glycitein and daidzein

Cancer is a public health problem worldwide, and one of the crucial steps within tumor progression is the invasion and metastasis of cancer cells, which are directly related to cancer-associated deaths in patients. Recognizing the molecular markers involved in invasion and metastasis is essential to find targeted therapies in cancer. Interestingly, about 50% of the discovered drugs used in chemotherapy have been obtained from natural sources such as plants, including isoflavonoids. Until now, most drugs are used in chemotherapy targeting proliferation and apoptosis-related molecules. Here, we review recent studies about the effect of isoflavonoids on molecular targets and signaling pathways related to invasion and metastasis in cancer cell cultures, in vivo assays, and clinical trials. This review also reports that glycitein, daidzein, and genistein are the isoflavonoids most studied in preclinical and clinical trials and displayed the most anticancer activity targeting invasion-related proteins such as MMP-2 and MMP-9 and also EMT-associated proteins. Therefore, the diversity of isoflavonoids is promising molecules to be used as chemotherapeutic in invasive cancer. In the future, more clinical trials are needed to validate the effectiveness of the various natural isoflavonoids in the treatment of invasive cancer.

Topics: Apoptosis; Biomarkers; Clinical Trials as Topic; Flavones; Genistein; Humans; Isoflavones; Neoplasms

Phytoestrogens are naturally occurring nonsteroidal phenolic plant compounds that, due to their molecular structure and size, resemble vertebrate steroids estrogens. This review is focused on plant flavonoids isoflavones, which are ranked among the most estrogenic compounds. The main dietary sources of isoflavones for humans are soybean and soybean products, which contain mainly daidzein and genistein. When they are consumed, they exert estrogenic and/or antiestrogenic effects. Isoflavones are considered chemoprotective and can be used as an alternative therapy for a wide range of hormonal disorders, including several cancer types, namely breast cancer and prostate cancer, cardiovascular diseases, osteoporosis, or menopausal symptoms. On the other hand, isoflavones may also be considered endocrine disruptors with possible negative influences on the state of health in a certain part of the population or on the environment. This review deals with isoflavone classification, structure, and occurrence, with their metabolism, biological, and health effects in humans and animals, and with their utilization and potential risks.

Topics: Animals; Equol; Genistein; Humans; Isoflavones; Phytoestrogens

Isoflavones (genistein, daidzein, and glycitein) are bioactive compounds with mildly estrogenic properties and often referred to as phytoestrogen. These are present in significant quantities (up to 4-5 mg·g

Topics: Biological Availability; Diet; Fermentation; Food Analysis; Food Handling; Food Irradiation; Food Storage; Genistein; Hot Temperature; Humans; Infant Food; Isoflavones; Nutritive Value; Phytoestrogens; Randomized Controlled Trials as Topic; Soy Foods

This updated meta-analysis was performed to clarify the relationship between phytoestrogens and prostate cancer risk. Twenty one case-control and two cohort studies were finally selected for this meta-analysis, totaling 11,346 cases and 140,177 controls. Analytical results showed that daidzein (OR = 0.85; 95% CI: 0.75-0.96), genistein (OR = 0.87; 95% CI: 0.78-0.98), and glycitein (OR = 0.89; 95% CI: 0.81-0.98) were associated with a reduction of prostate cancer risk, but total isoflavones (OR = 0.93; 95% CI: 0.84-1.04), equol (OR = 0.86; 95% CI: 0.66-1.14), total lignans (OROgna.05; 95% CI: 0.54-2.04), secoisolariciresinol (OR = 1.02; 95% CI: 0.83-1.24), matairesinol (OR = 0.91; 95% CI: 0.75-1.11), enterolactone (OR = 0.94; 95% CI: 0.73-1.20), and coumestrol (OR = 0.89; 95% CI: 0.76-1.06) were not. Sensitivity and publication bias analyses demonstrated that the pooled estimates were stable and reliable. The results support the notion that some phytoestrogens may have a role in decreasing the risk of prostate cancer. Additional large and well-designed cohort studies are needed to confirm these relationships.

