The NADPH oxidase enzyme system is the main source of superoxide anions in phagocytic and vascular cells. NADPH oxidase-dependent superoxide generation has been found to be abnormally enhanced in several chronic diseases. Evidence is accumulating that polyphenols may have the potential to improve cardiovascular health, although the mechanism is not fully established. Consumption of concentrated red grape juice, rich in polyphenols, has been recently shown to reduce NADPH oxidase activity in circulating neutrophils from human subjects. In the present work we studied whether red grape juice polyphenols affected NADPH oxidase subunit expression at the transcription level. For this, we used human neutrophils and mononuclear cells from peripheral blood, HL-60-derived neutrophils and the endothelial cell line EA.hy926.Superoxide production was measured with 2′7′-dichlorofluorescein diacetate or lucigenin, mRNA expression by real-time RT-PCR and protein expression by Western blot. Each experiment was performed at least three times. In all cell types tested, red grape juice, dealcoholised red wine and pure polyphenols decreased superoxide anion production. Red grape juice and dealcoholised red wine selectively reduced p47phox, p22phox and gp91phox expression at both mRNA and protein levels, without affecting the expression of p67phox. Pure polyphenols, particularly quercetin, also reduced NADPH oxidase subunit expression, especially p47phox, in all cell types tested. The present results showing that red grape juice polyphenols reduce superoxide anion production provide an alternative mechanism by which consumption of grape derivatives may account for a reduction of oxidative stress associated with cardiovascular and/or inflammatory diseases related to NADPH oxidase superoxide overproduction.

The aim of the study was to compare the effect of different whey protein-containing high-Ca diets on weight loss and weight regain in a model of diet-induced obesity. Obesity was induced in C57BL/6J mice with a high-fat (60 % of energy) diet. Weight loss by energy restriction was performed on four different high-Ca diets (1·8 % CaCO3) containing different whey proteins (18 % of energy): α-lactalbumin (ALA), β-lactoglobulin (BLG), lactoferrin (LF) and whey protein isolate (WPI). After 7 weeks of energy restriction some of the mice were killed and the rest were fed with the same diets ad libitum for 7 weeks. The mice on the LF diet lost significantly more weight than mice on the WPI diet. The body fat content in the ALA and LF groups was significantly lower than in the WPI group (P < 0·05) and the LF group differed significantly even from the BLG group (P < 0·05). Ad libitum feeding after weight loss resulted in weight regain in all groups and only the ALA diet significantly reduced fat accumulation during weight regain. The weight regain was most pronounced in the LF group, but the adipocyte size was still significantly smaller than in the other groups. There were no differences in food intake or apparent fat digestibility between the groups. It can be concluded that a high-Ca diet with ALA significantly improves the outcome of weight loss and subsequent weight regain during the feeding of a high-fat diet in C57BL/6J mice, in comparison with WPI.

Dietary long-chain PUFA, both n-3 and n-6, have unique benefits with respect to CVD risk. The aim of the present study was to determine the mechanisms by which n-3 PUFA (EPA, DHA) and n-6 PUFA (linoleic acid (LA), arachidonic acid (AA)) relative to SFA (myristic acid (MA), palmitic acid (PA)) alter markers of inflammation and cholesterol accumulation in macrophages (MΦ). Cells treated with AA and EPA elicited significantly less inflammatory response than control cells or those treated with MA, PA and LA, with intermediate effects for DHA, as indicated by lower levels of mRNA and secretion of TNFα, IL-6 and monocyte chemoattractant protein-1. Differences in cholesterol accumulation after exposure to minimally modified LDL were modest. AA and EPA resulted in significantly lower MΦ scavenger receptor 1 mRNA levels relative to control or MA-, PA-, LA- and DHA-treated cells, and ATP-binding cassette A1 mRNA levels relative to control or MA-, PA- and LA-treated cells. These data suggest changes in the rate of bidirectional cellular cholesterol flux. In summary, individual long-chain PUFA have differential effects on inflammatory response and markers of cholesterol flux in MΦ which are not related to the n position of the first double bond, chain length or degree of saturation.

