An experiment was conducted to determine the effects of supplementing different amounts of daidzein in a diet on the growth performance, blood biochemical parameters and meat quality of finishing beef cattle. Thirty finishing Xianan steers were distributed in three groups equilibrated by weight and fed three different dietary treatments (concentrate ratio = 80%): (1) control; (2) 500 mg/kg daidzein and (3) 1000 mg/kg daidzein, respectively. Steers were slaughtered after an 80-day feeding trial. Results showed that daidzein supplementation had no effect on the final body weight, average daily gain and feed conversion rate of steers. Steers fed with 1000 mg/kg daidzein had greater dry matter intake than those fed with control diets. Compared with the control group, the 1000 mg/kg daidzein group had a higher fat thickness, lower shear force and lightness. The pH, drip loss, cooking loss, redness (a*), yellowness (b*), moisture, ash, crude protein and intramuscular fat of the Longissimus dorsi muscle were unaffected by daidzein supplementation. Compared with the control group, the 1000 mg/kg daidzein group significantly increased the serum concentrations of insulin, free fatty acid and Glutamic-pyruvic transaminase. The 500 mg/kg daidzein group significantly increased the serum concentration of tetraiodothyronine compared with the control group. Supplemental daidzein did not affect the blood antioxidant ability and blood immune parameters in serum. In conclusion, daidzein supplementation above 500 mg/day modifies feed intake and metabolic and hormonal profile, with positive and negative effects on meat quality.

Genistein and daidzein are isoflavones which are reported to influence the reproductive system in a variety of mammalian species. This pilot study aimed to determine if dietary isoflavones could potentially influence reproductive parameters in domestic cats, when consumed during the postnatal development period. Cats (n = 12) were maintained on either a treatment (150 µg/g DM genistein and 150 µg/g DM daidzein, n = 4) or control (isoflavone free, n = 8) diet from weaning, up to 414 (±17.2) days post-weaning. Vaginal smears were taken thrice weekly and examined for oestrogen-induced cellular degradation in all cats. Behavioural indicators of oestrous were routinely scored for the presence or absence of six key behaviours. Genistein and daidzein did not alter puberty onset or oestrous cycle parameters in these cats (P > 0.05). Behavioural scores were higher in cats in the treatment group than control. Incidence of apparent spontaneous ovulation (inferred from extended inter-oestrous periods) was greater in treated cats than control cats, although serum hormone profiles were not available to confirm this observation. Further testing is warranted.

Genistein and daidzein are isoflavones which are reported to influence the reproductive system in a variety of mammalian species. This pilot study aimed to determine if dietary isoflavones could potentially influence reproductive tract histology or morphology in domestic cats, when consumed during the postnatal development period. Cats were maintained on either treatment (150 µg/g DM genistein and 150 µg/g DM daidzein, n = 4) or control (isoflavone free, n = 8) diets from weaning, up to 414 (±17.2) days post-weaning. Reproductive tissues were collected during routine ovario-hysterectomy and examined for histology and sex steroid receptor expression. Findings indicate that these dietary isoflavones influenced the expression of oestrogen receptor α (ERα) and oestrogen receptor β (ERβ), and progesterone receptor in feline reproductive tissues. One cat in the treatment group developed suppurative endometritis, but no evidence of uterotrophic or histological changes were found in any other cats. The potential to alter expression of hormone receptors in the reproductive tract of domestic cats exposed to genistein and daidzein warrants further investigation.

Genistein and daidzein are known to have both beneficial and adverse effects on human health due to their many biological actions at the cellular level. Both isoflavones have been shown to inhibit GLUT-mediated glucose transport across the plasma membrane of mammalian cells. Since lysosomal membrane transport is essential for maintaining cellular homeostasis, the present study examined the effects of genistein and daidzein on glucose and sulphate transport in isolated rat liver lysosomes. Both genistein and daidzein significantly inhibited lysosomal glucose uptake. Genistein was a more potent glucose transport inhibitor than daidzein, with a half-maximum inhibitory concentration (IC50) of 45 μmol/l compared with 71 μmol/l for daidzein. Uptake kinetics of d-glucose showed a significant decrease in Vmax (control:genistein treat = 1489 (sem 91):507 (sem 76) pmol/unit of β-hexosaminidase per 15 s) without a change in Km. The presence of 50 μm-genistein in the medium also reduced glucose efflux from lysosomes preloaded with 100 mm-d-glucose. Genistein also inhibited lysosomal sulphate transport. Similar to its effects on glucose uptake kinetics, genistein treatment caused a significant decrease in sulphate uptake Vmax (control:genistein treat = 87 (sem 4):59 (sem 5) pmol/unit of β-hexosaminidase per 30 s), while the Km was not affected. The evidence provided by the present study suggests that the most likely mechanism of lysosomal glucose transport inhibition by genistein is via direct interaction between genistein and the transporter, rather than mediation by tyrosine kinase inactivation. Genistein likely has a similar mechanism of directly inhibiting sulphate transporter.