Topics: Case-Control Studies; Cohort Studies; Diet, Healthy; Evidence-Based Medicine; Genistein; Humans; Isoflavones; Male; Men's Health; Phytoestrogens; Prostatic Neoplasms; Risk

The impact of soyfood intake on breast cancer risk has been investigated extensively. Much of this focus can be attributed to the soybean being a dietary source that is uniquely rich in isoflavones. The chemical structure of isoflavones is similar to that of estrogen, and isoflavones bind to both estrogen receptors (ER alpha and ER beta) (although they preferentially bind to and activate ER beta) and exert estrogen-like effects under some experimental conditions. Isoflavones also possess nonhormonal properties that are associated with the inhibition of cancer cell growth. Thus, there are several possible mechanisms by which soy may reduce the risk of breast cancer. However, the role of isoflavones in breast cancer has become controversial because, in contrast to the possible beneficial effects, some data from in vitro and animal studies suggest that isoflavones, especially genistein, the aglycone of the main soybean isoflavone genistin, may stimulate the growth of estrogen-sensitive tumors. Limited human data directly address the tumor-promoting effects of isoflavones and soy. Because the use of soyfoods and isoflavone supplements is increasing, it is important from a public health perspective to understand the impact of these products on breast cancer risk in women at high risk of the disease and on the survival of breast cancer patients. To this end, a workshop was held in November 2005 to review the existing literature and to make research recommendations. This paper summarizes the workshop findings and recommendations. The primary research recommendation is that the impact of isoflavones on breast tissue needs to be evaluated at the cellular level in women at high risk for breast cancer.

Topics: Animals; Anticarcinogenic Agents; Breast Neoplasms; Carcinogens; China; Clinical Trials as Topic; Congresses as Topic; Disease Models, Animal; Female; Genistein; Humans; Isoflavones; Los Angeles; Neoplasms, Hormone-Dependent; Phytoestrogens; Population Surveillance; Receptors, Estrogen; Risk Factors; Soy Foods

Postmenopausal estrogen depletion is a major contributing factor to bone loss. Soy isoflavones have variable effects on the prevention of postmenopausal bone loss, which is possibly related to the specific isoflavone content or the variable equol-producing capacity of individuals.. We aimed to determine the effects of the content of isoflavones in a soy supplement and the equol-producing ability of the individual on postmenopausal bone calcium retention.. The study was a blinded, randomized, crossover intervention trial in 24 postmenopausal women who were prescreened for their ability to convert daidzein to equol. Women were equilibrated with (41)Ca before the intervention. Interventions were 5 soy isoflavone oral supplements (2 doses of a genistein-rich soy supplement and 3 doses of mixed isoflavones in various proportions) and a bisphosphonate (risedronate). Each intervention was given sequentially for 50 d followed by a 50-d washout period. The percentage of bone calcium retention was determined from the change in urinary (41)Ca:calcium.. Interventions that ranged from 52 to 220 mg total isoflavones/d increased bone calcium retention between 3.4% and 7.6% (P < 0.05), which was a moderate effect compared with that of risedronate at 15.3% (95% CI: 7.1%, 22.7%; P = 0.0014). The most-effective soy intervention delivered 105.23 mg total isoflavones/d as genistein, daidzein, and glycitein in their natural ratios and increased bone calcium retention by 7.6% (95% CI: 4.9%, 10.2%; P < 0.0001). Genistein, at 52.85 mg/d, increased bone calcium retention by 3.4% (95% CI: 0.5%, 6.2%; P = 0.029); but there was no benefit at higher amounts (113.52 mg/d). There was no difference (P = 0.5) in bone calcium retention between equol producers and nonproducers.. Soy isoflavones, although not as potent as risedronate, are effective bone-preserving agents in postmenopausal women regardless of their equol-producing status, and mixed isoflavones in their natural ratios are more effective than enriched genistein. This trial was registered at clinicaltrials.gov as NCT00244907.

