In order to investigate the role of cholesteryl ester transfer protein (CETP) in the cholesterolaemic response to dietary fats, we analysed plasma lipid profiles of CETP-transgenic and control C57BL/6 mice fed standard chow (AIN-93G; AIN), a low-fat diet, and diets high in butter (saturated fatty acids; SFA), high-oleic acid safflower oil (monounsaturated fatty acids; MUFA), and safflower oil (polyunsaturated fatty acids; PUFA) for 5 weeks. Each group contained four or five mice. There were significant diet and diet×genotype effects on plasma total cholesterol (TC; P = 0·035 and P = 0·008 respectively), liver TC (P < 0·001 and P = 0·002 respectively), and esterified cholesterol (EC; P = 0·002 and P = 0·001 respectively); diet effects on plasma triacylglycerol (P = 0·007), liver free cholesterol (P < 0·001), and body weight (P = 0·027); a genotype effect on body-weight gain (P = 0·014); and a diet×genotype effect on energy intake (P = 0·006). In transgenic mice the SFA diet caused significantly higher plasma TC than the PUFA diet (P < 0·05). In control mice MUFA and PUFA diets, but not the SFA diet, caused significantly higher plasma TC than the low-fat and AIN diets (P < 0·05). Transgenic mice fed PUFA had lower plasma TC (P = 0·040), while transgenic mice fed MUFA had lower LDL+VLDL-cholesterol (P = 0·013) than controls in the same dietary groups. Transgenic mice fed MUFA and PUFA diets also had significantly higher liver TC (P = 0·020 and P = 0·002 respectively) and EC (P = 0·040 and P = 0·036 respectively) than controls fed the same diets. In the present study we showed that: (1) CETP transgenic mice had a cholesterolaemic response to dietary fats similar to that in human subjects; (2) CETP transgenic mice fed PUFA showed significantly lower plasma TC, while those fed MUFA had lower LDL+VLDL-cholesterol than controls; (3) hepatic accumulation of cholesterol, possibly resulting from the combination of the enhanced cholesteryl ester transfer to apolipoprotein B-containing lipoproteins and increased hepatic uptake of cholesterol, may contribute to the cholesterol-lowering effect of MUFA and PUFA in CETP-transgenic mice; (4) CETP may play a role in appetite and/or energy regulation.

The expressions of hexokinase IV (glucokinase, GK) and hexokinase (HK)-I genes were analysed during early ontogenesis of common carp (Cyprinus carpio). Unlike HK-I gene, which was expressed during all the stages of the development, GK was only induced by the first feeding with high levels of dextrin as a source of carbohydrate. This study confirms the high capacity of common carp to use glucose even at the very early stages of development.

The racemic mixture, [6RS]-5-formyltetrahydrofolate, is widely used clinically. In human subjects, orally-administered pure unnatural C-6 isomers, [6R]-5-formyltetrahydrofolate and [6S]-5,10-methenyltetrahydrofolate, were recently shown to be metabolized to the natural isomer, [6S]-5-methyltetrahydrofolate. We re-analysed the data from human studies published during the past four decades in which oral doses (≤10 mg) of racemic mixtures of these folates were used. We re-evaluated the data to determine whether these racemic mixtures are only 50 % bioactive or, as we now predict, more than 50 % bioactive. Our analyses indicate that, in human subjects, oral doses of the racemic mixture of the two formyltetrahydrofolates are 20–84 % more bioactive than would be predicted. These data are consistent with the following pathway: chemical conversion of these folates to 10-formyltetrahydrofolate; oxidation of 10-formyltetrahydrofolate to 10-formyldihydrofolate; subsequent enzymic conversion of 10-formyldihydrofolate to dihydrofolate by 5-amino-4-imidazolecarboxamide ribotide transformylase; and finally the well-established metabolism of dihydrofolate to [6S]-5-methyltetrahydrofolate. An additional review of the literature supports the in vivo oxidation of 10-formyltetrahydrofolate occurring to a certain extent, as 10-formyl-folic acid is rapidly formed after the administration of folic acid (pteroylglutamic acid) or 5-formyltetrahydrofolate in human subjects. The dogma that an oral dose of the unnatural C-6 isomer of 5-formyltetrahydrofolate is not bioactive in human subjects does not withstand scrutiny, most probably due to the previously unrecognized in vivo oxidation of 10-formyltetrahydrofolate. This discovery unveils new folate metabolism in human subjects.

