The objective of the present study was to determine whether the consumption of ≥ 250 v. < 250 mg of the long-chain n-3 fatty acids (n-3 LCFA) per d is associated with a reduction in the risk of fatal and non-fatal CHD in individuals with no prior history of CHD. A comprehensive and systematic review of the published scientific literature resulted in the identification of eight prospective studies (seven cohorts and one nested case–control study) that met predefined inclusion criteria. Relative to the consumption of < 250 mg n-3 LCFA per d, the consumption of ≥ 250 mg/d was associated with a significant 35·1 % reduction in the risk of sudden cardiac death and a near-significant 16·6 % reduction in the risk of total fatal coronary events, while the risk of non-fatal myocardial infarction was not significantly reduced. In several meta-analyses, which were based on US studies, risk of CHD death was found to be dose-dependently reduced by the n-3 LCFA, with further risk reductions observed with intakes in excess of 250 mg/d. Prospective observational and intervention data from Japan, where intake of fish is very high, suggest that n-3 LCFA intakes of 900 to 1000 mg/d and greater may confer protection against non-fatal myocardial infarction. Thus, the intake of 250 mg n-3 LCFA per d may, indeed, be a minimum target to be achieved by the general population for the promotion of cardiovascular health.

Recent studies have suggested that chito-oligosaccharides can have anti-adipogenic properties. The objectives of the present study were to evaluate the anti-adipogenic potential of four different chito-oligosaccharides (molecular weight (MW) < 1000, 1000–3000, 3000–5000 and 5000–10 000 Da) and to identify molecular mechanisms underlying the chito-oligosaccharide-mediated inhibition of adipogenesis. Mouse 3T3-L1 cells were allowed to differentiate in the presence of chito-oligosaccharide. At day 8 post-induction of differentiation, lipid accumulation, free glycerol release and the quantitative expression of adipogenic marker genes were evaluated. Chito-oligosaccharides had concentration- and MW-dependent inhibitory effects on lipid accumulation (P < 0·001 and < 0·05, respectively), as well as a concentration-dependent effect (P < 0·001) on free glycerol release and the expression of adipogenic marker genes. The 5000–10 000 Da chito-oligosaccharide was selected for subsequent molecular studies. A panel of forty-four lipid metabolic pathway-specific genes was analysed by quantitative real-time PCR. Chito-oligosaccharide-mediated inhibition of adipogenesis was associated with the up-regulation of the IL-6 gene at all concentrations of chito-oligosaccharide examined and the PG-endoperoxide synthase 2 (PTGS2) gene at higher concentrations of chito-oligosaccharide. The effect of chito-oligosaccharide on gene expression was validated by measuring IL-6 protein concentrations in the media. Finally, an IL-6 promoter assay was developed to characterise the effect of chito-oligosaccharide on the transcriptional activity of the IL-6 promoter, which was increased in a concentration-dependent manner (P < 0·001). We conclude that IL-6 is a candidate signalling molecule in the chito-oligosaccharide-mediated inhibition of adipogenesis in 3T3-L1 cells.

Prevention of cancer through nutritional intervention has gained significant recognition in recent years. Evidence revealed from mechanistic investigations coupled with molecular epidemiology show an inverse association of dietary flavonoids intake with cancer risk. The chemopreventive and anticarcinogenic potential of Selaginella bryopteris, a traditional Indian herb referred to as ‘Sanjeevani’ in the Ayurvedic system of medicine, was examined in the present study. Comprehensive in vitro and in vivo studies were conducted on the flavonoid-rich benzene fraction of the aqueous extract that demonstrated a significant cytoprotective activity. Biomarkers of chemoprevention such as proliferative index and status of cell-cycle regulatory proteins, antioxidant property, anti-inflammatory effect, reversal of stress-induced senescence and genoprotective effect were investigated in human and murine cell cultures. Chemopreventive potential was assessed in benzopyrene-induced lung carcinogenesis and 7,12-dimethyl benz(a)anthracene-mediated skin papillomagenesis test models. Inhibition of DNA fragmentation, unperturbed cell-cycle regulation, maintenance of intracellular antioxidant defence, anti-inflammatory activity, prevention of stress-induced senescence and genoprotective effects against methyl isocyanate carcinogenicity was observed. Medium-term anticarcinogenicity and two-stage skin papillomagenesis tests strongly substantiated our in vitro observations. Results from the present study provide evidence of anticarcinogenic and chemopreventive activities of S. bryopteris hitherto unreported and reaffirm the nutritional significance of flavonoids in cancer prevention.

