We previously demonstrated that the chronic consumption of a high-fat diet (HFD) promotes lung and liver metastases of 4T1 mammary carcinoma cells in obesity-resistant BALB/c mice. To examine early transcriptional responses to tumour progression in the liver and lungs of HFD-fed mice, 4-week-old female BALB/c mice were divided into four groups: sham-injected, control diet (CD)-fed; sham-injected, HFD-fed (SH); 4T1 cell-injected, CD-fed (TC); 4T1 cell-injected, HFD-fed (TH). Following 16 weeks of either a CD or HFD, 4T1 cells were injected into the mammary fat pads of mice in the TC and TH groups and all mice were continuously fed identical diets. At 14 d post-injection, RNA was isolated from hepatic and pulmonary tissues for microarray analysis of mRNA expression. Functional annotation and core network analyses were conducted for the TH/SH Unique gene set. Inflammation in hepatic tissues and cell mitosis in pulmonary tissues were the most significant biological functions in the TH/SH Unique gene set. The biological core networks of the hepatic TH/SH Unique gene set were characterised as those genes involved in the activation of acute inflammatory responses (Orm1, Lbp, Hp and Cfb), disordered lipid metabolism and deregulated cell cycle progression. Networks of the pulmonary Unique gene set displayed the deregulation of cell cycle progression (Cdc20, Cdk1 and Bub1b). These HFD-influenced alterations may have led to favourable conditions for the formation of both pro-inflammatory and pro-mitotic microenvironments in the target organs that promote immune cell infiltration and differentiation, as well as the infiltration and proliferation of metastatic tumour cells.

Subcutaneous adipose tissue (scAT) and peripheral blood mononuclear cells (PBMC) play a significant role in obesity-associated systemic low-grade inflammation. High-fat diet (HFD) is known to induce inflammatory changes in both scAT and PBMC. However, the time course of the effect of HFD on these systems is still unknown. The aim of the present study was to determine the time course of the effect of HFD on PBMC and scAT. New Zealand white rabbits were fed HFD for 5 or 10 weeks (i.e. HFD-5 and HFD-10) or regular chow (i.e. control (CNT)-5 and CNT-10). Thereafter, metabolic and inflammatory parameters of PBMC and scAT were quantified. HFD induced hyperfattyacidaemia in HFD-5 and HFD-10 groups, with the development of insulin resistance in HFD-10, while no changes were observed in scAT lipid metabolism and inflammatory status. HFD activated the inflammatory pathways in PBMC of HFD-5 group and induced modified autophagy in that of HFD-10. The rate of fat oxidation in PBMC was directly associated with the expression of inflammatory markers and tended to inversely associate with autophagosome formation markers in PBMC. HFD affected systemic substrate metabolism, and the metabolic, inflammatory and autophagy pathways in PBMC in the absence of metabolic and inflammatory changes in scAT. Dietary approaches or interventions to avert HFD-induced changes in PBMC could be essential to prevent metabolic and inflammatory complications of obesity and promote healthier living.

The reversibility of osteopenia secondary to isolated Ca deficiency (CaDef) is still not clear. We studied the effect of severe CaDef on Ca homeostasis and bone accrual in a ‘hypercalcaemic’ animal, the rabbit, during the post-weaning period and its reversibility on Ca supplementation. Male Belgian 5-week-old rabbit pups were fed CaDef diet (0·026% Ca) for 10 weeks. As compared with those fed with a normal chow diet (0·45% Ca), CaDef pups developed significant hypocalcaemia (P<0·05), hypocalciuria (urinary Ca 76 (sem 12) V 17 (sem 1) mg/l;P<0·005), hypophosphataemia (serum inorganic P 100 (sem 6) V. 65 (sem 4) mg/l; P<0·005), secondary hyperparathyroidism (SHPT) (serum intact parathyroid hormone human equivalent 18·2 (sem 1·4) V. 125·0 (sem 4·5) pg/ml; P<0·001) and elevated serum calcitriol levels (34·0 (sem 3·9) V. 91·0 (sem 1·0) pg/ml; v<0·005). Elevated urinary C-terminal telopeptide of class I collagen (P<0·005) and total serum alkaline phosphatase (P<0·005) suggested increased bone turnover. There was a significantly lower gain in bone mineral density (BMD) and bone mineral content (BMC) in the whole body and lumbar spine in vivo, and various sub-regions of the femur and tibia in vitro.Supplementation of adequate Ca (0·45% Ca) after 15 weeks on the normal diet resulted in rapid catch-up growth, and resolution of SHPT. Rapid gain in various BMD and BMC parameters continued at 30 weeks of age, and both were comparable with those in rabbits on a normal diet. We conclude that Ca deficiency-induced SHPT and poor bone accrual in growing rabbit pups are rapidly reversible with Ca supplementation. The present study indicates that early intervention may be a more appropriate window period for human nutritional corrective measures.

