To assess energy balance in very sick medical patients requiring prolonged acute mechanical ventilation and its possible impact on outcome, we conducted an observational study of the first 14 d of intensive care unit (ICU) stay in thirty-eight consecutive adult patients intubated at least 7 d. Exclusive enteral nutrition (EN) was started within 24 h of ICU admission and progressively increased, in absence of gastrointestinal intolerance, to the recommended energy of 125·5 kJ/kg per d. Calculated energy balance was defined as energy delivered − resting energy expenditure estimated by a predictive method based on static and dynamic biometric parameters. Mean energy balance was − 5439 (sem 222) kJ per d. EN was interrupted 23 % of the time and situations limiting feeding administration reached 64 % of survey time. ICU mortality was 72 %. Non-survivors had higher mean energy deficit than ICU survivors (P = 0·004). Multivariate analysis identified mean energy deficit as independently associated with ICU death (P = 0·02). Higher ICU mortality was observed with higher energy deficit (P = 0·003 comparing quartiles). Using receiver operating characteristic curve analysis, the best deficit threshold for predicting ICU mortality was 5021 kJ per d. Kaplan–Meier analysis showed that patients with mean energy deficit ≧5021 kJ per d had a higher ICU mortality rate than patients with lower mean energy deficit after the 14th ICU day (P = 0·01). The study suggests that large negative energy balance seems to be an independent determinant of ICU mortality in a very sick medical population requiring prolonged acute mechanical ventilation, especially when energy deficit exceeds 5021 kJ per d.

Sucrose increases postprandial blood glucose concentrations, and diets with a high glycaemic response may be associated with increased risk of obesity, type 2 diabetes and CVD. Previous studies have suggested that polyphenols may influence carbohydrate digestion and absorption and thereby postprandial glycaemia. Berries are rich sources of various polyphenols and berry products are typically consumed with sucrose. We investigated the glycaemic effect of a berry purée made of bilberries, blackcurrants, cranberries and strawberries, and sweetened with sucrose, in comparison to sucrose with adjustment of available carbohydrates. A total of twelve healthy subjects (eleven women and one man, aged 25–69 years) with normal fasting plasma glucose ingested 150 g of the berry purée with 35 g sucrose or a control sucrose load in a randomised, controlled cross-over design. After consumption of the berry meal, the plasma glucose concentrations were significantly lower at 15 and 30 min (P < 0·05, P < 0·01, respectively) and significantly higher at 150 min (P < 0·05) compared with the control meal. The peak glucose concentration was reached at 45 min after the berry meal and at 30 min after the control meal. The peak increase from the baseline was 1·0 mmol/l smaller (P = 0·002) after ingestion of the berry meal. There was no statistically significant difference in the 3 h area under the glucose response curve. These results show that berries rich in polyphenols decrease the postprandial glucose response of sucrose in healthy subjects. The delayed and attenuated glycaemic response indicates reduced digestion and/or absorption of sucrose from the berry meal.

Studies have shown that various antioxidants are decreased in different age-related degenerative diseases and thus, oxidative stress would have a central role in the pathogenesis of many disorders that involve neuronal degeneration, including Alzheimer's disease (AD). The present study aimed to assess the nutritional status of Se in AD patients and to compare with control subjects with normal cognitive function. The case–control study was carried out on a group of elderly with AD (n 28) and compared with a control group (n 29), both aged between 60 and 89 years. Se intake was evaluated by using a 3-d dietary food record. Se was evaluated in plasma, erythrocytes and nails by using the method of hydride generation atomic absorption spectroscopy. Deficient Se intake was largely observed in the AD group. AD patients showed significantly lower Se levels in plasma, erythrocytes and nails (32·59 μg/l, 43·74 μg/l and 0·302 μg/g) when compared with the control group (50·99 μg/l, 79·16 μg/l and 0·400 μg/g). The results allowed us to suggest that AD has an important relation with Se deficiency.

