The development of atherosclerosis is influenced by genetic, lifestyle and nutritional risk factors. Zn and metallothionein deficiency can enhance oxidative-stress-related signalling processes in endothelial cells, and since changes in available plasma Zn may affect the Zn status of the endothelium, Zn deficiency could be a risk factor for IHD. Although the association of Zn with many proteins is essential for their function, three key signalling processes are highlighted as being principal targets for the effect of Zn deficiency: the activation of NF-κB, the activation of caspase enzymes and the signalling of NO. The need to develop a reliable indicator of Zn status is critical to any epidemiological approach for studying the relationship between Zn status and disease incidence. Studies using appropriate animal models and investigating how the plasma Zn pool influences endothelial intracellular labile Zn would be helpful in appreciating the importance of Zn deficiency in atherogenesis.

In vitro studies with isolated sheep rumen epithelium have shown that an increase in the lumen K concentration induces an increase in the transmural potential difference across the rumen epithelium (serosal side: positive), which is associated with a decrease in Mg transport. However, at lumen K concentrations >80 mmol/l, Mg transport across the epithelium became independent of the lumen K concentration. The present study was carried out to determine whether this observation also occurs in vivo. Four ruminally fistulated wethers were fed four rations supplemented with KHCO3 (15·7, 37·6, 59·4 or 77·4 g K/kg DM) in a 4×4 Latin square design. Increased K intakes significantly increased the rumen K concentration. For all data combined, Mg absorption expressed as % intake was negatively correlated with the rumen K concentration. However, apparent Mg absorption either expressed in absolute terms (g/d) or as % intake was not significantly affected when the dietary K concentration was increased from 59·4 to 77·4 g/kg DM. Rumen K concentration was inversely correlated with the transmural potential difference (blood side: positive) (Pearson's r −0·709; R2adj 0·468, P=0·002, n 16). It is concluded that in wethers apparent Mg absorption becomes independent of the dietary K concentration when the K concentration is >60 g/kg DM or equivalent to a postprandial rumen K concentration of about 125 mmol/l.

The aim of our present study was to compare the efficiency of conjugated linoleic acids (CLA) and fish oil in modulating atherogenic risk markers. Adult male hamsters were given a cholesterol-rich diet (0·6 g/kg) for 8 weeks; the diet was supplemented with 5 g cis-9,trans-11-CLA isomer/kg, 12 g CLA mixture (CLA-mix)/kg, 12 g fish oil/kg or 12 g fish oil + 12 g CLA-mix/kg. The plasma cholesterol status was improved only with the cis-9,trans-11-CLA (HDL-cholesterol and HDL-cholesterol:LDL-cholesterol ratio, P<0·05), but was of borderline significance for CLA-mix (HDL-cholesterol:LDL-cholesterol ratio, P=0·06), with an increase (33–40 %) in the liver lipoprotein receptors (scavenger receptor-type I and LDL ApoB/E receptor) and HDL-binding protein 2 (P<0·05). A 100 % pigment gallstones incidence and a slight insulin resistance (homeostatic model assessment index) were observed in the CLA-mix-fed hamsters (P=−0·031). In comparison, fish-oil feeding alone improved merely the scavenger receptor-type I and HDL-binding protein 2 liver status and faeces sterol output. For most of our present observations, the concomitant intake of fish oil and CLA-mix gave dominant effects that were exclusive and specific to one or the other oil. In conclusion, part of the beneficial effects of CLA in the present study can be ascribed to the cis-9,trans-11-isomer, and these did not generally overlap with those of fish oil. In addition, the CLA-mix effects are clearly affected by the marine (n-3) fatty acids.