In the present study, fifteen growing pigs were used to determine the whole-body oxidation, retention efficiency (RE) and apparent conversion (AC) of α-linolenic acid (18 : 3n-3) to n-3 highly unsaturated fatty acids (HUFA), including EPA (20 : 5n-3) and DHA (22 : 6n-3). The pigs were fed a diet containing 10 % flaxseed for 30 d. Whole-body fatty acid composition was determined at initial (27·7 (se 1·9) kg), intermediate (day 15; 39·2 (se 1·4) kg) and final (45·7 (se 2·2) kg) body weight. On day 12, four pigs were fed 10 mg/kg of uniformly labelled 13C-18 : 3n-3 (single-bolus dose) to determine the oxidation of 18 : 3n-3. Expired $$CO_{2} $$$$ samples were collected for 24 h thereafter. The whole-body content of n-3 PUFA increased linearly (P< 0·0001) with time; however, the content of 22 : 6n-3 exhibited a quadratic response (P< 0·01) with a peak occurring at 15 h. As a proportion of intake, the RE of 18 : 3n-3 tended to reduce with time (P= 0·098). The AC of ingested 18 : 3n-3 to the sum of n-3 HUFA was reduced with time (P< 0·05; 12·2 v. 7·53 % for days 0–15 and days 15–30, respectively). The AC of 18 : 3n-3 to 20 : 5n-3 or 22 : 6n-3 was lower than that to 20 : 3n-3, both for days 0–15 (P< 0·05; 1·14 or 1·07 v. 7·06 %) and for days 15–30 (P< 0·05; 1·51 or 0·33 v. 4·29 %). The direct oxidation of 18 : 3n-3 was 7·91 (se 0·98) % and was similar to the calculated disappearance of 18 : 3n-3 between days 0 and 30 (8·81 (se 5·24) %). The oxidation of 18 : 3n-3 was much lower than that reported in other species. The AC of 18 : 3n-3 to n-3 HUFA was reduced over time and that to 20 : 3n-3 in the present study was much higher than that reported in other species and should be explored further.

Associations between meat consumption and heart disease have been assessed in several studies, but it has been suggested that other dietary factors influence these associations. The aim of the present study was to assess whether meat consumption is associated with ischaemic heart disease (IHD), and if the association is modified by dietary quality. The analyses were based on the cohort of adult participants in the Danish National Survey on Diet and Physical Activity in 2000–2002, 2003–2008 and 2011–2013. From these surveys, information on meat consumption and dietary quality was extracted. The cohort was followed in national registers to identify incident IHD. Associations were estimated using Cox regression analyses adjusting for socio-demographic and lifestyle factors. Analyses of associations between meat consumption and IHD stratified by dietary quality were subsequently evaluated. Among the 8007 participants, the median follow-up was 9·8 years and 439 cases of IHD were recorded. The results suggested a trend between consumption increments of 100 g/d of red meat (hazard ratio (HR) = 1·23; 95 % CI 0·99, 1·53) or of 50 g/d of processed meat (HR = 1·09; 95 % CI 0·93, 1·29) and higher risk of IHD. The trends were, however, not statistically significant. Stratification by dietary quality did not suggest that associations between meat consumption and risk of IHD were modified by dietary quality. This population-based cohort study with detailed dietary information suggested a trend with higher meat consumption being associated with higher risk of IHD, but the association was not statistically significant. Results did not indicate that dietary quality modifies such associations.

Analyses in nutritional epidemiology usually assume a uniform effect of a nutrient. Previously, four subgroups of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study of Finnish male smokers aged 50–69 years were identified in which vitamin E supplementation either significantly increased or decreased the risk of pneumonia. The purpose of this present study was to quantify the level of true heterogeneity in the effect of vitamin E on pneumonia incidence using the I2 statistic. The I2 value estimates the percentage of total variation across studies that is explained by true differences in the treatment effect rather than by chance, with a range from 0 to 100 %. The I2 statistic for the effect of vitamin E supplementation on pneumonia risk for five subgroups of the ATBC population was 89 % (95 % CI 78, 95 %), indicating that essentially all heterogeneity was true variation in vitamin E effect instead of chance variation. The I2 statistic for heterogeneity in vitamin E effects on pneumonia risk was 92 % (95 % CI 80, 97 %) for three other ATBC subgroups defined by smoking level and leisure-time exercise level. Vitamin E decreased pneumonia risk by 69 % among participants who had the least exposure to smoking and exercised during leisure time (7·6 % of the ATBC participants), and vitamin E increased pneumonia risk by 68 % among those who had the highest exposure to smoking and did not exercise (22 % of the ATBC participants). These findings refute there being a uniform effect of vitamin E supplementation on the risk of pneumonia.