The aim of the present study was to investigate the usefulness of abdominal diameters to indicate visceral fat, their relationship with serum lipids and their capability of detecting changes in visceral fat. Before and after weight loss, visceral and subcutaneous fat, and the sagittal and transverse diameters were assessed by magnetic resonance imaging (MRI) in forty-seven obese men and forty-seven premenopausal obese women with an initial body mass index of 31·0 (SD 2·4) kg/m2. In a subsample (n 21), diameters, were also measured by anthropometry in the standing and supine positions. They were strongly correlated with the diameters derived from the MRI scans. Serum levels of total and HDL-cholesterol and triacylglycerol were measured before weight loss. In women the sagittal diameter correlated less strongly with visceral fat than anthropometrically-assessed waist circumference and waist:hip ratio (WHR). In men these associations were comparable. Changes in visceral fat with weight loss were more strongly correlated with changes in the sagittal diameter and sagittal:transverse diameter ratio (STR) than with changes in waist circumference or WHR in men. In women, changes in the anthropometric variables and the separate diameters (except STR) were not associated with visceral fat loss. In men, but not in women, both the sagittal diameter and the visceral fat area were related to serum lipids. It is concluded that the sagittal diameter and STR may have advantages over waist circumference and WHR in men, particularly in assessing changes in visceral fat, but this could not be demonstrated in women. The ability to predict visceral fat from circumferences and diameters or their ratios is, however, limited in obese men and women.

Studies on the impact of single nutrients on the risk of CVD have often given inconclusive results. Recent research on dietary patterns has offered promising information on the effects of diet as a whole on the risk of CVD. The Cardiovascular Risk in Young Finns Study is an ongoing, prospective cohort study with a 21-year follow-up to date. The subjects were children and adolescents at baseline (3–18 years, n 1768) and adults at the latest follow-up study (24–39 years, n 1037). We investigated the associations between two major dietary patterns and several risk factors for CVD. In longitudinal analyses with repeated measurements, using multivariate mixed linear regression models, the traditional dietary pattern (characterised by high consumption of rye, potatoes, butter, sausages, milk and coffee) was independently associated with total and LDL cholesterol concentrations, apolipoprotein B and C-reactive protein concentrations among both genders, and also with systolic blood pressure and insulin levels among women and concentrations of homocysteine among men (P < 0·05 for all). A dietary pattern reflecting more health-conscious food choices (such as high consumption of vegetables, legumes and nuts, tea, rye, cheese and other dairy products, and alcoholic beverages) was inversely, but less strongly associated with cardiovascular risk factors. Our results support earlier findings that dietary patterns have a role in the development of CVD.

The objective of the present study was to determine the potential long-term impact of dietary habits on age-related decline among 4809 elderly women (born between 1925 and 1930) in the ‘Etude Epidémiologique de Femmes de la Mutuelle Générale de l'Education Nationale’ (E3N) study, a French epidemiological cohort. In 1993, an extensive diet history self-administered questionnaire was sent to all participants, and in 2006 another questionnaire on instrumental activities of daily living (IADL) and recent cognitive change was sent to a close relative or friend of each woman. Logistic models adjusted for socio-demographic, lifestyle and health factors were performed to evaluate associations between habitual dietary intakes and two outcomes of interest based on the informant response: recent cognitive decline and IADL impairment. Recent cognitive decline was associated with lower intakes of poultry, fish, and animal fats, as well as higher intakes of dairy desserts and ice-cream. IADL impairment was associated with a lower intake of vegetables. The odds of recent cognitive decline increased significantly with decreasing intake of soluble dietary fibre and n-3 fatty acids but with increasing intake of retinol. The odds of IADL impairment increased significantly with decreasing intakes of vitamins B2, B6 and B12. These results are consistent with a possible long-term neuroprotective effect of dietary fibre, n-3 polyunsaturated fats and B-group vitamins, and support dietary intervention to prevent cognitive decline.