Diabetes-prone BioBreeding (BBdp) rats often present an enteropathy that may precede the onset of autoimmune insulitis. The aim of the present study was to assess the influence of sex, the time course, the strain specificity, the distribution along the intestinal tract and the effect of diet for the changes in the activity of gut invertase, maltase and lactase found in BBdp rats, as compared with both Wistar–Furth (WF) and diabetes-resistant BioBreeding (BBc) rats. These hydrolases were measured, therefore, at day 10, 30, 45, 70, 95 and 120 in three intestinal segments of WF, BBc and BBdp rats fed, after weaning, either a protective high-casein diet, which decreases the incidence of diabetes in the BBdp rats, or one of two diabetogenic diets (National Toxicology Program; NTP or wheat-gluten-based; WG). Except for a somewhat lower lactase activity in the BBdp rats, no obvious difference in hydrolyase activity between the three strains of rats was observed at day 10. Between days 30 and 120, however, the activity of the hydrolases, especially that of invertase and lactase, was lower in the BBdp rats than in either the WF or BBc rats, at least when considering the animals fed either the NTP or WG diet. These findings support the view that BBdp rats exposed to a diabetogenic diet develop an enteropathy well before the onset of autoimmune insulitis, in a manner somehow comparable with the situation found in some type 1 diabetic patients, in whom coeliac disease may be diagnosed before diabetes onset.

Although garlic and onions have long been associated with putative cardiovascular health benefits, the effects of different commercially available onions and level of intake have not been studied. Therefore, the aim of the present study was to evaluate the potential health benefits of raw onions using the pig as a biomedical model. Twenty-five female (Large White×Landrace) pigs were used in a (2×2)+1 factorial experiment. Pigs were fed a standard grower diet supplemented with 100 g tallow/kg with the addition of Allium cepa var. cavalier or var. destiny at 0, 10 or 25 g/MJ digestible energy for 6 weeks. Overall, the consumption of onions resulted in significant reductions in plasma triacylglycerol; however, the reductions were most pronounced in pigs fed destiny onions (−26 %, P=0·042). Total plasma cholesterol and LDL:HDL ratios were not significantly different. Onion supplementation, regardless of the variety, resulted in dose-dependent reductions in erythrocyte counts and Hb levels, while the white blood cell concentrations, particularly lymphocytes, were increased in pigs that consumed onions. Furthermore, indices of blood clotting were largely unaffected by onion consumption. In conclusion, dietary supplementation with raw brown onions has moderate lipid-modulating and immunostimulatory properties. However, daily onion intake >25 g/MJ digestible energy could be detrimental to erythrocyte numbers.

The effects of structured medium- and long-chain triacylglycerols (MLCT) in diets containing 50–200 g fat/kg on body fat accumulation were compared with those of long-chain triacylglycerols (LCT) in rats. In rats fed ad libitum, weights of intra-abdominal adipose tissues and carcass fat contents were significantly smaller (P<0·05) in rats fed the 150–200 g MLCT/kg diet than in rats fed 150–200 g LCT/kg diet. Serum and liver triacylglycerol contents were significantly greater (P<0·05) in rats fed 200 g MLCT/kg diet, as were hepatic capacities of citrate synthase and cytochrome oxidase (P<0·05). The effects of MLCT on body fat were also examined in adult rats fed a limited amount of food (approximately 50 % of ad libitum intake). Reduction of body fat deposition during the food restriction was the same between in LCT and MLCT groups. These results suggest that accumulation of body fat was less efficient during long-term feeding of MLCT than LCT in rats fed high-fat diets ad libitum. The effect of MLCT on body fat might be influenced by the dietary fat content or by energy sufficiency.