Topics: Administration, Oral; Aged; Bone and Bones; Calcium; Cross-Over Studies; Dietary Supplements; Dose-Response Relationship, Drug; Equol; Female; Genistein; Glycine max; Humans; Isoflavones; Middle Aged; Osteoporosis, Postmenopausal; Phytoestrogens; Postmenopause; Risedronic Acid

Isoflavones are present in soy foods and soy-based supplements. Despite low plasma isoflavone concentrations in the general Western population, concentrations in supplement users exceed those suggested to be beneficial for health in Asian populations, raising concerns for adverse effects. To aid risk assessment, quantification of the relation between isoflavone intake and plasma concentrations is essential.. Plasma samples were collected from postmenopausal women in three placebo-controlled crossover studies with 8-week periods for supplements (two studies, ~100 mg isoflavones/day, n=88) or 4-week periods for soy foods (one study, ~48 mg isoflavones/day, n=15). Plasma isoflavone concentrations (daidzein, equol, genistein and glycitein) were quantified using high-performance liquid chromatography and electrochemical detection. The association between plasma concentrations and isoflavone intake, equol producer status, intake-producer interaction and background dietary intake was assessed based on the assumption of a log-linear relation.. Median plasma total isoflavone concentrations after the soy food and supplement interventions were respectively 2.16 and 3.47 μmol/l for equol producers and 1.30 and 2.39 μmol/l for non-producers. Regression analysis showed that doubling isoflavone intake increased plasma concentrations by 55-62% (±s.e. 1-2%, R(2)>0.87) for daidzein, genistein, equol (only for producers) and total isoflavones; for glycitein the association was weaker (15±1%, R(2)=0.48). Adjustments for energy, carbohydrate and fat intake did not affect these estimates. Inter-individual variation, estimated based on repeated measures in one of the studies, was 30-96%.. Although the relation between isoflavone intake and plasma concentrations was adequately quantified, the use of isoflavone intake data for risk assessment needs caution due to large inter-individual variation in plasma concentrations.

Topics: Aged; Asian People; Cross-Over Studies; Dietary Supplements; Equol; Female; Genistein; Humans; Isoflavones; Middle Aged; Postmenopause; Risk Assessment; Soy Foods

Isoflavone supplements, consumed by women experiencing menopausal symptoms, are suggested to have positive effects on menopause-related adiposity and cardiovascular disease risk profile, but discussions about their safety are still ongoing.. The objective was to study the effects of an 8-wk consumption of 2 different isoflavone supplements compared with placebo on whole-genome gene expression in the adipose tissue of postmenopausal women.. This double-blind, randomized, placebo-controlled crossover intervention consisted of 2 substudies, one with a low-genistein (LG) supplement (56% daidzein + daidzin, 16% genistein + genistin, and 28% glycitein + glycitin) and the other with a high-genistein (HG) supplement (49% daidzein + daidzin, 41% genistein + genistin, and 10% glycitein + glycitin). Both supplements provided ∼ 100 mg isoflavones/d (aglycone equivalents). After the 8-wk isoflavone and placebo period, whole-genome arrays were performed in subcutaneous adipose tissue of postmenopausal women (n = 26 after LG, n = 31 after HG). Participants were randomized by equol-producing phenotype, and data analysis was performed per substudy for equol producers and nonproducers separately.. Gene set enrichment analysis showed downregulation of expression of energy metabolism-related genes after LG supplementation (n = 24) in both equol-producing phenotypes and oppositely regulated expression for equol producers (down) and nonproducers (up) after HG supplementation (n = 31). Expression of inflammation-related genes was upregulated in equol producers but downregulated in nonproducers, independent of supplement type. Only 4.4-7.0% of the genes with significantly changed expression were estrogen responsive. Body weight, adipocyte size, and plasma lipid profile were not affected by isoflavone supplementation.. Effects of isoflavones on adipose tissue gene expression were influenced by supplement composition and equol-producing phenotype, whereas estrogen-responsive effects were lacking. LG isoflavone supplementation resulted in a caloric restriction-like gene expression profile for both producer phenotypes and pointed toward a potential beneficial effect, whereas both supplements induced anti-inflammatory gene expression in equol producers. The study was registered at clinicaltrials.gov as NCT01556737.

