This study demonstrates that feeding difructose anhydride III (DFAIII) improves bone strength and femoral mineral concentrations in a rat model of oestrogen deficiency. We showed the relationship between Ca, Mg and P absorption and bone characteristics in rats. Two groups of female Sprague-Dawley rats (6 weeks old) underwent bilateral ovariectomy (ovariectomized rats, OVX rats) or bilateral laparotomy (sham rats). At 10 weeks old, OVX and sham rats were divided into three subgroups and fed a control, 1·5 % DFAIII or 3 % DFAIII diet for 8 weeks, respectively. Ca but not Mg absorption rates were lowered by ovariectomy; however, ingestion of the 1·5 % and 3 % DFAIII diets similarly restored the reduced Ca absorption in OVX rats at 4 and 8 weeks after feeding of the test diets. DFAIII increased Mg absorption dose-dependently in sham and OVX rats. The bone strength, femoral Ca and Mg concentrations, and distal bone mineral density in the 3 % DFAIII group were higher than those in the control group in OVX rats. The absorption rates of Ca and Mg were significantly correlated with femoral Ca and Mg concentrations and strength, which suggests that increasing both Ca and Mg absorption improves bone characteristics in OVX rats. There were no differences in any of the variables in the femur between the 1·5 % and 3 % DFAIII groups in OVX rats. In conclusion, feeding of a low dose of DFAIII increased intestinal Ca and Mg absorption, and the promotive effect of DFAIII persisted for over 8 weeks. This effect was associated with prevention of ovariectomy-induced osteopenia.

Strenuous exercise is known to induce oxidative stress leading to the generation of free radicals. The purpose of the present study was to investigate the effects of lycopene, an antioxidant nutrient, at a relatively low dose (2·6 mg/kg per d) and a relatively high dose (7·8 mg/kg per d) on the antioxidant status of blood and skeletal muscle tissues in rats after exhaustive exercise. Rats were divided into six groups: sedentary control (C); sedentary control with low-dose lycopene (CLL); sedentary control with high-dose lycopene (CHL); exhaustive exercise (E); exhaustive exercise with low-dose lycopene (ELL); exhaustive exercise with high-dose lycopene (EHL). After 30 d, the rats in the three C groups were killed without exercise, but the rats in the three E groups were killed immediately after an exhaustive running test on a motorised treadmill. The results showed that xanthine oxidase (XO) activities of plasma and muscle, and muscular myeloperoxidase (MPO) activity in group E were significantly increased compared with group C. Compared with group E, the elevations of XO and MPO activities of muscle were significantly decreased in group EHL. The malondialdehyde concentrations of plasma and tissues in group E were significantly increased by 72 and 114 %, respectively, compared with those in group C. However, this phenomenon was prevented in rats of the ELL and EHL groups. There was no significant difference in the GSH concentrations of erythrocytes in each group; however, exhaustive exercise resulted in a significant decrease in the GSH content of muscle. In conclusion, these results suggested that lycopene protected muscle tissue from oxidative stress after exhaustive exercise.

It has been reported that green tea has a thermogenic effect, due to its caffeine content and probably also to the catechin, epigallocatechin-3-gallate (EGCG). The main aim of the present study was to compare the effect of a mixture of green tea and Guarana extracts containing a fixed dose of caffeine and variable doses of EGCG on 24 h energy expenditure and fat oxidation. Fourteen subjects took part to this randomized, placebo-controlled, double-blind, cross-over study. Each subject was tested five times in a metabolic chamber to measure 24 h energy expenditure, substrate oxidation and blood pressure. During each stay, the subjects ingested a capsule of placebo or capsules containing 200 mg caffeine and a variable dose of EGCG (90, 200, 300 or 400 mg) three times daily, 30 min before standardized meals. Twenty-four hour energy expenditure increased significantly by about 750 kJ with all EGCG–caffeine mixtures compared with placebo. No effect of the EGCG–caffeine mixture was observed for lipid oxidation. Systolic and diastolic blood pressure increased by about 7 and 5 mmHg, respectively, with the EGCG–caffeine mixtures compared with placebo. This increase was significant only for 24 h diastolic blood pressure. The main finding of the study was the increase in 24 h energy expenditure with the EGCG–caffeine mixtures. However, this increase was similar with all doses of EGCG in the mixtures.

