The shortcomings of the metabolic implications of the current protein requirements model are reviewed, and an alternative model, validated with [1-13C]leucine balance results in human adults, is presented and evaluated in the context of defining protein requirements. The model identifies metabolic demands for amino acids as comprising a small fixed component and a variable adaptive component that is relatively insensitive to acute food or protein intake, but which changes slowly with a sustained change in intake, enabling N equilibrium to be achieved. The model accounts for the apparent low efficiency of utilisation of animal proteins in N balance studies and enables more realistic efficiency values to be measured within an experimental framework that takes account of the adaptive metabolic demand. However, the complex relationship between the adaptive metabolic demand and habitual level and quality of protein intake prevents prediction of protein quality by amino acid scoring, which can markedly underestimate actual values. In contrast to the current model, for fully adapted individuals risk of deficiency (i.e. negative N balance after complete adaptation) will only start to increase when intakes fall below the range of the true minimum requirements, i.e. a value that is currently unknown, but likely to be between 0·40 and 0·50 g/kg per d at the lower end of the reported distribution of requirements. At intakes greater than this with additional metabolic demands varying directly with intake, deficiency is only likely as a short-term response to a change to a lower intake within the adaptive range. Thus, for adults satisfying energy needs on most mixed human diets, intakes will be within the adaptive range, and N equilibrium ceases to be a useful indicator of nutritional adequacy of protein. In the context of prescriptive dietary guidelines it may be expedient to retain current values until the benefits (and any risks) of protein intakes within the adaptive range can be quantified. However, from a diagnostic perspective, indicators other than N balance need to be identified, since maintenance of N balance can no longer be used as a surrogate of adequate protein-related health.

The objective of the present study was to investigate the effects of dietary macronutrient ratio on energy metabolism and on skeletal muscle mRNA expression of avian uncoupling protein (UCP), thought to be implicated in thermogenesis in birds. Broiler chickens from 2 to 6 weeks of age received one of three isoenergetic diets containing different macronutrient ratios (low-lipid (LL) 30 v. 77 g lipid/kg; low-protein (LP) 125 v. 197 g crude protein (N×6·25)/kg; low-carbohydrate (LC) 440 v. 520 g carbohydrate/kg). LP chickens were characterised by significantly lower body weights and food intakes compared with LL and LC chickens (−47 and −38 % respectively) but similar heat production/kg metabolic body weight, as measured by indirect calorimetry, in the three groups. However, heat production/g food ingested was higher in animals receiving the LP diet (+41 %, P<0·05). These chickens also deposited 57 % less energy as protein (P<0·05) and 33 % more as fat. No significant differences in energy and N balances were detected between LL and LC chickens. The diets with the higher fat contents (i.e. the LP and LC diets) induced slightly but significantly higher relative expressions of avian UCP mRNA in gastrocnemius muscle, measured by reverse transcription–polymerase chain reaction, than the LL diet (88 and 90 v. 78 % glyceraldehyde-3-phosphate dehydrogenase respectively, P<0·05). Our present results are consistent with the recent view that UCP homologues could be involved in the regulation of lipid utilisation as fuel substrate and provide evidence that the macronutrient content of the diet regulates energy metabolism and especially protein and fat deposition.

The utilisation of essential amino acids (EAA) by the mammary gland of lactating dairy cows fed fresh forages was studied to provide basic information useful in designing strategies to increase the production of milk protein from pasture-fed dairy cows. The relationship between the flux of EAA in the whole body and their uptake by the mammary gland was determined in four cows in early lactation (length of time in milk 44 (SD 14·5) d) producing 21 (SD 4·0) kg milk/d. The cows were maintained in metabolism stalls and fed fresh perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) pasture ad libitum or restricted to 75 % ad libitum intake. The whole-body fluxes of amino acids (AA) were measured using an arterio-venous infusion of universally 13C-labelled AA. Whole-body fluxes of fourteen AA were estimated. Isotope dilution indicated that mammary utilisation accounted for one-third of the whole-body flux of EAA, with individual AA ranging between 17 and 35 %. Isoleucine, leucine, valine and lysine were the EAA with the greatest partitioning towards the mammary gland (up to 36 % of the whole-body flux), which could reflect a potentially limiting effect on milk protein synthesis. In the case of AA with low partitioning to the mammary gland (for example, histidine), it is suggested that non-mammary tissues may have priority over the mammary gland and therefore the supply of this AA may also limit milk protein synthesis.