The objective of this project was to collect and evaluate data on nutrient intake and status across Europe and to ascertain whether any trends could be identified. Surveys of dietary intake and status were collected from across Europe by literature search and personal contact with country experts. Surveys that satisfied a defined set of criteria – published, based on individual intakes, post-1987, adequate information provided to enable its quality to be assessed, small age bands, data for sexes separated above 12 years, sample size over 25 and subjects representative of the population – were selected for further analysis. In a small number of cases, where no other data for a country were available or where status data were given, exceptions were made. Seventy-nine surveys from 23 countries were included, and from them data on energy, protein, fats, carbohydrates, alcohol, vitamins, minerals and trace elements were collected and tabulated. Data on energy, protein, total fat and carbohydrate were given in a large number of surveys, but information was very limited for some micronutrients. No surveys gave information on fluid intake and insufficient gave data on food patterns to be of value to this project. A variety of collection methods were used, there was no consistency in the ages of children surveyed or the age cut-off points, but most surveys gave data for males and females separately at all ages. Just under half of the surveys were nationally representative and most of the remainder were regional. Only a small number of local surveys could be included. Apart from anthropometric measurements, status data were collected in only seven countries. Males had higher energy intakes than females, energy intake increased with age but levelled off in adolescent girls. Intakes of other nutrients generally related to energy intakes. Some north–south geographical trends were noted in fat and carbohydrate intakes, but these were not apparent for other nutrients. Some other trends between countries were noted, but there were also wide variations within countries. A number of validation studies have shown that misreporting is a major problem in dietary surveys of children and adolescents and so all the dietary data collected for this project should be interpreted and evaluated with caution In addition, dietary studies rely on food composition tables for the conversion of food intake data to estimated nutrient intakes and each country uses a different set of food composition data which differ in definitions, analytical methods, units and modes of expression. This can make comparisons between countries difficult and inaccurate. Methods of measuring food intake are not standardised across Europe and intake data are generally poor, so there are uncertainties over the true nutrient intakes of children and adolescents across Europe. There are insufficient data on status to be able to be able to draw any conclusions about the nutritional quality of the diets of European children and adolescents.

Although gestational diabetes mellitus (GDM) is associated with an increased risk of maternal and neonatal morbidity, there is no consensus as to the optimal approach of nutritional management in these patients. The present study was designed to assess the effect of the Dietary Approaches to Stop Hypertension (DASH) eating plan on glucose tolerance and lipid profiles of pregnant women with GDM. The present randomised controlled clinical trial was performed among thirty-four women diagnosed with GDM at 24–28 weeks of gestation. Subjects were randomly assigned to consume either the control diet (n 17) or the DASH eating pattern (n 17) for 4 weeks. The control diet was designed to contain 45–55 % carbohydrates, 15–20 % protein and 25–30 % total fat. The macronutrient composition of the DASH diet was similar to the control diet; however, the DASH diet was rich in fruits, vegetables, whole grains and low-fat dairy products, and contained lower amounts of saturated fats, cholesterol and refined grains with a total of 2400 mg Na/d. Fasting blood samples were taken at baseline and after 4 weeks of intervention to measure fasting plasma glucose, glycated Hb (HbA1c) and lipid profiles. Participants underwent a 3 h oral glucose tolerance tests and blood samples were collected at 60, 120 and 180 min to measure plasma glucose levels. Adherence to the DASH eating pattern, compared with the control diet, resulted in improved glucose tolerance such that plasma glucose levels reduced at 60 ( − 1·86 v. − 0·45 mmol/l, Pgroup= 0·02), 120 ( − 2·3 v. 0·2 mmol/l, Pgroup= 0·001) and 180 min ( − 1·7 v. 0·22 mmol/l, Pgroup= 0·002) after the glucose load. Decreased HbA1c levels ( − 0·2 v. 0·05 %, Pgroup= 0·001) was also seen in the DASH group compared with the control group. Mean changes for serum total ( − 0·42 v. 0·31 mmol/l, Pgroup= 0·01) and LDL-cholesterol ( − 0·47 v. 0·22 mmol/l, Pgroup= 0·005), TAG ( − 0·17 v. 0·34 mmol/l, Pgroup= 0·01) and total:HDL-cholesterol ratio ( − 0·6 (sd 0·9) v. 0·3 (sd 0·8), Pgroup= 0·008) were significantly different between the two diets. Additionally, consumption of the DASH diet favourably influenced systolic blood pressure ( − 2·6 v. 1·7 mmHg, Pgroup= 0·001). Mean changes of fasting plasma glucose ( − 0·29 v. 0·15 mmol/l, Pgroup= 0·09) were non-significant comparing the DASH diet with the control diet. In conclusion, consumption of the DASH eating pattern for 4 weeks among pregnant women with GDM resulted in beneficial effects on glucose tolerance and lipid profiles compared with the control diet.