I in the chain sow diet → blood serum of sow → sow milk → piglet serum was investigated in two experiments with a total of eighty-one sows and their piglets. In experiments conducted during the last trimester of gravidity and the 28 d of lactation, diets with glucosinolates (1.9 mmol/kg diet via 100 g ground rapeseed/kg diet (Expt 1) and 2.1 and 4.2 mmol/kg diet via 75 and 150 g rapeseed press cake/kg diet (Expt 2)) were compared with control groups without rapeseed products. From 0 to 600 μg I/kg was added to sow diets during lactation. Diets without supplementary I decreased the I concentration particularly in milk and piglet serum. The presence of rapeseed and rapeseed press cake were indicated by a thiocyanate concentration increase, mainly in sow serum. The diets with glucosinolates decreased the milk and piglet serum I concentration. Spot urine and faeces samples from sows eating the rapeseed-press cake diets had increased I concentration. The sows’ serum I and thyroxine did not respond to glucosinolates (Expt 1) or these diets caused an increase in concentration (Expt 2). Both these criteria seem unsuitable for the diagnosis of I status of adult animals. Glucosinolates and their degradation compounds may affect the thyroid and the mammary glands resulting in lower I milk transfer and higher renal and intestinal I excretion.

The metabolic response to a 150 or 400 g 13C-labelled pasta meal was studied for 8 h following rest or exercise at low or moderate workload (n 6). Following rest, the 400 g meal totally suppressed fat oxidation (v. 14.1 g following the 150 g meal) and a small amount of glucose was converted into fat (4.6 g), but fat oxidation remained high in subjects who had exercised following both the small (21.8 and 34.1 g) and large meal (14.1 and 32.3 g). Exogenous glucose oxidation was significantly higher in subjects who had remained at rest both following the small (67.6 g v. 60.4 and 51.3 g in subjects who exercised at low and moderate workloads) and large meal (152.2 v. 123.0 and 127.2 g). Endogenous glucose oxidation was similar in the three groups following the 150 g meal (42.3–58.0 g), but was significantly lower following the 400 g meal in subjects who had exercised at low workload (24.2 v. 72.2 g following rest; P<0.05), and was totally suppressed in those who had exercised at moderate workload. As a consequence, a larger positive glycogen balance was observed in subjects who exercised before the large meal (182.8–205.1 g v. 92.4 g following rest; P<0.05). Total fat oxidation calculated from 08.00 hours to 20.00 hours was similar in subjects who exercised at low and moderate workloads. These results indicate that: (1) de novo lipogenesis, which plays only a minor role for the disposal of an acute dietary carbohydrate load, is totally suppressed following exercise, even when a very large carbohydrate load is ingested; (2) the reduction in glycogen turnover as well as a preferential conversion of glucose into glycogen are responsible for the increase in glycogen stores following exercise; (3) for a similar energy expenditure, exercise at low workload for a longer period does not favour fat oxidation when the post-exercise period is taken into account.

Large doses of ascorbic acid may mobilise Fe from Fe-binding proteins in vivo which in turn could catalyse lipid peroxidation, a process associated with degenerative diseases. This hypothesis was tested in vitro in the serum of Fe-loaded animals. Eighteen male guinea pigs weighing about 500 g on arrival were allocated to two groups of nine. Fe loading was induced in one group by two intraperitoneal injections of 200 mg iron dextran given on days 1 and 5. Blood (6 ml) was drawn from all animals on day 12 by cardiac puncture. Serum and LDL were separated. Serum was tested for loosely-bound Fe (bleomycin assay) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS) assay) and LDL for susceptibility to in vitro oxidation (TBARS and conjugated diene assays). On day 12, another intraperitoneal injection of 200 mg iron dextran was given to the animals in the Fe-loaded group. On day 19, all animals were given 75 mg ascorbic acid by intraperitoneal injection. Blood (6 ml) was drawn 4 h later by cardiac puncture. Serum and LDL assays were repeated. Ascorbic acid increased loosely-bound Fe and in vitro oxidation in the serum from animals of the Fe-loaded group but not in the serum from animals of the control group. Susceptibility of LDL to in vitro oxidation increased after the ascorbic acid injection in the control group but there was no further increase in the Fe-loaded group. These data suggest that large doses of ascorbic acid promote Fe mobilisation and in vitro oxidation in the serum of Fe-loaded animals.

Under conditions of chronic supply the liver removes most amino acids (AA) in excess of net anabolic needs. Little information is available, however, on how acute alterations in AA supply (as might occur with once-daily feeding regimens) are controlled by the liver. Are these also extracted completely in a ‘first-pass’ manner or are there limitations to hepatic uptake? Furthermore, is the rate of removal ‘saturable’ (by Michaelis–Menten kinetics) over the range of supply experienced under normal feeding conditions? These questions have been addressed in a study that involved acute (4.5 h) increases in AA supply. Four sheep were prepared with trans-hepatic vascular catheters and were offered a basal diet (equivalent to 1.6×energy maintenance) throughout. On four occasions, at 7 d intervals, they were infused with various amounts of an AA mixture into the mesenteric vein over a 4.5 h period. The mixture contained fourteen AA in the proportions present in rumen microbial protein. The amounts infused were calculated to provide an additional one, two, three and four times that absorbed from the basal diet. Continuous blood collections were removed over 2 h intervals before (basal diet only) and at 0.5–2.5 and 2.5–4.5 h of AA infusion. Transfers of AA, from the digestive tract and to the liver, were calculated for both plasma and total blood. The recovery of the infused AA across the portal-drained viscera (PDV) was quantitative (100%) only for histidine and proline, the remaining AA were recovered at 56–83 %. These losses correlated (P <0.001) with the arterial concentrations and were probably due to removal of AA from the systemic circulation by the tissues of the digestive tract. Despite the wide range of net PDV appearances (i.e. absorbed plus infused), the percentage of most AA removed by the liver remained constant, but the percentage varied with AA (from 34 for proline to 78 for tryptophan for blood transfers). Thus, even when supply was increased 5-fold over baseline there was no indication that the transport into the liver declined, indeed the absolute removals continued to increase. In contrast, the branched-chain AA (isoleucine, leucine and valine) did not exhibit constant percentage extractions. Their percentage extractions were always the lowest (16, 10 and 25 respectively) and tended to decline at the highest infusion rates, indicative of saturation in hepatic transport and/or metabolism. The arterial concentrations of all infused AA increased (P <0.001) with rate of infusion, again indicative that the liver did not extract all the net AA available across the PDV. Absolute amounts removed were similar between plasma and blood, indicating that most of the hepatic transfers occurred from plasma. The fractional rates of transfer from total inflow to the liver (i.e. with re-circulated AA included) were 3- to 4-fold lower than rates based on the amounts absorbed plus infused. The highest percentage extraction for total blood inflows was for serine (27), but most were between 6 and 16, except for the branched-chain AA, which were all <1. Use of percentage extractions based on total inflows are probably more appropriate for development of mathematical models of liver metabolism, and the current data suggest that constant values may be applied. The needs of the liver for specific mechanisms involving phenylalanine and histidine (plasma protein synthesis), glycine (detoxification of xenobiotics) and alanine (gluconeogenesis) probably also require to be included in such models.

To clarify the bioavailability of vitamin B12 in lyophylized purple laver (nori; Porphyra yezoensis), total vitamin B12 and vitamin B12 analogue contents in the laver were determined, and the effects of feeding the laver to vitamin B12-deficient rats were investigated. The amount of total vitamin B12 in the dried purple laver was estimated to be 54.5 and 58.6 (SE 5.3 and 7.5 respectively) ΜG/100 g dry weight by lactobacillus bioassay and chemiluminescent assay with hog intrinsic factor respectively. the purple laver contained five types of biologically active vitamin b12 compounds (cyano-, hydroxo-, sulfito-, adenosyl- and methylcobalamin), in which the vitamin b12 coezymes (adenosyl- and methylcobalamin) comprised about 60 % of the total vitamin b12. when 9-week-old vitamin b12-deficient rats, which excreted substantial amounts of methylmalonic acid (71.7(se 20.2) μmol/d) in urine, were fed the diet supplemented with dried purple laver (10 μg/kg diet) for 20 d, urinary methylmalonic acid excretion (as an index of vitamin B12 deficiency) became undetectable and hepatic vitamin B12 (especially adenosylcobalamin) levels were significantly increased. These results indicate that vitamin B12 in dried purple laver is bioavailable to rats.