Mg deficiency accelerates Fe accumulation in the liver, which may induce various metabolic disturbances. In the present study, we examined the gene expression of Hepcidin, a peptide hormone produced in the liver to regulate intestinal Fe absorption negatively, in Mg-deficient rats. Although liver Fe concentration was significantly higher in rats fed an Mg-deficient diet for 4 weeks than in rats fed a control diet, Hepcidin expression in the liver was comparable between the dietary groups. Previous studies revealed that Fe overload up-regulated Hepcidin expression through transcriptional activation by Fe-induced bone morphogenetic protein (Bmp) 6, a growth/differentiation factor belonging to the transforming growth factor-β family, in the liver. Mg deficiency up-regulated the expression of Bmp6 but did not affect the expression of inhibition of DNA binding 1, a sensitive Bmp-responsive gene. In addition, the expression of Bmp receptors such as activin receptor-like kinase 2 (Alk2), activin receptor type IIA (Actr2a), activin receptor type IIB (Actr2b) and Bmp type II receptor (Bmpr2) was lower in the liver of Mg-deficient rats than in that of control rats. The present study indicates that accumulation of hepatic Fe by Mg deficiency is a stimulant inducing Bmp6 expression but not Hepcidin expression by blunting Bmp signalling possibly resulting from down-regulation of the receptor expression. Unresponsive Hepcidin expression may have a role in Mg deficiency-induced changes related to increased liver Fe.

Recent evidence strongly supports the contention that grape seed extract (GSE) improves hyperglycaemia and hyperinsulinaemia in high-fructose-fed rats. To explore the underlying molecular mechanisms of action, we examined the effects of GSE on the expression of muscle proteins related to the insulin signalling pathway and of mRNA for genes involved in the adiponectin signalling pathway. Compared with rats fed on a normal diet, high-fructose-fed rats developed pathological changes, including insulin resistance, hyperinsulinaemia, hypertriacylglycerolaemia, a low level of plasma adiponectin and a high level of plasma fructosamine. These disorders were effectively attenuated in high-fructose-fed rats supplemented with GSE. A high-fructose diet causes insulin resistance by significantly reducing the protein expression of insulin receptor, insulin receptor substrate-1, Akt and GLUT4, and the mRNA expression of adiponectin, adiponectin receptor R1 (AdipoR1) and AMP-activated protein kinase (AMPK)-α in the skeletal muscle. Supplementation of GSE enhanced the expression of insulin signalling pathway-related proteins, including Akt and GLUT4. GSE also increased the mRNA expression of adiponectin, AdipoR1 and AMPK-α. In addition, GSE increased the mRNA levels of glycogen synthase and suppressed the mRNA expression of glycogen synthase kinase-3-α, causing an increase in glycogen accumulation in the skeletal muscle. These results suggest that GSE ameliorates the defective insulin and adiponectin signalling pathways in the skeletal muscle, resulting in improved insulin resistance in fructose-fed rats.

Rosemary (Rosmarinus officinalis L.) extracts (RE) are natural antioxidants that are used in food, food supplements and cosmetic applications; exert anti-inflammatory and anti-hyperglycaemic effects; and promote weight loss, which can be exploited to develop new preventive strategies against metabolic disorders. Therefore, the aim of the present study was to evaluate the preventive effects of rosemary leaf extract that was standardised to 20 % carnosic acid (RE) on weight gain, glucose levels and lipid homeostasis in mice that had begun a high-fat diet (HFD) as juveniles. The animals were given a low-fat diet, a HFD or a HFD that was supplemented with 500 mg RE/kg body weight per d (mpk). Physiological and biochemical parameters were monitored for 16 weeks. Body and epididymal fat weight in animals on the HFD that was supplemented with RE increased 69 and 79 % less than those in the HFD group. Treatment with RE was associated with increased faecal fat excretion but not with decreased food intake. The extract also reduced fasting glycaemia and plasma cholesterol levels. In addition, we evaluated the inhibitory effects of RE in vitro on pancreatic lipase and PPAR-γ agonist activity; the in vitro findings correlated with our observations in the animal experiments. Thus, the present results suggest that RE that is rich in carnosic acid can be used as a preventive treatment against metabolic disorders, which merits further examination at physiological doses in randomised controlled trials.