The present study measuring resting energy expenditure (REE; kJ/d) longitudinally using indirect calorimetry in six elderly women aged ≥70 years following surgery for hip fracture, describes changes over time (days 10, 42 and 84 post-injury) and compares measured values to those calculated from routinely applied predictive equations. REE was compared to REE predicted using the Harris Benedict and Schofield equations, with and without accounting for the theoretical increase in energy expenditure of 35 % secondary to physiological stress of injury and surgery. Mean (95 % CI) measured REE (kJ/d) was 4704 (4354, 5054), 4090 (3719, 4461) and 4145 (3908, 4382) for days 10, 42 and 84, respectively. A time effect was observed for measured REE, P=0·003. Without adjusting for stress the mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Harris Benedict equation were 1 (−9, 12), 10 (2, 18) and 9 (1, 17) for days 10, 42 and 84, respectively. The mean difference and 95 % limits of agreement for measured and predicted REE (kJ/kg per d) for the Schofield equation without adjusting for stress were 8 (−3, 19), 16 (6, 26) and 16 (10, 22) for days 10, 42 and 84, respectively. After adjusting for stress, REE predicted from the Harris Benedict or Schofield equations overestimated measured REE by between 38 and 69 %. Energy expenditure following fracture is poorly understood. Our data suggest REE was relatively elevated early in recovery but declined during the first 6 weeks. Using the Harris Benedict or Schofield equations adjusted for stress may lead to overestimation of REE in the clinical setting. Further work is required to evaluate total energy expenditure before recommendations can be made to alter current practice for calculating theoretical total energy requirements of hip fracture patients.

Evidence suggests that low folate status may be detrimental to mood and associated with depleted cerebrospinal fluid levels of the neurotransmitter serotonin (5-hydroxytryptamine; 5-HT). A placebo-controlled trial was carried out to determine the effect of folic acid supplementation (100 μg for 6 weeks followed by 200 μg for a further 6 weeks) upon subjective mood (Positive and Negative Affect Schedule) and biochemical markers of mood (5-HT) in healthy males (n 23). Blood samples were obtained at baseline (week 0) and during the intervention at week 6 and week 12. Subjective mood assessments were obtained at week 0 and week 12. The results showed an increase in serum and erythrocyte folate concentrations (P=0·02 and P=0·003, respectively) and a corresponding decrease in plasma homocysteine (P=0·015) in response to the folic acid intervention. Neither subjective mood nor 5-HT levels, however, were significantly altered in response to the change in folate status. Folic acid given at physiological doses did not appear to improve the mood of healthy folate-replete individuals over a 12-week period. Further research is needed to address the effect of folic acid supplementation or of longer duration or increased dose, particularly in the face of sub-optimal folate status.

The effects of conjugated linoleic acid (CLA), γ-linolenic acid (GLA), linoleic acid (LA), and their combinations, on skin composition in mice were investigated. Mice (8 weeks old) were orally administered with either LA, GLA, CLA, LA + GLA, LA + CLA, or CLA + GLA for 4 weeks. Then, the skin was analysed for triacylglycerol content, fatty acid composition and collagen content. Additionally, thicknesses of the dermis layer and subcutaneous tissue layer, and the size and number of adipocytes were measured histologically. The skin fatty acid composition was modified depending upon the fatty acid composition of supplemented oils. In each oil-alone group, skin triacylglycerol content was the highest in LA, followed by GLA and CLA treatments. Combinations with CLA had a similar triacylglycerol content compared with the CLA-alone group. No significant changes in collagen content were observed among any treatments. The effects on subcutaneous thickness were similar to the results obtained in the triacylglycerol contents, where groups supplemented with CLA alone or other fatty acids had significantly thinner subcutaneous tissue compared with the LA-alone group. However, no significant difference was detected in the thickness of the dermis layers. The number of adipocytes was highest in the LA + GLA group and tended to be reduced by CLA with or without the other fatty acids. These results suggest that CLA alone or in combination with other fatty acids strongly modifies skin composition in mice.