We investigated the gastrointestinal handling and post-absorptive metabolic handling of [1,1,1-13C]tripalmitin and [1-13C]glycocholate during recovery from severe childhood malnutrition. Eight children were studied on three occasions: at admission (phase 1), during rapid catch-up growth (phase 2) and when weight-for-height had reached 90 % of the reference (phase 3). Breath samples were obtained over a 24 h period and stools were collected over 3 d following the administration of each tracer. At admission, the lipid content of stool expressed as a percentage of ingested lipid was 6 (range 0.7–28.9) but less variation was shown between children at phase 2 (3.3 (range 0.9–4.1)) and phase 3 (1.4 (range 0.4–2.5)). The excretion of 13C in stool varied markedly between children at admission (11.1 (SD 5.4) % administered dose) and during rehabilitation (phase 2, 15.4 (sd 16.5) % administered dose; phase 3, 6.2 (sd 10.2) % administered dose). About 5 % of the absorbed label was recovered on breath at each stage (% absorbed dose; phase 1, 5.1 (sd 6.0); phase 2, 5.2 (sd 3.1); phase 3, 6.4 (sd 6.6)). None of the children exhibited significant bile salt malabsorption as a consequence of small intestinal overgrowth. Of the 13C measured in stool, more label was recovered in fatty acids than triacylglycerols during each of the three phases and this was interpreted to reflect a failure to absorb the products of digestion. The results show that not all the children had problems associated with the digestion and absorption of 13C-labelled tripalmitin in severe malnutrition and during recovery, which was not reflected in gross lipid balance across the gastrointestinal tract. Absorbed lipid was more likely to be deposited as adipose tissue than to satisfy the immediate needs for energy.

The present study was performed to further investigate the adaptive component of thermogenesis that appears during prolonged energy restriction. Fifteen obese men and twenty obese women underwent a 15-week weight-loss programme. During this programme, body weight and composition as well as resting energy expenditure (REE) were measured at baseline, after 2 and 8 weeks of energy restriction (-2929 kJ/d) and drug therapy (or placebo), and finally 2–4 weeks after the end of the 15-week drug therapy and energy restriction intervention, when subjects were weight stable. Regression equations were established in a control population of the same age. These equations were then used to predict REE in obese men and women at baseline, after 2 and 8 weeks, as well as after the completion of the programme. In both men and women body weight and fat mass were significantly reduced (P < 0.05 in all cases) while fat-free mass remained unchanged throughout the programme. At baseline, REE predicted from the regression equation was not significantly different from the measured REE in men, while in women the measured REE was 13 % greater than predicted. After 2 weeks of energy restriction, measured REE had fallen by 469 and 635 kJ/d more than predicted and this difference reached 963 and 614 kJ/d by week 8 of treatment in men and women respectively. Once body-weight stability was recovered at the end of the programme, changes in REE remained below predicted changes in men (-622 kJ/d). However, in women changes in predicted and measured REE were no longer different at this time, even if the women were maintaining a reduced body weight. In summary, the present results confirm the existence of adaptive thermogenesis and give objective measurements of this component during weight loss in obese men and women, while they also emphasize that in women this component seems to be essentially explained by the energy restriction.

The validity of 7 d weighed records of diet obtained for pre-menopausal Mexican women was assessed by two independent methods: the energy intake:BMR (EI:BMR) and the dietary N:urinary N (DN:UN). For the latter, complete urine collections are required and completeness was assessed from measurements of para-aminobenzoic acid (PABA) excretion. There were forty-six adult female subjects in the study, thirty-four were from Mexico City and twelve were from a rural population in the Central Highlands, Mexico. However, data were rejected from five urban women for whom the PABA excretion data suggested incomplete urine collection on four or more days. BMR was measured with Oxylog portable O2 consumption meters, and physical activity level was assessed from a self-completed activity diary. An approximate relationship between the EI:BMR ratio and the DN:UN ratio suggested that the rejection limits on the EI:BMR ratio recommended by are wider than the limits on the DN:UN ratio recommended by . Using the recommended cut-off points for EI:BMR but wider limits for DN:UN, twenty-one and twenty-five women respectively had acceptable intake records by the two methods, and sixteen of them by both methods. In conclusion the modification of the DN:UN limits to 0.92 and 1.70 to set acceptable intake values makes the use of measurements of N and energy balance comparable. Urine values with PABA recoveries greater than 100±15 % should be rejected, as should UN values validated by less than 3 d.