Phylloquinone is converted into menaquinone-4 and accumulates in extrahepatic tissues. Neither the route nor the function of the conversion is known. One possible metabolic route might be the release of menadione from phylloquinone by catabolic activity. In the present study we explored the presence of menadione in urine and the effect of vitamin K intake on its excretion. Menadione in urine was analysed by HPLC assay with fluorescence detection. Urine from healthy male volunteers was collected before and after administration of a single dose of K vitamins. Basal menadione excretion in non-supplemented subjects (n 6) was 5·4 (sd 3·2) μg/d. Urinary menadione excretion increased greatly after oral intake of the K vitamins, phylloquinone and menaquinone-4 and -7. This effect was apparent within 1–2h and peaked at about 3h after intake. Amounts of menadione excreted in 24h after vitamin K intake ranged, on a molar basis, from 1 to 5% of the administered dose, indicating that about 5–25% of the ingested K vitamins had been catabolized to menadione. Menadione excretion was not enhanced by phylloquinone administered subcutaneously or by 2′,3′-dihydrophylloquinone administered orally. In archived samples from a depletion/repletion study (Booth et al. (2001)Am J Clin Nutr74, 783–790), urinary menadione excretion mirrored dietary phylloquinone intake. The present study shows that menadione is a catabolic product of K vitamins formed after oral intake. The rapid appearance in urine after oral but not subcutaneous administration suggests that catabolism occurs during intestinal absorption. The observations make it likely that part of the menaquinone-4 in tissues results from uptake and prenylation of circulating menadione

1. Quantitative selenium metabolism has been studied in normal young New Zealand women by measuring total Se intake and urinary and faecal Se output, and by using values for absorption, excretion and turnover of 75Se determined after administration of [75Se]selenomethionine or [75Se]selenite.

2. In a period of 14 d when a normal ad lib. diet was being consumed, mean dietary Se for four women was 24.2 μg/d, mean urinary Se was 13.1 μg/d and mean faecal Se was 10.8 μg/d; mean Se balance during this time was +0.3 μg/d.

3. Intestinal absorption of food Se was 0.76−0.83 of intake (mean 0.79).

4. Whole-body Se was calculated in three different ways; (a) using the specific activity of urinary Se and retained whole-body 75Se; (b) using plasma Se and the occupancy of 75Se in whole-body and plasma; (c) using absorbed food Se and the occupancy of absorbed 75Se in whole-body.

5. Whole-body Se calculated from measurements obtained following the administration of [75Se]seleno-methionine was 4.7−10.0 mg (mean 6.9) using method (a), 4.1−7.2 mg (mean 5.2) using method (b) and 4.3−8.9 mg (mean 6.2) using method (c).

6. Whole-body Se calculated from results obtained after giving [75Se]selenite was 2.7−3.4 mg (mean 2.9) using method (a), 2.3−5.0 mg (mean 3.5) using method (b) and 2.1−3.0 mg (mean 2.6) using method (c).

7. The results of this study indicate that the minimum dietary requirement of Se for the maintenance of normal human health is probably not more than 20 μg/d.