Isoflavone intake is associated with various properties beneficial to human health which are related to their antioxidant activity, for example, to their ability to increase LDL oxidation resistance. However, the distribution of isoflavones among plasma lipoproteins has not yet been elucidated in vivo. Therefore, the objective of the present study was to investigate the association between daidzein (DAI) and lipoproteins in human plasma upon administration of the aglycone and glucoside form. Five men aged 22–30 years participated in a randomised, double-blind study in cross-over design. After ingestion of DAI and daidzein-7-O-β-d-glucoside (DG) (1 mg DAI aglycone equivalents/kg body weight) blood samples were drawn before isoflavone administration as well as 1, 2, 3, 4·5, 6, 8, 10, 12, 24 and 48 h post-dose. Concentrations of DAI in the different lipoprotein fractions (chylomicrons, VLDL, LDL, HDL) and in the non-lipoprotein fraction were analysed using isotope dilution capillary GC/MS. The lipoprotein fraction profiles were similar for all subjects and resembled those obtained for plasma in our previously published study. The lipoprotein distribution based on the area under the concentration–time profiles from 0 h to infinity in the different fractions were irrespective of the administered form: non-lipoprotein fraction (53 %) > LDL (20 %) > HDL (14 %) > VLDL (9·5 %) > chylomicrons (2·5 %). Of DAI present in plasma, 47 % was associated to lipoproteins. Concentrations in the different lipoprotein fractions as well as in the non-lipoprotein fraction were always higher after the ingestion of DG than of DAI. Taken together, these results demonstrate an association between isoflavones and plasma lipoproteins in vivo.

A consensus view of soyabean phyto-oestrogens in clinical interventions in post-menopausal women is presented that is based on data from the EU-funded project Phytohealth. The phyto-oestrogens, primarily genistein and daidzein, were given as soyabean-protein isolates, whole-soyabean foods or extracts, supplements or pure compounds. A comprehensive literature search was conducted with well-defined inclusion or exclusion criteria. For areas for which substantial research exists only placebo-controlled double-blind randomised controlled trials (RCT) conducted on healthy post-menopausal women were included. For emerging areas all available human studies in post-menopausal women were reviewed. In order to make cross comparisons between studies the doses of isoflavones were calculated as aglycone equivalents. There is a suggestion, but no conclusive evidence, that isoflavones from the sources studied so far have a beneficial effect on bone health. The consumption of whole-soyabean foods and soyabean-protein isolates has some beneficial effects on lipid markers of cardiovascular risk. The consumption of isolated isoflavones does not affect blood lipid levels or blood pressure, although it may improve endothelial function. For menopausal symptoms there is currently limited evidence that soyabean-protein isolates, soyabean foods or red-clover (Trifolium pratense L.) extract are effective but soyabean isoflavone extracts may be effective in reducing hot flushes. There are too few RCT studies to reach conclusions on the effects of isoflavones on breast cancer, colon cancer, diabetes or cognitive function. The health benefits of soyabean phyto-oestrogens in healthy post-menopausal women are subtle and even some well-designed studies do not show protective effects. Future studies should focus on high-risk post-menopausal women, especially in the areas of diabetes, CVD, breast cancer and bone health.

Phyto-oestrogens, naturally occurring hormone-like chemicals in plant food, may play a protective role against hormone-related chronic diseases. South Asian migrants in the UK have a lower incidence of hormone-related cancer than their hosts but the extent to which this difference may be due to phytoestrogen intake is not known. The aim was to compare habitual phytoestrogen intake in first-generation South Asian migrant women and native British women. South Asian (n 221) and native British women (n 50) were recruited from general practitioner lists and were asked to provide monthly 24h recalls for a period of 1 year. An enhanced phytoestrogen database was compiled using data from a literature search and unpublished data. A sub-sample of South Asian women (n 100) and the native British women (n 40) also provided blood samples every 3 months during the 1-year period. The median daily intakes (μg/d) of isoflavones (184·2 v. 333·9) and lignans (110·8 v. 148·8) were significantly lower in South Asians than in the native British (P<0·001, P=0·04 respectively).There were no significant differences in mean plasma isoflavone levels (nmol/l) but plasma enterolactone was significantly lower in the South Asians (13·9 (sd17·5) v. 28·5 (SD23·3),P<0·001). The main sources of phytoestrogens were bread and vegetables in both ethnic groups. Habitual phytoestrogen intake in South Asian and native British women was below 1mg/d and was higher in the native British diet. The present study does not support the hypothesis that differences in phytoestrogen intake, or in circulating levels, could explain differences in hormone-related cancer risks between these two populations.

