Ochratoxin A (OTA), a mycotoxin frequently present in food and feedstuffs, produces a wide range of toxic effects, including cell death via lipid peroxidation. In one human and four animal cell lines we determined the half lethal concentration (LC50) of OTA, its effect on reactive oxygen species (ROS) production, and its ability to induce cytochrome p450 activity. We also examined the protective effect of α-tocopherol and all-trans-retinol in the most sensitive cell lines (i.e. bovine mammary epithelia, for which LC50 was 0·8μg/ml (24h), and Madin Darby canine kidney, for which LC50 was 4·3μg/ml (48h)). Pre-incubation for 3h with either antioxidant significantly (P<0·05) ameliorated the OTA-induced reduction in cell viability and significantly decreased (P<0·05) ROS production. These findings indicate that oxidative stress is an important factor in OTA cytotoxicity. Supplementation with antioxidant molecules may counteract the short-term toxicity of this mycotoxin.

The incidence of hormone-dependent cancers, such as those of the breast and prostate, is much lower in Eastern countries such as China and Japan in comparison with the Western world. Diet is believed to have a major effect on disease risk and one group of compounds, the phyto-oestrogens, which are consumed in large amounts in Asian populations, have been implicated in cancer protection. This view follows the finding that plasma and urinary levels of phyto-oestrogens are much higher in areas where cancer incidence is low in comparison with areas of high cancer incidence. The phyto-oestrogens are comprised of two main groups; the isoflavones and lignans. Of the isoflavones, genistein and daidzein have been the most widely studied. These compounds have been shown to possess anticancer properties; however their precise mechanism of action remains to be elucidated. In comparison, few studies have investigated the effects of lignans in breast and prostate cancer. In vitro studies have shown that genistein exerts biphasic effects on cancer cell growth, stimulating growth at low concentrations (<10μM) and inhibiting growth at high concentrations (>10μM), which suggests that low phyto-oestrogen levels may stimulate cancer growth in vivo. Plasma phyto-oestrogen concentrations of >10μM cannot be achieved by dietary intake and therefore the timing of exposure to phyto-oestrogens may be of the utmost importance in determining their chemopreventive effects. The present paper reviews the effects of phyto-oestrogens on breast and prostate cancer in vivo and in vitro and discusses possible mechanisms of action via which these compounds may exert their effects.

In the present study we examined whether oral administration of bovine lactoferrin (bLF) reduces plasma or hepatic triacylglycerol and cholesterol in mice. When bLF mixed with a standard commercial diet (10g/kg) was given to mice for 4 weeks, plasma triacylglycerol and NEFA decreased, while plasma HDL-cholesterol levels increased (P<0·01). These changes in plasma lipid profiles were accompanied by significant decreases in hepatic cholesterol and triacylglycerol contents. When mice were fed a high-fat diet containing 300·0g lard, 10·0g cholesterol and 2·5g bovine bile powder/kg for 4 weeks, bovine LF did not have any significant effects on plasma or hepatic cholesterol and triacylglycerol concentrations. Furthermore, bLF had no significant effects on faecal excretion of total bile acids in mice. Interestingly, bLF showed a suppressive effect on the lymphatic triacylglycerol absorption in chronically treated rats. We conclude that bLF has a beneficial effect on plasma cholesterol levels and retards hepatic lipid accumulation in mice fed a standard diet.

The present study investigated the susceptibility of dietary n-3 PUFA to ruminal biohydrogenation, the stability of ingested vitamin E in the rumen and the subsequent uptake of PUFA and vitamin E into plasma. Six cannulated sheep were assigned to six diets over five 33d periods, in an incomplete 6×5 Latin square. The diets, based on dried grass, were formulated to supply 50g fatty acids/kg DM using three lipid sources: Megalac® (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK), linseed (formaldehyde-treated; Trouw Nutrition, Northwich, Ches., UK) and linseed–fish oil (formaldehyde-treated linseed+fish oil). The diets were supplemented with 100 or 500mg α-tocopheryl acetate/kg DM. Fat source or level of vitamin E in the diet did not alter microbial activity in the rumen. Biohydrogenation of linoleic acid (18:3n-6; 85–90%), linolenic acid (18:3n-3; 88–93%), docosahexaenoic acid (22:6n-3; 91%) and EPA (20:5n-3; 92%) was extensive. Feeding formaldehyde-treated linseed elevated concentrations of 18:3n-3 in plasma, whilst 22:6n-3 and 20:5n-3 were only increased by feeding the linseed–fish oil blend. Duodenal recovery of ingested vitamin E was high (range 0·79–0·92mg/mg fed). High dietary vitamin E was associated with increased plasma α-tocopherol (2·57 v. 1·46μg/ml for 500 and 100mg α-tocopheryl acetate/kg DM respectively), although all concentrations were low. Plasma vitamin E levels, however, tended to decrease as the type and quantity of PUFA in the diet increased. The present study illustrates that nutritionally beneficial PUFA in both fish and linseed oils are highly susceptible to biohydrogenation in the rumen. Although α-tocopheryl acetate resisted degradation in the rumen, plasma vitamin E status remained deficient to borderline, suggesting either that uptake may have been impaired or metabolism post-absorption increased.