To determine whether or not the weight (and fat) loss induced by oleoyl-oestrone treatment results only as a consequence of decreased food intake, we compared treated animals with a pair-fed model. To this end, Wistar female rats received daily oral gavages of 10μmol/kg per d oleoyl-oestrone in sunflower oil, or vehicle alone for 10 or 20d. A second group of rats received the gavage of sunflower oil and the same amount of food ingested as the oleoyl-oestrone-treated animals (pair-fed group). Rats treated with oleoyl-oestrone maintained glucidic metabolism homeostasis despite a marked decrease in adipose tissue weight (P<0·001). Pair-fed rats exhibited a different pattern, comparable to short-term starvation, with greatly decreased glycogen stores (P<0·0001). The most significant effects were detected in the 10d period groups. Oleoyl-oestrone affected the activity of the ponderostat system not only by decreasing appetite but also by modifying energy partition: treated animals maintained their glucose and energy homeostasis despite decreased food intake and the massive depletion of lipid stores.

This study was undertaken to investigate the growth performance and immune responses of broiler chicks fed diets supplemented with conjugated linoleic acid (CLA). Two hundred and forty day-old Arbor Acre male broiler chicks were randomly allotted into four dietary treatments with different inclusion levels of CLA (0, 2·5, 5·0 or 10·0g pure CLA/kg) for 6 weeks. Growth performance, lysozyme activity, peripheral blood mononuclear cell (PBMC) proliferation, prostaglandin E2 (PGE2) synthesis and antibody production were investigated. There were no significant differences in growth performance among treatments (P>0·05). Chicks fed 10·0g CLA/kg diet produced 40% and 49% more lysozyme activity in serum and spleen than the control group at 21d of age (P<0·05). Dietary CLA enhanced the PBMC proliferation in response to concanavalin A at the age of 21 and 42d (P<0·05). Systemic and peripheral blood lymphocytic synthesis of PGE2 in chicks fed 10·0g CLA/kg diet was significantly decreased by 57% and 42% compared to chicks fed control diet (P<0·05). Antibody production to sheep red blood cell and bovine serum albumin were elevated in either 2·5 or 10·0g CLA/kg dietary treatments (P<0·05). The results indicated dietary CLA could enhance the immune response in broiler chicks, but did not alter the growth performance.

Endogenous vitamin D deficiency (low serum 25(OH)D3) is a necessary but insufficient requirement for the genesis of vitamin D-deficiency rickets and osteomalacia. The magnitude of the independent contributions of dietary factors to rachitic and osteomalacic risk remains uncertain. We reanalysed two weighed dietary surveys of sixty-two cases of rickets and osteomalacia and 113 normal women and children. The independent associations of four dietary variables (vitamin D, Ca, fibre and meat intakes) and daylight outdoor exposure with rachitic and osteomalacic relative risk were estimated by multivariate logistic regression. Meat and fibre intakes showed significant negative and positive associations respectively with rachitic and osteomalacic relative risk (RR; zero meat intake: RR 29·8 (95 % CI 4·96, 181), P<0·001; fibre intake: RR 1·53 (95 % CI 1·01, 2·32), P+0·043). The negative association of meat intakes with rachitic and osteomalacic relative risk was curvilinear; relative risk did not fall further at meat intakes above 60 g daily. Daylight outdoor exposure showed a significant negative association with combined relative risk (RR 0·33 (95 % CI 0·17, 0·66), P<0·001). Operation of the meat and fibre risk factors was related to sex, age and dietary pattern (omnivore/lactovegetarian), mainly determined by religious affiliation. The mechanism by which meat reduces rachitic and osteomalacic risk is uncertain and appears independent of revised estimates of meat vitamin D content. The meat content of the omnivore Western diet may explain its high degree of protection against nutritional rickets and osteomalacia from infancy to old age in the presence of endogenous vitamin D deficiency.

Eating behaviour depends partly on food preference, which is itself determined by different types of emotions. Among the emotions generated by food, disgust with red meat is common in women and can lead to reduced meat consumption. We tested the hypothesis that low meat intake is related to different negative emotions towards meat but does not affect the emotions expressed towards other food categories. Food intake of sixty women was followed throughout each day for 1 week and allowed us to assign women to two groups (low v. high meat-eating women). They were then invited to assess the intensity of twenty-six emotions described by words and induced by thirty food pictures. We determined the number of necessary dimensions to describe the space created by the twenty-six words. The results showed differences in emotions between the low and high meat-eating women. As expected, there were overall differences in the emotions generated by the thirty food pictures. Six clusters of emotions were necessary and sufficient to summarise the emotional space. These dimensions were described by ‘disappointment’, ‘satisfaction’, ‘guilt’, ‘doubt’, ‘amused’ and ‘indifference’. As expected, the low meat-eating women felt more ‘disappointment’, ‘indifference’ and less ‘satisfaction’ towards meat than did the high meat-eating women. However, the low meat-eating women also stated other negative emotions such as ‘doubt’ towards some starchy foods. The only foods that they liked more than high meat-eating women were pears and French beans. In conclusion, low meat consumption was associated with specific negative emotions regarding meat and other foods.