Short-chain fructo-oligosaccharides (FOS) are known to have beneficial effects on health. However, the effects of FOS on insulin resistance have not been fully clarified. We observed the effects of FOS feeding on insulin sensitivity and adipocytokine release from abdominal adipocytes in weaning rats. Male Sprague–Dawley rats, 3 weeks old, were divided into three groups and fed a sucrose-based American Institute of Nutrition (AIN)-93 growth diet (control), the control diet containing 5 % FOS for 5 weeks (FOS-5wk) or the control diet for 2 weeks followed by the 5 % FOS diet for 3 weeks (FOS-3wk). Tail blood was collected after fasting for 9 h on day 33 of feeding, and glucose and insulin levels were measured. On the last day, rats were anaesthetised and killed after the collection of aortic blood. Small- and large-intestinal mesenteric fat tissues were immediately excised, and the release of adiponectin, leptin and TNF-α was evaluated from the subsequently isolated adipocytes. The weight of the large-intestinal mesenteric fat, fasting blood insulin level and homeostatic model assessment for insulin resistance decreased in a time-dependent manner, and were much lower in the FOS-5wk group than in the control group. These values were correlated with aortic blood leptin levels. The secretion rate of leptin from the isolated mesenteric adipocytes in the small intestine, but not in the large intestine, was lower in the FOS-fed groups than in the control group. In conclusion, FOS feeding improved insulin sensitivity accompanied by the reduction in large-intestinal fat mass and leptin secretion from the mesenteric adipocytes of the small intestine.

Intra-uterine growth retardation has been linked to the development of type 2 diabetes in later life. Mitochondrial changes have been suggested as a link between fetal malnutrition and adult insulin resistance. Taurine has been implicated in this process. We investigated whether protein malnutrition in early life alters mitochondria of the pancreatic islets in adulthood, and whether taurine supplementation restores these changes. Male offspring of rats fed a control diet, a low-protein diet or a low-protein diet supplemented with taurine during pregnancy and lactation were weaned onto the control diet. In each group, at 20 weeks of age, intravenous glucose tolerance tests, euglycaemic–hyperinsulinaemic clamp studies, morphometric analysis of the pancreatic islets and ultra-structural analysis of the mitochondria of the β-cells were performed. The expressions of cytochrome c oxidase (COX) I and mitochondrial respiratory chain complex II were also measured. Fetal protein-malnourished rats showed decreased pancreatic islet mass and reduced insulin-secretory responses to a glucose load. These rats also showed reduced mitochondrial DNA-encoded COX I gene expression in the islets. Electron microscopic examination showed abnormal mitochondrial shapes in the β-cells of fetal protein-malnourished rats. Taurine supplementation to the low-protein diet restored all these changes. Our findings indicate that a maternal protein-restriction diet causes long-lasting mitochondrial changes that may contribute to the development of type 2 diabetes later in life. The lack of taurine may be a key causative factor for these dysfunctional mitochondrial changes.

Periparturient relaxation of immunity (PPRI) to parasites in mammals results in higher worm burden and worm egg excretion and may have a nutritional basis. Nippostrongylus brasiliensis re-infected lactating rats fed low-crude protein (CP) diets show an augmented degree of PPRI compared with their high CP-fed counterparts. However, such effects of CP scarcity have been confounded by metabolisable energy (ME) scarcity due to increased intake of the high-CP foods. Here, we independently assessed the effects of dietary CP and ME scarcity on the degree of PPRI. Second, parity rats were infected with N. brasiliensis larvae before mating. Upon parturition, dams were allocated to one of six feeding treatments (1–6), consisting of two levels of dietary ME supply, each with three levels of CP supply. On day 2 of lactation, dams were either re-infected with 1600 N. brasiliensis larvae or sham-infected with PBS, while litter size was standardised at ten pups. Dams and litters were weighed daily until either day 8 or 11 of lactation, when worm burdens were assessed as a proxy for PPRI. Increased CP and ME supply independently improved lactational performance. While ME supply did not affect parasitism, increasing CP supply reduced worm burden and the percentage of female worms in the small intestine; the latter was especially pronounced at the lower level of ME supply. The present results support the view that PPRI to parasites may be sensitive to CP scarcity, but not to moderate ME scarcity.