Topics: Adipose Tissue; Adiposity; Aged; Cross-Over Studies; Dietary Supplements; Double-Blind Method; Equol; Female; Gene Expression; Genistein; Humans; Isoflavones; Middle Aged; Netherlands; Nutritional Status; Postmenopause; Surveys and Questionnaires

Seaweed and soy foods are consumed daily in Japan, where breast cancer rates for postmenopausal women are significantly lower than in the West. Likely mechanisms include differences in diet, especially soy consumption, and estrogen metabolism. Fifteen healthy postmenopausal women participated in this double-blind trial of seaweed supplementation with soy challenge. Participants were randomized to 7 wk of either 5 g/d seaweed (Alaria) or placebo (maltodextrin). During wk 7, participants also consumed a daily soy protein isolate (2 mg isoflavones/kg body weight). After a 3-wk washout period, participants were crossed over to the alternate supplement schedule. There was an inverse correlation between seaweed dose (mg/kg body weight) and serum estradiol (E2) (seaweed-placebo = y = -2.29 x dose + 172.3; r = -0.70; P = 0.003), [corrected] which was linear across the range of weights. Soy supplementation increased urinary daidzein, glycitein, genistein, and O-desmethylangolensin (P = 0.0001) and decreased matairesinol and enterolactone (P < 0.05). Soy and seaweed plus soy (SeaSoy) increased urinary excretion of 2-hydroxyestrogen (2-OHE) (P = 0.0001) and the ratio of 2-OHE:16alpha-hydroxyestrone (16alphaOHE(1)) (P = 0.01). For the 5 equol excretors, soy increased urinary equol excretion (P = 0.0001); the combination of SeaSoy further increased equol excretion by 58% (P = 0.0001). Equol producers also had a 315% increase in 2:16 ratio (P = 0.001) with SeaSoy. Seaweed favorably alters estrogen and phytoestrogen metabolism and these changes likely include modulation of colonic bacteria.

Topics: Aged; Cross-Over Studies; Diet; Dietary Supplements; Double-Blind Method; Equol; Estradiol; Estrogens; Female; Genistein; Humans; Isoflavones; Middle Aged; Phytoestrogens; Placebos; Postmenopause; Seaweed; Sex Hormone-Binding Globulin; Soybean Proteins

The aim of this study was to determine whether equol excretion status and plasma hormone and leptin concentrations can be influenced by consumption of a probiotic supplement. A secondary focus was to investigate whether male equol excretors have a hormone profile consistent with reduced prostate cancer risk.. The design was a randomized, single-blinded, placebo-controlled, parallel-arm trial.. Thirty-one (31) of the initially enrolled 39 subjects, 18 to 37 years old, completed all study requirements.. Subjects consumed either probiotic capsules (containing Lactobacillus acidophilus and Bifidobacterium longum) or placebo capsules for 2 months. Fasting plasma concentrations of testosterone (T), dihydrotestosterone (DHT), androstanediol glucuronide (AAG), androstenedione (A), dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG), and leptin were measured on days 1 and 57. Urinary excretion of genistein, glycitein, daidzein, O-desmethylangolensin (O-Dma), and equol was measured on days 4 and 61 following a 4-day soy challenge.. Probiotic consumption did not significantly alter equol excretor status, plasma hormone, or leptin concentrations in these subjects. At baseline, there were no differences in plasma hormone concentrations between equol excretors and nonexcretors; however, the low number of equol excretors included in this study limits the strength of this finding.. The 2-month intervention with probiotic capsules did not significantly alter equol excretion, plasma hormone, or leptin concentrations in these subjects. A secondary finding was that male equol excretors in this study did not exhibit a hormone profile consistent with reduced prostate cancer risk, although this result should be interpreted with caution.

