosis was similar in well nourished and malnourished animals. The greater levels of spontaneously apoptotic cells associated with malnutrition could be related to alterations of the microenvironment of the thymus and/or to an obstruction of early thymocyte maturation.

Elevated plasma total homocysteine is considered to be a graded risk factor for cardiovascular disease. Folate, through its homocysteine-lowering potential, may therefore be protective. Folate, however, may have protective effects independent of homocysteine-lowering. We have measured the effects of folate on Cu-catalysed oxidative damage to the unsaturated lipids in human LDL. Experiments were carried out in the presence of citrate, and followed increases in absorption at 234 nm, which measures the amount of conjugated diene produced. There is a lag time during which endogenous antioxidants are oxidised, followed by rapid oxidation of lipid. Addition of 0–6 μM-5-methyltetrahydrofolate produced a dose-dependent increase in the lag time, suggesting that folate may have a direct anti-oxidant role in vivo, which is independent of any indirect effects through lowering of homocysteine levels.

As the number of older people is growing rapidly worldwide and the fact that elderly people are also apparently living longer, dementia, the most common cause of cognitive impairment is getting to be a greater public health problem. Nutrition plays a role in the ageing process, but there is still a lack of knowledge about nutrition-related risk factors in cognitive impairment. Research in this area has been intensive during the last decade, and results indicate that subclinical deficiency in essential nutrients (antioxidants such as vitamins C, E and β-carotene, vitamin B12, vitamin B6, folate) and nutrition-related disorders, as hypercholesterolaemia, hypertriacylglycerolaemia, hypertension, and diabetes could be some of the nutrition-related risk factors, which can be present for a long time before cognitive impairment becomes evident. Large-scale clinical trials in high-risk populations are needed to determine whether lowering blood homocysteine levels reduces the risk of cognitive impairment and may delay the clinical onset of dementia and perhaps of Alzheimer's disease. A curative treatment of cognitive impairment, especially Alzheimer's disease, is currently impossible. Actual drug therapy, if started early enough, may slow down the progression of the disease. Longitudinal studies are required in order to establish the possible link of nutrient intake – nutritional status with cognitive impairment, and if it is possible, in fact, to inhibit or delay the onset of dementia.

One of the objectives of the present study was to investigate whether 1 % conjugated linoleic acid (CLA) in the diet reduced tumour incidence in the colon of 1,2-dimethylhydrazine (DMH)-treated rats. Colon cancer was induced by injecting 6-week-old, male, Sprague–Dawley rats with 15 mg/kg DMH twice per week for 6 weeks. They were fed either 1 % CLA or a control diet ad libitum for 30 weeks. Dietary CLA significantly decreased colon tumour incidence (P<0·05). Our second objective was to investigate whether apoptosis in the colon mucosa of DMH-treated rats was affected by the amount of dietary CLA and whether the changes in apoptosis were related to those in fatty acid-responsive biomarkers. For this purpose, rats were killed after being fed a diet containing 0 %, 0·5 %, 1 % or 1·5 % CLA for 14 weeks. CLA was undetected in the mucosa of rats fed the 0 % CLA diet and increased to 5·9 mg/g phospholipid in rats fed the 0·5 % diet. The apoptotic index estimated by the terminal deoxynucleotidyl transferase-mediated dUTP nick and labelling technique was increased by 251 % and the 1,2-diacylglycerol content was decreased by 57 % in rats fed 0·5 % CLA. No further changes in these variables were observed when CLA in the diet was raised to 1·0 % or 1·5 %. However, dietary CLA decreased mucosal levels of prostaglandin E2, thromboxane B2 and arachidonic acid in a dose-dependent manner. The present data indicate that dietary CLA can inhibit DMH-induced colon carcinogenesis by mechanisms probably involving increased apoptosis.

