Fe bioavailability can be manipulated by the nutritional composition of a meal. Ascorbic acid and unidentified components of meat, fish and poultry, but particularly beef, all appear to enhance the absorption of non-haem Fe. The aim of the present study is to identify whether extracts of green-lipped mussels (GLM; Perna canaliculus) enhance non-haem Fe absorption in Caco-2 cells and to compare the effect with that of beef. Raw GLM and raw beef homogenates were digested in vitro with pepsin at pH 2, and pancreatin and bile salts at pH 7. Tracer 55Fe was used to measure cellular Fe uptake. Ascorbic acid was used as a positive control and egg albumin, exposed to the same in vitro digestion process, was used as a negative control. Caco-2 cell monolayers were incubated with treatments for 60 min. All values were standardised per μg of GLM, egg albumin, beef or ascorbic acid. The results showed that ascorbic acid enhanced non-haem Fe absorption to the highest degree. Beef and GLM digestates both significantly enhanced Fe absorption compared with egg albumin. In conclusion, GLM digestate significantly enhances non-haem Fe uptake in Caco-2 cells with a similar magnitude to that of beef.

n-3 Highly unsaturated fatty acids (HUFA), are essential components of neuronal membranes and mediate a range of complex bioactive properties including gene expression, myelination, cell-signalling and dopaminergic function. Deficits in n-3 HUFA have been linked to increased risks for addictive disorders, thus we posited that lower fish consumption would be associated with greater risks for perinatal smoking among 9640 mothers enroled in the Avon Longitudinal Study of Parents and Children. We used univariable and multivariable regression models to examine relationships between self-reported prenatal dietary intakes of n-3 HUFA-rich foods (fish and shellfish) and maternal smoking; outcomes included cessation and the number of cigarettes smoked per d. Both before and during pregnancy, there was consistent evidence (P<0·001) of protective fish intake–smoking associations; relative to mothers reporting no fish consumption, those who reported some fish consumption (<340 g/week) and high fish consumption (340 g+/week) at 32 weeks of gestation showed lower likelihoods of smoking (adjusted P values <0·001). Respective OR for these relationships were 0·87 (95% CI 0·77, 0·97) and 0·73 (95% CI 0·61, 0·86). Although the prevalence of smoking diminished, from a high of 31·6% (pre-pregnancy) to a low of 18·7% (second trimester), the magnitude of fish intake–smoking associations remained stable following adjustment for confounders. These observations suggest that greater fish or n-3 HUFA consumption should be evaluated as an intervention to reduce or prevent smoking in randomised clinical trials.

There is increasing interest in modelling longitudinal dietary data and classifying individuals into subgroups (latent classes) who follow similar trajectories over time. These trajectories could identify population groups and time points amenable to dietary interventions. This paper aimed to provide a comparison and overview of two latent class methods: group-based trajectory modelling (GBTM) and growth mixture modelling (GMM). Data from 2963 mother–child dyads from the longitudinal Southampton Women’s Survey were analysed. Continuous diet quality indices (DQI) were derived using principal component analysis from interviewer-administered FFQ collected in mothers pre-pregnancy, at 11- and 34-week gestation, and in offspring at 6 and 12 months and 3, 6–7 and 8–9 years. A forward modelling approach from 1 to 6 classes was used to identify the optimal number of DQI latent classes. Models were assessed using the Akaike and Bayesian information criteria, probability of class assignment, ratio of the odds of correct classification, group membership and entropy. Both methods suggested that five classes were optimal, with a strong correlation (Spearman’s = 0·98) between class assignment for the two methods. The dietary trajectories were categorised as stable with horizontal lines and were defined as poor (GMM = 4 % and GBTM = 5 %), poor-medium (23 %, 23 %), medium (39 %, 39 %), medium-better (27 %, 28 %) and best (7 %, 6 %). Both GBTM and GMM are suitable for identifying dietary trajectories. GBTM is recommended as it is computationally less intensive, but results could be confirmed using GMM. The stability of the diet quality trajectories from pre-pregnancy underlines the importance of promotion of dietary improvements from preconception onwards.