Cu is an essential cofactor for at least twelve mammalian enzymes including dopamine β-mono-oxygenase (DBM), which converts dopamine (DA) to noradrenaline (NA). Previous studies reported that certain Cu-deficient (Cu−) rat tissues have lower NA and higher DA than Cu-adequate (Cu+) tissues, suggesting that DBM function was impaired. However, in vitro studies suggested that DBM activity is higher in Cu− tissue. Experiments were conducted on adrenal glands (AG), medulla oblongata/pons (MO), vas deferens (VD) and heart (HT) from a single rat experiment to provide data to help clarify this puzzling contradiction. In vitro DBM activity assays showed Cu− samples had significantly higher activity than Cu+ samples in both AG and MO, but not VD. Activity data were confirmed by Western immunoblots. Quantitative real-time PCR demonstrated higher DBM mRNA in Cu− tissues but unaltered levels of several other cuproenzymes and Cu-binding proteins. Previous pharmacological data implied that high DBM was associated with low NA. HPLC analyses confirmed that NA and DA levels in Cu− MO, VD and HT were significantly lower and higher, respectively, than in Cu+ tissues. However, the NA content of AG was not statistically lower. Furthermore there was no correlation between higher DBM mRNA and lower NA in four Cu−tissues. Adequate dietary Cu is essential to support DBM function in vivo but additional studies are needed to determine the mechanism for increased DBM transcription associated with Cu deficiency.

The objective of the present study was to analyse whether there is a relationship between the clinical severity at the time of starting transpyloric enteral nutrition (TEN) and the onset of digestive tract complications in critically ill children. Between May 2005 and December 2007, we performed a prospective, observational study with the participation of 209 critically ill children aged between 3 d and 17 years and who received TEN. The characteristics of the nutrition and its tolerance were compared with the paediatric risk of mortality (PRISM), the paediatric index of mortality (PIM) and the paediatric logistic organ dysfunction index (PELOD) at the time of starting the nutrition. Higher PRISM and PELOD scores correlated with a later time of starting enteral nutrition, a longer time to reach the maximum daily energy delivery and a longer duration of the TEN. However, the severity scores did not correlate with the maximum energy delivery achieved. Abdominal distension or excessive gastric residues were observed in 4·7 % of the patients and diarrhoea in 4·3 %. The ability of the severity scores to predict diarrhoea was of 0·67 for PRISM, 0·63 for PELOD and 0·60 for PIM-2.The severity scores were not able to predict other digestive tract complications. Higher scores of clinical severity at the time of starting enteral nutrition correlate with a later initiation of the nutrition, a longer time to reach the maximum energy delivery and a longer duration of TEN. However, their ability to predict digestive tract complications is low.

The PPARγ gene regulates insulin sensitivity and adipogenesis. The Pro12Ala polymorphism of this gene has been related to fat accumulation. Our aim was to analyse the effects of a 2-year nutritional intervention with Mediterranean-style diets on adiposity in high-cardiovascular risk patients depending on the Pro12Ala polymorphism of the PPARγ gene. The population consisted of a substudy (774 high-risk subjects aged 55–80 years) of the Prevención con Dieta Mediterránea (PREDIMED) randomised trial aimed at assessing the effect of the Mediterranean diet for CVD prevention. There were three nutritional intervention groups: two of them of a Mediterranean-style diet and the third was a control group advised to follow a conventional low-fat diet. All the participants were genotyped by PCR-restriction fragment length polymorphism (RFLP). The results showed that carriers of the 12Ala allele allocated to the control group had a statistically significant higher change in waist circumference (adjusted difference coefficient = 2·37 cm; P = 0·014) compared with wild-type subjects after 2 years of nutritional intervention. This adverse effect was not observed among 12Ala carriers allocated to both Mediterranean diet groups. In diabetic patients a statistically significant interaction between Mediterranean diet and the 12Ala allele regarding waist circumference change was observed ( − 5·85 cm; P = 0·003). In conclusion, the Mediterranean diet seems to be able to reduce waist circumference in a high-cardiovascular risk population, reversing the negative effect that the 12Ala allele carriers of the PPARγ gene appeared to have. The beneficial effect of this dietary pattern seems to be higher among type 2 diabetic subjects.

