Pomegranate is a rich source of polyphenols, which are believed to be responsible for the oestrogenic activities of extracts of this fruit in mice. One of these potential activities is the prevention of bone loss. The objectives of the present study were to determine the effects of pomegranate extract on chondrogenesis and osteogenesis in mouse embryos in vivo and limb bud cultures in vitro. A total of fifty pregnant Balb/c mice were given vehicle, pomegranate juice extract (PJE), pomegranate husk extract (PHE) or a mixture of husk and juice extract (PME). Their embryos were stained with alizarin red S and alcian blue, and the length of the femur, tibia and their ossification zones were measured on day 19 of gestation. Bone Ca content in pregnant mice was also measured. Mice treated with PJE showed an increase in bone Ca content. Dietary supplementation with all extracts significantly increased embryo femur length and osteogenesis index. Mesenchymal cells from fetal limb buds were cultured and exposed to 10, 100, 1000 and 10 000 μg/ml of PJE, PHE or PME. The number of viable cells was greater in cultures exposed to the extracts than in control cultures. The number of cartilage nodules and their diameters were greater in extract-treated cell cultures, a finding which reflected increased cell proliferation and differentiation rates. In conclusion, the findings of the present study suggest that pomegranate is able to enhance bone formation.

1. Diets of fresh kale (Brassica oleracea) and ryegrass (Loliumperenne) – elover (Trifolium repens) herbage were fed to growing sheep in three experiments. In Expts 1 and 3 the sheep were confined indoors and fed at hourly intervals, and all were given supplementary iodine to counteract kale goitrogens. Lambs grazed the two forages for 24 weeks in Expt 2, with and without intramuscular injections of iodized oil. The kale and herbage contained respectively 11 and <0.1 g S-methyl-L-cysteine sulphoxide (SMCO)/kg dry matter (DM) and values for readily fermentable: structural carbohydrate (CHO) were 3.1 and 0.8, respectively.

2. Blood samples were withdrawn from indwelling catheters (Expts 1 and 3) or venipuncture (Expt 2) and the plasma analysed for a range of hormones using radioimmunoassay procedures. Glucose irreversible loss (GIL) was measured in Expt 1 using primed continuous infusions of D-[U-14C]glucose. Samples of adipose tissue were removed from the shoulder area in Expt 3, and rates of D-[U-14C]glucose and [U-14C]acetate incoporation and oxidation were measured in vitro, together with the rate of glycerol release.

3. In the presence of supplementary I2, kale feeding was associated with an elevation in plasma concentration of free thyroxine (T4) Regardless of I2 supplementation, sheep fed on kale had much higher plasma growth hormone concentrations than sheep fed on ryegrass-clover herbage, and this was accompanied by reduced plasma somatostatin concentrations.

4. Plasma insulin and glucagon concentrations were similar for sheep fed on the two diets; GIL tended to be slightly but not significantly greater (9.4%) for sheep fed on kale than for those fed on ryegrass-clover herbage.

5. Kale feeding was associated with increased uptakes of acetate and glucose into adipose tissue, reduced rates of oxidation of both substrates and no difference in rate of glycerol release. Each 1 nmol increase in glucose uptake was associated with 8.7 nmol acetate uptake ( P < 0.001).

6. It is proposed that ruminants counteract protein inactivation, caused by production of dimethyl disulphide from SMCO in the rumen, through increasing circulating concentrations of growth hormone and T4, which then stimulate synthesis of replacement body proteins.