The effect of feeding n-3 PUFA on the fatty acid composition of muscle, adipose tissue and liver of lambs was investigated. Groups of eight ram lambs per breed, Suffolk×Lleyn (24kg live weight) and Scottish Blackface (18kg live weight), were each fed one of six diets containing one of three fat sources (50g fatty acids/kg DM; Megalac® (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK) and formaldehyde-treated whole linseed (Trouw Nutrition UK, Northwich, Ches., UK) either alone or with fish oil (1:1, w/w) and either 100 or 500mg α-tocopheryl acetate/kg DM. Feed was offered ad libitum until slaughter at approximately half breed mature live weight. The type of dietary fat had no effect on intake, growth rate or feed conversion ratio. The 3·0-fold higher concentration of 18:3n-3 in the linseed compared with the Megalac® diet approximately doubled (P<0·001) the concentration in the neutral and polar lipid fractions of musculus semimembranosus and liver, and in adipose tissue it increased 2·5-fold. Feeding protected linseed also increased (P<0·001) concentrations of 20:5n-3 and 22:5n-3 in muscle polar lipids and both lipid fractions of liver. The linseed–fish oil raised the 20:5n-3 concentrations above those for the linseed diet and also increased 22:6n-3. Scottish Blackface lambs had lower concentrations of 18:3n-3 in all lipids compared with Suffolk x Lleyn lambs, but more 20:5n-3 in the polar lipids of muscle and liver. High levels of dietary vitamin E were associated with small decreases in the concentration of monounsaturated fatty acids and increases in PUFA. Linseed raised the PUFA:saturated fatty acid ratios in liver and adipose tissue but not in muscle, and improved the n-6:n-3 fatty acid ratio, as did the linseed–fish oil. Different combinations of dietary fatty acids and better protection against rumen biohydrogenation are required to improve muscle PUFA:saturated fatty acids ratios.

Urinary isoflavone excretion is used to monitor compliance and examine biological effects. The present study determined if there were alterations in urinary isoflavone excretion following the ingestion of different soya foods and if age and gender potentially modified profiles. Twenty premenopausal women, seventeen post-menopausal women and twenty men received a defined single oral bolus dose (0·44mg isoflavones/kg body weight) of soya milk, textured vegetable protein (TVP) or tempeh on three separate occasions. Baseline and four consecutive complete 24h pooled urines were collected during each period. Urinary genistein recovery was influenced by gender and food matrix. For women the urinary genistein recovery was higher following soya-milk consumption compared with TVP (P<0·05). Tempeh consumption also resulted in an increased urinary genistein recovery relative to soya milk in premenopausal women (P<0·052). No differences in urinary genistein recoveries between soya foods were observed in the men. Although urinary daidzein excretion was similar across the foods studied and was not affected by age or gender, conversion to its intestinal metabolite, equol, resulted in potential matrix and chemical composition effects; urinary equol excretion was higher (P<0·01) following tempeh ingestion among equol producers. Together these data suggest that the fractional absorption of genistein is potentially different in men and women and is influenced by the food matrix and chemical composition. Furthermore, the data suggest that the metabolism of daidzein may be altered by the chemical composition of the isoflavones ingested. Further studies are required to examine the effect of higher intake and define the relative influence of these factors in elderly population groups.