1. The influence of a low-energy diet when associated with high-cholesterol intake was investigated in seventeen normal men during an 8-week cross-over study. The subjects were given a daily supplement of two whole eggs and two egg yolks (approximately 1 g cholesterol) either with their usual diet for 4 weeks or with a low-energy diet for 4 weeks. Each subject took part randomly in both dietary periods.

2. During the first part of the study, no changes occurred in the plasma cholesterol of the subjects with egg supplementation of the usual diet.

3. In contrast, the low-energy diet and associated weight loss markedly decreased the tolerance to high- cholesterol intake resulting in increased plasma cholesterol. The mean rise was 22.7% but with wide individual variations in the response. This was almost completely normalized when the subjects returned to their usual energy intake indicating the involvement of weight reduction in the increase observed.

4. Changes in low-density-lipoprotein (LDL) cholesterol were parallel to those of total plasma cholesterol with an increase following the low-energy diet and normalization after body-weight recovery.

5. The opposite effect was shown with the low-energy diet after previous adaptation to the consumption of four eggs per day. This dietary regimen resulted in a decrease in plasma cholesterol although it was not significant. Moreover, the lipoprotein profile was improved with a decrease in very-low-density-lipoprotein (VLDL) cholesterol and an increase in high-density-lipoprotein (HDL) cholesterol.

6. High-cholesterol intake induced significant changes in lipoprotein composition whatever the energy ration. LDL and HDL were enriched in cholesterol esters as early as the 1st month of egg supplementation of the diet.

7. Taken together, the results emphasize the possible adverse effect of slimming diets when associated with high-cholesterol intake. The existence of ‘high-responders’ to these dietary conditions calls for special attention to be paid to the cholesterol content of restricted diets.

Non-resolving inflammation is characteristic of tuberculosis (TB). Given their inflammation-resolving properties, n-3 long-chain PUFA (n-3 LCPUFA) may support TB treatment. This research aimed to investigate the effects of n-3 LCPUFA on clinical and inflammatory outcomes of Mycobacterium tuberculosis-infected C3HeB/FeJ mice with either normal or low n-3 PUFA status before infection. Using a two-by-two design, uninfected mice were conditioned on either an n-3 PUFA-sufficient (n-3FAS) or -deficient (n-3FAD) diet for 6 weeks. One week post-infection, mice were randomised to either n-3 LCPUFA supplemented (n-3FAS/n-3+ and n-3FAD/n-3+) or continued on n-3FAS or n-3FAD diets for 3 weeks. Mice were euthanised and fatty acid status, lung bacterial load and pathology, cytokine, lipid mediator and immune cell phenotype analysed. n-3 LCPUFA supplementation in n-3FAS mice lowered lung bacterial loads (P = 0·003), T cells (P = 0·019), CD4+ T cells (P = 0·014) and interferon (IFN)-γ (P < 0·001) and promoted a pro-resolving lung lipid mediator profile. Compared with n-3FAS mice, the n-3FAD group had lower bacterial loads (P = 0·037), significantly higher immune cell recruitment and a more pro-inflammatory lipid mediator profile, however, significantly lower lung IFN-γ, IL-1α, IL-1β and IL-17, and supplementation in the n-3FAD group provided no beneficial effect on lung bacterial load or inflammation. Our study provides the first evidence that n-3 LCPUFA supplementation has antibacterial and inflammation-resolving benefits in TB when provided 1 week after infection in the context of a sufficient n-3 PUFA status, whilst a low n-3 PUFA status may promote better bacterial control and lower lung inflammation not benefiting from n-3 LCPUFA supplementation.