Topics: Adrenal Cortex Hormones; Adult; Androstane-3,17-diol; Androstenedione; Bifidobacterium; Dehydroepiandrosterone Sulfate; Dihydrotestosterone; Equol; Genistein; Hormones; Humans; Isoflavones; Lactobacillus acidophilus; Leptin; Male; Phytoestrogens; Pilot Projects; Probiotics; Sex Hormone-Binding Globulin; Single-Blind Method; Testosterone

We investigated the effects of consuming an isoflavone aglycone-enriched soya milk containing viable bifidobacteria on urinary isoflavone excretion and percentage recovery. Sixteen postmenopausal women were randomly divided into two groups to consume either fermented or non-fermented soya milk. Each group participated in a double-blind, crossover study with three 14 d supplementation periods, separated by a 14 d washout. Subjects ingested three daily dosages of isoflavone via the soya milk and collected four 24 h pooled urine specimens per supplementation period. Soya milks were prepared with soya protein isolate and soya germ, followed by fermentation with bifidobacteria. Isoflavone levels were quantified using HPLC. Non-fermented soya milks at 20, 40 and 80 mg isoflavone/200 ml contained 10 %, 9 % and 7 % aglycone, respectively, with their fermented counterparts containing 69 %, 57 % and 36 % aglycone (P<0.001). A trend to a greater percentage urinary recovery of daidzein and glycitein was observed among women consuming fermented soya milk at a dosage of 40 mg isoflavone (P=0.13). A distinct linear dose response for the fermented soya milk group (R2=0.9993) compared with the non-fermented group (R2=0.8865) suggested less interindividual variation in isoflavone absorption. However, total urinary isoflavone excretion was similar for both groups (P>0.05), with urinary isoflavone recovery at approximately 31 %. Increasing the isoflavone dosage correlated positively with its urinary excretion, but urinary percentage recovery of isoflavone was inversely related to dosage level. Hence, a modest dosage ranging from 20 to 30 mg/d may provide the most bioavailable source of isoflavone, regardless of whether it is via an aglycone-rich fermented soya milk or a glucoside-rich soya milk.

Topics: Bifidobacterium; Biological Availability; Body Mass Index; Body Weight; Cross-Over Studies; Double-Blind Method; Eating; Enzyme Inhibitors; Female; Fermentation; Genistein; Humans; Isoflavones; Middle Aged; Phytoestrogens; Postmenopause; Probiotics; Soy Milk

To determine the effects of diets rich in soy and linseed compared with a control diet on biochemical markers of prostate cancer in men diagnosed with prostate cancer.. Twenty-nine men diagnosed with prostate cancer and scheduled to undergo a radical prostatectomy were randomized to one of three groups: soy (high phytoestrogen), soy and linseed (high phytoestrogen), or wheat (low phytoestrogen). A bread was specially manufactured to incorporate 50 g of heat-treated (HT) soy grits or 50 g of HT soy grits and 20 g of linseed as part of the study participant's daily diet. Baseline and preoperative levels of prostate-specific antigen (PSA), free PSA, testosterone, sex hormone-binding globulin, free androgen index, and dihydrotestosterone were measured.. Statistically significant differences were detected between the HT soy grits group and the control wheat group for the percentage of change in total PSA (-12.7% versus 40%, P = 0.02) and the percentage of change in free/total PSA ratio (27.4% versus -15.6%, P = 0.01); and between the HT soy grits group and the HT soy grits and linseed group for the percentage of change in free androgen index (16.4% versus -15.5%, P = 0.04) and the percentage of change in free/total PSA ratio (27.4% versus -10%, P = 0.007).. The data from this study indicate that a daily diet containing four slices of a bread rich in HT soy grits favorably influences the PSA level and the free/total PSA ratio in patients with prostate cancer. This work provides some evidence to support epidemiologic studies claiming that male populations who consume high phytoestrogen diets have a reduced risk of prostate cancer development and progression.

Topics: Adenocarcinoma; Aged; Androgens; Biomarkers, Tumor; Bread; Dihydrotestosterone; Double-Blind Method; Flax; Genistein; Gonadal Steroid Hormones; Humans; Isoflavones; Male; Middle Aged; Neoplasm Proteins; Phytoestrogens; Phytotherapy; Prostate-Specific Antigen; Prostatic Neoplasms; Sex Hormone-Binding Globulin; Soy Foods; Testosterone