The health value for man of lipids in bovine muscles can be improved by the addition of PUFA to the animals' diets, but such treatments can modify fluidity of plasma lipoproteins and therefore their metabolic functions. The aim of the present study was to analyse whether changes in chemical composition of lipoproteins in steers fed sunflower oil-rich diets altered lipoprotein fluidity, measured by fluorescence polarization and electron spin resonance. LDL, light HDL and heavy HDL fractions were isolated by ultracentrifugation from plasma of eighteen crossbred Charolais×Salers steers. For a period of 70d, animals were given a control diet (C, n 6) consisting of hay (540g/kg) and concentrate mixture (460g/kg) or the same basal diet supplemented with sunflower oil rich in n-6 PUFA (40g/kg diet DM), given either as crushed seeds (S, n 6) or as a free oil infused directly into the duodenum (O, n 6), thus avoiding ruminal hydrogenation of PUFA. We have shown that in bovine animals: (1) fluidity measurements by fluorescence polarization must be made at the bovine physiological temperature (38·5°C); (2) heavy HDL always appear as the less fluid lipoparticles; (3) electron spin resonance, which does not depend on lipoparticle size, is more appropriate to compare the fluidity of LDL with that of light HDL. The values for lipoprotein fluidity measured by both methods indicated that linoleate-rich diets did not have any effect when compared with diet C; however, chemical variables support a fluidification of lipoparticles, since in steers given the diet O, n-6 PUFA concentrations increased in polar (×1·8) and neutral (×1·6) lipids in lipoparticles (P=0·0001). The phospholipid:protein ratio increased in light (+20%, P=0·019) and heavy (+23%, P=0·06) HDL and especially in LDL (+46%, P=0·0001); the total cholesterol:phospholipid ratio decreased in the three lipoprotein classes (−15 to −30%, NS). Diet S led to similar but less pronounced effects. We concluded that linoleate-rich diets modified the chemical composition of plasma lipoproteins in steers, but did not alter their fluidity; this probably occurred as a result of ‘homeoviscous adaptation’, which ensured their functional capacity.

Two 8-week feeding trials were conducted to determine the dietary Na requirement for juvenile hybrid tilapia (Oreochromis niloticus × O. aureus) reared in fresh water and seawater. In each experiment, NaCl was added to the basal diet at 0, 0·5, 1, 2, 3, 5, or 7g Na/kg diet (fresh water) and at 0, 0·2, 0·5, 0·8, 1·2, 1·5, 2, or 3g Na/kg diet (seawater). Each diet was fed to three replicate groups of fish, individual fish initially weighing 0·69 (se 0·01) g, in a closed, recirculating rearing system. In fresh water, the tilapia fed the diet supplemented with 2g Na/kg diet had significantly (P<0·05) greater weight gain than the fish fed the diets supplemented with ≥3 and ≤0·5g Na/kg diet. Feed efficiency (FE) in fish generally followed the weight-gain pattern. Gill Na+–K+ ATPase activity was highest in the fish fed the diets supplemented with 1–3g Na/kg diet, followed by the fish fed the diet with 7g Na/kg diet and lowest in the fish fed the unsupplemented control diet. In seawater, the weight gain, FE and gill Na+–K+ ATPase activity in fish were not affected by the dietary treatment. Analysis by polynomial regression of weight gain, by broken-line regression of gill Na+–K+ ATPase activity and by linear regression of whole-body Na retention of the fish reared in fresh water, indicated that the adequate dietary Na concentration for tilapia is about 1·5g/kg diet. The present study also suggests that no dietary Na is required for tilapia reared in seawater.

The present study examined whether replacing fat with inulin or lupin-kernel fibre influenced palatability, perceptions of satiety, and food intake in thirty-three healthy men (mean age 52 years, BMI 27·4kg/m2), using a within-subject design. On separate occasions, after fasting overnight, the participants consumed a breakfast consisting primarily of either a full-fat sausage patty (FFP) or a reduced-fat patty containing inulin (INP) or lupin-kernel fibre (LKP). Breakfast variants were alike in mass, protein and carbohydrate content; however the INP and LKP breakfasts were 36 and 37% lower in fat and 15 and 17% lower in energy density respectively compared with the FFP breakfast. The participants rated their satiety before breakfast then evaluated patty acceptability. Satiety was rated immediately after consuming the breakfast, then over the subsequent 4·5h whilst fasting. Food consumed until the end of the following day was recorded. All patties were rated above ‘neither acceptable or unacceptable’, however the INP rated lower for general acceptability (P=0·039) and the LKP lower for flavour (P=0·023) than the FFP. The LKP breakfast rated more satiating than the INP (P=0·010) and FFP (P=0·016) breakfasts. Total fat intake was 18g lower on the day of the INP (P=0·035) and 26g lower on the day of the LKP breakfast (P=0·013) than the FFP breakfast day. Energy intake was lower (1521kJ) only on the day of the INP breakfast (P=0·039). Both inulin and lupin-kernel fibre appear to have potential as fat replacers in meat products and for reducing fat and energy intake in men.

