The inclusion of genetically modified (GM) foods in the human diet has caused considerable debate. There is concern that the transfer of plant-derived transgenes to the resident intestinal microflora could have safety implications. For these gene transfer events to occur, the nucleic acid would need to survive passage through the gastrointestinal tract. The aim of the present study was to evaluate the rate at which transgenes, contained within GM soya and maize, are degraded in gastric and small bowel simulations. The data showed that 80 % of the transgene in naked GM soya DNA was degraded in the gastric simulations, while no degradation of the transgenes contained within GM soya and maize were observed in these acidic conditions. In the small intestinal simulations, transgenes in naked soya DNA were degraded at a similar rate to the material in the soya protein. After incubation for 30 min, the transgenes remaining in soya protein and naked DNA were 52 (SEM 13·1) % AND 34 (sem 17·5) %, respectively, and at the completion of the experiment (3 h) these values were 5 % and 3 %, respectively. In contrast to the soya transgene, the maize nucleic acid was hydrolysed in the small intestinal simulations in a biphasic process in which approximately 85 % was rapidly degraded, while the rest of the DNA was cleaved at a rate similar to that in the soya material. Guar gum and tannic acid, molecules that are known to inhibit digestive enzymes, did not influence the rate of transgene degradation in soya protein. In contrast guar gum reduced the rate of transgene degradation in naked soya DNA in the initial stages, but the polysaccharide did not influence the amount of nucleic acid remaining at the end of the experiment. Tannic acid reduced the rate of DNA degradation throughout the small bowel simulations, with 21 (sem 5·4) % and 2 (sem 1·8) % of the naked soya DNA remaining in the presence and absence of the phenolic acid, respectively. These data indicate that some transgenes in GM foods may survive passage through the small intestine.

The aims of this study were to determine the total body phylloquinone and its metabolic turnover in human subjects using a tracer dose of [5-H3]phylloquinone containing 55·5×104 MBq/mmol. Seven subjects aged 22 to 49 years were given 0·3 μg isotopic phylloquinone intravenously on a control diet (75 μg phylloquinone/d) and blood, urine and faeces were sampled periodically for 6 d. Five of these subjects were studied a second time after 3–8 weeks on a low-vitamin K diet (8 μg/d). The changes in the radioactivity of plasma phylloquinone with time were analysed by the method of residuals and fitted to a curve composed of two exponential components. The size of the exchangeable body pool was calculated by isotope dilution. Plasma phylloquinone levels fell during vitamin K restriction but the vitamin K-dependent coagulation factors did not change. After injection the first exponential decay curve t1/2 was 1·0 (SD 0·47) H IN THE SUBJECTS ON THE CONTROL DIET AND 0·49 (sd 0·27) h after vitamin K restriction. On the control diet, the second exponential t1/2 was 27·6 (sd 124) h that did not change on the low-vitamin K diet (t1/2=25·1 (sd 13·5) h). These results indicate that the turnover time for phylloquinone in human subjects is about 1·5 d. Urinary excretion of 3H-metabolites ranged from 30 % of the administered dose on the control diet to 38 % on the restricted diet and had the same turnover rate as the second component of the plasma decay curves. The exchangeable body pool of phylloquinone declined from about 1·0 μg/kg before restriction to lower values after vitamin K restriction. The faecal excretion of phylloquinone and its metabolites fell from 32 % of the administered dose on the control diet to 13 % on the restricted diet.

Diet composition and energy content modulate free fatty acid (FFA) release. The aim of this study was to evaluate the dose–response effects of euenergetic variations in dietary carbohydrate and fat content on postabsorptive FFA release. The rate of appearance (Ra) of palmitate was measured by infusion of [2,2-2H2]palmitate after an overnight fast in six healthy men on three separate occasions, i.e. after 7 d on euenergetic control, high-carbohydrate and high-fat diets. The protein content and composition was identical for each diet. Postabsorptive plasma fatty acid concentrations were not different between the high-carbohydrate and control diets (0·36 (SE 0·07) V. 0·43 (se 0·04) mmol/l), but were increased after the high-fat diet (0·75 (se 0·09) mmol/l, (P<0·01 compared with the other diets). Ra palmitate was not different between the high-carbohydrate and control diets (1·36 (se 0·20) v. 1·47 (se 0·15) μmol/kg per min). However, Ra palmitate was increased to 2·36 (se 0·26) μmol/kg per min after the high-fat diet (P<0·01 compared with the other diets). The fatty acid flux and whole-body fat oxidation were not affected by the high-carbohydrate diet compared with the control diet, but were increased by 67 and 47 % respectively, on the high-fat diet (P<0·01 compared with the other diets). A euenergetic high-fat diet results in increased postabsorptive FFA release and fat oxidation, whereas a euenergetic high-carbohydrate diet does not affect these variables of fat metabolism.