Bifidobacterium spp. typical of the human intestinal microbiota are believed to influence the balance of immune responses in the intestinal mucosa. The aim of the present study was to investigate the effect of different bifidobacterial species and their mixtures in in vitro experiments with peripheral blood mononuclear cells (PBMC) and Caco-2 cells. Bifidobacterium adolescentis, B. angulatum, B. breve, B. catenulatum, B. infantis, B. longum and two combinations of these bifidobacteria simulating the species composition found in faecal samples from breast-fed (BF) and formula-fed (FF) infants were used. The levels of several cytokines were measured by direct stimulation of PBMC and by stimulation of a Caco-2/PBMC co-culture with bifidobacteria. B. catenulatum and B. breve were the strongest enhancers of interferon-γ (IFN-γ) production by direct stimulation of PBMC. B. longum was the highest inducer of IL-10 and the lowest TNF-α stimulus. In the Caco-2/PBMC system, B. breve was the highest inducer of IL-8 production by Caco-2 cells, significantly different from B. infantis, B. adolescentis and the FF mixture (P < 0·05). IFN-γ produced by PBMC stimulated with the BF mixture (containing 22 % B. breve, compared with 7 % in the FF mixture) was significantly higher compared with B. adolescentis, B. infantis and B. longum. B. adolescentis also inhibited IFN-γ production compared with the FF mixture and B. longum. The proportion of different Bifidobacterium strains seems to be an important determinant of the cytokine balance in the simulated intestinal environment studied. B. breve and the combination of the Bifidobacterium species typically found in the microbiota of BF infants have shown the most significant effects.

A 6-month experiment with nine dietary treatments was conducted to determine amounts of S plus Mo supplements required to maintain normal hepatic concentrations of Cu and Mo and to prevent chronic Cu toxicity in lambs fed palm kernel cake (PKC) diets. All diets consisted of PKC supplemented with minerals and vitamins, and with appropriate amounts per kg DM of S (level 0 or level 1 = 1 g) as sodium sulfate and/or Mo (level 0; level 1 = 4 mg; level 2 = 8 mg; level 3 = 16 mg; level 4 = 32 mg) as ammonium molybdate to form treatments S0Mo1, S0Mo2, S0Mo3, S0Mo4, S1Mo0, S1Mo1, S1Mo2, S1Mo3 and S1Mo4.There was no effect (P>0·05) of dietary treatments on the growth performance of the lambs. The dietary supplement of 1 g S plus 8 mg Mo per kg dietary DM (treatment S1Mo2) prevented accumulation of Cu in the liver without elevation of the concentration of Mo (P>0·05). The treatments S0Mo1, S0Mo2 and S0Mo3 increased (P < 0·05) hepatic Cu concentrations from 376 μg/g DM to between 1090 and 1294 μg/g DM. Also, the treatments S1Mo3 and S1Mo4 resulted in higher (P < 0·05) hepatic Mo concentrations compared with the treatment S1Mo0. It was concluded that the dietary supplement of 1 g S plus 8 mg Mo/kg PKC DM added to the PKC used is sufficient to maintain normal hepatic concentrations of Cu and Mo and to prevent chronic Cu toxicity in sheep fed diets containing any amount of PKC.

Salmon provides long-chain (LC) n-3 PUFA and Se, which are well recognised for their health benefits. The n-3 and Se status of the New Zealand population is marginal. The objective of the present study was to compare the effects of consuming salmon v. supplementation with salmon oil on LC n-3 and Se status. Healthy volunteers (n 44) were randomly assigned to one of four groups consuming 2 × 120 g servings of salmon/week or 2, 4 or 6 salmon oil capsules/d for 8 weeks. Linear regression analysis predictive models were fitted to the capsule data to predict changes in erythrocyte LC n-3 levels with intakes of LC n-3 from capsules in amounts equivalent to that consumed from salmon. Changes in Se status (plasma Se and whole-blood glutathione peroxidase) were compared between the groups consuming salmon and capsules (three groups combined). Salmon, 2, 4 and 6 capsules provided 0·82, 0·24, 0·47 and 0·69 g/d of LC n-3 fatty acids. Salmon provided 7 μg/d and capsules < 0·02 μg/d of Se. The predictive model (r2 0·31, P = 0·001) showed that increases in erythrocyte LC n-3 levels were similar when intakes of 0·82 g/d LC n-3 from salmon or capsules (1·92 (95 % CI 1·35, 2·49) v. 2·32 (95 % 1·76, 2·88) %) were consumed. Plasma Se increased significantly more with salmon than with capsules (12·2 (95 % CI 6·18, 18·12) v. 1·57 (95 % CI − 2·32, 5·45) μg/l, P = 0·01). LC n-3 status was similarly improved with consumption of salmon and capsules, while consuming salmon had the added benefit of increasing Se status. This is of particular relevance to the New Zealand population that has marginal LC n-3 and Se status.