High postprandial lipaemia increases cardiovascular risk. Algae consumption may affect postprandial lipoproteinaemia. The effects of dietary alga and cholesterol supplementation on postprandial lipaemia and lipoproteinaemia and arylesterase (AE) activity in growing male Wistar rats were tested in the present study. Six groups of ten rats were fed a casein-based diet for 3 weeks. Three of the diets contained 2·4 % cholesterol-raising agent (Chol), while the other three did not (NChol). Seven percentage of the control diets (NChol-C and Chol-C) consisted of a cellulose–wheat starch mix (35:65), while the Nori alga diets (NChol-N and Chol-N) and Konbu diets (NChol-K and Chol-K) contained 7 % of each respective freeze-dried alga. Postprandial plasma was obtained after a 3 h diet withdrawal. Supplementary cholesterol and alga type significantly affected (at least P < 0·05) the cholesterol, TAG, phospholipid and protein contents of the various lipoprotein fractions. AE enzyme activity increased (P < 0·05) in NChol rats given Nori and Konbu diets. NChol-K, but not NChol-N, rats displayed higher (P < 0·05) plasma cholesterol, TAG and phospholipid levels than NChol-C animals. NChol-K rats presented higher TAG, phospholipid, protein and lipoprotein mass values than their NChol-C counterparts. Inclusion of algae in Chol diets decreased (P < 0·001) the postprandial hypertriacylglycerolaemia. The Chol-N diet affected most lipoprotein fraction contents. Chol-N rats had lower postprandial cholesterolaemia and a better lipoprotein profile (fewer LDL and a tendency toward more HDL and fewer cholesterol-enriched VLDL) than Chol-K rats, suggesting that Nori is the alga of choice in dietary treatment of hypercholesterolaemia.

The purpose of the present study was to investigate whether water-insoluble fish protein (IFP) from Alaska pollock (Theragra chalcogramma) prevents hypercholesterolaemia induced by ovarian hormone deficiency. Wistar female rats, aged 6 months, were subjected to sham-operation or ovariectomy, and fed a cholesterol-free diet containing casein or IPF as a protein source for 28 d. Body-weight gain and food intake increased in the ovariectomised rats as compared with the sham-operated rats. Plasma total cholesterol concentration was decreased and faecal bile acid excretion was increased by IFP in the ovariectomised rats, but not in the sham-operated rats. Plasma homocysteine concentration was decreased by IFP in the ovariectomised rats, but not in the sham-operated rats. Liver lipids and liver cholesterol concentrations were increased and cholesterol 7α-hydroxylase (CYP7A1) activity was decreased by ovariectomy, but not by diet. Bile acid content and the ratio of cholic acid groups to chenodeoxycholic acid groups in bile were increased by ovariectomy, but decreased by IFP. Bile acid content in the small intestine was increased by IFP in the ovariectomised rats, but not in the sham-operated rats. 3-Hydroxy-3-methylglutaryl-CoA reductase and microsomal TAG transfer protein mRNA levels were decreased by ovariectomy and IFP, whereas LDL-receptor mRNA level was decreased by ovariectomy but unaffected by diet. Thus, the preventive effect of IFP on the ovarian hormone deficiency-associated increase in plasma cholesterol concentration seems to be mediated by accelerated faecal excretion of bile acids, coupled with an increase in the intestinal pool of bile acids.

