Current evidence strongly supports a role for the breast tumour suppressor genes, BRCA1 and BRCA2, in both normal development and carcinogenesis. In vitro observations reported that BRCA1 and BRCA2 are expressed in a cell cycle-dependent manner. Interestingly, differences in the actions of n-3 and n-6 polyunsaturated fatty acids have been observed: while the n-3 polyunsaturated fatty acids have been described to reduce pathological cell growth, the n-6 polyunsaturated fatty acids have been found to induce tumour proliferation. Here, we examined the expression of BRCA1 and BRCA2 in breast cell lines after treatment with polyunsaturated fatty acids. Real-time quantitative polymerase chain reaction determinations conclusively demonstrated increases in BRCA1 and BRCA2 mRNA expressions in MCF7 and MDA-MB 231 tumour cell lines after treatment with n-3 polyunsaturated fatty acids (eicosapentaenoic acid and docosahexaenoic acid), but no variation was noticed with the n-6 polyunsaturated fatty acid (arachidonic acid). On the other hand, no variation of the expression of BRCA1 and BRCA2 mRNA was detected in MCF10a normal breast cell line treated by polyunsaturated fatty acids. The level of BRCA1 and BRCA2 proteins quantified by affinity chromatography remained unchanged in tumour (MCF7, MDA-MB 231) and normal (MCF10a) breast cell lines. We suggest the presence of a possible transcriptional or post-transcriptional regulation of BRCA1 and BRCA2 after n-3 polyunsaturated fatty acid treatment in breast tumour cells.

Sugar-free confectionery products containing the low-energy, non-cariogenic sweetener isomalt are widely available in the market place and increasingly aimed at children. However, over-consumption of such products may lead to gastrointestinal symptoms and/or osmotic diarrhoea. Little is known about the gastrointestinal tolerance of children following consumption of isomalt. The aim of the present study was to assess gastrointestinal symptoms in children following consumption of sugar-free confectionery containing isomalt compared with sweets containing sucrose. In a double-blind, randomised, controlled, crossover study, sixty-seven children aged 6–9 years ingested 25 g hard-boiled sweets containing either sucrose or isomalt on two consecutive test days. Isomalt sweets were received as enthusiastically as sucrose sweets and, when given the choice, 97 % of children asked to be given the isomalt or the sucrose sweets on the second test day. Most children did not report multiple symptoms and few experienced symptoms on both days of isomalt consumption. However, significantly more children reported stomach-ache (P<0·01), abdominal rumbling (P<0·025) and passing watery faeces (P<0·001) following consumption of isomalt sweets compared with sucrose sweets. Consumption of 25 g isomalt-containing sweets by children is not associated with significant gastrointestinal effects graded as ‘considerably more than usual’ or multiple symptoms, but is associated with a laxative effect and increase in symptoms graded as ‘slightly more than usual’. For the majority of children in the present study, 25 g isomalt-containing sweets represents an acceptable level of consumption, although some children are sensitive to the effects of isomalt ingestion.

Flavanols are the most abundant flavonoids in the human diet where they exist as monomers, oligomers and polymers. In the present study, catechin, the procyanidin dimer B3 and a grapeseed extract containing catechin, epicatechin and a mixture of procyanidins were fed to rats in a single meal. After the meals, catechin and epicatechin were present in conjugated forms in both plasma and urine. In contrast, no procyanidins or conjugates were detected in the plasma or urine of any rats. Procyanidins were not cleaved into bioavailable monomers and had no significant effects on the plasma levels or urinary excretion of the monomers when supplied together in the grapeseed extract. We conclude that the nutritional effects of dietary procyanidins are unlikely to be due to procyanidins themselves or monomeric metabolites with the intact flavonoid-ring structure, as they do not exist at detectable concentrations in vivo. Future research should focus on other procyanidin metabolites such as phenolic acids and on the effects of the unabsorbed oligomers and polymers on the human gastrointestinal tract.