The aim of the present study was to evaluate Mg absorption from a test meal served with an oxalate-rich vegetable, spinach, as compared with a test meal served with a vegetable with a low oxalate content, kale. Mg absorption was measured by a stable-isotope technique based on extrinsic labelling of the test meals and faecal monitoring of the excreted isotope labels. Nine healthy adults participated in the study. The test meals were based on 100g phytate-free white bread, served with 300g spinach (6·6mmol oxalate; 0·7mmol 25Mg label added, 5·0mmol total Mg) or 300g kale (0·1mmol oxalate; 1·2mmol 26Mg label added, 4·8mmol total Mg). The test meals were served on days 1 and 3, at breakfast and lunch, using a cross-over design. The results from the present study demonstrated that apparent Mg absorption was significantly lower from the meal served with spinach (26·7 (sd 10·4) %) than the meal served with kale (36·5 (sd 11·8) %) (P=0·01). However, the lower fractional apparent Mg absorption from the test meal served with spinach can be assumed to be, at least partly, counterbalanced by the higher native Mg content of spinach as compared with kale. Although based on indirect evidence, i.e. not based on an evaluation of added (or removed) oxalic acid, the difference in Mg absorption observed in the present study is attributed to the difference in oxalic acid content between the two vegetables.

The urinary excretion of soya isoflavones and gut microflora metabolites was investigated in infants and children who had been fed soya-based infant formulas in early infancy. These infants and children were compared with cows'-milk formula-fed controls, to determine at what age gut microflora metabolism of daidzein to equol and/or O-desmethylangolensin (O-DMA) was established, and whether exposure to isoflavones in early infancy influences their metabolism at a later stage of development. Sixty infants and children (aged 4 months–7 years) participated in the study; thirty in each of the soya and control groups. There were four age groups. These were: 4–6 months (seven in the soya group and seven in the control group); 7–12 months (seven in the soya group and nine in the control group); 1–3 years (six in the soya group and eight in the control group); 3–7 years (ten in the soya group and six in the control group). Urine samples were collected to measure isoflavonoids by MS, and faecal samples were collected to measure gut-health-related bacterial composition, by fluorescent in situ hybridisation with oligonucleotide probes, and metabolic activity. A soya challenge (typically a soya yoghurt alternative product containing 4·8g soya protein and on average 22mg total isoflavones) was given to control-group infants (>6 months) and children, and also to soya-group children that were no longer consuming soya, to determine their ability to produce equol and/or O-DMA. Urinary genistein, daidzein and glycitein were detected in all infants (4–6 months) fed soya-based infant formula; O-DMA was detected in 75% of infants but equol was detected in only 25%. In the controls (4–6 months), urinary isoflavonoids were very low or not detected. In the older age groups (7 months–7 years), O-DMA was found in the urine samples of 75% of the soya group and 50% of the controls, after the soya challenge. Equol excretion was detected in 19% of the soya-group infants and children, and in only 5% of the controls. However, in the oldest (3–7 years) children, the proportion excreting O-DMA and equol was similar in both groups. Faecal bacterial numbers for bifidobacteria (P<0·001), bacteroides and clostridia (P<0·05) were significantly lower for the soya group compared with the control group. There appears to be no lasting effect of early-life isoflavone exposure on isoflavone metabolism.