Anthropometric indices of adiposity include BMI, waist circumference and waist:height ratio. In the recruitment phase of a prospective cohort study carried out between 1998 and 2002 we studied a population sample of 11 786 white Caucasian non-pregnant women in Southampton, UK aged 20–34 years, and explored the extent to which proposed cut-off points for the three indices identified the same or different women and how these indices related to adiposity. Height, weight and waist circumference were measured and fat mass was estimated from skinfold thicknesses; fat mass index was calculated as fat mass/height1·65. Of the subjects, 4869 (42 %) women were overweight (BMI ≥ 25 kg/m2) and 1849 (16 %) were obese (BMI ≥ 30 kg/m2). A total of 890 (8 %) subjects were not overweight but had a waist circumference ≥ 80 cm and 748 (6 %) subjects were overweight but had a waist circumference < 80 cm (6 %). Of the women, 50 % had a BMI ≥ 25 kg/m2 or a waist circumference ≥ 80 cm or a waist:height ratio ≥ 0·5. Of the variation in fat mass index, 85 % was explained by BMI, 76 % by waist circumference and 75 % by waist:height ratio. Our findings demonstrate that many women are differentially classified depending on which index of adiposity is used. As each index captures different aspects of size in terms of adiposity, there is the need to determine how the three indices relate to function and how they can be of use in defining risk of ill health in women.

Fish oil supplementation during pregnancy has been associated with lower levels of cord blood IL-13, suggesting that the administration of n-3 fatty acids may attenuate the development of allergic disease. The present study aimed to investigate the mechanism by which n-3 fatty acid administration influences the production of IL-13. Pregnant BALB/c mice were fed nutritionally complete high-fat diets (15 %, w/w) with an n-3 fatty acid-enriched (DHA 1 %, w/w) or control diet (0 % DHA) immediately following delivery. Pups were exposed during suckling and weaned to the maternal diet for the remainder of the study. The production of IL-13, IL-4, IL-10 and interferon-γ from the splenocytes of ovalbumin (ova)-sensitised animals was assessed following in vitro ova stimulation or unstimulated conditions. Human T helper type 2 (Th2) cells were mitogen-stimulated in the presence or absence of DHA (10 μm) and assessed for IL-13 and IL-4 expression using intracellular flow cytometry. The influence on transcriptional activation was studied using a human IL-13 promoter reporter construct and electromobility shift assay. Ova-activated splenocytes from DHA-fed mice produced less IL-13 (57·2 (se 21·7) pg/ml) and IL-4 (7·33 (se 3·4) pg/ml) compared with cells from the animals fed the control diet (161·5 (se 45·0), P< 0·05; 33·2 (se 11·8), P< 0·05). In vitro, DHA inhibited the expression of IL-13 protein from human Th2 cells as well as transcriptional activation and binding of the transcription factors cyclic AMP response element binding and activating transcription factor 2 to the human IL-13 promoter. These data indicate the potential of n-3 fatty acids to attenuate IL-13 expression, and suggest that they may subsequently reduce allergic sensitisation and the development of allergic disease.

Major surgery induces an immuno-inflammatory response accompanied by oxidative stress that may impair cellular function and delay recovery. The objective of the study was to investigate the effect of an enteral supplement, containing glutamine and antioxidants, on circulating levels of immuno-inflammatory markers after major gastrointestinal tract surgery. Patients (n 21) undergoing major gastrointestinal tract surgery were randomised in a single-centre, open-label study. The effects on circulating levels of immuno-inflammatory markers were determined on the day before surgery and on days 1, 3, 5 and 7 after surgery. Major gastrointestinal surgery increased IL-6, TNF receptor 55/60 (TNF-R55) and C-reactive protein (CRP). Surgery reduced human leucocyte antigen-DR (HLA-DR) expression on monocytes. CRP decrease was more pronounced in the first 7 d in the treatment group compared with the control group. In the treatment group, from the moment Module AOX was administered on day 1 after surgery, TNF receptor 75/80 (TNF-R75) level decreased until the third post-operative day and then stabilised, whereas in the control group the TNF-R75 level continued to increase. The results of the present pilot study suggest that enteral nutrition enriched with glutamine and antioxidants possibly moderates the immuno-inflammatory response (CRP, TNF-R75) after surgery.

Nutritional geometry (NG) is a novel dietary analysis approach that considers nutrient balance, rather than single nutrient effects, on health and behaviour. Through NG, recent animal experiments have found that lifespan and reproduction are differentially altered by dietary macronutrient distribution. Epidemiological research using NG reports similar findings for human ageing. Yet, the relation of macronutrient balance to human reproduction, especially reproductive maturation, remains undefined. We studied the impact of childhood macronutrient intake on pubertal maturation, by applying NG to an Australian longitudinal adolescent dataset. Food records, collected at age 8 years from 142 pre-pubertal children (females, 92; males, 50), were analysed for absolute energy, percentage energy and energy-adjusted residuals from protein, carbohydrate and fat. Pubertal stage change (assessed at 8, 13 and 15 years) was modelled to obtain individual mathematical estimates of pubertal timing and tempo. Timing of menarche was recorded. The association of macronutrients to pubertal timing/tempo was assessed via NG, involving generalised additive models and heat maps to aid interpretation. Results showed lower dietary protein (relative to carbohydrate and fat) in girls consistently predicted earlier pubertal timing and menarche, and was related to faster pubertal tempo (all P < 0·05). No significant associations were identified in boys for both timing and tempo. Results suggest a role of non-protein macronutrients in facilitating female maturation; corroborating feeding and reproductive behaviour patterns observed in earlier NG studies of primates. Application of NG to other adolescent datasets is required to confirm the present findings. Such work would advance understanding of how nutrient balance shapes human development and health.