The difference between vitamin K metabolism in the liver and that in the bone of vitamin K-deficient rats was examined. After 17 d administration of vitamin K-deficient food, vitamin K in the liver was almost depleted, and prothrombin time (PT) was prolonged. Serum total osteocalcin level was slightly decreased by vitamin K deficiency, whereas serum undercarboxylated osteocalcin level did not change. The level of menaquinone (MK)-4 as well as that of phylloquinone was decreased, but approximately 40 % of the initial level still existed in the femur after the 17 d period. A single-dose administration of vitamin K (250 nmol/kg body weight) markedly increased vitamin K level in the liver but not in the femur. These results suggest that the turnover of vitamin K in the bone is slower than that in the liver, and bone metabolism may be little affected by the short period of intake of vitamin K-deficient food. However, intake of a larger amount of vitamin K is required for its accumulation in the bone than in the liver. Furthermore, the counteracting effect of MK-7 on prolonged PT in vitamin K-deficient rats was found to be higher than phylloquinone or MK-4.

Sweet lupin (Lupinus albus), a protein-rich legume devoid of anti-nutritional factors, is considered to have a high potential for protein nutrition in man. Results concerning the nutritional value of lupin protein are, however, conflicting in animals and very scarce in human subjects. Furthermore, where fibre-rich protein sources are concerned, the long-term nutritional results are often obscured, particularly since fibre-promoted colonic fermentation may bias the energy supply and redistribute N flux. We therefore studied, during the postprandial phase, the bioavailability and utilisation of lupin-flour protein in nine healthy men who had ingested a mixed meal containing intrinsically 15N-labelled lupin flour as the protein source (Expt 1). The real ileal digestibility (RID) and ileal endogenous N losses (IENL) were assessed using a perfusion technique at the terminal ileum, and the N content and 15N enrichment of ileal samples. Lupin flour exhibited a high RID of 91 (SD 3) % AND LOW IENL (5·4 (sd 1·3) mmol N/h). Postprandial dietary deamination was also assessed from body dietary urea and urinary dietary N excretion, and compared with results in nine healthy men following an isoenergetic meal containing a 15N-soyabean-protein isolate with a similar RID, as a control (Expt 2). Postprandial dietary deamination was similar after lupin and soyabean meals (17 (sd 2) and 18 (sd 4) % ingested N respectively). We therefore conclude that lupin protein is highly bioavailable, even if included in fibre-rich flour, and that it can be used with the same efficiency as soyabean protein to achieve postprandial protein gain in healthy human subjects.

According to a 4×4 Latin square design, four adult ovariectomised sows fed at a similar energy level (516 kJ ME/kg body weight (BW)0·75 per d) received one of four diets successively: a control low-dietary-fibre (DF) diet (diet C, 100 g total DF/kg DM) and three fibre-rich diets (200 g total DF/kg DM) that corresponded to a combination of diet C and maize bran (diet MB), wheat bran (diet WB), or sugar-beet pulp (diet SBP). Sows were adapted to the diet for 12 d before an 8 d measurement period. Digestibility of energy and nutrients in the diets, and total heat production (HP) and its components (fasting HP, activity HP and thermic effect of feeding (TEF), were measured. The TEF was partitioned between a short-term component (TEFst) and a long-term component (TEFlt). Total tract digestibility of nutrients and energy was greater for diet C; among the three other diets, the digestibility coefficients were higher for diet SBP than for diets MB and WB. Energy losses from CH4 were linearly related to the digestible total DF intake (+1·4 kJ/g). Fasting HP at zero activity averaged 260 kJ/kg BW0·75 per d. Activity HP represented 20 % total HP, or 83 kJ/kg BW0·75 per d on average. Total TEF and TEFlt were higher (P<0·05) for diet WB than for the other diets. However, total HP (406 kJ/kg BW0·75 per d) was not significantly affected by diet characteristics. Our results suggest that metabolic utilisation of dietary energy is little affected by the addition and origin of DF, at least under the conditions of the present study.