Many people take dietary supplements, but information on characteristics associated with their use is lacking. The relationship between lifestyle behaviours, morbidity and use of dietary supplements has not been examined and earlier studies have limited applicability to a general population. These issues were addressed in the current study. Information was obtained by postal questionnaire sent to a sample of the general population. The questionnaire was completed by 70·5% of the sample (15465 from a total sample of 21923), with at least one-third (35·5%) taking dietary supplements. In adjusted analyses, supplement users were more likely to be women, white, home-owners, non-smokers and physically active. Use of vitamin, mineral and/or antioxidant supplements was associated with eating more fruits and vegetables, and taking fish-oil supplements was associated with eating oil-rich fish. A history of CVD or risk factors for CVD reduced the risk of taking vitamins, minerals and/or antioxidants or fish-oil supplements. Those reporting musculoskeletal disorders such as arthritis were more likely to take fish-oil supplements For the first time, we have shown that dietary supplement use is related to different types of morbidity. In particular, people at risk of primary or secondary CVD seem less likely to use dietary supplements, despite possible benefits shown in clinical trials. Public health organisations need to develop guidelines for the public and health professionals regarding the uncontrolled use of dietary supplements in the community.

It is generally accepted that supplemental Ca and/or vitamin D is effective in reducing the incidence of bone fractures; this is supported by numerous randomised controlled trials and meta-analyses. However, a question that has received much less attention is whether dietary Ca, i.e. Ca in physiological doses in normal food intake, also affects bone fracture risk. The present study aims to review the effect of dietary Ca on bone fractures at the hip, spine and radius in women >35 years old, and to compare these results with previous meta-analyses. MEDLINE (1966–1999) and reference lists in papers were searched for observational dietary Ca studies. Data were extracted in duplicate and separately. Heterogeneity and publication bias were tested. Observational studies failed to show any association between dietary Ca intake and risk of hip fracture (risk ratio 1·01, 95% CI 0·96, 1·07 for each increment of 300mg dietary Ca intake/d). There is a suggestion that either extremely low Ca intake may increase fracture risk, or that East Asian women may respond differently to increasing Ca intake.

Milk fat is high in saturated fatty acids (SFA) and high intakes of SFA are associated with cardiovascular diseases. The aim of the present study was to prospectively evaluate the potential risk of a first-ever acute myocardial infarction (AMI) in relation to the estimated milk-fat intake, reflected as the proportions of pentadecanoic acid (15:0) and heptadecanoic acid (17:0) in serum lipid esters. This was evaluated in a study population selected within the Västerbotten Intervention Program and the northern Sweden ‘Monitoring of Trends and Determinants in Cardiovascular disease’ survey populations. A prospective case–control design was used. The proportions of the biomarkers were lower in the cases (n78) than in the controls (n156), who were matched for age, sex, sampling time and geographical region. The standardised odds ratios of becoming an AMI case were between 0·7 and 0·8 for the biomarkers. The proportions of 15:0 and 17:0 in serum phospholipids were significantly and negatively correlated to serum concentrations of plasminogen activator inhibitor-1, tissue-type plasminogen activator, triacylglycerols, insulin, specific insulin, pro-insulin and leptin (all P<0·0001), suggesting a negative relationship to the insulin-resistance syndrome and the risk of CHD. Adjustment for BMI did not materially change the relationships. Although there seems to be a negative association between milk-fat intake as mirrored by the proportions of 15:0 and 17:0 in serum lipid esters and a first-ever AMI, adjustment for clinical risk factors removed this relationship.

The relationships between n-3 and n-6 fatty acids in subcutaneous fat, BMI and overweight status were investigated in eighty-eight children from Crete and Cyprus. Overweight status, BMI and serum lipid levels were similar in children at both locations, but Cretan children had higher levels of total MUFA than Cypriot children (62·2 (sd 2·8) v. 52·2 (sd 2·8)% area, respectively, P<0·001) and consequently Cypriot children had higher levels of total saturated, polyunsaturated, trans, n-3 and n-6 fatty acids. Cypriot children had also higher levels of individual n-3 and n-6 fatty acids, specifically linoleic, α-linolenic and dihomo-γ-linolenic acids. The variance of BMI was better explained (38·2%) by adipose tissue arachidonic acid content than any other n-3 and n-6 fatty acids. Mean levels of arachidonic acid, dihomo-γ-linolenic acid and docosahexaenoic acid were higher in overweight and obese subjects. All obese subjects fell in the 4th quartile of arachidonic acid levels, whereas 88·9% of overweight subjects fell in the 3rd and 4th quartile of arachidonic acid. These results indicate positive associations between adipose tissue arachidonic acid and BMI and overweight status. Further research could clarify whether this association is causal.