Breast cancer is the most common cancer in women and a significant cause of death. Mutations of the oncosuppressor genes BRCA1 and BRCA2 are associated with a hereditary risk of breast cancer, and dysregulation of their expression has been observed in sporadic cases. Soya isoflavones have been shown to inhibit breast cancer in studies in vitro, but associations between the consumption of isoflavone-containing foods and breast cancer risk have varied in epidemiological studies. Soya is a unique source of the phytoestrogens daidzein (4′,7-dihydroxyisoflavone) and genistein (4′,5,7-trihydroxyisoflavone), two molecules that are able to inhibit the proliferation of human breast cancer cells in vitro. The aim of the present study was to determine the effects of genistein (5μg/ml) and daidzein (20μg/ml) on transcription in three human breast cell lines (one dystrophic, MCF10a, and two malignant, MCF-7 and MDA-MB-231) after 72h treatment. The different genes involved in the BRCA1 and BRCA2 pathways (GADD45A, BARD1, JUN, BAX, RB1, ERα, ERβ, BAP1, TNFα, p53, p21Waf1/Cip1, p300, RAD51, pS2, Ki-67) were quantified by real-time quantitative RT-PCR, using the TaqMan method and an ABI Prism 7700 Sequence Detector (Applied Biosystems). We observed that, in response to treatment, many of these genes were overexpressed in the breast cancer cell lines (MCF-7 and MDA-MB-231) but not in the dystrophic cell line (MCF10a).

Epidemiological studies indicate that Asian women have a lower incidence of breast cancer compared with their counterparts in the West, and soya consumption has been suggested as a contributory factor. Clinical and animal studies have revealed that cyclooxygenase-2 (COX-2) expression is associated with a risk of breast cancer. In the present study, we investigated the effect of soya isoflavones on the expression of COX-2 in the breast cell line MCF-7. Genistein, daidzein and equol were found to inhibit COX-2 expression induced by phorbol 12-myristate 13-acetate (PMA). Similar findings were observed in the COX-2 protein analysis. In order to study transcriptional control, a fragment of the 5′-flanking region of the hCOX-2 gene was amplified and inserted into a firefly luciferase reporter plasmid. The reporter assay indicated that the transactivation of the hCOX-2 promoter was induced by PMA, and activity was inhibited with the co-administration of genistein, daidzein or equol. An activator protein-1 (AP-1)/cyclic AMP response element binding protein (CREB) binding site (−59/−53) was identified in hCOX-2 promoter, and this could be critical in PMA-induced COX-2 expression. Truncation reporter plasmids with (−70/−36) and without (−51/−36) AP-1/CREB were constructed for subsequent analysis. The results revealed that the hCOX-2 promoter transactivation suppressed by isoflavone could be dependent on AP-1/CREB binding. Nonetheless, this study illustrated that the soya isoflavones reduced COX-2 expression, which could be important in the post-initiation events of breast carcinogenesis.

Particular intestinal bacteria are capable of metabolizing the soya isoflavone daidzein to equol and/or O-desmethylangolensin (O-DMA), and the presence of these metabolites in urine after soya consumption are markers of particular intestinal bacteria profiles. Prevalences of equol producers and O-DMA producers are approximately 30–50 % and 80–90 %, respectively, and limited observations have suggested that these daidzein-metabolizing phenotypes are stable within individuals over time. Characterizing stability of these phenotypes is important to understand their potential as markers of long-term exposure to particular intestinal bacteria and their associations with disease risk. We evaluated concordance within an individual for the equol-producer and O-DMA-producer phenotypes measured at two time points (T1, T2), 1–3 years apart. Phenotypes were ascertained by analysing equol and O-DMA using GC-MS in a spot urine sample collected after 3 d soya (source of daidzein) supplementation. In ninety-two individuals without recent (within 3 months before phenotyping) or current antibiotics use, 41 % were equol producers at T1 and 45 % were equol producers at T2, and 90 % were O-DMA producers at T1 and 95 % were O-DMA producers at T2. The percentage agreement for the equol-producer phenotype was 82 and for the O-DMA-producer phenotype was 89. These results indicate that these phenotypes are stable in most individuals over time, suggesting that they provide a useful biomarker for evaluating disease risk associated with harbouring particular intestinal bacteria responsible for, or associated with, the metabolism of the soya isoflavone daidzein.