A study was carried out to examine whether the responsiveness of small intestinal epithelial cells to dietary carbohydrate varied during the daily 24 h cycle. The effect of sucrose on disaccharidase activities was compared during a period of decreasing disaccharidase activities, i.e. between 22.00 and 10.00 hours, and increasing disaccharidase activities, i.e. between 10.00 and 22.00 hours, in the jejunum of 7-week-old-rats. Rats were fed on a low-starch, high-fat diet (Lst; starch 5 and fat 73% of gross energy), or a high-starch, low-fat diet (Hst; starch 70 and fat 7% of gross energy). Both dietary groups exhibited typical diurnal variations in jejunal sucrase (EC 3.2.1.48), maltase (EC 3.2.1.20) and lactase (EC 3.2.1.23) activities, exhibiting a peak around 22.00 hours and a trough at approximately 10.00 hours. When rats were fed on diet Lst for 7 d and then force-fed on an isoenergetic sucrose diet (S; sucrose 40 and fat 37% of gross energy) for 6 or 12 h they exhibited increased sucrase, maltase and lactase activities compared with rats fed on diet Lst. The absolute increase in disaccharidase activities was similar regardless of the time diet S was given or whether rats were killed at 10.00 hours or at 22.00 hours. Analyses of sucrase and lactase activities along the villus–crypt columns showed that the distribution of cell cohorts that responded to diet S was not influenced by the time of introduction of diet S. These findings suggest that small intestinal epithelial cells possess the ability to respond to dietary carbohydrate throughout the daily 24 h cycle.

The precision with which glycaemic responses, expressed as incremental area under the curve (AUC), can be measured may be improved by using the average of several measures of fasting blood glucose (FBG). To see if taking two fasting blood samples would increase the precision of AUC, the glycaemic responses elicited by four test meals (50 g glucose; 50 g glucose plus 10 g fat and 10 g protein; 100 g white bread; 100 g white bread plus 10 g fat and 10 g protein) were determined in thirteen overnight-fasted healthy subjects. Two fasting blood samples were taken 5 min apart (−5 min and 0 min before starting to eat) with glucose measured three times in each sample. AUC was calculated using different estimates of FBG derived from the three measures of glucose in the two fasting blood samples and each set of AUC values subjected to ANOVA. Unexpectedly, the results were more precise when AUC was calculated from mean glucose in the 0 min blood sample (FBG0) than from mean glucose in the two different fasting blood samples. The 95 % CI of the AUC calculated using FBG0 in thirteen subjects was ±29·8; to obtain the same CI using the mean of the two fasting blood samples would require fourteen subjects. These results suggest that taking two fasting blood samples does not necessarily improve, and may even reduce, the precision of AUC as a measure of glycaemic response. Further studies are needed before requiring that two fasting blood samples be taken for determining glycaemic index.

Non-alcoholic fatty liver disease (NAFLD) is closely associated with insulin resistance and obesity. Hence, carbohydrate quality could be of relevance to the risk of NAFLD, but prospective data are lacking. The aim of the present study was to investigate longitudinal associations between carbohydrate quality (including dietary glycaemic index (GI) and intakes of sugar, starch and fibre) and markers of liver function in an older Australian population. The analysis was based on 866 participants ( ≥ 49 years) of the Blue Mountains Eye Study with fasting blood specimens and dietary intake data at baseline and 5-year follow-up. Multi-level mixed regression analysis was used to relate dietary GI and sugar, starch and fibre intake to the liver enzymes alanine aminotransferase (ALT) and γ-glutamyltransferase (GGT), as well as fasting TAG and HDL-cholesterol (HDL-C). After adjustment for potential confounding factors, a lower fibre intake was cross-sectionally related to higher GGT (P= 0·02) and fasting TAG (P= 0·002) levels, with fruit fibre being the most relevant fibre source (P= 0·095 for GGT; P= 0·003 for TAG). A higher dietary GI was associated with lower HDL-C (P= 0·046). Changes in carbohydrate quality during 5 years were not related to changes in ALT, GGT, TAG or HDL-C (P≥ 0·08). In conclusion, the absence of longitudinal associations between carbohydrate quality and liver enzymes and serum lipids in this older population does not support a major role of carbohydrate nutrition in liver function among the elderly.