Genistein may be useful in the prevention or treatment of prostate cancer; however, it causes genetic damage in cultured human cells.. The objective was to assess the potential genotoxicity of a purified soy unconjugated isoflavone mixture in men with prostate cancer.. Twenty patients with prostate cancer were treated with 300 mg genistein/d for 28 d and then with 600 mg/d for another 56 d. In peripheral lymphocytes, DNA strand breaks were assessed as nuclear tail moment, chromosomal damage was assessed as micronucleus frequency (MF), and translocations of the MLL gene (11q23) were assessed by using fluorescence in situ hybridization. Values are also reported for 6 healthy men. The studies were performed under Investigational New Drug application no. 54 137 at a tertiary referral academic medical center.. No changes in group average or individual nuclear tail moment and MF were observed. We observed a single elevated MF value in one subject that exceeded a clinical threshold set before we initiated the study. A significant decrease in average COMET tail moment was observed on day 28 relative to day 0. We detected no genistein-induced rearrangements of the MLL gene in the 3 subjects we studied with this technique. MF increased significantly in lymphocytes exposed in vitro to unconjugated genistein at concentrations > or = 100 micromol/L. Total genistein never exceeded a peak concentration of 27.1 micro mol/L, and unconjugated genistein never exceeded a peak concentration of 0.32 micromol/L.. Although isoflavones are capable of inducing genetic damage in vitro, a similar effect was not observed in subjects treated with a purified soy unconjugated isoflavone mixture.

Topics: Adult; Cells, Cultured; Comet Assay; DNA Damage; DNA-Binding Proteins; Gene Deletion; Genistein; Glycine max; Histone-Lysine N-Methyltransferase; Humans; In Situ Hybridization, Fluorescence; Isoflavones; Lymphocytes; Male; Micronuclei, Chromosome-Defective; Myeloid-Lymphoid Leukemia Protein; Prostatic Neoplasms; Proto-Oncogenes; Transcription Factors; Translocation, Genetic

To determine the effect of soy-derived isoflavones on hot flushes, endometrial thickness, and the vascular reactivity of uterine and cerebral arteries.. Double-blind, randomized, placebo-controlled trial.. Healthy volunteers in an academic research environment.. Sixty-two postmenopausal women aged 45-60 years attending the Outpatient Menopause Clinic of our gynecological departments.. The patients were administered 72 mg of soy-derived isoflavones or placebo under double-blind conditions. The daily number of hot flushes was recorded in a diary. Endometrial thickness was measured by means of transvaginal ultrasound; the uterine, internal carotid, and middle cerebral arteries were evaluated using Doppler ultrasound.. The daily number of hot flushes, endometrial thickness, and arterial pulsatility index (PI).. Both treatments led to a 40% reduction in the number of hot flushes. Soy-derived isoflavones had no effect on endometrial thickness or the PI of the uterine and cerebral arteries.. The daily administration of 72 mg of soy-derived isoflavones is no more effective than placebo in reducing hot flushes in postmenopausal women. It also has no effect on endometrial thickness or the PI of the uterine and cerebral arteries.

Topics: Cerebral Arteries; Double-Blind Method; Endometrium; Female; Genistein; Glycine max; Hot Flashes; Humans; Isoflavones; Middle Aged; Regional Blood Flow; Uterus

Soy isoflavones are being evaluated as chemopreventive agents for breast and other cancers.. The objective was to perform safety and pharmacokinetic studies of purified unconjugated isoflavone preparations containing genistein, daidzein, and glycitein in postmenopausal women.. Twenty-four healthy postmenopausal women ingested a single dose of 1 of 2 purified (from soybeans) isoflavone preparations that delivered a genistein dose of 2, 4, 8, or 16 mg/kg body wt. These doses were higher than those previously administered to human females. Toxicity studies were performed 24 h and 3, 6, 14, and 30 d after isoflavone administration. Kinetic studies were performed during the first 24 h.. We observed a 7% decrease in systolic and diastolic blood pressure and a 32% decrease in the neutrophil count 24 h after treatment with formulation A. Isolated episodes of nausea, pedal edema, and breast tenderness were judged to be possibly related to the study treatment. The terminal plasma half-lives for free genistein, daidzein, and glycitein averaged 3.8, 7.7, and 3.4 h, respectively. The terminal pseudo half-lives for total genistein and total daidzein in plasma averaged 10.1 and 10.8 h, respectively. The estimated bioavailabilities of both total genistein and total daidzein from each of the 2 formulations were not significantly different.. A single-dose administration of purified unconjugated isoflavones at amounts that exceed normal dietary intakes had minimal clinical toxicity in healthy postmenopausal women. The pharmacokinetic data suggest that chronic dosing at 12-24-h intervals would not lead to progressive accumulation of these isoflavones.

