The mink is a seasonal breeder with a propensity for seasonal fatness, and it is very responsive to changes in energy supply. The objectives of the present study were first, to validate a multi-species leptin assay for mink (Mustela vison) plasma, and second, to evaluate how plasma leptin and insulin concentrations responded to energy restriction and body-weight loss and refeeding with restoration of body reserves. The study was performed with six very fat yearling females (initial mean body weight 1451 (SD 119) g, i.e. approximately 300 g more than for a female in normal body condition). The animals were fed in restricted amounts (about 35 % metabolizable energy requirement for maintenance) in order to reach a very lean body condition. The target weight of 800 g was reached after about 1 month of restriction. The animals were then refed ad libitum until almost complete weight recovery. Blood samples were taken on days 1, 24, 34 (end of restriction), 44, 55 and 71 (end of experiment) and analysed for plasma concentrations of leptin and insulin. Three females were mated on day 44. Leptin and insulin concentrations mirrored each other and clearly reflected changes in body weight. Significant (P<0·001) Pearson correlation coefficients of 0·75 (leptin–insulin), 0·72 (leptin–body weight) and 0·59 (insulin–body weight) were found. Two of the three females that were mated gave birth to normal litters. It was concluded that the leptin assay yielded acceptable results for animals with body weight:fat content within the range investigated here, and that plasma leptin reflected body fat mass.

Long-chain n-3 fatty acids (FA) consistently inhibit the growth of human breast cancer (BC) cells both in culture and in grafts in immunosuppressed mice. Large cohort studies have, however, failed to confirm a protective effect for fish oils rich in n-3 FA against BC risk. The present review examines new evidence on biological mechanisms which may be involved in the inhibition of mammary carcinogenesis by long-chain n-3 FA, focusing on an apoptotic effect by its lipid peroxidation products. Dietary intake of n-3 FA leads to their incorporation into cell membrane lipids. Increased apoptosis in human BC cells following exposure to long-chain n-3 FA such as eicosapentaenoic and docosahexaenoic acids is generally ascribed to their inhibition of cyclooxygenase 2 which promotes mammary carcinogenesis. In addition however, long-chain n-3 FA are particularly likely to activate peroxisome proliferator-activated receptor (PPAR)-γ, a key regulator of lipid metabolism but also capable of modulating proliferative activity in a variety of cells including mammary cells. Expression of PPAR-γ in the nucleus is activated by second messengers such as J series prostaglandins and the latter have been shown to cause apoptosis in vivo in explants of human BC cells in immunosuppressed mice. In mammary tumours, it is observed that long-chain FA not only increase apoptosis, but also increase lipid peroxidation, and the apoptotic effect can be reversed by antioxidants. The rationale for use of n-3 FA dietary supplements in counteracting BC progression needs to be tested clinically in a phase 2 pilot study, while at the same time, the effect on whole-body lipid peroxidation needs to be monitored. Dietary supplements of fish oil rich in n-3 FA are proposed for premenopausal women over the age of 40 years who are shown to be at increased BC risk. Biological markers in breast tissue of BC progression will be monitored, and observed changes related to serial plasma levels of isoprostanes as a measure of whole-body lipid peroxidation.

A food can be regarded as functional if it is satisfactorily demonstrated to affect beneficially one or more target functions in the body, beyond adequate nutritional effects, in a way which is relevant to either the state of well-being and health or the reduction of the risk of a disease. Health claims are expected to be authorized for functional foods based either on enhanced function (type A claim) or disease risk reduction (type B claim). Their development is a unique opportunity to contribute to the improvement of the quality of the food offered to consumer's choice for the benefit of his well-being and health. But only a rigorous scientific approach producing sound data will garantee its success. The functional food components that are discussed in the proceedings of the 3rd ORAFTI Research Conference are the inulin-type fructans, natural food components found in miscellaneous edible plants. They are non-digestible oligosaccharides that are classified as dietary fiber. The targets for their functional effects are the colonic microflora that use them as selective ‘fertilizers’; the gastrointestinal physiology; the immune functions; the bioavailability of minerals; and the metabolism of lipids. Potential health benefits may also concern reduction of the risk of some diseases like intestinal infections, constipation, non-insulin dependent diabetes, obesity, osteoporosis or colon cancer. The present proceedings review the scientific data available and, by reference to the concepts in functional food science, they assess the scientific evidence which will be used to substantiate health claims.