Maternal nutrition and growth hormone (GH) treatment during early- to mid-pregnancy can each alter the subsequent growth and differentiation of muscle in progeny. We have investigated the effects of varying maternal nutrition and maternal treatment with porcine (p) GH during the second quarter of pregnancy in gilts on semitendinosus muscle cross-sectional area and fibre composition of progeny, and relationships between maternal and progeny measures and progeny muscularity. Fifty-three Large White×Landrace gilts, pregnant to Large White×Duroc boars, were fed either 2·2 kg (about 35 % ad libitum intake) or 3·0 kg commercial ration (13·5 MJ digestible energy, 150 g crude protein (N×6·25)/kg DM)/d and injected with 0, 4 or 8 mg pGH/d from day 25 to 50 of pregnancy, then all were fed 2·2 kg/d for the remainder of pregnancy. The higher maternal feed allowance from day 25 to 50 of pregnancy increased the densities of total and secondary fibres and the secondary:primary fibre ratio in semitendinosus muscles of their female progeny at 61 d of age postnatally. The densities of secondary and total muscle fibres in semitendinosus muscles of progeny were predicted by maternal weight before treatment and maternal plasma insulin-like growth factor-II during treatment. Maternal pGH treatment from day 25 to day 50 of pregnancy did not alter fibre densities, but increased the cross-sectional area of the semitendinosus muscle; this may be partially explained by increased maternal plasma glucose. Thus, maternal nutrition and pGH treatment during the second quarter of pregnancy in pigs independently alter muscle characteristics in progeny.

Alkalizing beverages are highly effective in preventing the recurrence of calcium oxalate (Ox), uric acid and cystine lithiasis. The aim of the present study was to evaluate the influence of grapefruit-juice and apple-juice consumption on the excretion of urinary variables and the risk of crystallization in comparison with orange juice. All investigations were carried out on nine healthy female subjects without any history of stone formation and aged 26–35 years. Each juice was tested in a 5 d study. During the study, the subjects received a standardized diet. Fluid intake of 2·75 litres was composed of 2·25 litres neutral mineral water, 0·4 litre coffee and 0·1 litre milk. On the fourth and fifth day 0·5 litre mineral water was partly substituted by 0·5 or 1·0 litre juice under investigation respectively. The influence on urinary variables was evaluated in 24 h urine samples. In addition, the BONN risk index of CaOx, relative supersaturation (RS)CaOx crystallization was determined. Due to an increased pH value and an increased citric acid excretion after consumption of each juice, the RSCaOx decreased statistically significantly (P<0·05) for grapefruit juice, but not significantly for orange and apple juice. The BONN risk index yielded a distinct decrease in the crystallization risk. We showed that both grapefruit juice and apple juice reduce the risk of CaOx stone formation at a magnitude comparable with the effects obtained from orange juice.

Raw or germinated faba bean (Vicia faba minor var. Alameda) and chickpea (Cicer arietinum kabuli var. Athenas) seed meals were incorporated in essential amino acid-supplemented and energy-equalized diets for growing (65 (SD 1) g) male Wistar rats as the only sources of dietary protein. A lactalbumin-based diet was used as the control. Faecal dry weight and N excretion of animals fed legume-containing diets were greater (P<0·01) and DM digestibility lower (P<0·01) than controls. Apparent faecal digestibilities of amino acids were found to be not different or lower (P<0·01) than controls in rats fed diets containing legume seeds, either germinated or not. Total diaminopimelic acid (DAPA) and purine bases excretion were significantly (P<0·01) higher than controls in rats fed both legume seed meals. Faecal bacterial N calculated according to DAPA or purine bases values was similar and significantly higher (P<0·01) than controls in rats fed legume seed meals. Bacterial N accounted for 50 to 80 % of total faecal N in rats fed legume diets. Apparent faecal N digestibility values (53–65 %) of rats fed legume-based diets were lower (P<0·01) than controls, but became substantially higher (85–92 %) when calculated taking into account bacterial N values. Faecal NSP digestibility values for legume diets were 40–57 g/100 g ingested. Germination decreased starch faecal excretion (P<0·05) and increased (P<0·05) faecal starch and NSP digestibilities of faba bean seeds.

The effect of increased dietary intakes of α-linolenic acid (ALNA) or eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for 2 months upon plasma lipid composition and capacity for conversion of ALNA to longer-chain metabolites was investigated in healthy men (52 (SD 12) years). After a 4-week baseline period when the subjects substituted a control spread, a test meal containing [U-13C]ALNA (700 mg) was consumed to measure conversion to EPA, docosapentaenoic acid (DPA) and DHA over 48 h. Subjects were then randomised to one of three groups for 8 weeks before repeating the tracer study: (1) continued on same intake (control, n 5); (2) increased ALNA intake (10 g/d, n 4); (3) increased EPA+DHA intake (1·5 g/d, n 5). At baseline, apparent fractional conversion of labelled ALNA was: EPA 2·80, DPA 1·20 and DHA 0·04 %. After 8 weeks on the control diet, plasma lipid composition and [13C]ALNA conversion remained unchanged compared with baseline. The high-ALNA diet resulted in raised plasma triacylglycerol-EPA and -DPA concentrations and phosphatidylcholine-EPA concentration, whilst [13C]ALNA conversion was similar to baseline. The high-(EPA+DHA) diet raised plasma phosphatidylcholine-EPA and -DHA concentrations, decreased [13C]ALNA conversion to EPA (2-fold) and DPA (4-fold), whilst [13C]ALNA conversion to DHA was unchanged. The dietary interventions did not alter partitioning of ALNA towards β-oxidation. The present results indicate ALNA conversion was down-regulated by increased product (EPA+DHA) availability, but was not up-regulated by increased substrate (ALNA) consumption. This suggests regulation of ALNA conversion may limit the influence of variations in dietary n-3 fatty acid intake on plasma lipid compositions.

The present study examined the ontogeny of mitochondrial protein abundance in adipose tissue and lungs over the first month of life in the sheep and the extent to which this may be altered by maternal undernutrition during the final month of gestation. The ontogeny of uncoupling protein (UCP), voltage-dependent anion channel (VDAC) and cytochrome c abundance were determined in adipose tissue and lungs sampled from near-term fetuses and young sheep aged 4 h, 1, 7 and 30 d. In adipose tissue, the abundance of UCP1, VDAC and cytochrome c all peaked at 1 d of age and then decreased by 30 d of age, at which stage the brown adipose tissue-specific UCP1 was no longer detectable but UCP2 was clearly abundant. For the lungs, however, UCP2 and VDAC abundance both peaked 7 d after birth and then decreased by 30 d of age. During postnatal development, therefore, a marked change in mitochondrial protein abundance occurs within both adipose tissue and lungs. Maternal nutrient restriction had no effect on lamb growth or tissue weights at 30 d of age but was associated with increased abundance of UCP2 and VDAC but not cytochrome c in both adipose tissue and lungs. These mitochondrial adaptations within both adipose tissue and the lungs of offspring born to previously nutrient-restricted mothers may compromise adipose tissue and lung function during periods of environmental stress.

Dietary fat influences plasma levels of coagulation factor VII (FVII) and serum phospholipids (PL). It is, however, unknown if the fat-mediated changes in FVII are linked to PL. The present study aimed to investigate the effects of dietary fat on fasting and postprandial levels of activated FVII (FVIIa), FVII coagulant activity (FVIIc), FVII protein (FVIIag) and choline-containing PL (PC). In a randomized single-blinded crossover-designed study a high-fat diet (HSAFA), a low-fat diet (LSAFA), both rich in saturated fatty acids, and a high-fat diet rich in unsaturated fatty acids (HUFA) were consumed for 3 weeks. Twenty-five healthy females, in which postprandial responses were studied in a subset of twelve, were included. The HSAFA diet resulted in higher levels of fasting FVIIa and PC compared with the LSAFA and the HUFA diets (all comparisons P≤0·01). The fasting PC levels after the LSAFA diet were also higher than after the HUFA diet (P<0·001). Postprandial levels of FVIIa and PC were highest on the HSAFA diet and different from LSAFA and HUFA (all comparisons P≤0·05). Postprandial FVIIa was higher on the HUFA compared with the LSAFA diet (P<0·03), whereas the HUFA diet resulted in lower postprandial levels of PC than the LSAFA diet (P<0·001). Significant correlations between fasting levels of PC and FVIIc were found on all diets, whereas FVIIag was correlated to PC on the HSAFA and HUFA diet. The present results indicate that dietary fat, both quality and quantity, influences fasting and postprandial levels of FVIIa and PC. Although significant associations between fasting FVII and PC levels were found, our results do not support the assumption that postprandial FVII activation is linked to serum PC.

Retinyl palmitate added to Fe-fortified maize bread has been reported to enhance Fe absorption in adult Venezuelan subjects but not in Western Europeans. It is not known to what extent these results were influenced by differences in vitamin A status of the study subjects. The objective of the present study was to evaluate the influence of retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe in children with vitamin A deficiency, before and after vitamin A supplementation. Erythrocyte incorporation of Fe-stable isotopes was measured 14 d after intake of maize porridge (2·0 mg Fe added as ferrous sulfate) with and without added retinyl palmitate (3·5 μmol; 3300 IU). The study was repeated 3 weeks after vitamin A supplementation (intake of a single dose of 210 μmol retinyl palmitate; ‘vitamin A capsule’). Vitamin A status was evaluated by the modified relative dose–response (MRDR) technique. Retinyl palmitate added to the test meal reduced the geometric mean erythrocyte incorporation of Fe at baseline from 4·0 to 2·6 % (P=0·008, n 13; paired t test). At 3 weeks after vitamin A supplementation, geometric mean erythrocyte incorporation was 1·9 and 2·3 % respectively from the test meal with and without added retinyl palmitate (P=0·283). Mean dehydroretinol:retinol molar ratios were 0·156 and 0·125 before and after intake of the single dose of 210 μmol retinyl palmitate; ‘vitamin A capsule’ (P=0·15). In conclusion, retinyl palmitate added to the labelled test meals significantly decreased erythrocyte incorporation of Fe in children with vitamin A deficiency at baseline but had no statistically significant effect 3 weeks after vitamin A supplementation. The difference in response to retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe before and after intake of the vitamin A capsule indicates, indirectly, changes in vitamin A status not measurable by the MRDR technique. The lack of conclusive data on the effect of retinyl palmitate on Fe absorption indicates the complexity of the interactions between vitamin A status, dietary vitamin A and Fe metabolism.

Reduced protein intake during pregnancy decreased maternal hepatic and plasma docosahexaenoic acid concentrations and impaired docosahexaenoic acid accumulation into fetal brain in the rat. The present study investigated whether restriction of maternal protein intake during pregnancy in the rat alters membrane phospholipid fatty acid composition in the offspring after weaning. Female rats (six per group) were mated and fed diets containing either 180 or 90 g protein/kg throughout pregnancy. Mothers were transferred to standard chow after delivery and the litters reduced to eight pups. Weaning was at 28 d and pups were killed 5 to 6 d later. Tissue weights or membrane total phosphatidylcholine (PC) and phosphatidylethanolamine (PE) concentrations in the offspring did not differ between dietary groups. There were significant differences between the 180 and 90 g/kg groups in liver, brain, lung and heart fatty acid composition that differed between tissues and phospholipid classes. For example, docosahexaenoic and arachidonic acid concentrations were 23 and 10 % lower respectively in hepatic PC, but not PE, in the 90 g/kg group. In brain, docosahexaenoic acid concentration was 17 % lower in PC, but not PE, while arachidonic acid content was 21 % greater in PE but unchanged in PC. The greatest differences were in unsaturated fatty acids, which suggests alterations to desaturase activities and/or the specificity of phospholipid biosynthesis. These results suggest that restricted maternal protein intake during pregnancy results in persistent alterations to membrane fatty acid content.

Using the World Health Organization criterion, the prevalence of sub-clinical vitamin A deficiency can be assessed using plasma retinol concentrations <0·7 μmol/l. However, plasma retinol can be depressed by infection; thus, the use of this criterion alone may overestimate deficiency. In the present study, we investigated the usefulness of the acute-phase proteins (APP) α1-antichymotrypsin (ACT) and α1-acid glycoprotein (AGP), plasma carotenoids and anthropometric and gestational indices to interpret plasma retinol in the blood of 192 apparently healthy Nigerian neonates collected randomly during days 1–20 postpartum. The mean weight (2·64 kg) and length (0·458 m) of the neonates and plasma concentrations (geometric mean, μmol/l) of retinol (0·54), α-carotene (0·072), ß-carotene (0·076) and lutein (0·080) were low. The prevalence of vitamin A deficiency was 72 %, indicating a severe public health problem. Babies who were of low birth weight (P<0·003) or premature and low birth weight (P<0·023) had significantly lower retinol concentrations than full-term normal weight babies. Thirty-two neonates had abnormal ACT and forty-four abnormal AGP concentrations. Positive correlations between retinol and ACT (r 0·186, P=0·05) and AGP (r 0·31, P=0·0001) during days 1–5 may be due to the increasing plasma retinol from maternal milk and a coincidental increasing capacity to synthesise APP. Subsequently, negative correlations between retinol and ACT (r −0·28, P=0·02) and AGP (r −0·29, P=0·018) from day 6 onwards reflected the continuing increase in plasma retinol, but no further increase in the APP. Overall, weight, ACT, lutein and age explained 30 % of the variance in retinol, but lutein was the most significant (r2 0·18, P<0·0001). Hence, the distribution of plasma retinol concentrations in this group of neonates was more strongly linked with nutrition (via the surrogate marker lutein) than infection.

Single 13C6-labelled doses of pteroylmonoglutamic acid (PteGlu; 634 nmol) or 5-formyltetrahydrofolic acid (431–569 nmol) were given to fasted adult volunteers, and the rise in total and 13C-labelled plasma 5-methyltetrahydrofolic acid metabolite monitored over 8 h by HPLC and liquid chromatography–MS. The dose-adjusted area under the curve (AUC) for total (labelled plus unlabelled) plasma 5-methyltetrahydrofolic acid following a 5-formyltetrahydrofolic acid test dose was 155 % that obtained following a PteGlu test dose. Surprisingly, an average 60 and 40 % of the total plasma 5-methyltetrahydrofolic acid response to [13C6]PteGlu and [13C6]5-formyltetrahydrofolic acid, respectively, was unlabelled; an observation never before reported. Short-term kinetics of plasma [13C6]5-methyltetrahydrofolic acid showed a slower initial rate of increase in plasma concentration and longer time to peak following an oral dose of [13C6]PteGlu compared with that for an oral dose of [13C6]5-formyltetrahydrofolic acid, while the [13C6]5-methyltetrahydrofolic acid AUC for [13C6]5-formyltetrahydrofolic acid was 221 % that for [13C6]PteGlu. These data indicate that PteGlu and 5-formyltetrahydrofolic acid, which are thought to be well absorbed (about 90 %) at physiological doses, exhibit dramatically different rates and patterns of plasma response. A limitation in the rate of reduction of PteGlu before methylation could result in slower mucosal transfer of [13C6]5-methyltetrahydrofolic acid derived from [13C6]PteGlu into the plasma. This, when coupled with an observed similar plasma clearance rate for [13C6]5-methyltetrahydrofolic acid metabolite derived from either folate test dose, would yield a comparatively smaller AUC. These findings suggest potential problems in interpretation of absorption studies using unlabelled or labelled folates where the rate of increase, the maximum increase, or the AUC, of plasma folate is employed for test foods (mainly reduced folates) v. a ‘reference dose’ of PteGlu.

The present investigation was undertaken to study whether conventional male Wistar rats could be used as a model for pigs with regard to total tract digestibilities of NSP and macronutrients and whether nebacitin-treated rats could be used as a model for small intestinal digestibility in pigs. Nineteen experimental diets prepared from different fractions of wheat and oats, and which all had been evaluated in experiments with ileal cannulated pigs, were used for the present study. There was a close relationship between the total tract digestibilities of organic matter in the two species. The same was the case with regard to the digestibility of total NSP and arabinoxylans, but the values were on average 6 % lower in rats than in pigs. On average, there were no significant differences between rats and pigs with regard to faecal protein digestibility. However, protein in oat-based diets was significantly better digested in the rat than the pig. The digestibility of fat was consistently higher in rats than in pigs, with the biggest difference being found in oat-based diets, in which most of the fat was locked in cell structures. For the wheat-based diets, in which a large proportion of the fat was present as added fat, there was a greater similarity between the two species. In nebacitin-treated rats the digestibility of organic matter, starch, protein and fat was negatively related to the dietary level of NSP, but this model could not be used to predict the small intestinal digestibility of NSP and macronutrients in ileal-cannulated pigs.

In previous studies, we showed that docosahexaenoic acid (DHA) ingestion enhanced the susceptibility of rat liver and kidney to lipid peroxidation, but did not increase lipid peroxide formation to the level expected from the relative peroxidizability index (P-index) of the total tissue lipids. The results suggested the existence of some suppressive mechanisms against DHA-induced tissue lipid peroxide formation, as increased tissue ascorbic acid (AsA) and glutathione levels were observed. Therefore, we focused initially on the role of AsA for the suppressive mechanisms. For this purpose, we examined the influence of different levels of dietary AsA (low, moderate, high and excessive levels were 100, 300 (control), 600 and 3000 mg/kg diet respectively) on the tissue lipid peroxide and antioxidant levels in AsA-requiring Osteogenic Disorder Shionogi/Shi-od/od (ODS) rats fed DHA (6·4 % total energy) for 32 or 33 d. Diets were pair-fed to the DHA- and 100 mg AsA/kg diet-fed group. We found that the lipid peroxide concentrations of liver and kidney in the DHA-fed group receiving 100 mg AsA/kg diet were significantly higher or tended to be higher than those of the DHA-fed groups with AsA at more than the usual control level of 300 mg/kg diet. Contrary to this, the liver α-tocopherol concentration was significantly lower or tended to be lower in the DHA and 100 mg AsA/kg diet-fed group than those of the other DHA-fed groups. However, tissue lipid peroxide formation and α-tocopherol consumption were not suppressed further, even after animals received higher doses of AsA. The present results suggest that higher than normal concentrations of tissue AsA are not necessarily associated with the suppressive mechanisms against dietary DHA-induced tissue lipid peroxide formation.

The breakdown of glucosinolates, a group of thioglucoside compounds found in cruciferous plants, is catalysed by dietary or microbial myrosinase. This hydrolysis releases a range of breakdown products among which are the isothiocyanates, which have been implicated in the cancer-protective effects of cruciferous vegetables. The respective involvement of plant myrosinase and gut bacterial myrosinase in the conversion, in vivo, of glucosinolates into isothiocyanates was investigated in sixteen Fischer 344 rats. Glucosinolate hydrolysis in gnotobiotic rats harbouring a whole human faecal flora (Flora+) was compared with that in germ-free rats (Flora−). Rats were offered a diet where plant myrosinase was either active (Myro+) or inactive (Myro−). The conversion of prop-2-enyl glucosinolate and benzyl glucosinolate to their related isothiocyanates, allyl isothiocyanate and benzyl isothiocyanate, was estimated using urinary mercapturic acids, which are endproducts of isothiocyanate metabolism. The highest excretion of urinary mercapturic acids was found when only plant myrosinase was active (Flora−, Myro+ treatment). Lower excretion was observed when both plant and microbial myrosinases were active (Flora+, Myro+ treatment). Excretion of urinary mercapturic acids when only microbial myrosinase was active (Flora+, Myro− treatment) was low and comparable with the levels in the absence of myrosinase (Flora−, Myro− treatment). No intact glucosinolates were detected in the faeces of rats from the Flora+ treatments confirming the strong capacity of the microflora to break down glucosinolates. The results confirm that plant myrosinase can catalyse substantial release of isothiocyanates in vivo. The results also suggest that the human microflora may, in some circumstances, reduce the proportion of isothiocyanates available for intestinal absorption.