Topics: Aged; Blood Pressure; Dose-Response Relationship, Drug; Drug Combinations; Female; Genistein; Glycine max; Half-Life; Humans; Isoflavones; Middle Aged; Postmenopause

Soy isoflavones are potential cancer chemoprevention treatments.. We conducted safety studies of purified unconjugated genistein, daidzein, and glycitein, and defined pharmacokinetic parameters for their absorption and metabolism.. Thirty healthy men ingested a single dose of 1 of 2 isoflavone preparations purified from soy. The delivered doses of genistein (1, 2, 4, 8, or 16 mg/kg body wt) were higher than those previously administered to humans. Formulation A was composed of 90 +/- 5% genistein, 10% daidzein, and 1% glycitein. Formulation B was composed of 43% genistein, 21% daidzein, and 2% glycitein.. We observed no clinically significant behavioral or physical changes after treatment. We observed elevations in lipoprotein lipase and hypophosphatemia that were possibly related to the treatment but that were associated with no clinical toxicity. Considerable quantities of isoflavones were excreted in urine as conjugates. The terminal elimination rate, elimination half-life, area under the curve, maximum plasma concentration, apparent systemic clearance, and volume of distribution were estimated for genistein and daidzein. The mean elimination half-lives with both formulations were 3.2 h for free genistein and 4.2 h for free daidzein. The mean pseudo half-lives were 9.2 h for total genistein and 8.2 h for total daidzein.. Dietary supplements of purified unconjugated isoflavones administered to humans in single doses exceeding normal dietary intake manyfold resulted in minimal clinical toxicity. Genistein and daidzein (free and total) were rapidly cleared from plasma and excreted in urine.

Topics: Adult; Anticarcinogenic Agents; Estrogens, Non-Steroidal; Genistein; Glycine max; Half-Life; Humans; Isoflavones; Male

Soybean products containing isoflavones are widely consumed in Western and Asian diets for putative health benefits, but adverse effects are also possible. The conjugated forms of isoflavones present in a soy nutritional supplement (predominately acetyl glucosides) and in blood from two human volunteers after consuming the supplement (7- and 4'-glucuronides and sulfates) were identified using liquid chromatography coupled with electrospray/tandem mass spectrometry. Circulating conjugates of genistein and daidzein were quantified using selective enzymatic hydrolysis and deuterated internal standards for liquid chromatography-electrospray/mass spectrometry. The levels of isoflavone glucuronides were much greater than the corresponding sulfates or aglycones. The substrate activities of genistein and daidzein were evaluated with recombinant human UDP glucuronosyl transferase (UGT) and sulfotransferase (SULT) by using enzyme kinetics. The SULTs 1A1*2, 1E, and 2A1 catalyzed formation of a single genistein sulfate; however, SULTs 1A2*1 and 1A3 had no observed activity. None of the SULTs showed activity with daidzein. Although several UGTs (1A1, 1A4, 1A6, 1A7, 1A9, and 1A10) catalyzed 7- and 4'-glucuronidation of genistein or daidzein, the UGT 1A10 isoform, which is found in human colon but not liver, was found to be specific for genistein. Glucuronidation of only genistein was observed in human colon microsomes, although nearly equal activity was observed for daidzein in human liver and kidney microsomes. These findings suggest a prominent role for glucuronidation of genistein in the intestine concomitant with absorption, although hepatic glucuronidation of absorbed genistein and daidzein aglycones is also likely.