The effects of dietary lipids on the fatty acid composition of rat pancreatic membranes and acinar cell function were investigated. Weaning rats were fed for 8 weeks on one of two diets which contained 100 g virgin olive oil (OO) or sunflowerseed oil (SO)/kg. Pancreatic plasma membranes were isolated and fatty acids determined. Amylase secretion and cytosolic concentrations of Ca2+ and Mg2+ were measured in pancreatic acini. Membrane fatty acids were profoundly affected by the diets; the rats fed OO had higher levels of 18:1n-9 (42·86 (sem 1·99) %) and total MUFA compared with the animals fed SO (25·37 (sem 1·11) %). Reciprocally, the SO diet resulted in greater levels of total and n-6 PUFA than the OO diet. The most striking effect was observed for 18:2n-6 (SO 17·88 (sem 1·32) %; OO 4·45 (sem 0·60) %), although the levels of 20:4n-6 were also different. The proportion of total saturated fatty acids was similar in both groups, and there was only a slight, not significant (P=0·098), effect on the unsaturation index. Compared with the OO group, acinar cells from the rats fed SO secreted more amylase at rest but less in response to cholecystokinin octapeptide, and this was paralleled by reduced Ca2+ responses to the secretagogue. The results confirm that rat pancreatic cell membranes are strongly influenced by the type of dietary fat consumed and this is accompanied by a modulation of the secretory activity of pancreatic acinar cells that involves, at least in part, Ca2+ signalling.

Assessing the total energy expenditure (TEE) and the levels of physical activity in free-living conditions with non-invasive techniques remains a challenge. The purpose of the present study was to investigate the accuracy of a new uniaxial accelerometer for assessing TEE and physical-activity-related energy expenditure (PAEE) over a 24 h period in a respiratory chamber, and to establish activity levels based on the accelerometry ranges corresponding to the operationally defined metabolic equivalent (MET) categories. In study 1, measurement of the 24 h energy expenditure of seventy-nine Japanese subjects (40 (SD 12) years old) was performed in a large respiratory chamber. During the measurements, the subjects wore a uniaxial accelerometer (Lifecorder; Suzuken Co. Ltd, Nagoya, Japan) on their belt. Two moderate walking exercises of 30 min each were performed on a horizontal treadmill. In study 2, ten male subjects walked at six different speeds and ran at three different speeds on a treadmill for 4 min, with the same accelerometer. O2 consumption was measured during the last minute of each stage and was expressed in MET. The measured TEE was 8447 (SD 1337) kJ/d. The accelerometer significantly underestimated TEE and PAEE (91·9 (SD 5·4) and 92·7 (SD 17·8) % chamber value respectively); however, there was a significant correlation between the two values (r 0·928 and 0·564 respectively; P<0·001). There was a strong correlation between the activity levels and the measured MET while walking (r2 0·93; P<0·001). Although TEE and PAEE were systematically underestimated during the 24 h period, the accelerometer assessed energy expenditure well during both the exercise period and the non-structured activities. Individual calibration factors may help to improve the accuracy of TEE estimation, but the average calibration factor for the group is probably sufficient for epidemiological research. This method is also important for assessing the diurnal profile of physical activity.

The influence of the source of dietary fat on postprandial thermogenesis and substrate oxidation rates, was examined in twelve postmenopausal women aged 57–73 years, with BMI 21·9–38·3 kg/m2. A single blind, randomised, paired comparison of two high-fat, isoenergetic, mixed test meals was conducted. The major source of fat was either cream (CREAM) or extra virgin olive oil (EVOO). RMR, diet-induced thermogenesis (DIT) and substrate oxidation rates over 5 h were measured by indirect calorimetry. There were no differences in body weight, RMR, fasting carbohydrate or fat oxidation rates between the two occasions. DIT (EVOO 97 (sd 46) v. CREAM 76 (sd 69) kJ/5 h and EVOO 5·2 (sd 2·5) v. CREAM 4·1 (sd 3·7)% energy) did not differ between the two test meals. The postprandial increase in carbohydrate oxidation rates, relative to their respective fasting values (ΔCOX), was significantly lower following the EVOO meal (EVOO 10·6 (sd 8·3) v. CREAM 17·5 (sd 10) g/5 h; paired t test, P=0·023), while postprandial fat oxidation rates (ΔFOX) were significantly higher (EVOO 0·0 (sd 4·4) v. CREAM -3·6 (sd 4·0) g/5 h; P=0·028). In the eight obese subjects, however, DIT was significantly higher following the EVOO meal (EVOO 5·1 (sd 2·0) v. CREAM 2·5 (sd 2·9) %; P=0·01). This was accompanied by a significantly lower ΔCOX (EVOO 10·9 (sd 9·9) v. CREAM 17·3 (sd 10·5) g/5 h; P=0·03) and significantly higher ΔFOX (EVOO 0·11 (sd 4·4) v. CREAM −4·1 (sd 4·5) g/5 h, P=0·034). The present study showed that olive oil significantly promoted postprandial fat oxidation and stimulated DIT in abdominally obese postmenopausal women.