Cu appears to have many important functional roles in the body that apparently relate, among others, to the maintenance of immune function, bone health and haemostasis. Some have suggested a role for long-term marginal Cu deficiency in the aetiology of a number of degenerative diseases. Accurate diagnosis of marginal Cu deficiency, however, has remained elusive despite an increased understanding of the biochemistry of Cu and its physiological roles in the body. Traditional markers of Cu status, such as serum Cu and caeruloplasmin protein concentrations are insensitive to subtle changes in Cu status. Cu-containing enzymes, such as Cu–Zn-superoxide dismutase, cytochrome c oxidase and diamine oxidase, may be more reliable but evidence to date is not conclusive. Development of markers sensitive to marginal Cu status is essential before conclusions can be drawn concerning the risks of long-term intake of suboptimal dietary Cu. As Cu appears to be essential for maintenance of immune function, activities of specific immunological markers, altered in Cu deficiency, offer alternatives. This review evaluates a selection of immunological markers that could be considered potentially sensitive markers of marginal Cu status. The indices of immune function reviewed are neutrophil function, interleukin 2 production, blastogenic response to mitogens and lymphocyte subset phenotyping.

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.

The defining effect of prebiotics is to stimulate selectively the growth of bifidobacteria and lactobacilli in the gut and, thereby, increase the body's natural resistance to invading pathogens. Prebiotic carbohydrates may also have additional, less specific, benefits because they are fermented in the large intestine. The prebiotic carbohydrates that have been evaluated in humans at the present time largely consist of fructans or galactans. There is consistent evidence from in vitro and in vivo studies that these are not digested by normal human enzymes, but are readily fermented by anaerobic bacteria in the large intestine. There are no reports of faecal recovery of measurable quantities of prebiotic carbohydrates. Through fermentation in the large intestine, prebiotic carbohydrates yield short-chain fatty acids, stimulate the growth of many bacterial species in addition to the selective effects on lactobacilli and bifidobacteria, they can also produce gas. Along with other fermented carbohydrates, prebiotics have mild laxative effects, although this has proved difficult to demonstrate in human studies because the magnitude of laxation is small. Potentially, the most important effect of prebiotic carbohydrates is to strengthen the body's resistance to invading pathogens and, thereby, prevent episodes of diarrhoea. At the present time, this effect has not been convincingly demonstrated in either adults or children, although there have been attempts to ameliorate the diarrhoea associated with antibiotics and travel, but without success. However, prebiotic carbohydrates clearly have significant and distinctive physiological effects in the human large intestine, and on the basis of this it is likely that they will ultimately be shown to be beneficial to health.

Reactive oxygen species have been implicated in Helicobacter pylori-mediated gastric carcinogenesis, whereas diets high in antioxidant vitamins C and E are protective. We have examined the effect of vitamin C and E supplements in combination with H. pylori eradication on reactive oxygen species activity in H. pylori gastritis. H. pylori-positive patients were randomized into four groups: triple therapy alone (Bismuth chelate, tetracycline, and metronidazole for 2 weeks), vitamins alone (200 mg vitamin C and 50 mg vitamin E, both twice per day for 4 weeks), both treatments or neither. Plasma and mucosal ascorbic acid, malondialdehyde and reactive oxygen species were determined before and after treatment. Compared with normal controls (n 61), H. pylori-positive patients (n 117) had higher mucosal reactive oxygen species and malondialdehyde levels and lower plasma ascorbic acid. Plasma ascorbic acid doubled in both groups of patients receiving vitamins and mucosal levels also increased. Malondialdehyde and reactive oxygen species fell in patients in whom H. pylori was eradicated but vitamin supplements were not effective either alone or in combination with H. pylori eradication. Supplements of vitamins C and E do not significantly reduce mucosal reactive oxygen species damage in H. pylori gastritis.

Diet, among other environmental and genetic factors, is currently recognised to have an important role in health and disease. There is increasing evidence that the human colonic microbiota can contribute positively towards host nutrition and health. As such, dietary modulation has been proposed as important for improved gut health, especially during the highly sensitive stage of infancy. Differences in gut microflora composition and incidence of infection occur between breast- and formula-fed infants. Human milk components that cannot be duplicated in infant formulae could possibly account for these differences. However, various functional food ingredients such as oligosaccharides, prebiotics, proteins and probiotics could effect a beneficial modification in the composition and activities of gut microflora of infants. The aim of the present review is to describe existing knowledge on the composition and metabolic activities of the gastrointestinal microflora of human infants and discuss various possibilities and opportunities for its nutritional modulation.