The objectives of the present study were to investigate the recovery of [14C]allantoin in urine of sheep dosed intravenously and degradation of allantoin by rumen micro-organisms. The recovery of [14C]allantoin in the urine of eight sheep was measured during three periods in two experiments. Individual values of [14C]allantoin recovery varied from 66 to 95 % (mean value 83 (SE 1·6) %). The recovery of [14C]allantoin showed no relation to the level of feed intake. There was some evidence that glomerular filtration rate was an important factor affecting the amount of urinary allantoin recovered in one experiment. Incomplete recovery of plasma [14C]allantoin in the urine indicated losses of plasma [14C]allantoin via non-renal routes. This is supported by the disappearance of 14C from rumen contents incubated in vitro with [14C]allantoin for 48 h (88 %) and the presence of 14C in saliva in vivo from sheep sampled after dosing with [14C]allantoin. However, the amount of 14C activity in the saliva was very low (equivalent to only 1·5 % of the total dose in sheep producing saliva at a rate of 15 litres/d). The proportion of renal and non-renal excretion of purine derivatives was found to be unpredictable both between and within individual animals. The factors responsible for this variability need to be identified, and existing models of excretion of purine derivatives may need to be modified accordingly to improve their accuracy of prediction. A single intravenous injection of [4,5-14C]allantoin provides a simple alternative to infusion methods used to measure the proportion of plasma allantoin excreted in the urine of sheep. Using this method it may be feasible to validate PD excretion models in other ruminant livestock.

In the UK, starch contributes up to 25 % of energy intake in adults (). The present study investigated the acute response to a starchy meal on whole-body glucose metabolism and assessed insulin sensitivity in men compared with women. Low insulin sensitivity has been postulated to pre-dispose individuals to a cluster of associated abnormalities known to increase the risk of CHD. Metabolic responses to a 13C-labelled meal were determined in conjunction with a primed continuous infusion of D-[6,6-2H]glucose in groups of healthy age- and BMI-matched men and women. Peripheral plasma glucose disposal (Gd) was computed using non-steady state kinetics in a single compartment model, simultaneously with determination of whole-body net glucose oxidation by indirect calorimetry. Insulin sensitivity was derived using cumulative Gd as the dependent variable, and time and the integrated insulin concentration as independent variables. The female group had the higher fractional rate of glucose appearance in plasma from starch (P=0·019) immediately after ingestion. Females also had a higher rate of plasma Gd and a significantly higher insulin-dependent Gd (6·8 v. 5·6 μg glucose/(min.kg) per pmol insulin, P<0·05) compared with the males. A smaller absolute pool of endogenous glucose in females allowed the rate of exogenous 13CO2 production to be significantly higher in the females (P=0·007) corresponding also to a significantly higher (P<0·05) carbohydrate oxidation rate obtained by indirect calorimetry. The present study suggests that during the ingestion of a starchy meal, females exhibit higher glucose flux and greater whole-body insulin sensitivity than males.

The effect of dietary vitamin E on immunoglobulin A (IgA) antibody production, which acts as the first line of defence at the intestinal mucosa, has not been evaluated in chickens. In the present study the impact of the inclusion of supplementary levels of vitamin E to the diet, on total and antigen-specific IgA antibody titres, T-cell subsets and Ia+ cells, was assessed. From hatching, chickens received a maize-based diet which was supplemented with either 25, 250, 2500 or 5000 mg dl-α-tocopherol acetate/kg. Primary immunisation with tetanus toxoid (T. toxoid) emulsified in a vegetable oil-in-water adjuvant was administered by the intraperitoneal route at 21 d of age. At 35 d of age all birds received an oral booster vaccination of T. toxoid. Significantly higher total IgA antibody titres were present in the day 42 intestinal scrapings of birds receiving the 5000 mg/kg vitamin E-supplemented diet (VESD) (P=0·05) and a notable increase was observed in birds receiving the 250 mg/kg VESD (P=0·06). At days 21 and 42 total serum IgA antibody titres of birds receiving the 250 mg/kg VESD was significantly higher (P<0·05) than the control birds. Following immunisation with T. toxoid, birds receiving the 250 and the 5000 mg/kg VESD had elevated anti-T. toxoid IgA antibody titres in final day intestinal scrapings, which, for the latter group was statistically significant (P=0·02). Both of these groups also demonstrated increased titres of anti-T. toxoid IgA in the serum at day 42. Birds receiving the 250 mg/kg VESD exhibited a notable increase in the percentage of T-helper cells and Ia+ cells in peripheral blood on day 26. The results illustrate the potential for some levels of dietary vitamin E supplementation to act as an immunomodulator of total and antigen-specific IgA antibody.