The incidence of childhood obesity is rising dramatically throughout industrialised countries. To evaluate and study the impact of childhood obesity on lipoprotein metabolism, we developed a new animal model of premature obesity. Yucatan mini-pigs aged 4 months were studied over a 12-month period from childhood to adulthood. Animals were divided into two groups: the first group were overfed a Western misbalanced diet; the second group were normally fed a recommended human-type diet. Cholesterol and triacylglycerol concentrations in VLDL-, LDL- and HDL-lipoproteins were followed from baseline to adulthood by fast protein liquid chromatography. At 10 (the end of sexual maturation) and 16 months old (adulthood), liver, visceral and subcutaneous adipose tissues were sampled. Real-time RT-PCR was performed in order to compare apo AI, apo B, apo C-III, PPAR-α, insulin receptor and lipoprotein lipase gene expression between groups and ages. Differences between groups were observed only after sexual maturity. Adult overfed mini-pigs had a higher LDL-cholesterol:HDL-cholesterol ratio (P<0·05; 0·55 (SE 0·06) for overfed v. 0·42 (SE 0·04) for normally fed pigs at the tenth month of the study). In both groups, VLDL-triacylglycerol decreased (P<0·05). VLDL-triacylglycerol evolution in the overfed group was associated with an increase in LDL-triacylglycerol plasma concentrations (P<0·05) after sexual maturation. LDL-triacylglycerol concentration in overfed mini-pigs went from an average of 0·28 mmol/l before sexual maturation to reach an average concentration of 0·56 mmol/l afterwards. This phenomenon has never been observed in similar studies when obesity is induced in adult mini-pigs and may represent a specific hallmark of an obesity induced during sexual maturity.

Anthropometric parameters and catch-up growth were prospectively evaluated in fifty late-diagnosed children with coeliac disease aged 2·25–10 years after 1–4 years of adhering to a strict gluten-free diet (GFD). The anthropometric parameters were expressed as Z scores relative to National Centre for Health Statistics standards using Epi Info 2000 (weight-for-height Z score (WHZ) and height-for-age Z score (HAZ)). Catch-up growth was evaluated by repeated measures. ANOVA, overall significance by an F test and pair-wise comparisons for estimated marginal means using the least significant difference. At the time of enrolment, no significant difference was observed in WHZ and HAZ between children diagnosed before (group 1) or after (group 2) 4 years of age. On follow-up, HAZ was significantly higher in group 1 after the first and third years of the GFD (P=0·04 and 0·02, respectively), with a non-significant increase after completing 4 years of the GFD (P=0·22). Feeding the GFD resulted in an overall significant (F=3·99, P=0·011) increase in HAZ up to 4 years of follow-up. However, the catch-up in height was incomplete, with stunting in sixteen (55·4 %) of twenty-nine children after 3 years and in seven (46·6 %) of fifteen children after 4 years on the GFD. Pair-wise comparisons demonstrated a linear catch-up growth during the initial follow-up on GFD. Treatment with the GFD did not result in an overall significant increase in WHZ up to 4 years of follow-up (F=1·01, P=0·42). Our results suggest that, in children with late-diagnosed coeliac disease, treatment with a GFD leads to a normalisation of body mass and a significant but incomplete recovery in HAZ during 4 years of follow-up.