Nutrition support is an important part of care management in critically ill patients, not only to prevent and treat malnutrition but also it has a significant impact on recovery from illness and overall outcome. There is little information available about present nutritional support practice for patients in intensive care units (ICU) in the UK. This survey was designed to evaluate the present nutrition support practice in ICU and high dependency units (HDU) in England. Data were gathered by a 72 h phone survey from 245 ICU and HDU in 196 hospitals in England. A questionnaire was completed over the telephone, including general information, nutrition support and teams involved in the nutrition management in the ICU. Of 1286 total patients in the ICU, 703 (54·6 %) were receiving nasogastric feeding, two (1·5 %) were receiving feeding via a percutaneous endoscopically placed gastrostomy tube and two (1·5 %) were receiving nasojejunal feeding. One hundred and forty-seven (11·4 %) patients were on parenteral feeding during the study period. A nutrition support team was not available in 158 (83·1 %) ICU and there was no dietitian or specialist nutrition nurse to cover ICU in nine (4·7 %) hospitals. In conclusion, the present survey reported an increased trend in usage of enteral feeding in ICU in England, and a reduction in the use of parenteral nutrition compared with previous surveys. However, we are still far from integrating nutrition into care management in the ICU.

Intake of trans-fatty acids (TFA), especially industrially produced TFA (I-TFA), has been associated with the risk of CHD through influence on serum lipid levels. Other causal pathways remain less investigated. In the present cross-sectional study of middle-aged men representing a broad range of BMI, the association between intake of TFA, I-TFA and ruminant TFA (R-TFA) and obesity-associated risk markers of CHD was assessed. The study comprised 393 Danish men (median age 49 years) with a median BMI of 28·4 kg/m2. Intake of TFA was estimated based on 7 d dietary records, whereas outcomes of interest (waist circumference, sagittal abdominal diameter, percentage of truncal fat, C-reactive protein, IL-6, blood lipids, blood pressure, HbA1c and insulin sensitivity index) were obtained through clinical examination. The associations were assessed by linear regression analysis. The median intake of total TFA among the 393 men was 1·3 g/d, covering a daily I-TFA intake of 0·4 g (10–90th percentile 0·0–1·0) and R-TFA intake of 0·9 g (10–90th percentile 0·4–1·8). Intake of these amounts of TFA showed no significant associations with abdominal fatness, inflammatory markers, blood lipids, blood pressure and insulin homeostasis. Among middle-aged men with a generally low intake of TFA, neither I-TFA nor R-TFA was significantly related to obesity-associated risk markers of CHD. The decreased average intake of I-TFA in Denmark since 1995 is suggested to effectively prevent occurrence of the adverse metabolic changes and health consequences, which have formerly been observed in relation to, especially, I-TFA intake.

Studies that investigated complex actual eating behaviours of the general population and their relation to cardiometabolic risk markers are sparse. We aimed to identify dietary patterns within a nationally representative sample of 4025 German adults by factor analysis based on validated dietary history interviews. Furthermore, we evaluated associations of the derived dietary patterns with abnormalities clustered within the metabolic syndrome and related metabolic markers by logistic regression models and ANCOVA. A high adherence to the ‘processed foods’ pattern reflected a high intake of refined grains, processed meat, red meat, high-sugar beverages, eggs, potatoes, beer, sweets and cakes, snacks and butter, whereas a high adherence to the ‘health-conscious’ pattern represented a high intake of vegetables, vegetable oils, legumes, fruits, fish and whole grains. For subjects in the highest compared with those in the lowest quintile of the processed foods pattern, the occurrence of abdominal obesity was 88 (95 % CI 31, 169) % higher, hypertension was 34 (95 % CI − 4, 86) % higher, hypertriacylglycerolaemia was 59 (95 % CI 11, 128 ) % higher and the metabolic syndrome was 64 (95 % CI 10, 143) % higher when adjusted for age, sex, energy intake, socio-economic status, sport activity and smoking. Furthermore, subjects in the highest quintile had statistically significantly higher uric acid concentrations and lower folate concentrations (P for trend < 0·05). In contrast, subjects in the highest quintile of the health-conscious pattern had a 30 (95 % CI 10, 46) % lower occurrence of hypertension, higher folate concentrations and lower homocysteine and fibrinogen concentrations (P for trend < 0·05). These data strengthen the findings from non-representative studies and emphasise the importance of healthy overall food patterns for preventing metabolic disturbances.