Studies of Fe absorption in pregnancy often make unfounded assumptions of erythrocyte incorporation. Therefore, we measured the absorption and utilisation of Fe during early and late pregnancy by the erythrocyte incorporation of two stable isotopes. 8·5 mg 57Fe (oral) and 0·5 mg 58Fe (intravenous) were given to five non-pregnant women, to five women in early gestation (12 weeks) and five women in late gestation (36 weeks). The stable isotope ratios in whole blood 14 d later were measured by MS. Together with estimation of body Fe mass, this enabled the calculation of Fe absorption and erythrocyte incorporation. In non-pregnant women, Fe absorption averaged 20·3 (range 10·2–34·3) %. It was not significantly different in early pregnancy (11·8 (range, 4·4–24·8) %), but during late pregnancy Fe absorption increased to 59·0 (range 38·2–77·2) %. All non-pregnant and early-pregnancy subjects had normal Fe status, but two women in late pregnancy had evidence of Fe insufficiency. During early and late pregnancy, mean erythrocyte incorporation was 63·4 (SD 12·1) % AND 71·0 (sd 10·4) % respectively, significantly reduced (P=0·003) compared with non-pregnant subjects (90·1 (sd 6·0) %). Decreased erythrocyte incorporation of absorbed Fe in early pregnancy is compatible with reduced Fe demand and low oral absorption. However, during late pregnancy decreased erythrocyte incorporation associated with high absorption and Fe insufficiency is different from the high erythrocyte incorporation which occurs in non-pregnant Fe-deficient women. This suggests that part of the aetiology of Fe deficiency during pregnancy may be the reduction of Fe utilisation.

A low resting metabolic rate (RMR) has been proposed as a possible cause for the increased body fat commonly seen in women compared with men. Absolute RMR is higher in men, but whether RMR adjusted for lean body mass (LBM) remains higher is unresolved. The objective of the present study was to determine whether RMR adjusted for various body composition factors differed between healthy adult men and women. Thirty men (28·3±8·0 years, BMI 23·7±2·1 kg/m2) and twenty-eight women (28·7±6·9 years, BMI 22·2±1·9 kg/m2) were included in the analyses. RMR was measured by open-circuit indirect calorimetry for 60 min. Extracellular water (ECW) was measured by corrected Br- space and total body water (TBW) by 2H dilution. LBM was estimated as TBW/0·732. Intracellular water (ICW) was calculated as TBW-ECW, and body cell mass (BCM) as ICW/0·732. Men were heavier and had higher BMI, LBM, BCM and ECW, but less fat mass. Absolute RMR was higher in men than women (7280±844 v. 5485±537 kJ/d, P<0·0001). This difference became non-significant when RMR was adjusted for LBM by ANCOVA (6536±630 v. 6282±641 kJ/d, P=0·2191), but remained significant when adjusted for BCM (6680±744 v. 6128±756 kJ/d, P=0·0249). Fat mass explained a significant amount of variation in RMR in women (r2 0·28, P=0·0038), but not in men (r2 0·03, P=0·3301). The relationships between body fat and the various subcompartments of BCM and RMR require further elucidation.

The present study evaluates the influence of different amounts of fat added to starch on postprandial glucose metabolism (exogenous and endogenous). Nine women (24 (SE 2) YEARS OLD, BMI 20·4 (se 0·7) kg/m2) ingested 1 week apart 75 g glucose equivalent of 13C-labelled starch in the form of pasta without (low fat; LF) or with 15 (medium fat; MF) or 40 (high fat; HF) g sunflower oil. During the 7 h following meal consumption, plasma glucose, non-esterified fatty acids, triacylglycerols (TG) and insulin concentrations, and endogenous (using [6,6-2H2]glucose) and exogenous glucose turnover were determined. With MF and HF meals, a lower postprandial glucose peak was observed, but with a secondary recovery. A decrease in exogenous glucose appearance explained lower glycaemia in HF. At 4 h after the HF meal the insulin, insulin:glucose and postprandial blood TG were higher than those measured after the LF and MF meals. Despite higher insulinaemia, total glucose disappearance was similar and endogenous glucose production was suppressed less than after the LF and MF meals, suggesting insulin resistance. Thus, the addition of a large amount of fat appears to be unfavourable to glucose metabolism because it leads to a feature of insulin resistance. On the contrary, the MF meal did not have these adverse effects, but it was able to decrease the initial glycaemic peak.