The effects of Lactobacillus gasseri JCM 1131T on isoflavonoid levels within the caecum and plasma were assessed in adult mice. Male 5-week-old mice were fed an AIN 93M diet for 30 d. Two groups of mice were administered either L. gasseri JCM 1131T (the LGI group) or physiological saline solution (the control (CI) group) daily for 5 d before dissection. The plasma daidzein concentration was significantly higher in the LGI group, however, their plasma equol concentration was significantly less than in the CI group. The total amount of equol present as aglycone in the caecum was significantly greater in the CI group, but there was no significant difference in the total daidzein present as caecal aglycone. In an in vitro incubation of daidzin with the faecal flora of mice, the equol concentration was significantly higher in the CI group. The numbers of lactobacilli present were significantly higher in the LGI group. The present data suggest that the administration of L. gasseri is likely to influence the effect of isoflavonoids on the host via changes in the gastrointestinal environment.

Dietary isoflavones are currently receiving much attention because of their potential role in preventing coronary artery disease and other chronic diseases. Accumulating evidence from cell culture and laboratory animal experiments indicates that isoflavones have the potential to prevent or delay atherogenesis. Suggested mechanisms of action include: a reduction in low-density lipoprotein (LDL) cholesterol and a potential reduction in the susceptibility of the LDL particle to oxidation; (2) an improvement in vascular reactivity; (3) an inhibition of pro-inflammatory cytokines, cell adhesion proteins and nitric oxide (NO) production; and (4) an inhibition of platelet aggregation. These mechanisms are consistent with the epidemiological evidence that a high consumption of isoflavone-rich soy products is associated with a reduced incidence of coronary artery disease. Biological effects of isoflavones are dependent on many factors, including dose consumed, duration of use, protein-binding affinity, and an individual's metabolism or intrinsic oestrogenic state. Further clinical studies are necessary to determine the potential health effects of isoflavones in specific population groups as we currently know little about age-related differences in exposure to these compounds and there are few guidelines on optimal dose for cardiovascular health benefits.

The aim of this study was to identify the level of isoflavone intake (total isoflavones, daidzein and genistein) in four European countries: Ireland, Italy, The Netherlands and the UK. For this purpose national food composition databases of isoflavone content were created in a comparable way, using the Vegetal Estrogens in Nutrition and the Skeleton (VENUS) analytical data base as a common basis, and appropriate food consumption data were selected. The isoflavone intake in Ireland, Italy, The Netherlands and the UK is on average less than 1 mg/d. Small groups of consumers of soya foods could be identified in Ireland, The Netherlands and the UK. The estimated intake levels are low compared with those found in typical Asian diets (∼20–100 mg/d) and also low compared with levels where physiological effects are expected (60–100 mg/d). The results (including a subgroup analysis of soya product consumers) showed that such levels are difficult to achieve with the European diets studied here.

There has been much recent interest in the cardiovascular benefits of dietary isoflavones. The aim of the present in vitro studies was to investigate potential anti-thrombogenic and anti-atherogenic effects of the isoflavones genistein and daidzein in platelets, macrophages and endothelial cells. Pre-treatment with either isoflavone inhibited collagen-induced platelet aggregation in a dose-dependent manner. In a macrophage cell line (RAW 264·7) activated with interferon γ plus lipopolysaccharide, both isoflavones were found to inhibit NO production and tumour necrosis factor α (TNF-α) secretion dose-dependently, but they did not affect mRNA levels for inducible nitric oxide synthase and cyclo-oxygenase-2. Both isoflavones also dose-dependently decreased monocyte chemoattractant protein-1 secretion induced by TNF-α in human umbilical vein endothelial cells. Compared with daidzein, genistein exerted greater inhibitory effects for all parameters studied. The present data contributes to our knowledge on the molecular mechanisms by which isoflavones may protect against coronary artery disease. Further studies are required to determine whether the effects of isoflavones observed in the current in vitro studies are relevant to the aetiology of coronary artery disease in vivo.

Food samples (n 114) were prepared from vegetables commonly eaten in Europe. The glycosidic forms of the phyto-oestrogens daidzein and genistein were extracted from the dried foods into aqueous methanol. The isoflavones were quantified by GC–MS after hydrolytic removal of any conjugated carbohydrate. Completeness of extraction and any procedural losses of the isoflavones were accounted for using synthetic daidzin (7-O-glucosyl-4′-hydroxyisoflavone) and genistin (7-O-glucosyl-4′5-dihydroxyisoflavone) as internal standards. Of the 114 foods assayed, at a limit of quantification of 0·1 μg/kg dry weight, forty-eight contained no detectable daidzein or genistein, forty-one contained less than 100 μg/kg of the two isoflavones combined and the remaining twenty-five contained more than this amount. Soyabean products contained between 470 and 1420 mg (average of 960 mg) daidzein and genistein combined per kg wet weight of food, and legumes contained between 20 and 5750 μg/kg wet weight of food, with an average of 620 μg/kg. Cooking by boiling in water caused a decrease in the daidzein and genistein content of food in twenty-four of twenty-eight foods. The extent of the decrease was variable and warrants further investigation. The present paper comprises the first comprehensive description of the content of daidzein and genistein in vegetables.