Earlier studies in animals have suggested an essential role for Si in connective tissues, but such works have not been replicated per se. Nonetheless, a study conducted in 2000 has reported that Si may be essential during pregnancy for the growing fetus, since serum Si concentrations in infants were approximately 300 % higher than those in older children and adults and serum Si concentrations in pregnant women were approximately 300 % lower than those in age-matched non-pregnant controls. To reproduce these potentially important findings, in the present study, serum Si concentrations were measured in fourteen pregnant women (15–24 weeks of gestation) and compared with those of seventeen non-pregnant, non-lactating female controls. Serum Si concentrations were also measured in fourteen full-term mothers at the time of delivery and in the umbilical cord (UC) vein and artery where possible. Fasting serum Si concentrations in pregnant women were not significantly different from those of the female controls and showed little change with advancing gestation (r 0·2). Mean serum Si concentrations in the UC vein samples were 52 % higher, while those in the UC artery samples were 235 % higher than those in the maternal forearm vein samples, although data were widely spread and differences were not significant. Mean maternal forearm vein Si concentrations at delivery were 50 % lower than those of pregnant women and female controls, but, again, these were not significant. Overall, we note that there are significant analytical challenges in comparing baseline Si levels between different groups; notwithstanding, our findings cannot confirm a reduction in fasting serum Si levels during pregnancy, but, equally, we cannot rule out higher serum Si levels in newborns than in their mothers, and further work is required.

There is a dearth of data on the iodine balance studies of Chinese population. In the present study, we aimed to explore the appropriate recommended nutrient intake (RNI) of iodine based on healthy Chinese women. A 4-week study was conducted in twenty-five Chinese euthyroid women. Uniform diets with different iodine contents were provided in two different periods, in which non-iodised salt was given in the first 3 weeks, followed by 1 week of iodised salt administration. The total iodine intake from diet, water and air as well as the total iodine excretion through urine, faeces and respiration were monitored and determined. The sweat iodine loss was also considered. Moreover, the regression curve model was established between the 24 h iodine intake and 24 h iodine excretion. The 24 h iodine intake in the two periods was 194·8 (sd 62·9) and 487·1 (sd 177·3) μg/d, respectively. The 24 h iodine excretion was 130·9 (sd 39·5) and 265·4 (sd 71·8) μg/d, respectively. Both 24 h iodine intake and 24 h iodine excretion of the two periods were significantly different (all P<0·05). The iodised salt contributed approximately 62·7 % of the total daily iodine intake. Moreover, 92·3 % (277/300) of samples were in positive balance, while twenty-three cases were in negative balance. Our data show that the estimated average requirement for iodine was 110·5 μg/d. Therefore, the RNI for iodine to non-pregnant, non-lactating Chinese women was 154·7 μg/d.

Long-Evans Cinnamon (LEC) rats spontaneously develop fulminant hepatitis, associated with excess Cu accumulation in the liver: thus, they are considered an animal model of Wilson's disease. In the present study, we investigated the ability of excess dietary histidine to reduce the excess accumulation of liver Cu in LEC rats by comparing them with Fischer rats. The results clearly showed that the excess-histidine diet markedly stimulated the Cu excretion in urine, and significantly decreased the liver Cu content in LEC rats by 47·5%. The serum Cu content in LEC rats was not influenced by excess dietary histidine. We also compared the effects of excess dietary histidine on some liver antioxidant enzyme activities, liver and serum lipid levels and serum alanine aminotransferase activity of LEC and Fischer rats. Dietary histidine decreased the activities of total and Cu, Zn-superoxide dismutase in the liver of both strains. In LEC rats, the liver cholesterol content decreased, and serum cholesterol and phospholipids levels increased on feeding the excess-histidine diet. When fed on the basal diet, the serum alanine aminotransferase activity was higher in LEC rats than in Fischer rats, but a significant decrease in serum alanine aminotransferase activity of LEC rats was observed on feeding the excess-histidine diet. These results suggest that excess dietary histidine is effective in removing Cu ions from the liver of LEC rats. Thus, it may be of benefit in the prevention or treatment of liver injury in LEC rats and in patients with Wilson's disease.

The potential to reduce cardiovascular morbidity through dietary modification remains an area of intense clinical and scientific interest. Any putatively beneficial intervention should be tested within a randomised controlled trial which records appropriate endpoints, ideally incident CVD and death. However, the large sample sizes required for these endpoints and associated high costs mean that the majority of dietary intervention research is conducted over short periods among either healthy volunteers or those at only slightly increased risk, with investigators using a diverse range of surrogate measures to estimate arterial health in these studies. The present review identifies commonly employed techniques, discusses the relative merits of each and highlights emerging approaches.