Phytosterols contain an unsaturated ring structure and therefore are susceptible to oxidation under certain conditions. Whilst the cytotoxicity of the analogous cholesterol oxidation products (COP) has been well documented, the biological effects of phytosterol oxidation products (POP)have not yet been fully ascertained. The objective of the present study was to examine the cytotoxicity of β-sitosterol oxides and their corresponding COP in a human monocytic cell line (U937), a colonic adenocarcinoma cell line (CaCo-2) and a hepatoma liver cell line (HepG2). 7β-Hydroxysitosterol, 7-ketositosterol, sitosterol-3β,5α,6β-triol and a sitosterol-5α,6α-epoxide–sitosterol-5β,6β-epoxide (6:1) mixture were found to be cytotoxic to all three cell lines employed; the mode of cell death was by apoptosis in the U937 cell line and necrosis in the CaCo-2 and HepG2 cells. 7β-Hydroxysitosterol was the only β-sitosterol oxide to cause depletion in glutathione, indicating that POP-induced apoptosis may not be dependent on the generation of an oxidative stress. A further objective of this study was to assess the ability of the antioxidants α-tocopherol, γ-tocopherol and β-carotene to modulate POP-induced cytotoxicity in U937 cells. Whilst α/γ-tocopherol protected against 7β-hydroxycholesterol-induced apoptosis, they did not confer protection against 7β-hydroxysitosterol-or 7-ketositosterol-induced toxicity, indicating that perhaps COP provoke different apoptotic pathways than POP. β-Carotene did not protect against COP- or POP-induced toxicity. In general, results indicate that POP have qualitatively similar toxic effects to COP. However, higher concentrations of POP are required to elicit comparable levels of toxicity.

The aim of this study was to describe dietary glycaemic index (GI) and glycaemic load (GL) values in the diets of Danish children, and to examine the associations between dietary GI, GL and body fatness. Data were collected during 1997–8 as part of the European Youth Heart Study. The study population comprised 485 children aged 10 years and 364 children aged 16 years from Odense County, Denmark. Dietary GI and GL were estimated using international food tables, and the associations between energy-adjusted dietary GI, GL and body fatness were analysed by multiple linear regression. The mean daily dietary GI value was 85 (SD 6·9) with a range of 62–111. No significant differences were found between age groups and gender. The daily dietary GL was higher among boys aged 16, with a GL of 330 (sd 95) (P<0·05), compared with girls or younger boys. Dietary GL was higher among 10-year-old boys than girls (250 (sd 81) v. 230 (sd 66) P<0·05), whereas dietary GL among 16-year-old girls was 230 (sd 56). Neither dietary GI nor GL was associated with the sum of four skinfolds (ΣSF) among girls or among 10-year-old boys. Among 16-year-old boys, significant associations were observed between dietary GI and ΣSF (β+0·60, SE+0·21, P=0·006), and between dietary GL and ΣSF (β+0·15, SE+0.06, P=0·009). In conclusion, dietary GI and GL were positively associated with body fatness among Danish boys aged 16 years, whereas no associations were found among girls or younger boys.

A 6-month intervention study with a quasi-experimental design was conducted to evaluate the effectiveness of an Internet-based, stepwise, tailored-feedback intervention promoting four key components of the Mediterranean diet. Fifty-three (intervention group) and nineteen (control group) healthy females were recruited from the Universities of Glasgow and Glasgow Caledonian, Scotland, respectively. Participants in the intervention group received tailored dietary and psychosocial feedback and Internet nutrition education over a 6-month period, while participants in the control group were provided with minimal dietary feedback and general healthy-eating brochures. Internet education was provided via an innovative Mediterranean Eating Website. Dietary changes were assessed with 7 d estimated food diaries at baseline and 6 months, and data were analysed to calculate the Mediterranean Diet Score, a composite score based on the consumption of eight components of the traditional Mediterranean diet. The ‘intention-to-treat’ analyses showed that, at 6 months, participants in the intervention group had significantly increased their intake of vegetables, fruits and legumes, as well as the MUFA:saturated fatty acid ratio in their diet, and had significantly increased plasma HDL-cholesterol levels and a reduced ratio of total:HDL-cholesterol. Participants in the control group increased their intake of legumes but showed no other favourable significant changes compared with baseline. This Internet-based, tailored-feedback intervention promoting components of the Mediterranean diet holds promise in encouraging a greater consumption of plant foods, as well as increasing monounsaturated fat and decreasing saturated fat in the Scottish diet; it also shows that the Mediterranean diet can be adopted by healthy individuals in northern European countries.