Delayed peak response of plasma retinyl esters (RE) relative to plasma triacylglycerols (TAG) and apolipoprotein (Apo) B-48 responses following a fat load supplemented with vitamin A raised doubts about the use of vitamin A to label dietary-derived lipids and lipoproteins. The present study compared the use of water-miscible and oil-soluble retinyl palmitate (RP) as markers of dietary-derived lipoproteins in healthy subjects along with the measurements of postprandial plasma TAG and ApoB-48 responses to investigate whether the delayed peak response observed was due to delayed intestinal output of RE from oil-based solutions. Nine healthy female subjects were given a standard test meal containing a dose (112 mg) of RP in either water-miscible or oil-soluble form in random order, on two separate occasions after a 12 h overnight fast. The results showed that the mean plasma RE concentrations reached a peak significantly later than mean plasma TAG and ApoB-48 concentrations when oil-soluble RP was consumed, whereas plasma RE peaked earlier relative to plasma TAG and ApoB-48 responses when water-miscible RP was used. The results suggested a more rapid absorption with a significantly higher and earlier peak response of plasma RE when water-miscible RP was consumed. This was in contrast to the delayed initial appearance and later sustained higher concentrations of plasma RE during the late postprandial period when oil-soluble RP was consumed. The RE response to the water-miscible RP showed better concordance with plasma TAG response than that of oil-soluble RP.

Conventional models of energy utilization by animals, based on partitioning metabolizable energy (ME) intake or net energy (NE), are reviewed. The limitations of these methods are discussed, including various experimental, analytical and conceptual problems. Variation in the marginal efficiency of utilizing energy can be attributed to various factors: diet nutrient composition; animal effects on diet ME content; diet and animal effects on ME for maintenance (MEm); experimental methodology; and important statistical issues. ME partitioning can account for some of the variation due to animal factors, but not that related to nutrient source. In addition to many of the problems associated with ME, problems with NE pertain to: estimation of NE for maintenance (NEm); experimental and analytical methodology; and an inability to reflect variation in the metabolic use of NE. A conceptual framework is described for a new model of energy utilization by animals, based on representing explicit flows of the main nutrients and the important biochemical and biological transformations associated with their utilization. Differences in energetic efficiency from either dietary or animal factors can be predicted with this model.

Incorporation of coconut oil (CO) rich in lauric acid into the milk diet induces a lipid infiltration of the liver (steatosis) in 1-month-old calves. Among possible steps involved in diet-induced liver steatosis, the ability of the calf liver to synthesize apolipoprotein (Apo) B and to secrete it as part of VLDL particles was investigated. Liver samples were taken from calves fed for 17 d on a conventional milk replacer containing CO (n 5) and beef tallow (BT, n 4) as reference. Samples were cut into slices 0·5 mm thick and subsequently incubated for 12 h in a medium containing a [35S]methionine–[35S]cysteine mix and 0·8 mM-sodium laurate or oleate, the major fatty acids of CO and BT diets respectively. Concentrations of total [35S]proteins, [35S]albumin and [35S]ApoB in liver cells were 2-fold lower (P=0·08, 0·0004 and 0·03 respectively) in CO- than in BT-fed calves. Although the total amount of proteins secreted (including albumin) was similar in both groups of calves, the amount of VLDL-[35S]Apo secreted was 2-fold lower (P=0·004) in CO- than in BT-fed calves. These results suggest that a CO-enriched milk diet induces in preruminant calves a lipid infiltration of the liver by decreasing ApoB synthesis, leading to a reduction in secretion of VLDL particles.

The aim of the present work was to estimate volatile fatty acid (VFA) production rate in the rumen of sheep fed two levels of intake using both a tracer (TM; by isotope dilution) and a non-tracer method (NTM; by supplementary infusion) in steady-state conditions. Six wethers received a diet containing 700 g lucerne hay and 300 g ground maize/kg in eight equal meals at 3 h intervals per d. The diet (9·8 MJ metabolizable energy (ME)/kg DM) was offered at 90 % ad libitum consumption (high intake, HI) or 45 % ad libitum consumption (low intake, LI) in a 2×2 crossover design. Each sheep received five intrarumen VFA solutions infused continuously for 24 h at rates of 250 ml and 165 ml/h for the HI and LI respectively. The first infusion, considered as a control treatment (Con), consisted of a solution of [1-13C]propionate (7 mmol/d). The four other solutions were isoenergetic (1·9 MJ ME/kg DM intake) mixtures of unlabelled propionate (C3) and butyrate (C4) at different levels: 0·90 mol C4/kg DM intake; 0·60 mol C4+0·45 mol C3/kg DM intake; 0·30 mol C4+0·90 mol C3/kg DM intake; 1·35 mol C3/kg DM intake. The VFA infusions did not affect rumen fermentation of the basal diet (pH, osmotic pressure, protozoa numbers), and comparable DM digestibility of the diet among the different treatments was observed. Both estimation methods demonstrated a similar increase (1·7-fold) in the rumen VFA production rate of sheep fed at intakes varying between 0·9 to 1·7 times maintenance. Irrespective of the intake level, the rumen production rate of individual VFA was on average 1·5-fold higher when estimated by the TM compared with the NTM. Rumen VFA production rates estimated by the NTM and TM represented 80 % and 120 % ME intake respectively. The difference between NTM and TM estimates seems likely to be caused mainly by overestimation of the VFA production rates by the TM.

During the past few years, there have been two major developments, if not revolutions, in the field of energy balance and weight regulation. The first at the molecular level, which was catalysed by developments in DNA screening technology together with the mapping of the human genome, has been the tremendous advances made in the identification of molecules that play a role in the control of food intake and metabolic rate. The second, at the systemic level, which centered upon the use of modern technologies or more robust analytical techniques for assessing human energy expenditure in response to starvation and overfeeding, has been the publication of several papers providing strong evidence that adaptive thermogenesis plays a much more important role in the regulation of body weight and body composition than previously thought. Within these same few years, several new members of the mitochondrial carrier protein family have been identified in a variety of tissues and organs. All apparently possess uncoupling properties in genetically-modified systems, with two of them (uncoupling protein (UCP) 2 and UCP3) being expressed in adipose tissues and skeletal muscles, which are generally recognised as important sites for variations in thermogenesis and/or in substrate oxidation. Considered as breakthrough discoveries, the cloning of these genes has generated considerable optimism for rapid advances in our molecular understanding of adaptive thermogenesis, and for the identification of new targets for pharmacological management of obesity and cachexia. The present paper traces first, from a historical perspective, the landmark events in the field of thermogenesis that led to the identification of these genes encoding candidate UCP, and then addresses the controversies and on-going debate about their physiological importance in adaptive thermogenesis, in lipid oxidation or in oxidative stress. The general conclusion is that UCP2 and UCP3 may have distinct primary functions, with UCP3 implicated in regulating the flux of lipid substrates across the mitochondria and UCP2 in the control of mitochondrial generation of reactive oxygen species. The distinct functions of these two UCP1 homologues have been incorporated in a conceptual model to illustrate how UCP2 and UCP3 may act in concert in the overall regulation of lipid oxidation concomitant to the prevention of lipid-induced oxidative damage.

The nutritional and physiological modifications that occur during the weaning period induce adaptations of tissue metabolism in all mammal species. Among the adaptations due to weaning in ruminants, the regulation of lipoprotein lipase (LPL) activity, one of the rate-limiting steps of fatty acid utilization by tissues, was still unknown. The present study aimed at comparing LPL activity and gene expression in the heart, seven skeletal muscles and three adipose tissue sites between two groups of seven preruminant (PR) or ruminant (R) calves having a similar age (170 d), similar empty body weight (194 kg) at slaughter, and similar net energy intake from birth onwards. Triacylglycerol content of adipose tissues was 16 % lower in R than in PR calves, (P<0·01). This could be partly the result from a lower LPL activity (-57 %, P<0·01). LPL mRNA levels were also lower in R calves (-48 % to -68 %, P<0·01) suggesting a pretranslational regulation of LPL activity. Activity and mRNA levels of LPL did not differ significantly in the heart and skeletal muscles except in the masseter in which LPL activity and mRNA levels were higher (+50 % and +120 % respectively, P<0·01) in the R calves. Regulation of LPL in masseter could be explained by the high contractile activity of this muscle after weaning due to solid food chewing. In conclusion, weaning in the calf affects LPL activity and expression in adipose tissues, but not in skeletal muscles except the masseter.