Epigenetic studies suggest that diseases that develop in adulthood are related to certain conditions to which the individual is exposed during the initial stages of life. Experimental evidence has demonstrated that offspring born to mothers maintained on high-Na diets during pregnancy have higher mean arterial pressure (MAP) in adulthood. Although these studies have demonstrated the importance of prenatal phases to hypertension development, no evidence regarding the role of high Na intake during postnatal phases in the development of this pathology has been reported. Therefore, in the present study, the effects of Na overload during childhood on induced water and Na intakes and on cardiovascular parameters in adulthood were evaluated. Experiments were carried out in two groups of 21-d-old rats: experimental group, maintained on hypertonic saline (0·3 m-NaCl) solution and food for 60 d, and control group, maintained on tap water and food. Later, both groups were given water and food for 15 d (recovery period). After the recovery period, chronic cannulation of the right femoral artery was performed in unanaesthetised rats to record baseline MAP and heart rate (HR). The experimental group was found to have increased basal MAP (98·6 (sem 2·6) v. 118·3 (sem 2·7) mmHg, P< 0·05) and HR (365·4 (sem 12·2) v. 398·2 (sem 7·5) beats per min, P< 0·05). There was a decrease in the baroreflex index in the experimental group when compared with that in the control group. A water and Na intake test was performed using furosemide. Na depletion was found to induce an increase in Na intake in both the control and experimental groups (12·1 (sem 0·6) ml and 7·8 (sem 1·1), respectively, P< 0·05); however, this increase was of lower magnitude in the experimental group. These results demonstrate that postnatal Na overload alters behavioural and cardiovascular regulation in adulthood.

1. Eighteen males (31–62 years) who habitually consumed significant amounts of table sucrose (approximately 25% of total carbohydrate intake) were supplied with their usual intake of sucrose for consumption in conjunction with their normal diet for 1 year, and a record kept of the amount consumed. The sucrose was then replaced isoenergetically by dried glucose syrup (55 D. E.) which contained saccharin to equate the sweetness to that of sucrose.

2. Fasting blood samples were taken every 4 weeks during the 2 years, and the plasma analysed for glucose, cholesterol, triglycerides and phospholipid-P by automated colorimetric methods. Dietary questionnaries were issued every 3 months to confirm that subjects were not substantially altering their diets.

3. In subjects whose weight remained unchanged and in those who lost weight there was a significant fall in cholesterol (P < 0.025) and phospholipid (P < 0.025) in the glucose-syrup period compared with the sucrose period; triglycerides did not change. In subjects who gained weight there was a significant increase in triglycerides (P < 0.05), but no change in cholesterol; phospholipid-P fell significantly (P < 0.0005).

4. The dietary modification in this experiment was sufficiently long to ensure that subjects had adapted, and the results obtained show stable changes in blood lipids which may be attributed to the isoenergetic replacement of table sucrose by glucose syrup.

Mechanisms responsible for derangements in Fe homeostasis in chronic inflammatory conditions are not entirely clear. The aim of the present study was to test the hypothesis that inflammation affects the expression of Fe-related proteins in the duodenum and monocytes of patients with chronic inflammatory disorders, thus contributing to dysregulated Fe homeostasis. Duodenal mucosal samples and peripheral blood monocytes obtained from patients with chronic inflammatory disorders, namely ulcerative colitis (UC), Crohn's disease (CD) and rheumatoid arthritis, were used for gene and protein expression studies. Hb levels were significantly lower and serum C-reactive protein levels were significantly higher in patients in the disease groups. The gene expression of several Fe-related proteins in the duodenum was significantly up-regulated in patients with UC and CD. In patients with UC, the protein expression of divalent metal transporter 1 and ferroportin, which are involved in the absorption of dietary non-haem Fe, was also found to be significantly higher in the duodenal mucosa. The gene expression of the duodenal proteins of interest correlated positively with one another and negatively with Hb. In patients with UC, the gene expression of Fe-related proteins in monocytes was found to be unaffected. In a separate group of patients with UC, serum hepcidin levels were found to be significantly lower than those in the control group. In conclusion, the expression of Fe-related proteins was up-regulated in the duodenum of patients with chronic inflammatory conditions in the present study. The effects appeared to be secondary to anaemia and the consequent erythropoietic drive.