Topics: Animals; Antineoplastic Agents; Cattle; Chromatography, Liquid; Female; Genistein; Glucuronides; Glucuronosyltransferase; Glycine max; Glycosides; Humans; Isoflavones; Kinetics; Male; Mass Spectrometry; Microsomes; Substrate Specificity; Sulfates; Sulfotransferases

Effects of the isoflavone supplement on hormonal states in young premenopausal women were studied by cross-over study design. Administration of 20 mg or 40 mg isoflavones (IF) by tablets, of which 1 g contained 43.5 mg daidzein, 6.0 mg genistein, 24.0 mg glycitein, to 40 young female students for one month caused a prolonged menstruation in 60% of young women, shortened menstruation in 20% of young women, 17% remained unchanged and 3% became irregular. Larger dose tended to elongate more, but 17beta-estradiol levels in both follicular and luteal phages were not different between 20 mg and 40 mg isoflavone intake. Equor excreters tended to show low plasma progesteron level in the luteal phase. Detailed hormonal analysis on 3 of students by a cross over study design showed decreased level of 17beta-estradiol throughout the menstruation cycle. SHBG significantly increased about 10% in all three. DEAS, androstendione, and testosterone showed different responses according to the follicular or luteal phase. T3 and T4 increased as a result of isoflavone tablet administration in the follicular phase, but it decreased in the luteal phase. These changes suggest that isoflavones influence not only estrogen receptor-related functions but the hypothalamo-hypophysis-gonadal axis.

Topics: Androstenedione; Cross-Over Studies; Dehydroepiandrosterone Sulfate; Diet Records; Dietary Supplements; Estradiol; Female; Follicular Phase; Genistein; Glycine max; Hormones; Humans; Hypocotyl; Isoflavones; Luteal Phase; Menstrual Cycle; Progesterone; Testosterone; Thyroxine; Triiodothyronine

To analyze the impact of soy protein dietary supplements containing phytoestrogens on menopausal symptoms in healthy postmenopausal women.. A double-blind, placebo-controlled trial was conducted in 94 healthy postmenopausal women aged 50-75 years, with 44 randomized to soy supplements containing 118 mg of isoflavones (daidzein, genistein, glycitein and their respective glycosides), and 50 to an identically presented casein placebo. A validated questionnaire on menopausal symptoms was administered at baseline and at 3 months of treatment. Compliance was assessed by high-performance liquid chromatography assay of urinary phytoestrogens. Statistical analysis was completed using non-parametric statistical methods and multivariate analysis.. At baseline 80% of women recruited were experiencing menopausal symptoms, although symptom severity was mild. Those consuming phytoestrogen supplements had 13- and 17-fold increases in urinary excretion of genistein and daidzein, respectively, with no change in the placebo group. Active soy supplements did not significantly alter either individual symptoms or specific symptom category scores when compared to placebo. Within-group comparisons revealed that the active group reported a significant improvement in vaginal dryness (p = 0.01), libido (p = 0.009), facial hair (p = 0.04) and dry skin (p = 0.027). However, similarly, those on placebo reported an improvement in libido (p = 0.015), facial hair (p = 0.014) and dry skin (p = 0.011) but not vaginal dryness.. In this group of 94 older postmenopausal women with a high frequency of mild menopausal symptoms, 3 months of soy supplements containing phytoestrogens did not provide symptomatic relief compared with placebo.

Topics: Aged; Double-Blind Method; Estrogens, Non-Steroidal; Female; Genistein; Hirsutism; Humans; Isoflavones; Libido; Menopause; Middle Aged; Phytoestrogens; Placebos; Plant Preparations; Postmenopause; Skin Diseases; Soybean Proteins; Surveys and Questionnaires; Treatment Outcome; Vaginal Diseases

Isoflavones in soybeans are well-known phytoestrogens. Soy isoflavones present in conjugated forms are converted to aglycone forms during processing and storage. Isoflavone aglycones (IFAs) of soybeans in human diets have poor solubility in water, resulting in low bioavailability and bioactivity. Enzyme-mediated glycosylation is an efficient and environmentally friendly way to modify the physicochemical properties of soy IFAs. In this study, we determined the optimal reaction conditions for

Topics: beta-Glucans; Biological Availability; Chromatography, High Pressure Liquid; Deinococcus; Escherichia coli; Genetic Vectors; Genistein; Glucosyltransferases; Glycine max; Glycosylation; Isoflavones; Mass Spectrometry; