Raised plasma triacylglycerol is an independent risk factor for cardiovascular disease, and an understanding of factors which regulate the synthesis and degradation of lipoproteins which carry triacylglycerol in the blood may lead to novel approaches to the treatment of hypertriacylglycerolaemia. An active microsomal triacylglycerol transfer protein (MTP) is essential for the assembly of particles which transport triacylglycerol through the circulation. After absorption in the intestine, dietary fat and fat-soluble vitamins are incorporated into chylomicrons in the intestinal epithelial cells, and these lipoproteins reach the bloodstream via the lymphatic system. Patients with the rare genetic disorder, abetalipoproteinaemia, in which MTP activity is absent, present clinically with fat-soluble vitamin and essential fatty acid deficiency, indicating a key role for MTP in the movement of fat into the body. The triacylglycerol-rich lipoprotein found in fasting blood, VLDL, is assembled in the liver by an MTP-dependent process similar to chylomicron assembly, and transports triacylglycerol to extra-hepatic tissues such as adipose tissue and heart. In the absence of MTP activity, VLDL are not synthesized and only extremely low levels of triacylglycerol are present in the blood. Dietary components, including fat, cholesterol and ethanol, can modify the expression of the MTP gene and, hence, MTP activity. The present review summarizes current knowledge of the role of MTP in the assembly and secretion of triacylglycerol-rich lipoproteins, and the regulation of its activity in both animal and cell systems.

The aim of the present work was to investigate the influence of the intestinal microflora on the changes in hepatic cytochrome P450 apoproteins induced by dietary glucosinolates. Ten rats harbouring a conventional digestive microflora were offered either a diet containing 390g myrosinase-free rapeseed meal/kg (n 5) or a control diet devoid of glucosinolates (n 5). A similar trial was performed using germ-free rats. After 4 weeks of exposure to the dietary regimens, animals were slaughtered and their livers removed for preparation of microsomes and analysis of cytochrome P450 (EC 1.14.14.1). The glucosinolate-rich diet decreased the concentration of total cytochrome P450 in conventional rats only (-34%). The bacterial status did not modify the concentration of apoproteins CYP1A2 and CYP2B1/B2, but greatly decreased the concentration of the male constitutive isoform CYP2C11 (-53 and -45% respectively in conventional and germ-free rats). Germ-free rats fed on the glucosinolate-rich diet had a greater concentration of CYP3A (+139%) and a lower concentration of CYP2E1 (-32%) than their counterparts fed on the control diet. However, these differences were absent in conventional animals. On the whole, the influence of the intestinal microflora on the changes in hepatic cytochrome P450 due to the consumption of cruciferous vegetables is very complex and obviously involves different mechanisms according to the apoprotein.

Previous epidemiological studies have suggested an association between low levels of physical activity, fitness and the metabolic cardiovascular syndrome. However, many studies have used subjective non-quantitative questionnaire-based methods for assessing physical activity which do not distinguish between the different dimensions of this complex exposure, and in which measurement error in the exposure has not been estimated. These deficiencies in the measurement of this exposure complicate the interpretation of the results of epidemiological studies, and consequently make it difficult to design appropriate interventions and to estimate the expected benefit which would result from intervention. In particular, it is unclear whether public health advice should be to increase total energy expenditure, or to attempt to raise fitness by recommending periods of vigorous activity. To separate the effects of fitness and total energy expenditure in the aetiology of the metabolic cardiovascular syndrome, we measured the physical activity level (PAL), defined as total energy expenditure: BMR, and fitness (maximum O2 consumption (Vo2max per kg), measured in a sub-maximal test) in a cross-sectional population-based study of 162 adults aged 30–40 years. Heart-rate monitoring with individual calibration was used to measure total energy expenditure using the HRFlex method (Ceesay et al. 1989) which has been validated previously against doubly-labelled water and whole-body calorimetry. The relationship between a single measure of PAL, Vo2max per kg and the usual or habitual level for each exposure was measured in a sub-study of twenty-two subjects who undertook four repeated measures over the course of 1 year. This study design allows the reliability coefficient to be computed, which is used to adjust the observed associations for measurement error in the exposure. Twelve men (16.4%) and sixteen women (18.0%) were defined as having one or more features of the metabolic cardiovascular syndrome. The univariate odds ratio for each increasing quartile for PAL was 0.64 (95 % CI 0.43–0.94) and was 0.49 (95 % CI 0.32–0.74) for Vo2max per kg, suggesting that the association with the metabolic cardiovascular syndrome was stronger for fitness than for PAL. However, after adjustment for obesity and sex, and correction for exposure measurement error, the odds ratio per quartile for PAL was 0.32 (95 % CI 0.13–0.83) and 0.44 (95 % CI 0.24–0.78) for Vo2max per kg. Thus, although univariate analysis would suggest that fitness has a stronger association with the metabolic cardiovascular syndrome than PAL, this conclusion is reversed once confounding and the differences in measurement error are considered. We conclude from the present study that the metabolic cardiovascular syndrome is strongly associated with reduced habitual energy expenditure. The method employed to assess the exposure in the present study demonstrates the utility of assessing a known dimension of physical activity using a physiologically-based and objective measure with repeated estimation to adjust for measurement error. Such quantitative epidemiological data provide the basis for planning and evaluating the expected benefit of population-level interventions.

We have compared the effects on lipoproteins and haemostatic variables of two hard margarines with similar functional properties, one traditional margarine containing partially hydrogenated fish oil (PHFO), and one experimental margarine based on vegetable oil (VO). Both were all-purpose cooking margarines with nearly identical functional properties. Trans fatty acids from PHFO in the traditional margarine were replaced mostly by saturated, monounsaturated and trans fatty acids of vegetable origin in the new formulation. Both test margarines contained approximately the same amount of cis polyunsaturated fatty acids. Sixteen female normolipidaemic students consumed each diet with the two test margarines for 14 d in random order (crossover design). The amount of fat was 31 % energy in the PHFO diet and 32 % energy in the VO diet. The test margarines provided approximately 26% energy in both diets. In the PHFO diet 7.8 % of the energy was derived from trans fatty acids and 9.2 % from saturated fatty acids (12:0, 14:0 and 16:0) while in the VO diet, 1.1 % energy was derived from trans fatty acids and 13.3% from saturated fatty acids (12:0, 14:0 and 16:0). The natural content of cholesterol in PHFO was deliberately not balanced by addition of cholesterol to the VO diet, thus the PHFO diet contained 215 mg and the VO diet 86 mg cholesterol per 8.5 MJ. LDL-cholesterol concentration was 19 % higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio LDL-cholesterol:HDL-cholesterol was 12.6 % higher in subjects on the PHFO diet compared with the VO diet (P < 0.01). The level of apolipoprotein (apo)A-I was 6 % lower in subjects on the PHFO diet compared with the VO diet (P < 0.01). The ratio apoB:apoA-I was 10.4 % higher in subjects on the PHFO diet than on the VO diet (P < 0.01). There were no significant differences in total cholesterol, HDL-cholesterol, triacylglycerols, apoB, lipoprotein(a) and haemostatic variables between the diets. Our results demonstrate that PHFO, with its unfavourable effects on plasma lipids, can be replaced by vegetable oils in margarine without appreciable loss of functional properties but with significant improvement in the effects on plasma lipoproteins.

The present study investigated whether the extent of fermentation of NSP in human subjects could be predicted by an in vitro batch system. Fibre sources studied were five mixed diets containing different amounts and types of fibre and three single fibre sources (citrus fibre concentrate, coarse and fine wholemeal rye bread). Fermentation in human subjects was determined in balance experiments in women who were also donors of the faecal inocula. In vitro fermentations were performed with fibre residues prepared from duplicates of the fibre-containing foods consumed during the balance trials. Fermentation of total NSP in vivo was between 65.8 and 88.6% for the mixed diets and 54.4, 58.0 and 96.9 % for the coarse and fine wholemeal rye breads and the citrus fibre concentrate respectively. For the mixed diets and the citrus fibre concentrate, mean differences between the extent of NSP degradation after 24 h in vitro incubation and that in vivo were between −0.7 and 5.0 %. Differences were significant for one diet (P < 0.05). For the wholemeal rye breads, the fermentation in vitro exceeded that in vivo significantly, but the magnitude of the difference in each case was small and without physiological importance. Particle size of breads had no influence on the extent of NSP degradation. These results indicate that the in vitro batch system used could provide quantitative data on the fermentation in vivo of NSP in mixed diets and some single fibre sources. An in vitro incubation time of 24 h was sufficient to mimic the NSP degradation in vivo.

Energy expenditure was estimated using the doubly-labelled water (DLW) method in summer in five free-living adult, non-pregnant, non-lactating, red deer (Cervus elaphus) hinds (weight 107.3 (se 0.9) kg; age 6 (se 1) years) on lowland pasture under typical farming conditions. Climatic conditions were monitored throughout the experiment. Errors due to 2H losses in CH4 and faeces were calculated from previous estimates of stoichiometries. CH4 production, fractionated water loss, urinary N and O2 consumption were estimated using an iterative approach. The water flux (rH2O) in these animals consuming only fresh grass was 12 (se 0.5) kg/d, the CO2 production (rCO2) was 1271 (se 4.0) litres/d and the mean energy expenditure was 25 (se 0.8) MJ/d. There were no significant differences in the isotope distribution spaces and flux rates, rH2O, rCO2 or energy expenditure using the multi-point or two-point approaches to calculation. The DLW-derived energy expenditure of 25 MJ/d is approximately 20% higher than the recommended intake of 21 MJ/d for adult hinds kept outdoors (Adam, 1986) and, at 757 kJ/kg0.75 per d, one third higher than the value of 570 kJ/kg0.75 per d for stags penned indoors (Key et al. 1984).

To investigate whether long-chain n-3 polyunsaturated fatty acids could cross the porcine placenta in late pregnancy and alter neonatal piglet tissue composition, multiparous sows (seven per diet) were fed on diets containing a supplement (30 g/kg) of either soyabean oil or tuna oil for the last 21 d of pregnancy and the first 7 d of lactation. The proportions of all fatty acids, except 18:1n-7, differed between diets: in particular, the tuna-oil-containing diet supplied more 22:6n-3 and less 18:2n-6 fatty acids than the soyabean-oil-containing diet. The proportions of n-3 fatty acids, particularly 22:6n-3 (g/100 g total fatty acids) in sow plasma, colostrum and milk were increased and the proportion of 18:2n-6 was decreased by feeding tuna oil. Piglet tissue n-3 fatty acid proportions (particularly 22:6n-3), obtained shortly after birth, were increased in piglets born to tuna-oil-fed sows compared with progeny of soyabean-oil-fed sows. The increase in the proportion of n-3 fatty acids (g/100 g total fatty acids) in piglet tissues as a result of tuna-oil feeding, compared with soyabean-oil-feeding, was in the order plasma>liver>erythrocytes>spleen>brain>retina. Piglets born to tuna-oil-fed sows had a lower viability score at birth than the progeny of soyabean-oil-fed sows. The proportions of long-chain n-3 fatty acid in tissues of new-born piglets were increased by feeding tuna oil to the sow in late pregnancy; however no improvements in piglet viability were observed.

The effects of dietary fish oil and digestible protein (DP) levels on muscle fatty acid composition and susceptibility to lipid peroxidation were studied in two representative fish species for human nutrition, from fresh and seawater, rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax). In rainbow trout, higher concentrations of dietary fat and DP led to higher weight gain (g/d) (P = 0.001 and P = 0.043 respectively). Additionally, an interaction effect was observed in this species, since the effect of DP was only evident when the dietary fat concentration was low (P = 0.043). A similar tendency was also observed in European sea bass, although with less marked differences among nutritional treatments. Trout fed on diets with a higher concentration of dietary fat had higher concentrations of intramuscular total and neutral lipids in the dorsal muscle (P = 0.005). Increased levels of dietary DP led to significantly lower concentrations of polar lipids in the dorsal muscle of both rainbow trout (P = 0.005) and European sea bass (P = 0.006). In the neutral fraction of intramuscular lipids of dorsal muscle the concentration of n-3 fatty acids was positively affected by the dietary fat concentration in both rainbow trout (P = 0.04) and sea bass (P = 0.001). Muscle homogenates from trout and sea bass fed on diets rich in fish oil showed a significantly higher susceptibility to oxidation than muscle homogenates from fish fed on low-fat diets (P = 0.001). The higher DP concentration also increased susceptibility to oxidation. Moreover, in rainbow trout an interaction effect was observed where the pro-oxidant effect was of higher magnitude when the dietary concentration of both nutrients, fat and protein, was high (P = 0.004).

We studied glucose homeostasis in rat pups from dams fed on a normal-protein (170 g/kg) (NP) diet or a diet containing 60 g protein/kg (LP) during fetal life and the suckling period. At birth, total serum protein, serum albumin and serum insulin levels were similar in both groups. However, body weight and serum glucose levels in LP rats were lower than those in NP rats. At the end of the suckling period (28 d of age), total serum protein, serum albumin and serum insulin were significantly lower and the liver glycogen and serum free fatty acid levels were significantly higher in LP rats compared with NP rats. Although the fasting serum glucose level was similar in both groups, the area under the blood glucose concentration curve after a glucose load was higher for NP rats (859 (sem 58) mmol/l per 120 min for NP rats v. 607 (sem 52) mmol/l per 120 min for LP rats; P < 0.005). The mean post-glucose increase in insulin was higher for NP rats (30 (sem 4.7) nmol/l per 120 min for NP rats v. 17 (sem 3.9) nmol/l per 120 min for LP rats; P < 0.05). The glucose disappearance rate for NP rats (0.7 (sem 0.1) %/min) was lower than that for LP rats (1.6 (sem 0.2)%/min; P < 0.001). Insulin secretion from isolated islets (1 h incubation) in response to 16.7mmol glucose/l was augmented 14-fold in NP rats but only 2.6-fold in LP rats compared with the respective basal secretion (2.8 mmol/l; P < 0.001). These results indicate that in vivo as well as in vitro insulin secretion in pups from dams maintained on a LP diet is reduced. This defect may be counteracted by an increase in the sensitivity of target tissues to insulin.