Epidemiological investigation and animal studies have shown that dietary n-3 PUFA prevent the development and progression of certain types of cancer. However, conflicting results have been reported by the few studies that focused on the effect of dietary n-3 PUFA on the development of metastases. In the present study, we investigated the metastatic dissemination of murine T lymphoma lines with different metastatic potential transplanted into mice fed a fish oil diet, compared with mice fed a maize oil diet. Transplantation of highly metastatic S11 cells into animals fed a fish oil diet induced a large lymphomatoid infiltration in the spleen, associated with an eight-fold increase in spleen weight, compared with normal animals on the same diet. In contrast, only a limited increase in spleen weight was found in animals transplanted with S11 cells while fed a maize oil diet. No significant increase in spleen weight was found in animals transplanted with low-metastatic 164T2 cells regardless of whether they were fed a fish oil or a maize oil diet. At the end of experiment, an overt cachexia was shown by animals fed a fish oil diet transplanted with S11 cells, but not by those transplanted with 164T2 cells. The particularly high pro-metastatic effect of dietary n-3 PUFA on S11 cells rules out the generalisation that dietary n-3 PUFA inhibit tumour growth and progression.

Solving complex problems in large-scale research programmes requires cooperation and division of labour. Simultaneously, large-scale problem solving also gives rise to unintended side effects. Based upon 5 years of researching two large-scale nutrigenomic research programmes, we argue that problems are fragmented in order to be solved. These sub-problems are given priority for practical reasons and in the process of solving them, various changes are introduced in each sub-problem. Combined with additional diversity as a result of interdisciplinarity, this makes reassembling the original and overall goal of the research programme less likely. In the case of nutrigenomics and health, this produces a diversification of health. As a result, the public health goal of contemporary nutrition science is not reached in the large-scale research programmes we studied. Large-scale research programmes are very successful in producing scientific publications and new knowledge; however, in reaching their political goals they often are less successful.

Oleate has been shown to protect against palmitate-induced insulin resistance. The present study investigates mechanisms involved in the interaction between oleate and palmitate on insulin-stimulated glucose uptake by L6 skeletal muscle cells. L6 myotubes were cultured for 6 h with palmitate or oleate alone, and combinations of palmitate with oleate, with and without phosphatidylinositol 3-kinase (PI3-kinase) inhibition. Insulin-stimulated glucose uptake, measured by uptake of 2-deoxy-d-[3H]glucose, was almost completely prevented by 300 μm-palmitate. Cells incubated with oleate up to 750 μmol/l maintained a significant increase in insulin-stimulated glucose uptake. Co-incubation of 50–300 μm-oleate with 300 μm-palmitate partially prevented the decrease in insulin-stimulated glucose uptake associated with palmitate. Adding the PI3-kinase inhibitors wortmannin (10− 7 mol/l) or LY294002 (25 μmol/l) to 50 μm-oleate plus 300 μm-palmitate significantly reduced the beneficial effect of oleate against palmitate-induced insulin resistance, indicating that activation of PI3-kinase is involved in the protective effect of oleate. Thus, the prevention of palmitate-induced insulin resistance by oleate in L6 muscle cells is associated with the ability of oleate to maintain insulin signalling through PI3-kinase.

The EURopean micronutrient RECommendations Aligned (EURRECA) Network of Excellence is working towards developing aligned micronutrient recommendations across Europe. The purpose of the present study was to conduct a review of methods used in validation studies carried out in adults assessing dietary intake of EURRECA priority minerals. A search strategy and inclusion criteria were defined and a scoring system was developed to rate the quality of each validation study that produced a quality index with possible scores obtained ranging from 0·5 to 7. A MEDLINE and EMBASE literature review was conducted. Articles/validation studies meeting the inclusion criteria included: 79/88 for Fe; 95/104 for Ca; 13/15 for Se; 29/30 for Zn; 7/9 for iodine. The most frequently used method to ascertain dietary intake was the Food Frequency Questionnaire (FFQ), whereas dietary records (DR) and 24 h recalls were the most used reference methods. The correlation coefficients (CC) between study mineral intakes estimated by FFQ and the reference method were weighted according to the study's quality index and obtained acceptable to good ratings, ranging from 0·36 to 0·60 when the reference method was DR and from 0·41 to 0·58 when the reference was 24 h recalls. A minority of studies (n 9) used biomarkers for validation and among these, five included iodine obtaining a CC of 0·47. The FFQ was seen as a valid method for assessing mineral intake, particularly for Ca and, to a lower extent, for iodine and Zn. Se and Fe showed only acceptable correlations. The present review provides new insights regarding the characteristics that assessment methods for dietary mineral intakes should fulfil.

