Several studies have investigated the potential health benefits, including those associated with neurological function, of the n-3 fatty acid DHA. This has arisen in part because of the association between higher intakes of fish, which is a major dietary source of DHA, and reduced disease risk. In addition to DHA, fish also provides choline and vitamin D. The objective of the present study was to assess whether women in the first half of pregnancy with low fish intake also had low blood concentrations of vitamin D, choline and DHA. A total of 222 pregnant women at 16 weeks of gestation were examined for dietary intake, erythrocyte (phosphatidylethanolamine PE) DHA, plasma free choline and 25-hydroxyvitamin D (25(OH)D). Women who consumed ≤ 75 g fish/week (n 56) compared to ≥ 150 g fish/week (n 116) had lower dietary intake of DHA, total choline and vitamin D (P< 0·001), and lower erythrocyte PE DHA (5·25 (sd 1·27), 6·83 (sd 1·62) g/100 g total fatty acid, respectively, P< 0·01), plasma free choline (6·59 (sd 1·65), 7·40 (sd 2·05) μmol/l, respectively, P= 0·023) and 25(OH)D (50·3 (sd 20·0), 62·5 (sd 29·8) nmol/l, respectively, P< 0·01). DHA intake was positively related to the intake of vitamin D from foods (ρ 0·47, P< 0·001) and total choline (ρ 0·32, P< 0·001). Dietary intakes and biomarkers of DHA, choline and vitamin D status were assessed to be linked. This raises the possibility that unidentified concurrent nutrient inadequacies might have an impact on the results of studies addressing the benefits of supplemental DHA.

The aim of the present study was to investigate in human subjects whether or not the ingestion of two liquid meals that differed only in their fatty acid composition (due to the addition of olive oil (group O) or sunflowerseed oil (group S) as the source of dietary fat) would lead to differences in the pancreatic enzyme activities secreted into the duodenum. The experiments were performed in eighteen cholecystectomized subjects who, during the 30d period immediately before surgery, modified their habitual diets in such a way that their fat composition would reflect, as far as possible, that of the experimental meals. Lipase (EC 3.1.1.3), colipase, amylase (EC 3.2.1.1), chymotrypsin (EC 3.4.21.1) and trypsin (EC 3.4.21.4) activities were measured in duodenal contents aspirated before and after the ingestion of the test meals. The plasma levels of secretin and cholecystokinin (CCK) were also examined. Duodenal enzyme activities were similar in resting conditions. No significant differences were revealed in postprandial enzyme activities, except for lipase activity, which was higher in group O, probably in relation to the greater plasma CCK concentrations observed in this group. In the absence of enzyme output data, we should not exclude the possibility that the type of dietary fat will affect human pancreatic enzyme secretion to a greater extent than is evident from the present study, for instance through a flow-mediated effect, as we previously observed in dogs.

β-Ionone (BI), a precursor for carotenoids, is widely distributed in fruit and vegetables. Recent in vitro studies have demonstrated the potential anti-metastatic effects of BI, but the mechanisms underlying such actions are not clear. Because liver cancer is the most endemic cancer in Taiwan and in a large region of the world, we hereby investigate the anti-metastatic effects of BI and its mechanisms of actions in a highly metastatic human hepatocarcinoma SK-Hep-1 cells. We show that incubation of cells with BI (1–50 μm) for 24 and 48 h significantly inhibited cell invasion, migration and adhesion. Mechanistically, incubation of cells with BI (1–50 μm) for 24 h resulted in the following: (1) significant inhibition of matrix metalloproteinase (MMP)-2, MMP-9 and urokinase-type plasminogen activator activities, (2) up-regulation of protein expression of the tissue inhibitor of matrix metalloproteinase (TIMP)-1, TIMP-2 and plasminogen activator inhibitor-1, (3) down-regulation of the expression of migration-related proteins, including focal adhesion kinase (FAK), phosphorylated form of FAK, Rho, Rac1 and Cdc42 and (4) up-regulation of the expression of nm23-H1 protein (P < 0·05). Overall, the results show that BI effectively inhibits the metastasis of SK-Hep-1 cells, and this effect involves the regulation of gene expression and signal pathways related to invasion and migration.