Increased dietary consumption of the n-3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (20:5n-3; EPA) and docosahexaenoic acid (22:6n-6; DHA) is associated with their incorporation into circulating phospholipid and increased production of lipid peroxide metabolites. The relationship between peripheral blood mononuclear cell (PBMC) function, n-3 PUFA intake and antioxidant co-supplementation is poorly defined. We therefore investigated tumour necrosis factor (TNF)-α and interleukin (IL) 6 production by PBMC and phospholipid fatty acid composition in plasma and erythrocytes of healthy male subjects (n 16) receiving supplemental intakes of 0·3, 1·0 and 2·0 g EPA+DHA/d, as consecutive 4-week courses. All subjects were randomised in a double-blind manner to receive a concurrent antioxidant supplement (200 μg Se, 3 mg Mn, 30 mg D-α-tocopheryl succinate, 90 mg ascorbic acid, 450 μg vitamin A (β-carotene and retinol)) or placebo. There was a positive dose-dependent relationship between dietary n-3 PUFA intake and EPA and DHA incorporation into plasma phosphatidylcholine and erythrocyte phosphatidylethanolamine, with a tendency towards a plateau at higher levels of intake. Production of TNF-α and IL-6 by PBMC decreased with increasing n-3 PUFA intake but tended towards a ‘U-shaped’ dose response. Both responses appeared to be augmented by antioxidant co-supplementation at intermediate supplementary n-3 PUFA intakes. Thus, increased dietary n-3 PUFA consumption resulted in defined but contrasting patterns of modulation of phospholipid fatty acid composition and PBMC function, which were further influenced by antioxidant intake.

The effect of diet composition on post-absorptive (15 h fast) fatty acid and glucose metabolism was investigated in five healthy men after 2 weeks on a low-carbohydrate (Low-CHO) diet (30% energy intake from carbohydrates, 55% from fat, 15% from protein) and after 2 weeks on a high-carbohydrate (High-CHO) diet (energy intake 75, 10 and 15% from carbohydrates, fat and protein respectively). The diets were isoenergetic and comprised real foods. Stable-isotope tracer methodology and indirect calorimetry were employed to measure glucose and fatty acid kinetics and oxidation. The relative contribution of carbohydrate to the total energy expenditure was significantly higher after the High-CHO diet. After the High-CHO diet, total and plasma fatty oxidation (2·4 (SE 0·7) and 2·1 (SE 0·4) μmol/kg per min respectively) were significantly lower than after the Low-CHO diet (4·8 (SE 0·5) and 4·6 (SE 0·8) μmol/kg per min for total and plasma fatty oxidation respectively). The rate of appearance (Ra) of non-esterified fatty acids (NEFA) in plasma and the arterial NEFA concentration were both significantly lower following the High-CHO than the Low-CHO diet. However, even after the High-CHO diet, NEFA Ra was threefold higher than plasma fatty acid oxidation. Thus, the decrease in fatty acid oxidation after consumption of a high-carbohydrate diet for 2 weeks in healthy men is unlikely to result from decreased fatty acid delivery to the tissues. Glucose Ra and arterial plasma glucose concentration were similar after the two diets. After the High-CHO diet, arterial lactate concentration was higher and total carbohydrate oxidation rate well exceeded glucose Ra in plasma. Therefore, alterations in intracellular mechanisms may limit fatty acid oxidation after high-carbohydrate diets.

The present study determined the intra-individual variation of BMR measurements, using a standard out-patient protocol, with the subjects transporting themselves to the laboratory for the BMR measurements after spending the night at home. The effect of a non-fasting state and variation in daily habitual physical activity the day before testing was evaluated. Eight male and eleven female subjects participated in three BMR measurements with 2-week intervals. Physical activity was estimated with a tri-axial accelerometer for movement registration, during the 3 d before each BMR measurement. There were no significant differences in estimated BMR (ANOVA repeated measures, P=0·88) and in physical activity (ANOVA repeated measures, P=0·21). Mean within-subject CV in BMR was found to be 3·3 (SD 2·1) %, ranging from 0·4 to 7·2 %. Differences between BMR measurements could not be explained by differences in physical activity the day before; however the mean within-subject CV in BMR changed from 5·7 to 5·2 % after correcting for within-machine variability and from 5·2 to 3·3 % after excluding five measurements because of non-compliance to the protocol including fasting. In conclusion, BMR values measured with a standard out-patient protocol are sufficiently reproducible for most practical purposes despite the within-subject variability in physical activity the day before the measurement. For this purpose, however, non-fasting subjects must be excluded and a regular function check of the ventilated-hood system is recommendable.