The aims of the present study were to assess dietary intake of saccharides in middle-aged adults, and to determine whether intakes of these sugar nutrients were related to self-reported memory performance. A population-based sample of 1183 men and women (aged 40–60 years) completed questionnaires assessing everyday memory function. Dietary intake status of saccharides was estimated using a self-completed, quantified FFQ. After controlling for demographic and health measures (for example, time spent exercising, smoking and alcohol consumption), saccharide intake was related to better self-reported memory functioning. Thus, longer-term intakes of saccharides through the usual diet may be positively related to perceived memory performance in mid-life.

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

1. Allantoin excretion was determined in forty-four rats; twenty-four rats were 21 d old and twenty were 90 d old. The animals received ad lib. for 11 d diets that contained varying amounts of protein from casein and from gluten. Urinary allantoin excretion was determined during the last 3 d of the period.

2. The amount of allantoin excreted in both young and adult animals was independent of the diet consumed and was related to metabolic weight (W in kg0·73) and rate of gain in body-weight (ΔW in g/d), also to body nitrogen content (N in g0·73) and rate of gain in body N (ΔN in mg/d), according to the equations allantoin (mg/d) = 60·54 W0·73 + 2·12ΔW, and allantoin (mg/d) = 5·45 N0·73 + 0·071ΔN.

BMI, waist circumference (WC) and waist-to-height ratio (WHtR) can be used for discriminating children and adolescents at risk of CVD. However, consensus on how to use these anthropometric indicators is lacking for children and adolescents in Asia. Discrete criteria are promoted internationally, but continuous variables could be used. Data from a survey of 10 949 Vietnamese school-aged children (6–18 years) were used to evaluate the performance of anthropometric indicators to identify elevated blood pressure (BP), dyslipidaemia or at least three cardiovascular risk factors (CVRF). Weight, height, WC and BP were measured using standardised protocols; 1009 participants who had blood lipids were analysed. AUC was used to assess the performance, and the Youden index to identify optimal cut-offs. The prevalence of elevated BP, dyslipidaemia and CVRF was 26·5, 49·3 and 12·2 %, respectively. BMI, WC and WHtR had low capacity to identify elevated BP and dyslipidaemia (AUC range 0·61–0·66) but moderate capacity to identify CVRF (0·72–0·74). Optimal BMIZ cut-offs to identify elevated BP, dyslipidaemia and CVRF were 0·40, 1·01 and 1·1 sd; for WC z-score, they were 0·06, 0·49 and 0·62 sd; for WHtR, optimal cut-offs were close to 0·5. A BMIZ cut-off of 1·0 sd and a WHtR cut-off of 0·5 would, therefore, be useful criteria to identify Vietnamese children who are likely to have CVRF. However, further validation of these criteria in other studies of Asian children and adolescents is needed.

The interaction between nutrition and infection is a key determinant of human health. Traditionally the interaction has centered on the role of nutrients in defining host defenses and the impact of infection in defining nutritional needs and status. Over the past decades the interaction has expanded its scope to encompass the role of specific nutrients in defining acquired immune function, in the modulation of inflammatory processes and on the virulence of the infectious agent itself. More recently the role of micronutrients and fatty acids on the response of cells and tissues to hypoxic and toxic damage has been recognized suggesting a fourth dimension to the interaction. The list of nutrients affecting infection, immunity, inflammation and cell injury has expanded from traditional protein-energy supply to several vitamins, multiple minerals and more recently specific lipid components of the diet. The promise of nutrition in the defense against infection, inflammation and tissue injury has spawned a thriving pharma-nutritional supplement industry and the development of novel foods that require appropriate evaluation of efficacy, safety and effectiveness relative to costs. Academics need to aware of the ethics and the pitfalls in the interaction with industry; conversely industry has to define its role in the process of bringing new knowledge to useful products. The process needs to be interactive, transparent and clearly place public interest above all other considerations.