Shark-liver oil (SLO) contains two bioactive lipids: alkylglycerols and n-3 PUFA. Alkylglycerols have immunostimulating and haematopoietic properties, while n-3 PUFA are essential for optimal neonatal development. We investigated the beneficial effects of dietary supplementation with 32g SLO/d to twelve pregnant and then lactating sows (from day 80 of pregnancy to weaning) on the growth and immune status of their offspring, compared with a control group. Sows were vaccinated against Aujeszky's disease 21d before term. Blood samples were collected from sows before treatment, on delivery and 14d later, and from five piglets per litter on days 2, 21 and 36 after birth; colostrum and milk samples were collected 12h, 14 and 28d postpartum. Compared with controls, supplemented sows had higher levels of both erythrocytes and Hb in their blood, and higher concentrations of IgG, alkylglycerols and n-3 PUFA in their mammary secretions. In piglets from supplemented sows, leucocytes and IgG were higher. Supplementation with SLO resulted in an increase in Aujeszky antibodies in both blood and colostrum of sows after vaccination, together with an increase in Aujeszky antibodies in piglet blood. Our findings demonstrate that improvement of both passive and active immune status in piglets is related to the consumption of alkylglycerols associated with n-3 PUFA in the sow diet. The overall improvement in offspring health status by SLO supplementation to the mother could be of interest for optimisation of the lipid diet during and after pregnancy.

To study the absorption of microbial lysine in growing rabbits, a labelled diet (supplemented with 15NH4Cl) was administered to six animals (group ISOT); a control group (CTRL, four rabbits) received a similar, but unlabelled, diet. Diets were administered for 30d. An additional group of six animals were fed the unlabelled diet for 20d and then the labelled diet for 10d while wearing a neck collar to avoid caecotrophy (group COLL), in order to discriminate it from direct intestinal absorption. At day 30 animals were slaughtered and caecal bacteria and liver samples taken. The 15N enrichment in amino acids of caecal bacteria and liver were determined by GC–combustion/isotope ratio MS. Lysine showed a higher enrichment in caecal microflora (0·925 atom% excess, APE) than liver (0·215 APE) in group ISOT animals, confirming the double origin of body lysine: microbial and dietary. The COLL group showed a much lower enrichment in tissue lysine (0·007 (se 0·0029) APE for liver). Any enrichment in the latter animals was due to direct absorption of microbial lysine along the digestive tract, since recycling of microbial protein (caecotrophy) was avoided. In such conditions liver enrichment was low, indicating a small direct intestinal absorption. From the ratio of [15N]lysine enrichment between liver and bacteria the contribution of microbes to body lysine was estimated at 23%, with 97% of this arising through caecotrophy. Absorption of microbial lysine through caecotrophy was 119 (se 4·0) mg/d, compared with 406 (se 1·8) mg/d available from the diet. This study confirms the importance of caecotrophy in rabbit nutrition (15% of total protein intake).

Normal functioning of the immune system is crucial to the health of man, and diet is one of the major exogenous factors modulating individual immunocompetence. Recently, nutrition research has focused on the role of foods or specific food components in enhancing immune system responsiveness to challenges and thereby improving health and reducing disease risks. Assessing diet-induced changes of immune function, however, requires a thorough methodological approach targeting a large spectrum of immune system parameters. Currently, no single marker is available to predict the outcome of a dietary intervention on the resistance to infection or to other immune system-related diseases. The present review summarises the immune function assays commonly used as markers in human intervention studies and evaluates their biological relevance (e.g. known correlation with clinically relevant endpoints), sensitivity (e.g. within- and between-subject variation), and practical feasibility. Based on these criteria markers were classified into three categories with high, medium or low suitability. Vaccine-specific serum antibody production, delayed-type hypersensitivity response, vaccine-specific or total secretory IgA in saliva and the response to attenuated pathogens, were classified as markers with high suitability. Markers with medium suitability include natural killer cell cytotoxicity, oxidative burst of phagocytes, lymphocyte proliferation and the cytokine pattern produced by activated immune cells. Since no single marker allows conclusions to be drawn about the modulation of the whole immune system, except for the clinical outcome of infection itself, combining markers with high and medium suitability is currently the best approach to measure immunomodulation in human nutrition intervention studies. It would be valuable to include several immune markers in addition to clinical outcome in future clinical trials in this area, as there is too little evidence that correlates markers with global health improvement.