The present study measured changes in antioxidant enzyme activity in, and the phospholipid fatty acid composition of the ageing rat brain and tested whether dietary supplementation with thyme oil or thymol could provide beneficial effects. There were significant declines in superoxide dismutase (EC 1.15.1.1) and glutathione peroxidase (EC 1.11.1.9) activities and the total antioxidant status in the untreated rats with age, while thyme-oil- and thymol-fed rats maintained significantly higher antioxidant enzyme activities and total antioxidant status. The proportions of 18: 2n−6, 20: 1n−9, 22: 4n−6 and 22: 5n−3 in the brain phospholipids resulting from all three dietary treatments were significantly higher in 28-month-old rats than in 7-month-old rats. Only 20: 1n−9 levels in 28-month-old thyme-oil- and thymol-treated rats were significantly higher than in the age-matched control. The proportion of 22: 6n−3 in brain phospholipids, which declined with age in control rats, was also significantly higher in rats given either supplement. This latter finding is particularly important as optimum levels of 22: 6n−3 are required for normal brain function. These results highlight the potential benefit of thyme oil as a dietary antioxidant.

1. Examination of the digesta from all regions of the avian digestive tract showed that volatile fatty acids (VFAs) were present in greatest concentration in the caeca and that they comprised mainly acetic, propionic and butyric acids.

2. All droppings contained VFAs but they were present in highest concentration in those of caecal origin. Caecectomy was followed by a marked reduction in the total output of VFAs.

3. Birds 14–20 weeks of age had similar concentrations of VFAs along the tract and similar numbers and distribution of micro-organisms.

4. Portal blood contained all the VFAs found in the digestive tract whilst peripheral blood contained only acetic and formic acids.

5. The almost complete absence of VFAs from the tract contents of germ-free birds showed that the VFAs normally present in the tract were of microbial origin.

6. The presence of similar levels of acetate in the peripheral blood of conventional and germfree birds indicated that circulating acetate was mainly of endogenous and not microbial origin.

7. The significance of VFAs as an energy source is discussed.

The effect of feeding live Lactobacillus reuteri cells containing active bile salt hydrolase (BSH) on plasma cholesterol levels was studied in pigs. During an experiment lasting 13 weeks, twenty pigs were fed on a high-fat, high-cholesterol, low-fibre diet for the first 10 weeks, and a regular pig diet for the last 3 weeks. One group of animals received, twice daily, 11·25 (SD 0·16) log10 colony forming units of the potential probiotic bacteria for 4 weeks (from week 3 until week 7). From week 8 onwards, the treated group was again fed on the same diet as the control group without additions. The total faecal Lactobacillus counts were only significantly higher in the treated pigs during the first 2 weeks of L. reuteri feeding. Based on limited data, it was suggested that the administered Lactobacillus species had caused a temporary shift within the indigenous Lactobacillus population rather than permanently colonizing the intestinal tract. The probiotic feeding brought about significant lowering (P ≤ 0·05) of total and LDL-cholesterol concentrations in the treated pigs compared with the control pigs, while no change in HDL-cholesterol concentration was observed. The data for faecal output of neutral sterols and bile salts were highly variable between the animals of each group, yet they indicated an increased output in the treated pigs. Although the blood cholesterol levels went up in both groups during the 3 weeks following the Lactobacillus administration period, significantly lower serum total and LDL-cholesterol levels were observed in the treated pigs. During the final 3 weeks of normalization to the regular diet, cholesterol concentrations significantly decreased in both animal groups and the differences in total and LDL-cholesterol concentrations between the groups largely disappeared.

We tested the validity and reliability of the BioSpace InBody 320, Omron and Bod-eComm body composition devices in men and women (n 254; 21–80 years) and boys and girls (n 117; 10–17 years). We analysed percentage body fat (%BF) and compared the results with dual-energy X-ray absorptiometry (DEXA) in adults and compared the results of the InBody with underwater weighing (UW) in children. All body composition devices were correlated (r 0·54–0·97; P ≤ 0·010) to DEXA except the Bod-eComm in women aged 71–80 years (r 0·54; P = 0·106). In girls, the InBody %BF was correlated with UW (r 0·79; P ≤ 0·010); however, a more moderate correlation (r 0·69; P ≤ 0·010) existed in boys. Bland–Altman plots indicated that all body composition devices underestimated %BF in adults (1·0–4·8 %) and overestimated %BF in children (0·3–2·3 %). Lastly, independent t tests revealed that the mean %BF assessed by the Bod-eComm in women (aged 51–60 and 71–80 years) and in the Omron (age 18–35 years) were significantly different compared with DEXA (P ≤ 0·010). In men, the Omron (aged 18–35 years), and the InBody (aged 36–50 years) were significantly different compared with DEXA (P = 0·025; P = 0·040 respectively). In addition, independent t tests indicated that the InBody mean %BF in girls aged 10–17 years was significantly different from UW (P = 0·001). Pearson's correlation analyses demonstrated that the Bod-eComm (men and women) and Omron (women) had significant mean differences compared with the reference criterion; therefore, the %BF output from these two devices should be interpreted with caution. The repeatability of each body composition device was supported by small CV ( < 3·0 %).