The influence of myristic acid in a narrow physiological range (0·5 to 2·4 % of total dietary energy) on the plasma and hepatic cholesterol metabolism was investigated in the hamster. The hamsters were fed on a diet containing 12·5 g fat/100 g and 0·05 g cholesterol/100 g with 0·5 % myristic acid (LA diet) for 3 weeks (pre-period). During the following 3 weeks (test period), they were divided into four dietary groups with 0·5 % (LA), 1·2 % (LM), 1·8 % (ML) or 2·4 % (M) myristic acid. Finally, half the hamsters in each group were again fed the LA diet for another 3 weeks (post-period). At the end of the test period, the hepatic expression of the scavenger receptor BI (SR-BI) was lower in the LM, ML and M groups than in the LA group whereas the hepatic cholesteryl ester concentration was higher. Cholesterol 7α hydroxylase activity was lower in the ML and M groups than in the LA and LM groups while the sterol 27 hydroxylase and 3-hydroxy-3-methyl glutaryl coenzyme A reductase activities were not modulated by dietary myristic acid. This is the first time a negative correlation has been observed between the HDL-cholesterol concentration and the hepatic mass of SR-BI (r -0·69; P<0·0001) under physiological conditions. An inverse linear regression was also shown between SR-BI and the percentage of myristic acid in the diet (r -0·75; P<0·0001). The hepatic mass of SR-BI in the M group had increased at the end of the post-period compared with the test-period values. The present investigation shows that myristic acid modulates HDL-cholesterol via a regulation of the SR-BI expression.

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.

We previously described the effects of intake of dairy products on plasmid dissemination in the digestive tract of gnotobiotic mice associated with human faecal flora (HFF) and found that yoghurt, heat-treated yoghurt (HTY) and milk reduced population levels of transconjugants compared with findings in mice fed a standard mouse diet. In the case of lactose intake, transconjugants were not detected. The aim of the present study was to assess the possible interrelationships between these observations and other variables (bacterial ecology, pH, moisture, enzyme activities, short-chain fatty acid (SCFA) contents, lactic acid contents). Much of the interest of the present comparison lies in the fact that the animals were homogeneous in terms of age, gender, food and intestinal microflora, owing to the gnotobiotic mouse model maintained in sterile isolators. We observed no variation in SCFA and lactic acid contents or in the population levels of strictly anaerobic strains of Bacteroides and Bifidobacterium, and of the facultative anaerobic recipient Escherichia coli PG1 strain. The main modifications were the reduction of population levels of transconjugants in mice receiving yoghurt, HTY and milk, and concomitantly an increase of β-galactosidase and a decrease of β-glucosidase activities, compared with control mice fed a standard diet. Total inhibition of plasmid transfer was observed in HFF mice consuming lactose, and concomitantly the two enzyme activities (β-glucosidase and β-galactosidase) were increased, compared with the findings in control mice fed a standard diet. In axenic mice consuming lactose, plasmid transfer occurred, β-galactosidase was not detected and β-glucosidase was decreased. It is therefore proposed that these two enzyme activities influence plasmid transfer and persistence of transconjugants in the digestive tract of HFF associated mice. When both activities were increased there was a total inhibition of plasmid transfer (case of lactose intake). When β-galactosidase increased and β-glucosidase decreased (case of yoghurt, HTY and milk), plasmid transfer occurred at a lower efficiency than in the control group, resulting in lower population levels of transconjugants.