Protein synthesis and turnover in ruminal micro-organisms were assessed by stable-isotope methods in order to follow independently the fate of amino acid (AA)-C and -N in different AA. Rumen fluid taken from sheep receiving a grass hay–concentrate diet were strained and incubated in vitro with starch–cellobiose–xylose in the presence of NH3 and 5 g algal protein hydrolysate (APH)/l, in incubations where the labels were 15NH3, [15N]APH or [13C]APH. Total 15N incorporation was calculated from separate incubations with 15NH3 and [15N]APH, and net N synthesis from the increase in AA in protein-bound material. The large difference between total and net AA synthesis indicated that substantial turnover of microbial protein occurred, averaging 3·5 %/h. Soluble AA-N was incorporated on average more extensively than soluble AA-C (70 v. 50 % respectively, P=0·001); however, incorporation of individual AA varied. Ninety percent of phenylalanine-C was derived from the C-skeleton of soluble AA, whereas the incorporation of phenylalanine-N was 72 %. In contrast, only 15 % aspartate-C + asparagine-C was incorporated, while 45 % aspartate-N+asparagine-N was incorporated. Deconvolution analysis of mass spectra indicated substantial exchange of carboxyl groups in several AA before incorporation and a condensation of unidentified C2 and C4 intermediates during isoleucine metabolism. The present results demonstrate that differential labelling with stable isotopes is a way in which fluxes of AA synthesis and degradation, their biosynthetic routes, and separate fates of AA-C and -N can be determined in a mixed microbial population.

Lingzhi (Ganoderma lucidum) is a woody mushroom highly regarded in traditional medicine and is widely consumed in the belief that it promotes health and longevity, lowers the risk of cancer and heart disease and boosts the immune system. However, objective scientific validation of the putative health benefits of Lingzhi in human subjects is lacking, and issues of possible toxicity must be addressed. The present double-blinded, placebo-controlled, cross-over intervention study investigated the effects of 4 weeks Lingzhi supplementation on a range of biomarkers for antioxidant status, CHD risk, DNA damage, immune status, and inflammation, as well as markers of liver and renal toxicity. It was performed as a follow-up to a study that showed that antioxidant power in plasma increased after Lingzhi ingestion, and that 10 d supplementation was associated with a trend towards an improved CHD biomarker profile. In the present study, fasting blood and urine from healthy, consenting adults (n 18; aged 22–52 years) was collected before and after 4 weeks supplementation with a commercially available encapsulated Lingzhi preparation (1·44 g Lingzhi/d; equivalent to 13·2 g fresh mushroom/d) or placebo. No significant change in any of the variables was found, although a slight trend toward lower lipids was again seen, and antioxidant capacity in urine increased. The results showed no evidence of liver, renal or DNA toxicity with Lingzhi intake, and this is reassuring. The present study of the effects in healthy, well-nourished subjects provides useful, new scientific data that will support controlled intervention trials using at-risk subjects in order to assess the therapeutic effect of Lingzhi in the promotion of healthy ageing.