The prevalence of goitre in areas with mildly excessive iodine in drinking-water is still under debate. The aim of the present study was to explore the prevalence of goitre and its epidemiological characteristics in these areas using WHO criteria. To this end, three towns with a median water iodine (MWI) of 150–300 μg/l were selected randomly in Hengshui City of Hebei Province of China, and one town with a MWI of 105 μg/l was chosen as a control. A total of 452 children in the three towns and 120 children in the control town were randomly selected to measure thyroid volume (Tvol) by ultrasound. The goitre status of these children was judged using the criteria recommended by the WHO. The overall goitre prevalences in the three towns were 24·6 % (111/452) by age-specific Tvol and 33·0 % (149/452) by body surface area (BSA)-adjusted Tvol, which were significantly higher than that in the control town by both age-specific Tvol (14·0 % (17/120), P= 0·015) and BSA-adjusted Tvol (17·5 % (21/120), P= 0·001). Significant differences were found neither in goitre prevalences across sex by both age-specific Tvol (P= 0·078) and BSA-adjusted Tvol (P= 0·692) nor in that across age group by both BSA-adjusted Tvol (P= 0·461) and age-specific Tvol (P= 0·183). The present study found a high prevalence of goitre in children living in areas with mildly excessive iodine in drinking-water in Hebei Province. No significant differences were found in goitre prevalences across sex and age group. These results suggest that the Tvol reference values recommended by the WHO could be too low for Chinese children.

1. The vitamin B12 in sows' milk is strongly attached to a specific ‘binder’ protein, which is present in excess. The influence of this ‘binder’ on the uptake and retention of cyanocobalamin and two natural analogues(cobinamide and Co-α-[2-methyladenyl]cobamide) was investigated with neonatal piglets.

2. Retention of a single oral dose of cyano[58Co]cobalamin given before 7 d of age was consistently higher with suckled than with early-weaned piglets, as determined by measurement of whole-body radioactivity.

3. Efficiency of retention declined with age, more rapidly in early-weaned than in suckled animals; when the dose was given at 14 d approximately 30% was retained by both groups.

4. Distribution of the retained cyano[58Co]cobalamin within the body of the piglets was the same in both groups; about half was present in the liver.

5. Foraging piglets may ingest adventitious vitamin B12 and its analogues, which are present in the sow's faeces and in contaminated litter. The influence of the vitamin B12-binder in sows' milk on the uptake and retention of two non-cobalamin analogues, and the effects of the analogues on the uptake and retention of vitamin B12 from 2 to 14 d after parturition, were investigated with early-weaned piglets.

6. The analogues were detected in the liver but not in the body organs. They were also present in blood plasma, urine and bile, in high concentration relative to that of vitamin B12. The content of analogues in the liver was very small in relation to the amounts ingested, and much less than that of vitamin B12. There was no indication that the vitamin B12-binder in sows' milk influenced uptake and retention of the analogues, or that ingestion of analogues affected the content of vitamin B12 in the body organs and fluids examined.

Ageing women may choose to drink soya milk to reduce menopausal symptoms. As fermentation enriches soya milk with isoflavone aglycones, its beneficial qualities may improve. To reduce osteoporotic risk, however, soya milk must be Ca enriched, and it is not known how fermentation affects Ca bioavailability. A randomised crossover pilot study was undertaken to compare the Ca absorption of fortified soya milk with that of fermented and fortified soya milk in twelve Australian osteopenic post-menopausal women. The fortified soya milk was inoculated with Lactobacillus acidophilus American Type Culture Collection (ATCC) 4962 and fermented for 24 h at 37°C. Ca absorption from soya milk samples was measured using a single isotope radiocalcium method. Participants had a mean age of 54·8 (sd 12·3) years, with mean BMI of 26·5 (sd 5·5) kg/m2 and subnormal to normal serum 25-hydroxyvitamin D (mean 62·5 (sd 19·1) nmol/l). Participants consumed 185 kBq of 45Ca in 44 mg of Ca carrier. The mean fractional Ca absorption (α) from soya milk and fermented soya milk was 0·64 (sd 0·23) and 0·71 (sd 0·29), respectively, a difference not of statistical significance (P = 0·122). Although fermentation of soya milk may provide other health benefits, fermentation had little effect on acute Ca absorption.