The inclusion of genetically modified (GM) foods in the human diet has caused considerable debate. There is concern that the transfer of plant-derived transgenes to the resident intestinal microflora could have safety implications. For these gene transfer events to occur, the nucleic acid would need to survive passage through the gastrointestinal tract. The aim of the present study was to evaluate the rate at which transgenes, contained within GM soya and maize, are degraded in gastric and small bowel simulations. The data showed that 80 % of the transgene in naked GM soya DNA was degraded in the gastric simulations, while no degradation of the transgenes contained within GM soya and maize were observed in these acidic conditions. In the small intestinal simulations, transgenes in naked soya DNA were degraded at a similar rate to the material in the soya protein. After incubation for 30 min, the transgenes remaining in soya protein and naked DNA were 52 (SEM 13·1) % AND 34 (sem 17·5) %, respectively, and at the completion of the experiment (3 h) these values were 5 % and 3 %, respectively. In contrast to the soya transgene, the maize nucleic acid was hydrolysed in the small intestinal simulations in a biphasic process in which approximately 85 % was rapidly degraded, while the rest of the DNA was cleaved at a rate similar to that in the soya material. Guar gum and tannic acid, molecules that are known to inhibit digestive enzymes, did not influence the rate of transgene degradation in soya protein. In contrast guar gum reduced the rate of transgene degradation in naked soya DNA in the initial stages, but the polysaccharide did not influence the amount of nucleic acid remaining at the end of the experiment. Tannic acid reduced the rate of DNA degradation throughout the small bowel simulations, with 21 (sem 5·4) % and 2 (sem 1·8) % of the naked soya DNA remaining in the presence and absence of the phenolic acid, respectively. These data indicate that some transgenes in GM foods may survive passage through the small intestine.

Tannins have received considerable attention from animal nutritionists as potential agents for modifying ruminal fermentation patterns, or for exploring new feed resources. This group of secondary plant compounds is defined by their ability to form complexes with proteins. A widely accepted method for assaying the biological activity of extracted tannins is the precipitation of bovine serum albumin. The protein carries a radioactive label (125I) to allow direct quantification from the precipitate. Tannin–protein complexes dissolve in sodium dodecylsulfate solution. A dot-blot assay for protein determination, which is based on the reversible binding of a fluorochrome, benzoxanthene yellow, to the protein spots and is not disturbed by the presence of detergents, can replace the radioactive method by a fluorimetric measurement. A novel alternative to the last part of the dot-blot assay is to scan the stained protein spots in situ using a video camera and computer image analysis. Several filter sets were tested and, within a concentration range of 0·1–2·0 mg protein/ml, each of them yielded results identical to the original method while the time required was only 30 % of the working time consumed by the original procedure. The modified dot-blot assay should be applicable to the evaluation of tannin activity in all shrub and tree foliages considered as animal feed.

The potential ‘nutritional advantages’ of probiotics and prebiotics consist of preventive, and sometimes curative, effects against certain diseases. The evidence supporting such advantages, which requires randomised controlled trials and consistency of results from study to study, is rapidly increasing. This article summarizes the effects against diseases of intestinal origin. There is a high level of evidence for positive effects of some prebiotics to alleviate constipation and treat hepatic encephalopathy. Interesting aspects, but with a lower level of evidence at the present time, include prevention of colon cancer, intestinal infection, and recurrence of inflammatory bowel disease. There is a high level of evidence for positive effects of some probiotics in the alleviation of lactose intolerance, antibiotic-associated intestinal disorders and gastroenteritis. Evidence is rapidly growing regarding the prevention of recurrence of inflammatory bowel diseases. Positive trials have suggested preventive effects against intestinal colonization with specific gut pathogens including Clostridium difficile and Helicobacter pylori.

The bioavailability of Se from raw and cured selenomethionine-enriched (Se-enriched) salmon fillets was assessed in Se-deficient male albino rats (Mol: Wist). A low-Se Torula yeast feed was supplemented with 0, 50, 100, 150 or 200 μg Se/kg as sodium selenite or as Se from raw or cured Se-enriched salmon. The diets were fed to weanling rats for 10 and 30 d. Bioavailability of Se was assessed by metabolic balance, Se accumulation in femur, muscle, liver and plasma, and induction of Se-dependent glutathione peroxidase (EC 1·11·1.9; GSHpx) in plasma as response parameters. Except for the metabolic balance results, the slope-ratio method was used when calculating Se bioavailability from raw or cured Se-enriched fish fillets (test food) relative to sodium selenite (standard). The data for fractional apparent absorption and fractional retention showed differences (P<0·05) among all three Se sources in the order raw salmon > cured salmon > selenite. At 10 d, Se from raw and cured Se-enriched fish fillets tended to be more bioavailable than selenite. This was supported by the observations for Se accumulation in femur and muscle and induction of GSHpx activity. At 30 d, all response parameters showed a higher bioavailability of Se from raw and cured Se-enriched fish fillets compared with selenite. Differences (P<0·05) in Se accumulation in muscle at 10 and 30 d, and differences (P<0·05) in fractional apparent absorption and fractional retention suggested that curing salmon altered the utilisation of Se. The experimental results showed that enrichment of fish fillets with selenomethionine yields fillets with high Se bioavailability.