The present study was conducted to determine the pattern of incorporation of dietary EPA and docosahexaenoic acid (DHA) into milk, and to evaluate consequent changes in milk fat composition and sensory characteristics. Fourteen multiparous cows in early lactation were divided into two groups and were offered supplements for 10 d. While individual stalls after each morning milking, one group was offered a mixture of rumen-protected tuna oil (RPTO)–soyabean supplement (2 kg; 30:70, w/w; +RPTO) and the second group was offered the basal ration without RPTO (−RPTO). Both groups grazed together on a spring pasture after supplementation. Feeding supplemental RPTO increased the concentrations of EPA and DHA in milk fat from undetectable levels in −RPTO cows to 6·9 and 10·1 g/kg milk fat respectively. Total n-3 PUFA concentration in milk fat was increased three- to fourfold by tuna-oil supplementation (8·4 to 32·0 g/kg milk fat). There were no significant effects on milk production (35·4 v. 33·9 l/d), milk protein (28·2 v. 30·1 g/kg) or milk fat (36·2 v. 40·4 g/kg for −RPTO and +RPTO respectively). The concentration of total saturated fatty acids in milk fat was significantly reduced (568 v. 520 g/kg total fatty acids) and there was a 17 % reduction in the atherosclerotic index of milk after tuna-oil supplementation. Untrained consumer panellists (n 61) rated milk from both groups of cows similarly for taste and smell. We conclude that it is possible to enrich milk with n-3 PUFA without deleterious effects on yield, milk composition or sensory characteristics.

Maternal nutrition is important for fetal development, but its impact on the functional outcome of infants is still unclear. The present study investigated the effects of vitamin A and Fe supplementation during gestation on infant mental and psychomotor development. Mothers of infants from five villages in Indonesia were randomly assigned to supervised, double-blind supplementation once per week from approximately 18 weeks of pregnancy until delivery. Supplementation comprised 120 mg Fe + 500 μg folic acid with (n 94) or without (n 94) 4800 μg retinol in the form of retinyl acetate. Mothers of infants who participated in the national Fe+folic acid supplementation programme, but whose intake of supplements was not supervised, were recruited from four other villages (n 88). The mental and psychomotor development of infants was assessed, either at 6 or 12 months of age, using the Bayley Scales of Infant Development (BSID). We found no impact of vitamin A supplementation on mental or psychomotor development of infants. In addition, infants whose mothers had received weekly Fe supplementation had similar mental and psychomotor indices as those whose mothers had participated in the governmental Fe supplementation programme. The study population was moderately Fe and vitamin A deficient. The size of the treatment groups was large enough to detect a mean difference of 10 points on the BSID, which is less than 1 sd (15 points) of the average performance of an infant on the BSID. In conclusion, the present study did not find an impact of weekly supplementation of 4800 RE vitamin A in addition to Fe during gestation on functional development of Indonesian infants. However, smaller improvements in development may be seen if studied in a larger and/or more deficient population.

Infant formulas based on soyabean protein isolate are often used as an alternative to cows'-based formulas. However, the presence of phytic acid in soya formulas has raised concern about the absorption of trace elements and minerals from these products. The aim of the present study was to evaluate mineral and trace element absorption from regular and dephytinised soya formula in healthy infants. Soyabean protein isolate with a relatively low native content of phytic acid was used for production of a regular soya formula (300 mg phytic acid/kg liquid formula) and an experimental formula was based on dephytinised soya protein isolate (<6 mg phytic acid/kg liquid formula). Using a crossover study design, apparent mineral and trace element absorptions were measured by a stable isotope technique based on 72 h faecal excretion of non-absorbed stable isotopes (Zn, Fe, Cu and Ca) and by the chemical balance technique (Mn, Zn, Cu and Ca) in nine infants (69–191 d old). Fe absorption was also measured by erythrocyte incorporation 14 d after intake. The results from the present study demonstrated that Zn absorption, measured by a stable isotope technique, was significantly greater after dephytinisation (mean value 16·7 v. 22·6 %; P=0·03). No other statistically significant differences between the two formulas were observed. The nutritional benefit of dephytinisation was marginal in the present study. Based on these results, the use of soyabean protein isolate with low native content of phytic acid should be promoted for production of soya formulas and adequate addition of ascorbic acid to enhance Fe absorption should be ensured in the products.