The H+-coupled transporter, peptide transporter 1 (PepT1), is responsible for the uptake of dietary di- and tripeptides in the intestine. Using an in vivo continuously perfused gut loop model in Yucatan miniature pigs, we measured dipeptide disappearance from four 10 cm segments placed at equidistant sites along the length of the small intestine. Pigs were studied at 1, 2, 3 (suckling) and 6 weeks (post-weaning) postnatal age. Transport capability across the PepT1 transporter was assessed by measuring the disappearance of 3H-glycylsarcosine; real-time RT-PCR was also used to quantify PepT1 mRNA. Each of the regions of intestine studied demonstrated the capacity for dipeptide transport. There were no differences among age groups in transport rates measured in the most proximal intestine segment. Transport of 3H-glycylsarcosine was significantly higher in the ileal section in the youngest age group (1 week) compared with the other the suckling groups; however, all suckling piglet groups demonstrated lower ileal transport compared with the post-weaned pigs. Colonic PepT1 mRNA was maximal in the earliest weeks of development and decreased to its lowest point by week 6. These results suggest that peptide transport in the small intestine may be of importance during the first week of suckling and again with diet transition following weaning.

Endothelial function is a key mechanism in the development of CVD. Arginine and exercise are important non-pharmacological strategies for mitigating the impact of metabolic changes in the metabolic syndrome, but the effect of their combined administration is unknown. Thus, the aim of this study was to investigate the isolated and combined effects of aerobic training and arginine supplementation on metabolic variables and vascular reactivity in rats at high risk for developing the metabolic syndrome. Wistar rats were divided into two groups: control and fructose (F – water with 10 % fructose). After 2 weeks, the F group was divided into four groups: F, fructose+arginine (FA, 880 mg/kg per d of l-arginine), fructose+training (FT) and fructose+arginine+training (FTA); treatments lasted for 8 weeks, and no difference was observed in body mass gain. Arginine did not improve the body protein content, and both the FA and FT groups show a reversal of the increase in adipose tissue. Insulin increase was prevented by training and arginine, without additive effect, and the increase in serum TAG was prevented only by training. The F group showed impaired endothelium-dependent vasodilation and hyperreactivity to phenylephrine, but arginine and training were capable of preventing these effects, even separately. Higher nitric oxide level was observed in the FA and FT groups, and no potentiating effect was detected. Thus, only training was able to prevent the increase in TAG and improve the protein mass, and training and arginine exert similar effects on fat content, insulin and endothelial function, but these effects are not additive.

Vitamin D deficiency is a global public health concern. Studies of serum 25-hydroxyvitamin D (25(OH)D) determinants in young women are limited and few include objective covariates. Our aims were to define the prevalence of vitamin D deficiency and examine serum 25(OH)D correlates in an exploratory study of women aged 16–25 years. We studied 348 healthy females living in Victoria, Australia, recruited through Facebook. Data collected included serum 25(OH)D assayed by liquid chromatography-tandem MS, relevant serum biochemistry, soft tissue composition by dual-energy X-ray absorptiometry, skin melanin density, Fitzpatrick skin type, sun exposure using UV dosimeters and lifestyle factors. Mean serum 25(OH)D was 68 (sd 27) nmol/l and 26 % were vitamin D deficient (25(OH)D <50 nmol/l). The final model explained 56 % of 25(OH)D variance. Serum sex hormone-binding globulin levels, creatinine levels, sun exposure measured by UV dosimeters, a positive attitude towards sun tanning, typically spending >2 h in the sun in summer daily, holidaying in the most recent summer period, serum Fe levels, height and multivitamin use were positively associated with 25(OH)D. Fat mass and a blood draw in any season except summer was inversely associated with 25(OH)D. Vitamin D deficiency is common in young women. Factors such as hormonal contraception, sun exposure and sun-related attitudes, as well as dietary supplement use are essential to consider when assessing vitamin D status. Further investigation into methods to safely optimise vitamin D status and to improve understanding of the impact of vitamin D status on long-term health outcomes is required.