Homocysteine (Hcy) is recognised as a risk factor for IHD. Serum Hcy is negatively correlated with serum folate levels, the main sources of which are fruits, vegetables and legumes. The present case–control study was designed to examine the relationship between serum Hcy levels and IHD and to assess the role of dietary factors in the southern Mediterranean population of Crete, Greece. Serum Hcy, folate, vitamin B12, creatinine and glucose levels and a full lipid profile were measured in 152 patients with established IHD, median age 64 (range 33–77) years, and 152 healthy control subjects, age- and sex-matched. Dietary data were assessed using a 3 d food intake record. Compared with controls, patients with IHD had significantly higher daily intakes of vitamin B12 and MUFA and significantly lower intakes of carbohydrate, fibre, folate, cholesterol, n-3 fatty acids and total trans unsaturated fatty acids. Moreover, patients had significantly higher serum Hcy, vitamin B12 and creatinine levels, but significantly lower folate. Serum folate concentrations in both groups had a significant positive correlation with dietary fibre consumption and a significant inverse correlation with vitamin B12 intake. IHD patients should be encouraged to increase their daily dietary intake of fibre, folate and n-3 fatty acids, which are significant components of the traditional Cretan Mediterranean diet. Where dietary folate intake is inadequate, folate supplements are recommended to reduce elevated Hcy levels.

1. Changes in serum creatine kinase activity, creatinine excretion, mid-upper arm and thigh muscle circumference and the ‘X-ray muscle shadow weight’ of the upper arm and thigh muscles have been compared in children recovering from kwashiorkor.

2. Creatine kinase activity rose as the health of the child improved but was subject to large fluctuations mainly associated with the Occurrence of infections. For this reason it is unlikely to be of value in assessing nutritional status on the basis of changes in musculature.

3. Creatinine excretion also increased with treatment and was more consistent. This measurement should be of much greater value than serum creatine kinase.

4. The anthropometric measurements were relatively insensitive to quite large changes in limb musculature.

5. Assessment of musculature by soft-tissue X-rays of the limbs appeared to be a sensitive method of potential value.

1. A comparison was made of the composition of milk from front and rear teats in four sows. There were small and not significant differences in fat, protein and lactose contents, and in the fatty-acid composition of the milk fat with the exception of the 18:3 acid where the difference was also small but significant.

2. The effects of intravenous infusions of glucose and insulin in lactating sows on milk secretion and blood composition were investigated in two sows.

3. Intravenous infusion of glucose had no effect on blood plasma glucose concentration but increased the yields of lactose, protein and water.

4. Intravenous infusion of insulin depressed plasma glucose concentration and the yields of lactose and water. The yield of protein was unaffected.

5. It is concluded that differences between the non-ruminant (the sow) and the ruminant in the responses in milk secretion to glucose infusion may be related to differences in the sensitivity to insulin of mammary tissue.

Low-grade metabolic acidosis, consecutive to excessive catabolism of sulfur amino acids and a high dietary Na:K ratio, is a common feature of Western food habits. This metabolic alteration may exert various adverse physiological effects, especially on bone, muscle and kidneys. To assess the actual effects of various K salts, a model of the Westernised diet has been developed in rats: slight protein excess (20 % casein); cations provided as non-alkalinising salts; high Na:K ratio. This diet resulted in acidic urine (pH 5·5) together with a high rate of divalent cation excretion in urine, especially Mg. Compared with controls, K supplementation as KCl accentuated Ca excretion, whereas potassium bicarbonate or malate reduced Mg and Ca excretion and alkalinised urine pH (up to 8). In parallel, citraturia was strongly increased, together with 2-ketoglutarate excretion, by potassium bicarbonate or malate in the diet. Basal sulfate excretion, in the range of 1 mmol/d, was slightly enhanced in rats fed the potassium malate diet. The present model of low-grade metabolic acidosis indicates that potassium malate may be as effective as KHCO3 to counteract urine acidification, to limit divalent cation excretion and to ensure high citrate concentration in urine.