Fractured neck of femur occurs mostly in the older female population and is generally caused by falls. Malnutrition has been postulated as a factor that increases the tendency to suffer falls. Nutritional status of older female hospital patients admitted for emergency surgery for fractured neck of femur recruited (n 75), was compared with an age-matched independent-living group of females attending one of three local day centres (n 50). Dietary assessment was undertaken using three consecutive 24 h dietary recalls and, in the hip fracture group, completed menu cards were used as memory prompts. Data concerning key lifestyle characteristics were obtained using a face-to-face administered questionnaire. Blood samples were taken to determine levels of plasma albumin, transferrin, C-reactive protein (CRP), cholesterol, vitamin C, Se, Zn and total antioxidant status. Haemolysate samples were analysed for Se-dependent glutathione peroxidase activity. There were no significant differences in age between the two groups, but the hip fracture patients had lower mean values for body weight (59.6 v. 67.5 kg; p = 0.005), mindex (weight/demispan) (83· v. 94.4 kg/m; P < 0·0001), calculated BMI (24·1 v. 27·5 kg/m2P<0·0001), mid-upper arm circumference; 27·1 v. 31·3 cm, p = 0.001) and triceps skinfold thickness; 17·0 v. 18.9 mm, p = 0·005) than the home-living group. The hip fracture patients had lower intakes of energy (4·3 v. 5·4 MJ, p = 0·001), fat (p = 0·025), carbohydrate (p = 0·002), protein (p = 0·006), thiamine (p = 0·017), vitamin B6 (p = 0·001), calcium (p = 0·01), K (p = 0·001), Mg (p = 0·001), P (p = 0·001), Fe (p = 0·007), Se (p = 0·008) and NSP (p = 0·001). Mean intakes of both groups were below the estimated average requirement for energy and below the reference nutrient intakes for folate, Ca, vitamin D, Mg, K, Se and Zn. In a high percentage of the hip fracture group the dietary intake of particular nutrients fell below the lower reference nutrient intake for Se (73 %), Mg (54 %) and Fe (19 %). As expected, the fracture patients had reduced plasma albumin (P<0·0001) and increased CRP (P<0·001) values. They had higher plasma vitamin C levels (P<0·001) and lower cholesterol levels (p = 0.04) than the day centre attendees. There were no significant differences in plasma levels of Se, Zn, transferrin or haemolysate glutathione peroxidase activity between the two groups. However, there was evidence of under-nutrition in both groups as key anthropometric values were low, plasma nutrient and metabolite levels were below the standard reference ranges and many individuals had low dietary intakes for specified nutrients.

A prospective epidemiological study was carried out over 1 year to evaluate vitamin B complex dietary intake and status in Cuba, 2 years after the Cuban neuropathy epidemic of 1993. Of the 199 healthy middle-aged men selected, 141 completed the study. Volunteers were followed up every 3 months for 1 year. Dietary intake and status of thiamin, riboflavin, vitamin B6, folate and vitamin B12 were assessed each time. The dietary intake of vitamin B complex was low, particularly in June and July (folate), and October (thiamin). A deficient status was observed for vitamin B complex, except for vitamin B6. Vitamin B complex intake and status varied over the year. However, dietary intake and status were poorly related. The results prove that healthy Cuban men represent a vulnerable population in terms of vitamin B complex status and stress the necessity to both promote preventive multivitamin supplementation and produce local food rich in vitamin B complex.

A number of epidemiological studies have shown an association between β-carotene and the risk of cardiovascular diseases, whereas only a few studies are available concerning the association of lycopene with the risk of coronary events, and no studies have been undertaken concerning lycopene and stroke. Thus, we tested the hypothesis that low serum levels of lycopene are associated with increased risk of acute coronary events and stroke in middle-aged men previously free of CHD and stroke. The subjects were 725 men aged 46–64 years examined in 1991–3 in the Kuopio Ischaemic Heart Disease Risk Factor Study. Forty-one men had either a fatal or a non-fatal acute coronary event or a stroke by December 1997. In a Cox' proportional hazard's model adjusting for examination years, age, systolic blood pressure and three nutritional factors (serum folate, β-carotene and plasma vitamin C), men in the lowest quarter of serum lycopene levels (≤0·07 μmol/l) had a 3·3-fold (95 % CI 1.7, 6.4, P < 0·001) risk of acute coronary events or stroke compared with the others. Our study suggests that a low serum level of lycopene is associated with an increased risk of atherosclerotic vascular events in middle-aged men previously free of CHD and stroke.

The relationship between ponderal, linear and lower leg growth in children recovering from severe malnutrition remains unclear. We report on the early growth of 141 severely malnourished Bangladeshi children aged 6 to 36 months of age who were followed for 90 d. Mean (SD) weight for height (WHZ) and height for age (HAZ) catch-up growth Z scores over the 90 d were 1.6 (0.85) and 0.47 (0.325) respectively. mean (SD) lower leg length growth was 10.35 (4.5) mm. Change in HAZ was significantly associated with initial WHZ, but linear growth occurred in the presence of severe wasting and no threshold WHZ score was identified. Lower leg length gain correlated throughout with ponderal indices but with change in HAZ score only after day 45. Only initial WHZ score and maternal height predicted for linear growth and only accounted for 20 % of total variance. We conclude that linear growth occurs early in severely malnourished children but that knemometry behaves as a ponderal index acutely.