In the past, food-based dietary guidelines (FBDGs) were derived nearly exclusively by using systematic reviews on diet–health relationships and translating dietary reference values for nutrient intake into foods. This approach neglects many other implications that dietary recommendations have on society, the economy and environment. In view of pressing challenges, such as climate change and the rising burden of diet-related diseases, the simultaneous integration of evidence-based findings from different dimensions into FBDGs is required. Consequently, mathematical methods and data processing are evolving as powerful tools in nutritional sciences. The possibilities and reasons for the derivation of FBDGs via mathematical approaches were the subject of a joint workshop hosted by the German Nutrition Society (DGE) and the Federation of European Nutrition Societies (FENS) in September 2019 in Bonn, Germany. European scientists were invited to discuss and exchange on the topics of mathematical optimisation for the development of FBDGs and different approaches to integrate various dimensions into FBDGs. We concluded that mathematical optimisation is a suitable tool to formulate FBDGs finding trade-offs between conflicting goals and taking several dimensions into account. We identified a lack of evidence for the extent to which constraints and weights for different dimensions are set and the challenge to compile diverse data that suit the demands of optimisation models. We also found that individualisation via mathematical optimisation is one perspective of FBDGs to increase consumer acceptance, but the application of mathematical optimisation for population-based and individual FBDGs requires more experience and evaluation for further improvements.

1. Analbuminaemic and Sprague-Dawley (control) rats were fed on low- (60 g/kg) protein and control (200 g protein/kg) diets ad lib. from weaning. Males and females were studied separately. Body-weight and plasma protein concentrations were determined at 10 d intervals from 25 to 75 d of age. Electrophoresis of plasma proteins was performed in samples from day 75. Extracellular fluid volume was measured at 10 d intervals from day 45 onwards. Colloid osmotic pressure was measured in plasma and interstitial fluid (wick technique) at the start and end of the trial.

2. Body-weight increased much less on the low-protein diet than on the normal diet in both strains and sexes. The growth retardation was slightly more pronounced in the male analbuminaemic rats than in the male Sprague-Dawley controls.

3. Plasma protein concentration increased during normal growth in all groups, particularly in the female analbuminaemic rats. This increase was reduced by the 60 g protein/kg diet in all groups, with the exception of the male analbuminaemic rats.

4. Differences in plasma colloid osmotic pressure were similar to those seen in plasma protein concentration. Interstitial colloid osmotic pressure was higher in the control rats than in the analbuminaemic ones. The interstitial colloid osmotic pressure increased during growth in the control but not in the analbuminaemic rats. The difference in interstitial colloid osmotic pressure between the strains was maintained during low-protein intake, but at a lower level than during normal protein intake.

5. Subtracting interstitial from plasma colloid osmotic pressure, resulted in a rather similar transcapillary oncotic gradient in the various groups at 75 d, both on the control protein diet (11–14 mmHg), and on the lowprotein diet (9–11 mmHg).

6. All protein fractions were reduced to a similar extent by the low-protein diet in the control rats, whereas in the analbuminaemic rats protein fractions produced in the liver were more severely depressed.

7. Extracellular fluid volume as a percentage of body-weight was similar in all groups, and decreased with increasing age.

8. In conclusion, the analbuminaemic rats were able to maintain the transcapillary oncotic gradient on both diets by reducing the interstitial colloid osmotic pressure. Oedema was not observed.

9. Despite the absence of albumin, the protein-malnourished analbuminaemic rat is no more susceptible to hypoproteinaemia and oedema than its normal counterpart.