To evaluate the suitability for glycaemic index (GI) calculations of using blood sampling schedules and methods of calculating area under the curve (AUC) different from those recommended, the GI values of five foods were determined by recommended methods (capillary blood glucose measured seven times over 2·0 h) in forty-seven normal subjects and different calculations performed on the same data set. The AUC was calculated in four ways: incremental AUC (iAUC; recommended method), iAUC above the minimum blood glucose value (AUCmin), net AUC (netAUC) and iAUC including area only before the glycaemic response curve cuts the baseline (AUCcut). In addition, iAUC was calculated using four different sets of less than seven blood samples. GI values were derived using each AUC calculation. The mean GI values of the foods varied significantly according to the method of calculating GI. The standard deviation of GI values calculating using iAUC (20·4), was lower than six of the seven other methods, and significantly less (P<0·05) than that using netAUC (24·0). To be a valid index of food glycaemic response independent of subject characteristics, GI values in subjects should not be related to their AUC after oral glucose. However, calculating GI using AUCmin or less than seven blood samples resulted in significant (P<0·05) relationships between GI and mean AUC. It is concluded that, in subjects without diabetes, the recommended blood sampling schedule and method of AUC calculation yields more valid and/or more precise GI values than the seven other methods tested here. The only method whose results agreed reasonably well with the recommended method (ie. within ±5 %) was AUCcut.

Tea and coffee are rich in polyphenols with a variety of biological activities. Many of the demonstrated activities are consistent with favourable effects on the risk of chronic diseases. 4-O-methylgallic acid (4OMGA) and isoferulic acid are potential biomarkers of exposure to polyphenols derived from tea and coffee respectively. 4OMGA is derived from gallic acid in tea, and isoferulic acid is derived from chlorogenic acid in coffee. Our major objective was to explore the relationships of tea and coffee intake with 24 h urinary excretion of 4OMGA and isoferulic acid in human subjects. The relationships of long-term usual (111 participants) and contemporaneously recorded current (344 participants) tea and coffee intake with 24 h urinary excretion of 4OMGA and isoferulic acid were assessed in two populations. 4OMGA was related to usual (r 0·50, P<0·001) and current (r 0·57, P<0·001) tea intake, and isoferulic acid was related to usual (r 0·26, P=0·008) and current (r 0·18, P<0·001) coffee intake. Overall, our present results are consistent with the proposal that 4OMGA is a good biomarker for black tea-derived polyphenol exposure, but isoferulic acid may be of limited usefulness as a biomarker for coffee-derived polyphenol exposure.

In 1997, the prevalence of anaemia was 40·9 % among children aged 6–59 months in the State of Pernambuco, north-east Brazil. Using the same sample of children, we have investigated possible reasons for this high prevalence. A representative sample was selected through a three-stage process: proportional systematic random sampling of municipalities in the State, systematic random sampling of census sectors within these municipalities, and finally, simple random sampling of households with children aged 6–59 months to obtain the sample of 650 children. Data collection included demographic, environmental, socio-economic and maternal variables, and nutritional status and dietary intakes of the children. Multiple linear regression analysis was based on a hierarchical model of factors associated with Hb concentration. The mean Hb concentration of children aged 6–23 months was 10 g/l lower than that of older children. In the regression analysis, child age explained 8·3 % of the variance in Hb concentration. The intake of bioavailable Fe explained a further 3·3, serum retinol 2·7, diarrhoea 2·4, water treatment 1·7, sanitation 1·3 and low birth-weight 0·5 %. The final model explained 23·4 % of the variance in Hb concentration. We conclude that child age, bioavailable-Fe intake, serum retinol concentration, diarrhoea, water treatment, sanitation and low birth-weight are independently associated with Hb concentration. In north-east Brazil, anaemia prevention programmes among children should focus on those aged <2 years and should consider feasible strategies to improve intakes of bioavailable Fe and vitamin A, and reduce infection. Supplemental Fe should be given to low birth-weight infants.

The UK Food Standards Agency convened a group of expert scientists to review current research investigating emerging diet-related surrogate end points for colorectal cancer (CRC). The workshop aimed to overview current research and establish priorities for future research. The workshop considered that the validation of current putative diet-related surrogate end points for CRC and the development of novel ones, particularly in the emerging fields of proteomics, genomics and epigenomics, should be a high priority for future research.