Rapeseed oil (RSO) is a novel source of plant sterols, containing the unique brassicasterol in concentrations higher than allowed for plant sterol blends in food products in the European Union. Effects of RSO sterols and stanols on aortic atherosclerosis were studied in cholesterol-fed heterozygous Watanabe heritable hyperlipidaemic (Hh-WHHL) rabbits. Four groups (n 18 per group) received a cholesterol-added (2 g/kg) standard chow or this diet with added RSO stanol esters (17 g/kg), RSO stanol esters (34 g/kg) or RSO sterol esters (34 g/kg) for 18 weeks. Feeding RSO stanol esters increased plasma campestanol (P < 0·001) and sitostanol (P < 0·001) and aortic campestanol (P < 0·05) compared with controls. Feeding RSO sterol esters increased concentrations of plasma campesterol (P < 0·001), sitosterol (P < 0·001) and brassicasterol (P < 0·001) and aortic campesterol (P < 0·01). Significantly lower plasma cholesterol (P < 0·001) was recorded in the treated groups after 3 weeks and throughout the study. LDL-cholesterol was reduced 50 % in the high-dose RSO sterol ester (P < 0·01) and high-dose RSO stanol ester (P < 0·001) groups compared with controls. Atherosclerotic lesions were found in three rabbits in each of the RSO stanol ester groups and in one in the RSO sterol ester group. Aortic cholesterol was decreased in the treated groups (P < 0·001) in response to lowering of plasma cholesterol induced by RSO sterol and stanol esters. In conclusion, RSO stanol and sterol esters with a high concentration of brassicasterol were well tolerated. They were hypocholesterolaemic and inhibited experimental atherosclerosis in cholesterol-fed Hh-WHHL rabbits. A significant uptake of plant sterols into the blood and incorporation of campesterol and campestanol into aortic tissue was recorded.

Dietary Si (orthosilicic acid; OSA) appears important in connective tissue health, and although the sources and intakes of Si are well established, its absorption is not. Si absorption was measured from eight high-Si-containing sources: alcohol-free beer; OSA solution (positive control); bananas; green beans; supplemental choline-stabilised OSA (ChOSA); supplemental monomethyl silanetriol (MMST); supplemental colloidal silica (CS); magnesium trisilicate British Pharmacopoeia antacid (MTBP). Two of the supplements and the antacid were pre-selected following an in vitro dissolution assay. Fasting, healthy subjects (CS, n 3; others, n ≥ 5) each ingested two of the sources separated by a 1-week wash-out period. Blood and urine were collected and measured for total Si concentrations by inductively coupled plasma optical emission spectrometry. Absorption, based on urinary Si excretion, was highest for MMST and alcohol-free beer (64 % of dose), followed by green beans (44 %), OSA (43 %), ChOSA (17 %), bananas and MTBP (4 %) and CS (1 %). Peak serum concentrations occurred by 0.5 h for MMST and green beans, 1.5 h for OSA and alcohol-free beer, 2 h for ChOSA and CS, and 4 h for MTBP. Area under the serum curves correlated positively with urinary Si output (r 0·82; P < 0·0001). Absorption of Si from supplements and antacids was consistent with their known chemical speciation and kinetics of dissolution under simulated gastrointestinal conditions. Monomeric silicates were readily absorbed, while particulate silicates were decreasingly well absorbed with increasing polymerisation. The present results highlight the need to allow for relative absorption of Si from different foods or supplements in subsequent epidemiological and intervention studies.