The underlying mechanisms for the detrimental consequences of a high-fructose diet in animal models are not clear. However, the possibility exists that fructose feeding facilitates oxidative damage. Thus, the aim of the present study was to assess, in weaning rats, the effect of a high-sucrose diet v. starch diet for 2 weeks on oxidative stress variables. Plasma lipid levels were measured and lipid peroxidation was evaluated by urine and plasma thiobarbituric acid-reactive substances (TBARS). The susceptibilities of several tissues to peroxidation were determined in tissue homogenates after in vitro lipid peroxidation. Antioxidant defence variables were evaluated by measuring plasma and heart vitamin E levels, and heart superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities. Higher plasma triacylglycerol (P<0·01) and TBARS (P<0·01) levels were found in rats fed the sucrose diet as compared with the starch-fed group, whereas plasma α-tocopherol levels were significantly decreased in the sucrose-fed group compared with the starch-fed group (P<0·01). Higher urine TBARS (P<0·01) were found in the sucrose-fed group compared with the starch-fed group, suggesting increased production of these substances from lipid peroxidation in vivo. Higher susceptibility to peroxidation in heart, thymus and pancreas was also found in the sucrose-fed group v. the starch-fed group. No statistical differences were observed for liver TBARS level between the two groups. Heart SOD activity was significantly decreased (P<0·001) in the sucrose-fed group compared with the starch-fed group, whereas heart vitamin E level and GPX activity were not different between the groups. However, the in vitro generation of superoxide radical in heart homogenate, measured by electron spin resonance detection and spin trapping, was not increased in the sucrose-fed group compared with starch-fed rats. Altogether, the results indicate that a short-term consumption of a high-sucrose diet negatively affects the balance of free radical production and antioxidant defence in rats, leading to increased lipid susceptibility to peroxidation.

Epidemiological studies suggest that foods rich in flavonoids might reduce the risk of cardiovascular disease and cancer. The objective of the present study was to investigate the effect of green tea extract (GTE) used as a food antioxidant on markers of oxidative status after dietary depletion of flavonoids and catechins. The study was designed as a 2×3 weeks blinded human cross-over intervention study (eight smokers, eight non-smokers) with GTE corresponding to a daily intake of 18·6 mg catechins/d. The GTE was incorporated into meat patties and consumed with a strictly controlled diet otherwise low in flavonoids. GTE intervention increased plasma antioxidant capacity from 1·35 to 1·56 (P<0·02) in postprandially collected plasma, most prominently in smokers. The intervention did not significantly affect markers in fasting blood samples, including plasma or haemoglobin protein oxidation, plasma oxidation lagtime, or activities of the erythrocyte superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase. Neither were fasting plasma triacylglycerol, cholesterol, α-tocopherol, retinol, β-carotene, or ascorbic acid affected by intervention. Urinary 8-oxo-deoxyguanosine excretion was also unaffected. Catechins from the extract were excreted into urine with a half-life of less than 2 h in accordance with the short-term effects on plasma antioxidant capacity. Since no long-term effects of GTE were observed, the study essentially served as a fruit and vegetables depletion study. The overall effect of the 10-week period without dietary fruits and vegetables was a decrease in oxidative damage to DNA, blood proteins, and plasma lipids, concomitantly with marked changes in antioxidative defence.

This study investigates the effects of increased NH3 or amino acid supply on glutamine utilisation and production by the splanchnic tissues of fed sheep. Six sheep, prepared with vascular catheters in the aorta, mesenteric, portal and hepatic veins, were fed grass pellets to 1·1×energy maintenance requirements. Each treatment involved a 4 d abomasal infusion, of either ammonium bicarbonate (AMM; 23·4 μmol/kg0·75 per min), water (CONT), or a mixture of amino acids that excluded glutamine and glutamate (AA; 46·8 μmol amino acid-N/kg0·75 per min). The treatments simulated nutritional extremes in terms of the balance of absorbed N. Kinetics across the whole gut and the liver were monitored during an intra-jugular infusion of [5-15N]glutamine. Blood flow across the whole gut or liver were unaffected by treatment. Both AMM and AA infusions doubled the hepatic release of urea-N compared with CONT (P<0·02). AA infusion decreased arterial glutamine concentration by 26 % (P<0·01) and 23 % (P<0·05) compared with AMM and CONT respectively. Despite this, whole-body glutamine flux was not affected by treatment. In contrast, AMM infusion increased hepatic glutamine production by 40 % compared with CONT (P<0·02). This provided a mechanism to ensure NH3 supply to the periphery was maintained within the normal low physiological levels. Hepatic glutamine utilisation tended to increase during AA infusion, probably to ensure equal inflows of N to the ornithine cycle. Between 6 and 10 % of NH3 absorbed across the digestive tract was derived from the amido-N of glutamine. Overall, splanchnic glutamine utilisation accounted for 45–70 % of whole-body glutamine flux.