In the present study, the aim was to investigate the correlation between the acute and habitual dietary intake of flavanones, their main food sources and the concentrations of aglycones naringenin and hesperetin in 24 h urine in a European population. A 24-h dietary recall (24-HDR) and a 24-h urine sample were collected the same day from a subsample of 475 people from four different countries of the European Prospective Investigation into Cancer and Nutrition study. Acute and habitual dietary data were captured through a standardised 24-HDR and a country/centre-specific validated dietary questionnaire (DQ). The intake of dietary flavanones was estimated using the Phenol-Explorer database. Urinary flavanones (naringenin and hesperetin) were analysed using tandem MS with a previous enzymatic hydrolysis. Weak partial correlation coefficients were found between urinary flavanone concentrations and both acute and habitual dietary flavanone intakes (Rpartial = 0·14–0·17). Partial correlations were stronger between urinary excretions and acute intakes of citrus fruit and juices (Rpartial ∼ 0·6) than with habitual intakes of citrus fruit and juices (Rpartial ∼ 0·24). In conclusion, according to our results, urinary excretion of flavanones can be considered a good biomarker of acute citrus intake. However, low associations between habitual flavanone intake and urinary excretion suggest a possible inaccurate estimation of their intake or a too sporadic intake. For assessing habitual exposures, multiple urinary collections may be needed. These results show that none of the approaches tested is ideal, and the use of both DQ and biomarkers can be recommended.

Accurate and easy-to-use methods to assess free-living energy expenditure in response to physical activity in young children are scarce. In the present study, we evaluated the capacity of (1) 4 d recordings obtained using the Actiheart (mean heart rate (mHR) and mean activity counts (mAC)) to provide assessments of total energy expenditure (TEE) and activity energy expenditure (AEE) and (2) a 7 d activity diary to provide assessments of physical activity levels (PAL) using three sets of metabolic equivalent (MET) values (PALTorun, PALAdolph and PALAinsworth) in forty-four and thirty-one healthy Swedish children aged 1·5 and 3 years, respectively. Reference TEE, PALref and AEE were measured using criterion methods, i.e. the doubly labelled water method and indirect calorimetry. At 1·5 years of age, mHR explained 8 % (P= 0·006) of the variation in TEE above that explained by fat mass and fat-free mass. At 3 years of age, mHR and mAC explained 8 (P= 0·004) and 6 (P= 0·03) % of the variation in TEE and AEE, respectively, above that explained by fat mass and fat-free mass. At 1·5 and 3 years of age, average PALAinsworth values were 1·44 and 1·59, respectively, and not significantly different from PALref values (1·39 and 1·61, respectively). By contrast, average PALTorun (1·5 and 3 years) and PALAdolph (3 years) values were lower (P< 0·05) than the corresponding PALref values. In conclusion, at both ages, Actiheart recordings explained a small but significant fraction of free-living energy expenditure above that explained by body composition variables, and our activity diary produced mean PAL values in agreement with reference values when using MET values published by Ainsworth.

Leafy vegetables are important sources of provitamin A carotenoids. Information on their ability to provide vitamin A is often misleading because of the methodology used to assess bioefficacy. Mongolian gerbils were used to evaluate the bioefficacy of provitamin A carotenoids in tropical leafy vegetables (i.e. Solanum nigrum, Moringa oleifera, Vernonia calvoana and Hibiscus cannabinus) that are indigenous to Africa. Gerbils (n 67) were vitamin A-depleted for 5 weeks. After a baseline kill (n 7), the gerbils were weight-matched and assigned to six treatment groups (n 10; four vegetable groups; negative and positive controls). For 4 weeks, the treatments included 35 nmol vitamin A (theoretical concentrations based on 100 % bioefficacy) in the form of vegetables or retinyl acetate. In addition to their diets, the control and vegetable groups received daily doses of oil, while the vitamin A group received retinyl acetate in oil matched to prior day intake. Serum and livers were analysed for vitamin A using HPLC. Serum retinol concentrations did not differ among groups, but total liver vitamin A of the vitamin A and vegetable groups were higher than that of the negative control group (P < 0·0001). Liver β-carotene 15,15′-monooxygenase-1 expression levels were determined for two vegetable groups and were similar to the positive and negative controls. Conversion factors for the different leafy vegetables were between 1·9 and 2·3 μg β-carotene equivalents to 1 μg retinol. Small quantities of these vegetables maintained vitamin A status in gerbils through efficient bioconversion of β-carotene to retinol.