Although there are indications for modulatory effects of PUFA on associations between SNP and obesity risk, scientific evidence in human subjects is still scarce. The present analyses investigated interaction effects between SNP in candidate genes for obesity and PUFA in erythrocyte membranes on obesity risk. Within the second Bavarian Food Consumption Survey (cross-sectional, population-based), 568 adults provided blood samples. Fatty acid composition of erythrocyte membranes was analysed by means of GC. Genotyping was performed for twenty-one genes, including cytokines, adipokines, neurotransmitters and transcription factors. In addition, plasma IL-6 concentrations were analysed. For the statistical analysis, a logistic regression model assuming additive genetic effects was chosen. About 20 % of the study participants were classified as obese (BMI ≥ 30 kg/m2). Several significant gene–PUFA interactions were found, indicating regulatory effects of PUFA by gene variants of IL-2, IL-6, IL-18, TNF receptor family member 1B and 21, leptin receptor and adiponectin on obesity risk. After stratification by genotype, the strongest effects were found for rs2069779 (IL-2) and all tested PUFA as well as for rs1800795 (IL-6) and linoleic or arachidonic acid. The obesity risk of minor allele carriers significantly decreased with increasing fatty acid content. The genetic PUFA–IL-6 interaction was also reflected in plasma IL-6 concentrations. If replicated in a prospective study with sufficient statistical power, the results would indicate a beneficial effect of high PUFA supply for a substantial proportion of the population with respect to obesity risk.

There are no published data regarding the overall dietary glycaemic index (GI) and glycaemic load (GL) of Australian children and adolescents. We therefore aim to describe the dietary GI and GL of participants of the 2007 Australian National Children's Nutrition and Physical Activity Survey (2007ANCNPAS), and to identify the main foods contributing to their GL. Children, aged 2–16 years, who provided two 24 h recalls in the 2007ANCNPAS were included. A final dataset of 4184 participants was analysed. GI of each food item was assigned using a previously published method. GL was calculated, and food groups contributing to the GL were described by age group and sex. The weighted mean dietary GI and GL of the participants were 54 (sd 5) and 136 (sd 44), respectively. Among the nutrients examined, Ca had the highest inverse relationship with GI (P < 0·001), while percentage energy from starch was most positively associated with GI. The association between fibre density and GI was modest, and percentage energy from sugar had an inverse relationship with GI. Daily dietary GL contributed by energy-dense and/or nutrient-poor (EDNP) items in subjects aged 14–16 years was more than doubled that of subjects aged 2–3 years. To conclude, Australian children and adolescents were having a high-GI dietary pattern characterised by high-starchy food intake and low Ca intake. A significant proportion of their dietary GL was from EDNP foods. Efforts to reduce dietary GI and GL in children and adolescents should focus on energy-dense starchy foods.

Oestrogen replacement in ovariectomised (OVX) rats has been reported to attenuate food intake, especially during the light phase. To gain better insight into the central mechanism of oestrogen-induced reduction of food intake, we examined the effect of chronic oestrogen replacement in OVX rats on c-Fos expression in the suprachiasmatic nucleus (SCN) and on food intake during the light and dark phases. Eight-week-old female rats were ovariectomised and implanted with either an oestradiol (E2) or a vehicle pellet (Veh) subcutaneously. The animals were housed in an environment with a 12 h light–12 h dark cycle with the lights on at 07.00 hours. The amount of spontaneous food intake relative to each animal's body weight was significantly less for the E2 group than for the Veh group during the light phase, but there were no differences shown between these groups during the dark phase. There were no differences shown in the number of c-Fos-immunoreactive cells in the SCN in the E2 group compared with the Veh group during the early dark phase (22.00 hours; Zeitgeber time 15.00 (ZT15)), but the number was significantly higher than in the Veh group during the early light phase (10.00 hours; ZT3). This finding suggests that chronic oestrogen replacement chronically enhances SCN activity, specifically during the light phase. The oestrogen-induced enhancement of SCN activity during the light phase is possibly involved in the light phase-specific attenuation of food intake by oestrogen replacement in OVX rats.