The effect of feeding lactic-acid bacteria on indices of functions of lymphocytes obtained from Peyer's patches, peripheral blood and spleen from inbred Wistar-Furth rats were studied. Rats were fed on purified diets supplemented with 350 g milk or yoghurt/kg diet for 4 weeks. At the end of the feeding period, immune cells from the three sites were isolated and proliferation, interferon-γ production and lymphocyte subset composition were studied. Rats consuming yoghurt had a greater in vitro proliferative response to yoghurt bacteria in the three lymphoid compartments, a greater interferon-γ production in response to bacteria and concanavalin A in Peyer's patches and spleen, and a greater number of Peyer's patches B lymphocytes than milk-fed rats. Macrophage and T lymphocyte proportions and lymphocyte subset composition in the three sites were unaffected by yoghurt. These results indicate that feeding live bacteria contained in yoghurt may interact with the intestinal immune system, and influence the systemic immune system.

The effects of feeding beef suet (mostly saturated and monoenoic fatty acids), sunflower oil (rich in n-6 fatty acids) and fish oil (rich in n-3 fatty acids) on the response of mesenteric, omental, popliteal and perirenal adipocytes to experimentally-induced local inflammation were studied in adult guinea pigs. After 6 weeks on the experimental diets, the animals were fed standard chow, and lipopolysaccharide was injected unilaterally daily for 4 d to induce swelling of one popliteal lymph node. Basal lipolysis in the perinodal adipocytes of all depots studied was higher in the sunflower oil-fed animals than in the controls fed on standard chow, and lower in those fed on suet or fish oil. Dietary lipids altered rates of lipolysis during incubation with 10-5 M noradrenaline in all samples studied from the locally-activated popliteal depot, but only in adipocytes within 5 mm of a large lymph node in the other depots. The fish-oil diet attenuated the spread of increased lipolysis within the locally-activated popliteal adipose tissue, and from this depot to other node-containing depots. These experiments show that n-6 polyunsaturated fatty acids promote and n-3 fatty acids suppress the spread of immune activation to adipocytes within and between depots, and alter the sensitivity of perinodal adipocytes to noradrenaline. Dietary effects are reduced or absent in adipocytes in sites remote from lymph nodes, and thus such samples do not adequately represent processes in perinodal adipose tissue. These results are consistent with the hypothesis that perinodal adipocytes interact with adjacent lymphoid cells during immune responses.

A short quantitative food frequency questionnaire (FFQ) to assess folate intake was developed and validated against a 7-d weighed food intake record (7d-WR) and biochemical indices of folate status. Thirty-six men and women completed the self-administered FFQ on two occasions a month apart, kept a 7d-WR and gave two fasting blood samples at the beginning and end of the study for measuring serum and erythrocyte folate, respectively. Mean folate intakes were similar by repeat FFQ and correlated strongly (r 0·77 and r 0·72, P<0·001, for men and women, respectively). All other comparisons were done using the results of the FFQ administered on the first occasion. Men reported similar folate intakes on the FFQ and 7d-WR, but women reported greater intakes on the FFQ compared with the 7d-WR (P<0·05). There was a statistically significant correlation (partial, controlling for gender) between folate intakes reported by FFQ and 7d-WR (r 0·53, P<0·01). Folate intakes estimated by FFQ correlated significantly with serum (r 0·47, P<0·01), but not erythrocyte folate (r 0·25, P>0·05); the strength of the association was greater in men than in women. Validity coefficients estimated using the method of triads were higher for the FFQ than for the 7d-WR when serum folate was used as the biomarker. Overall, these results suggest that this short FFQ is a useful method for assessing folate intake, particularly in men.