Soya isoflavones (SIF) and folic acid (FA) both confer the biological properties of antioxidation; however, the mechanism of their antioxidant effect on nervous system development is unclear. Our purpose is to investigate the neuroprotective effects of SIF, FA or co-administration of SIF with FA against β-amyloid 1-40 (Aβ1-40)-induced learning and memory impairment in rats. In the present study, the learning and memory ability of rats and the amount of amyloid-positive neurons in the cerebral cortex and hippocampal CA1 area were measured. The levels of total antioxidant capacity (T-AOC), glutathione (GSH) and glutathione peroxidase (GSH-Px) in serum and brain tissue were also measured. The results showed that intracerebroventricular administration of Aβ1-40 resulted in a dramatic prolongation of the escape latency; however, in the SIF, FA and SIF+FA treatment groups, the functional deficits of learning and memory were significantly improved. Moreover, after Aβ1-40 injection, the levels of T-AOC and GSH were profoundly decreased, suggesting a decline of antioxidant activity in the rats. However, intragastric pre-treatment with SIF, or FA, or SIF+FA resulted in a significant increase of antioxidative activity. SIF, or FA, or SIF+FA treatments also reversed the Aβ1-40-induced increase in the amount of amyloid-positive neurons. These results suggest that: (1) learning or memory impairment in experimental rats was caused by Aβ1-40, which is probably attributed to Aβ-induced oxidative damage and deposition of β-amyloid peptides in the brain; (2) pre-administration of SIF and/or FA may prevent the pathological alterations caused by Aβ1-40 treatment and the neuroprotective effects of SIF and/or FA are indicated.

Isoflavonoids and fish oil may be protective against colorectal cancer, but the evidence in relation to breast cancer risk is ambiguous. In the present study, we have investigated the impact of soya-derived isoflavonoids and n-3 fatty acids from fish oil, both individually and in combination, on apoptosis, cell proliferation and oestrogen receptor (ER) expression in the colon and mammary gland of the rat. Female rats were fed diets high in n-3 fatty acids (80 g/kg diet) or soya protein (765 mg/kg diet isoflavones) for 2 weeks, and then killed before the removal of the colon and mammary glands. Cell proliferation and apoptosis were quantified morphologically in whole crypts and terminal end buds. The expressions of ERα and ERβ were measured in colon tissue scrapes and the mammary gland. Fish oil significantly increased apoptosis and decreased mitosis in both tissues, an effect associated with a decrease in the expressions of ERα and ERβ. Soya had no effect on apoptosis in either tissue, but reduced mitosis in the colon (P < 0·001) while increasing it in the mammary gland (P = 0·001). The changes in proliferation were associated with contrasting changes in the ER expression such that fish oil significantly decreased both ERβ and ERα, while soya increased ERα and decreased ERβ. The results may provide a novel mechanism by which n-3 fatty acids could reduce cancer risk, but the interpretation of the results in relation to soya consumption and breast cancer risk requires further investigation.

Dehydroepiandrosterone (DHEA), a steroid hormone that is secreted by the adrenal cortex in mammals, has an array of biological actions, including inhibition of fat synthesis, decreasing the number of adipocytes, and a reduction in mammalian metabolic efficiency. Recent studies showed that DHEA may decrease fat deposition in poultry, but the mechanism of action is unclear. In the present study, we demonstrate that DHEA stimulates intracellular cyclic adenosine 3′,5′-monophosphate (cAMP) accumulation in chicken hepatocytes during a 30 min incubation period. Increases in intracellular cAMP are evoked by as low as 0·1 μm-DHEA. The cAMP induced by DHEA, while suppressing cAMP-specific phosphodiesterase activity, also activates cAMP-dependent protein kinase A (PKA) in chicken hepatocytes. In addition, the activation of PKA leads to down-regulation of sterol regulatory element-binding protein-1 (SREBP-1). These findings demonstrate that direct action by DHEA leads to activation of the cAMP/PKA signalling system in the modulation of lipid metabolism by repressing SREBP-1, thereby providing a novel explanation for some of the underlying effects proposed for DHEA in the prevention of fat deposition in poultry.