Exogenous insulin-like growth factor (IGF)-I has been shown to increase growth rate in neonatal pigs while an analogue of IGF-I, long arginine (LR3) IGF-I, has been shown to be more potent than IGF-I in the rat. Therefore, two studies were conducted to determine whether IGF-I and LR3IGF-I increase growth in the artificially-reared neonatal pig. Expt 1 involved forty-two (2 kg initial weight) pigs infused with either control, IGF-I (2, 4 or 8 μg/h) or LR3IGF-I (2, 4 or 8 μg/h) infusions for 8 d. Pigs were weighed and then offered 1·7 MJ (gross energy) milk replacer/kg0·75 per d. Expt 2 involved eighteen pigs (2 kg initial weight) treated with control saline, IGF-I (8 μg/h) or LR3IGF-I (8 μg/h) infusions. After 9 d an additional pump was inserted to increase the infusion rates of each of the growth factors (16 μg/h) for a further 9 d. Cows' milk was provided ad libitum. In Expt 1 there was no overall effect of growth factors on daily weight gain or slaughter weight. However, milk intake was greater in pigs infused with growth factors (909 v. 867 g/d, P=0·027), with an apparently greater milk intake by the pigs infused with IGF-I compared with LR3IGF-I (920 v. 898 g/d, P=0·12). Infusion of LR3IGF-I decreased plasma IGF-I concentrations, but had no effect on plasma IGF-II concentrations. In Expt 2, neither IGF-I nor LR3IGF-I infusion had any effect upon daily weight gain over the first 9 d of the study. However, over the second 9 d of the study, daily weight gain was increased in LR3IGF-I-infused pigs (457 v. 386 g/d, P<0·01), but not in pigs infused with IGF-I (413 v. 386 g/d, P=0·15). Milk intake was not different during the first 9 d of the study but was significantly greater in pigs infused with growth factors over the second half of the study (3407 v. 2905 g/d, P<0·01). Plasma IGF-binding protein-3 concentrations were highly correlated (R=0·85) with average daily gain over the 3 d preceding blood sampling. In conclusion, exogenous IGF-I and particularly LR3IGF-I can increase growth rate and milk intake in artificially-reared pigs fed ad libitum but not in limit-fed piglets.

The DONALD study (Dortmund Nutritional and Anthropometric Longitudinally Designed study) gives the opportunity to evaluate long-term food and nutrient intake data on the basis of 3 d weighed dietary records of infants, children and adolescents since 1985. In this paper, we examine changes in energy and macronutrient intakes (protein, fat, saturated, mono- and polyunsaturated fatty acids, carbohydrates and added sugars) of 795 2–18-year-old subjects between 1985 and 2000 (4483 records). No significant changes in intakes of energy and of protein, polyunsaturated fatty acids and added sugars (as % energy intake, E %) were found. Fat intake decreased significantly in all age groups (between -0·20 and -0·26 E %/year), as well as intake of saturated fatty acids (between -0·11 and -0·14 E %/year) and monounsaturated fatty acids (between -0·07 and -0·014 E %/year). This decline was compensated for by a significant increase in carbohydrate intake (between +0·18 and +0·27 E %/year). The changes in macronutrient intake were mainly due to a decreased consumption of fats–oils (between -0·29 and -1·26 g/year) and meat–fish–eggs (between -0·21 and -2·92 g/year), whereas consumption of bread–cereals (between +0·12 and +2·42 g/year) and potatoes–pasta–rice (between +0·15 and +2·26 g/year) increased slightly. However, since recommended fat intake and fatty acid composition was not reached at the end of the study period by far, further efforts will be necessary to improve macronutrient composition and to stabilize favourable dietary habits.