Oranges are rich sources of flavonoids that are bioactive and may protect against age-related diseases. The absorption of orange flavanones may be affected by factors such as processing and subject anthropometric variables, and the bioactivity of the absorbed phytochemicals depends on how they are metabolised during absorption. In a randomised cross-over study, twenty subjects consumed a single portion of orange fruit (150 g) or juice (300 g) that contained the flavanones narirutin and hesperidin, and an additional 109 subjects across a broad age range (18–80 years) consumed the juice. Flavanone metabolites were measured in regularly collected samples of plasma and urine. After consumption of fruit or juice, flavanone conjugates, but not the aglycones, were detected in plasma and urine. The flavanone conjugates were shown to include the 7- and 4′-O-monoglucuronides of naringenin, the 7- and 3′-O-monoglucuronides of hesperetin, two hesperetin diglucuronides and a hesperetin sulfo-glucuronide, but no aglycones or rutinosides. Analysis of the plasma pharmacokinetic and urinary excretion data on a dose-adjusted basis indicated no difference in absorption or excretion of either flavanone between the fruit and juice matrices. In the extended urinary excretion dataset the individual variation was very large (range 0–59 % urinary yield). There was a small but significant (P < 0·05) decrease in the excretion of hesperetin (but not naringenin) with increasing age (P < 0·05), but the effects of sex, BMI and contraceptive pill use were shown not to be associated with the variation in flavanone excretion.

Lactoferrin has antimicrobial activity associated with peptide fragments lactoferricin (LFC) and lactoferrampin (LFA) released on digestion. These two fragments have been expressed in Photorhabdus luminescens as a fusion peptide linked to protein cipB. The construct cipB–LFC–LFA was tested as an alternative to antimicrobial growth promoters in pig production. Sixty piglets with an average live body weight of 5·42 (sem 0·59) kg were challenged with enterotoxigenic Escherichia coli and randomly assigned to four treatment groups fed a maize–soyabean meal diet containing either no addition (C), cipB at 100 mg/kg (C+B), cipB–LFC–LFA at 100 mg/kg (C+L) or colistin sulfate at 100 mg/kg (C+CS) for 3 weeks. Compared with C, dietary supplementation with C+L for 3 weeks increased daily weight gain by 21 %, increased recovery from diarrhoea, enhanced serum glutathione peroxidase (GPx), peroxidase (POD) and total antioxidant content (T-AOC), liver GPx, POD, superoxide dismutase and T-AOC, Fe, total Fe-binding capacity, IgA, IgG and IgM levels (P < 0·05), decreased the concentration of E. coli in the ileum, caecum and colon (P < 0·05), increased the concentration of lactobacilli and bifidobacteria in the ileum, caecum and colon (P < 0·05), and promoted development of the villus–crypt architecture of the small intestine. Growth performance was similar between C+L- and C+CS-supplemented pigs. The present results indicate that LFC–LFA is an effective alternative to the feed antibiotic CS for enhancing growth performance in piglets weaned at age 21 d.