Deoxynivalenol (DON) is a toxic fungal metabolite found on wheat, maize and barley. We previously reported a significant association between the amount of DON in a single 24 h urine sample and the average cereal intake over 7 d for 300 UK adults. In this more detailed analysis of the data, food diary information (n 255) for the day of urine collection (model I), the previous 24 h period (model II) and the day of urine collection plus the previous 24 h combined (model III) were further examined to assess whether the recent intake of cereal correlated more strongly with urinary DON, compared with the longer-term assessment of usual cereal intake from 7 d food diaries (model IV). DON was detected in 254/255 (99·6 %) urine samples (mean 12·0 μg/d; range not detected–66 μg/d). For all the models, total cereal intake was positively associated with urinary DON (P < 0·001) in each model. The goodness of fit (adjusted R2 value) was used to assess how well each model explained the variation in urinary DON. Model I provided a better goodness of fit (adjusted R2 0·22) than model IV (adjusted R2 0·19), whereas model III provided the best fit (adjusted R2 0·27). These data suggest that the inter-individual variation in urinary DON was somewhat better explained by recent cereal intake compared with usual cereal intake assessed over 7 d.

In order to assess nutritional adequacy, valid estimates of nutrient intake are required. Specifically, the EURopean micronutrient RECommendations Aligned (EURRECA) Network of Excellence needs clear guidelines for assessing the validity of reported micronutrient intakes and n-3 fatty acid (FA) intakes. The aim of the present study was to review the validity of methods used to measure the usual n-3 FA intake of a population. A systematic literature search was conducted for studies validating the methodology used for measuring the dietary intake of n-3 FA. The quality of the validation studies and the quality of the different dietary assessment methods were assessed using scoring systems developed by EURRECA. Fourteen papers, describing twenty studies, were identified for inclusion. According to the score system developed by EURRECA, all the studies were ranked as average, except two that were ranked as poor. The correlation coefficients between FA in subcutaneous fat and dietary intake of n-3 FA from four FFQ, one weighed record and one 24-h recall ranged between 0·40 and 0·60. Correlations between intake of n-3 FA from five FFQ, one dietary history and three weighed records and blood lipids were similar to the ones observed for subcutaneous fat. The summarised quality of the n-3 FA estimates derived from the FFQ was judged as good or acceptable according to the EURRECA scoring system. The literature describes subcutaneous fat as the best reference method, and the studies where this was used had moderate correlation coefficients and no dietary intake method was superior to any other.

The objective of the present study was to measure the differences in body composition, adipose tissue gene expression, blood metabolite and hormone concentrations, and insulin sensitivity in kittens exposed to high-protein (HP) or high-carbohydrate (HC) nutrition in utero and through the growth period. Eight dams were randomised onto two test diets, and fed the diets throughout gestation and lactation. Male offspring were evaluated for 9 months. Kittens were weaned at 2 months of age onto the same treatment diet as the dam and were allowed to consume diets ad libitum. The HC diet contained 34·3 % crude protein (CP), 19·2 % fat and 30·8 % digestible carbohydrate, while the HP diet contained 52·9 % CP, 23·5 % fat and 10·8 % digestible carbohydrate. Blood samples were collected at 6 months after birth. Body composition was determined at 2 and 8 months of age and an intravenous glucose tolerance test, neutering and adipose tissue biopsy conducted at 8 months of age. Physical activity was quantified at 6 and 9 months. Energy intake, DM intake and body weight were not different between groups. At 2 months, blood TAG were greater (P < 0·05) in kittens fed the HP diet. At 8 months, blood leptin was higher (P < 0·05) in kittens fed the HC diet, while chemokine receptor 5, hormone-sensitive lipase, uncoupling protein 2, leptin and insulin receptor mRNA were greater (P < 0·05) in kittens fed the HP diet. The present results demonstrate some of the changes in blood metabolites and hormones, physical activity and mRNA abundance that occur with feeding high protein levels to kittens.