The present study elucidates the association of intrinsic sugars and free sugars (FS) from all relevant sources with all-cause mortality in the prospective UK Biobank cohort. Sugar intake was assessed in 186 811 UK Biobank participants who completed at least one web-based 24-h dietary recall (Oxford WebQ). Cox proportional hazard regression models for all-cause mortality were used with sugar intake from different sources included as penalised cubic splines to allow non-linear predictor effects. Over a mean follow-up of 12·3 years, 8576 (4·6 %) deaths occurred. FS but not intrinsic sugars were significantly and dose-dependently associated with hazard ratio (HR) for all-cause mortality. The association with all-cause mortality was significant and dose dependent for FS in beverages, but not in solids with the mean (CI) HR at 50 g/d v. 0 g/d consumption at 1·10, 95 % CI (1·07, 1·14) and 1·01, 95 % CI (0·98, 1·03), respectively. Within the beverages subcategories, a significant dose-dependent association with mortality was detected for FS in soda/fruit drinks and milk-based drinks whereas this relation was NS for FS in pure juice and tea/coffee. FS in four different subtypes of solids, i.e. treats, cereals, toppings and sauces, were not positively associated with all-cause mortality. Major findings were robust in sensitivity analyses. In conclusion, only some FS sources were associated with all-cause mortality. Interventions targeting FS subtypes might be most effective concerning mortality if focused on the reduction of soda/fruit drinks and milk-based sugary drinks; however, the present results need to be confirmed by independent studies.

Higher milk intake has been associated with a lower stroke risk, but not with risk of CHD. Residual confounding or reverse causation cannot be excluded. Therefore, we estimated the causal association of milk consumption with stroke and CHD risk through instrumental variable (IV) and gene-outcome analyses. IV analysis included 29 328 participants (4611 stroke; 9828 CHD) of the European Prospective Investigation into Cancer and Nutrition (EPIC)-CVD (eight European countries) and European Prospective Investigation into Cancer and Nutrition-Netherlands (EPIC-NL) case-cohort studies. rs4988235, a lactase persistence (LP) SNP which enables digestion of lactose in adulthood was used as genetic instrument. Intake of milk was first regressed on rs4988235 in a linear regression model. Next, associations of genetically predicted milk consumption with stroke and CHD were estimated using Prentice-weighted Cox regression. Gene-outcome analysis included 777 024 participants (50 804 cases) from MEGASTROKE (including EPIC-CVD), UK Biobank and EPIC-NL for stroke, and 483 966 participants (61 612 cases) from CARDIoGRAM, UK Biobank, EPIC-CVD and EPIC-NL for CHD. In IV analyses, each additional LP allele was associated with a higher intake of milk in EPIC-CVD (β = 13·7 g/d; 95 % CI 8·4, 19·1) and EPIC-NL (36·8 g/d; 95 % CI 20·0, 53·5). Genetically predicted milk intake was not associated with stroke (HR per 25 g/d 1·05; 95 % CI 0·94, 1·16) or CHD (1·02; 95 % CI 0·96, 1·08). In gene-outcome analyses, there was no association of rs4988235 with risk of stroke (OR 1·02; 95 % CI 0·99, 1·05) or CHD (OR 0·99; 95 % CI 0·95, 1·03). Current Mendelian randomisation analysis does not provide evidence for a causal inverse relationship between milk consumption and stroke or CHD risk.

1. The concentration of 2-aminoethylphosphonic acid (AEP) in 96 h cultures of Tetrahymena pyriformis W was studied in order to apply it as an indicator in the assay of the relative nutritive value (RNV; protozoa population with test protein: protozoa population with whole-egg powder) of protein. Foodstuffs and food mixtures of different protein contents and qualities were used as test samples.

2. RNV values based on AEP determination (RNVAEP) were compared with corresponding values calculated from protozoa counts (RNVpc), as well as with biological value (bv) and net protein utilization (npu) of the same proteins assayed on rats.

3. Both for foodstuffs and food mixtures highly significant correlations were found between RNVAEP and RNVpc, RNVAEP and both bv and npu, and RNVpc, and both bv and npu.

4. AEP content in the protozoal suspension was preferred to cell count as a measure of growth response, since it took into account large differences in cell dimensions that were observed between cultures grown with different test proteins.

1. Pangola grass (Digitaria decumbens) grown with and without calcium fertilizer was cut at three stages of regrowth to measure voluntary intake of dry matter (DM) and digestibility of various components of the dried-grass diet by sheep kept in metabolism crates. To determine the extent of a simple Ca deficiency half the sheep on each diet were supplemented with 1·4g Ca/d. Retention times of the various dietary components in the reticulo-rumen were also determined.

2. Feeding a Ca supplement had no effect on voluntary intake or digestibility.

3. Ca fertilizer increased the Ca content of the grass from 2·2 to 3·8 g/kg DM and DM digestibility from 0·455 to 0·476 (P < 0·01) due to an increase in the digestibility of the hemicellulose.

4. Voluntary intake was increased from 38·8 to 43·2 g/kg body-weight0·75 per d by Ca fertilizer due to an 18% reduction in the period of time the DM was retained in the reticulo-rumen.

5. Regressions relating voluntary intake to DM digestibility for the Ca-fertilized and control grass were significantly different (P < 0·01). When compared at the same DM digestibility the voluntary intake of the Ca-fertilized grass was 2·6 g/kg body-weight0·75 per d higher than that of the control.