Intake and sources of phylloquinone (vitamin K1) were examined according to socio-demographic and lifestyle factors in free-living British people aged 65 years and over, from the 1994–5 National Diet and Nutrition Survey. Complete 4-d weighed dietary records were obtained from 1152 participants living in private households. Using newly-available, mainly UK-specific food content data, the weighted geometric mean intake of phylloquinone was estimated at 65 (95 % CI 62, 67) μg/d for all participants, with higher intakes in men than in women (70 v. 61 μg/d respectively, P<0·01). The mean nutrient densities of phylloquinone intake were 9·3 and 10·5 μg/MJ for men and women respectively (P<0·01), after adjusting for age group, region and smoking status. Of all the participants, 59 % had phylloquinone intakes below the current guideline for adequacy of 1 μg/kg body weight per d. Participants aged 85 years and over, formerly in manual occupations, or living in Scotland or in northern England reported lower phylloquinone intakes than their comparative groups. Overall, vegetables contributed 60 % of total phylloquinone intake, with cooked green vegetables providing around 28 % of the total. Dietary supplements contributed less than 0·5 % of phylloquinone intake. Participants living in northern England or in Scotland, in particular, derived less phylloquinone from vegetables than those living in southern England.

Plasma phylloquinone (vitamin K1) concentration was examined according to season, socio-demographic and lifestyle factors and phylloquinone intake in a nationally representative sample of British people aged 65 years and over from the 1994–5 National Diet and Nutrition Survey. Values for both plasma phylloquinone concentration and phylloquinone intake were available from 1076 participants (561 men, 515 women). Eight hundred and thirty-four were living in private households, 242 in residential or nursing homes. Weighted geometric mean plasma phylloquinone concentrations were 0·36 (95 % CI 0·06, 2·01) and 0·24 (95 % CI 0·06, 0·96) nmol/l in free-living and institution samples respectively. Plasma phylloquinone concentrations did not generally differ between men and women, although values in free-living people were significantly lower during autumn and winter (October to March). Plasma phylloquinone concentration was not significantly associated with age. Plasma phylloquinone concentrations were positively correlated with phylloquinone intake in free-living men and women (r 0·18 and 0·30 respectively, both P<0·001). Stepwise multiple regression analysis found that 11 % of the variation in plasma phylloquinone concentration was explained by phylloquinone intake, season and plasma triacylglycerol concentration. After adjustment for age and corresponding nutrient intakes, plasma phylloquinone concentration was significantly associated (each P<0·01) with plasma concentrations of triacylglycerol, cholesterol, retinol and 25-hydroxyvitamin D in free-living women but not men, and with plasma concentrations of carotenes, α- and γ-tocopherols and lutein in free-living men and women. The possibility of concurrent low fat-soluble vitamin status in elderly populations may be a cause for concern.

Determining total energy expenditure (EE) in children under free-living conditions has become of increasingly clinical interest. The aim of this study was to compare three different methods to assess EE triaxial accelerometry (TriTrac-R3D; Professional Products, Division of Reining International, Madison, WI, USA), activity diary and heart-rate (HR) monitoring combined with indirect calorimetry (IC). Twenty non-obese children and adolescents, aged 5.5 to 16.0 years, participated in this study. Results from the three methods were collected simultaneously under free-living conditions during the same 24 h schoolday period. Neither activity diary (5904 (SD 1756) KJ) NOR THE TRITRAC-R3D (6389 (sd 979) kJ) showed statistical differences in 24 h total EE compared with HR monitoring (5965 (sd 1911) kJ). When considering different physical activity (PA) periods, compared with HR monitoring, activity diary underestimates total EE during sedentary periods (P<0·001) and overestimates total EE and PA-EE during PA periods (P<0·001) because of the high energy cost equivalence of activity levels. The TriTrac-R3D, compared with HR monitoring, shows good agreement for assessing PA-EE during PA periods (mean difference +0·25 (sd 1·9) kJ/min; 95 % CI for the bias -0·08, 0·58), but underestimates PA-EE and it does not show good precision during sedentary periods (-0·87 (sd 1·4) kJ/min, P<0·001). Correlation between the vector magnitude generated by the TriTrac-R3D accelerometer and EE of activities derived from HR monitoring is high. When compared with the HR method, the TriTrac-R3D and activity diary are not systematically accurate and must be carefully used for the assessment of children's EE depending on the purpose of each study.