Dietary long-chain n-3 PUFA (n-3 LCPUFA) in infancy may have long-term effects on lifestyle disease risk. The present follow-up study investigated whether maternal fish oil (FO) supplementation during lactation affected growth and blood pressure in adolescents and whether the effects differed between boys and girls. Mother–infant pairs (n 103) completed a randomised controlled trial with FO (1·5 g/d n-3 LCPUFA) or olive oil (OO) supplements during the first 4 months of lactation; forty-seven mother–infant pairs with high fish intake were followed-up for 4 months as the reference group. We also followed-up 100 children with assessment of growth, blood pressure, diet by FFQ and physical activity by 7-d accelerometry at 13·5 (sd 0·4) years of age. Dried whole-blood fatty acid composition was analysed in a subgroup (n 49). At 13 years of age, whole-blood n-3 LCPUFA, diet, physical activity and body composition did not differ between the three groups. The children from the FO group were 3·4 (95 % CI 0·2, 6·6) cm shorter (P=0·035) than those from the OO group, and tended to have less advanced puberty (P=0·068), which explained the difference in height. There was a sex-specific effect on diastolic blood pressure (Psex×group=0·020), which was driven by a 3·9 (95 % CI 0·2, 7·5) mmHg higher diastolic blood pressure in the FO compared with the OO group among boys only (P=0·041). Our results indicate that early n-3 LCPUFA intake may reduce height in early adolescence due to a delay in pubertal maturation and increase blood pressure specifically in boys, thereby tending to counteract existing sex differences.

A short-term N balance study was conducted in twelve healthy female adults aged 21–32 years to determine their protein requirement. Four dietary protein levels (0·3, 0·4, 0·5 and 0·6 g protein/kg per d) were used. Energy intake of the subjects was kept constant at 0·18 MJ/kg per d. All subjects maintained their normal activity throughout the study period. N excretion was determined from the measurements of N in a total collection of urine, faeces, sweat and menstrual fluid for each dietary period. N balance during the four protein levels were −15·15 (SD 5·95), −5·53 (SD 6·71), + 6·15 (SD 4·76) and +12·05 (SD 8·63) mg N/kg per d for 0·3, 0·4, 0·5 and 0·6 g protein/kg per d respectively. The calculated average N requirements from regression analysis was 76·0 (sd 3·37) mg N/kg per d (0·48 g protein/kg per d). The estimate of allowance for individual variation to cover the 97·5 % population was 95 mg N/kg per d (0·6 g protein/kg per d). The net protein utilization (NPU) of the diet was 0·55. When compared with a similar study with men, there was a significant difference in the protein requirement between sexes. Thus, the unjustifiable sex difference in the protein allowance recommended by the Food and Agriculture Organization/World Health Organization/United Nations University (1985) Expert Consultation group must be reviewed.

Since the food habits of many elderly people are inadequate, the first experiment of the present study tested whether flavour amplification induces changes in preferences for and consumption of food and thus might result in a nutritional benefit. Two panels, one of 260 and one of 120 subjects, aged 19–98 years, took part in the study in which preferences for flavour-amplified yoghurt and Quorn® were measured. For both products, only a few of the young subjects (20 %) preferred the high flavour level; the percentage of subjects preferring the high flavour levels increased with age. These changes were highly significant. In a second experiment, participants received, under ad libitum conditions over 2 d in random order, a dish of yoghurt with either a high or a low flavour level. When adjusted for total consumption quantity, consumption of the highly flavoured yoghurt was not significantly correlated with age (r -0·03, P = 0·35). In a third experiment, odour perception was measured by determining the detection threshold for isoamylacetate. BMI values were obtained and the relationships between BMI and odour perception, age, preference and consumption were assessed in the age group 40–65 years. A significant correlation was observed between age and BMI (r 0·51, P < 0·0005). No significant correlation was observed between BMI and relative consumption of highly flavoured yoghurt (r -0·14, P = 0·14). A significant correlation was observed between BMI and preference for flavour-amplified yoghurt (r 0·35, P < 0·001). However, no significant correlation was observed between BMI and odour perception (r 0·07, P = 0·32). With increasing age, a combined influence of age, sex, BMI and odour perception on food preference is to be expected. According to our multiple regression analysis, BMI showed a significant partial regression coefficient (standardized β 0·36, P = 0·03). In conclusion, flavour amplification of food for older adults deserves attention, but specific approaches, which are tailored to the candidate food systems and older adult target groups, are needed.