6. It was concluded that Ca fertilizer increased both DM digestibility and voluntary intake as a result of changes in the structural composition of the grass and not by a simple increase in the Ca content of the diet.

The relationships between both dietary and empty body fatty acid composition and the morphometry, densitometry, geometry and biomechanical properties of the femur of growing pigs were analysed. A total of thirty-two pigs aged 115 d were divided into four groups (n 8 per group). The pigs were fed either a control diet (group C) or a diet supplemented with linseed oil (rich in α-linolenic acid (C18 : 3n-3), group L), fish oil (rich in EPA (C20 : 5n-3) and DHA (C22 : 6n-3), group F) and beef tallow (rich in SFA, group T). The diets differed in n-3 PUFA contents (0·63–18·52 g/kg) and n-6:n-3 PUFA ratios (0·91–14·51). At 165 d of age, the pigs were slaughtered and the fatty acids in the empty body were determined. Moreover, the left femur was dissected. The cortical wall thickness, cross-sectional area, cortical index, bone mineral content, bone mineral density, maximum elastic strength and maximum strength were lower (P<0·05) in the femurs of pigs from groups C and T than in those from groups F and L. Significant positive correlations were found between the densitometry, geometry and biomechanical properties of the femur and both dietary and empty body n-3 PUFA content, whereas significant negative correlations were observed between the same properties and both dietary and empty body n-6:n-3 PUFA ratio. The results of the present study suggest that in growing pigs α-linolenic acid has a similar positive effect on bone health to that of EPA and DHA.

Blood examination was conducted for the four Gidra-speaking village groups in Papua New Guinea, who were characterized by high Fe intake and high malaria prevalence with marked inter-village differences. The northern riverine villagers, whose Fe intake was higher than the other three village groups, did not suffer from Fe-deficiency anaemia in their malaria-endemic environment; nor did the inland villagers, with their second highest Fe intake and their malaria-free environment, suffer from Fe-deficiency anaemia. However, several individuals of the southern riverine village suffered from anaemia in a malaria-endemic environment, although their Fe intake was almost the same as the inland villagers’. A considerable proportion of the coastal villagers were anaemic, reflecting the lowest Fe intake and the highest malaria prevalence. An inter-village comparison of the relationships between haemoglobiin levels and transferrin saturation revealed that the southern riverine villagers needed smaller amounts of circulating Fe for erythropoiesis than the northern riverine and inland villagers, reflecting the long-term human-environment conditions such as the density of malaria vectors and the people’s dietary habits. Fe supplementation was not judged effective against hypoferraemia and/or anaemia in such a population. As the incidence of malaria had no significant long-lasting effect on Fe stores or circulating Fe concentration, but did have an effect on anaemia, the hypothesis that malaria causes a transfer of Fe from the blood to parenchymal tissues as a defence against infectious diseases was not supported.

To analyse the association of socio-demographic and health factors with vitamin D insufficiency and 25-hydroxyvitamin D (25(OH)D) concentration in Brazilian children aged 6–59 months. Data from 8145 children from the Brazilian National Survey on Child Nutrition (ENANI-2019) were analysed. The serum concentration of 25(OHD)D was measured using a chemiluminescent immunoassay. The prevalence of vitamin D insufficiency (25(OH)D < 50 nmol/l) and 95 % CI was calculated. Logistic and linear regression models were used to identify the variables associated with vitamin D insufficiency and serum 25(OH)D concentrations, respectively. The mean 25(OH)D concentration was 98·6 ± 36·0 nmol/l, and 4·3 % of the children presented vitamin D insufficiency. Children aged 6–23 months (OR = 2·23; 95 % CI 1·52, 3·26); belonging to Southeast (OR = 5·55; 95 % CI 2·34, 13·17) and South (OR = 4·57; 95 % CI 1·77, 11·84) regions; the second tertile of the National Wealth Score (OR = 2·14; 95 % CI 1·16, 3·91) and winter (OR = 5·82; 95 % CI 2·67, 12·71) and spring (OR = 4·84; 95 % CI 2·17, 10·80) seasons of blood collection were associated with a higher chance of vitamin D insufficiency. Female sex (β = −5·66, 95 % CI − 7·81, −3·51), urban location (β = −14·19, 95 % CI −21·0, −7·22) and no vitamin D supplement use (β = −6·01, 95 % CI −9·64, −2·39) were inversely associated with serum 25(OH)D concentration. The age of children and the Brazilian geographical region of household location were the main predictors of vitamin D insufficiency. In Brazil, vitamin D insufficiency among children aged 6–59 months is low and is not a relevant public health problem.