1. Nine experiments, each with one of six sheep with cannulated rumens given a constant diet of dried grass, were made in which oleic, linoleic or linolenic acid was infused into the rumen and energy and lipid metabolism were measured. One experiment was made in which palmitic acid was given. 2. Judged by changes in the composition of isolated fatty acids, the unsaturated fatty acids were hydrogenated in the rumen. An increase in the excretion of lipid in the faeces occurred when the unsaturated acids were given. The heat of combustion of the faeces increased by 12.6±3.0 kcal/100 kcal fatty acid, of which 94% was accounted for by the additional lipid. 3. Methane production fell when the unsaturated fatty acids were infused, the decreases being 13.8±1.6 kcal CH4;/I00 kcal oleic acid, 14.2±1.5 kcal CH4/100 kcal linoIeic acid and 16.4±1.3 kcal CH4/100 kcal Iinolenic acid. The introduction of a double bond into an n-alkyl acid was calculated to reduce methane production by 0.24±0.09 moles/mole double bond. 4. Because the depression of methane production on infusing the fatty acids exceeded the increase in the heat of combustion of the faeces, the metabolizable energy of the fatty acids was 104.1±5.3% of their heat of combustion. 5. The efficiencies with which the fatty acids were used to promote energy retention were 74.6±5.7% for oleic acid, 79.2±2.0 % for linoleic acid and 82.5±3.0% for linolenic acid. These efficiencies agreed with those noted in experiments by others with rats, horses and pigs given glycerides, but were higher than those noted by others when glycerides were added to the diets of ruminants.

The lack of robust measures of dietary exposure hinders a quantitative understanding of causal relationships between diet and health. Non-targeted metabolite fingerprinting was used to explore the relationships between citrus exposure in free-living human subjects, estimated by a FFQ, and the chemical content of urine. Volunteers (study 1, n 12; study 2, n 11) were classified into high-, medium- and low-frequency citrus consumption groups. Overnight and spot fasting urine samples were obtained after exposure to a standardised citrus-free evening meal. The urine samples were analysed by flow injection electrospray-ionisation MS followed by supervised multivariate data classification analysis to discover discriminatory features associated with the level of citrus exposure. Good separation of high and low citrus consumption classes was achieved. Deeper exploration of high-ranked explanatory mass signals revealed several correlated signals derived from proline betaine. Targeted analysis of the relative levels of proline betaine in both fasting and overnight urine samples demonstrated good correlation with FFQ exposure data. Acute exposure of volunteers to orange juice resulted in the appearance of proline betaine and several biotransformed products in postprandial urine samples. Biomarker validation showed sensitivities of 80·8–92·2 % and specificities of 74·2–94·1 % (false discovery rate-adjusted P values < 0·05) for elevated proline betaine in participants who reported high citrus consumption. Proline betaine biotransformation products displayed weaker quantitative relationships with habitual citrus exposure. Targeted screening for the presence of biotransformation products of hesperidin and narirutin, known to be abundant in oranges, revealed that they were relatively poor indicators of citrus exposure.

The human vitamin K requirement is not known precisely, but the minimal requirement is often assumed to be between 0·5 and 1 x 10−6g/kg body weight. In the present study we addressed the question to what extent circulating vitamin K concentrations are influenced by the form in which the vitamer is consumed. The experimental group consisted of five healthy volunteers who received phylloquinone after an overnight fast. On the first day of three successive weeks the participants consumed 1 mg (2·2 µmol) phylloquinone, either in the form of a pharmaceutical preparation (Konakion®), or in the form of spinach + butter, or as spinach without added fat. Circulating phylloquinone levels after spinach with and without butter were substantially lower (7·5- and 24·3-fold respectively) than those after taking the pharmaceutical concentrate. Moreover, the absorption of phylloquinone from the vegetables was 1·5 times slower than from Konakion. In a second experiment in the same five volunteers it was shown that relatively high amounts of menaquinone-4 enter the circulation after the consumption of butter enriched with this vitamer. It is concluded that the bioavailability of membrane-bound phylloquinone is extremely poor and may depend on other food components, notably fat. The bioavailability of dietary vitamin K (phylloquinone+menaquinones) is lower than generally assumed, and depends on the form in which the vitamin is ingested. These new insights may lead to a revision of the recommended daily intake for vitamin K.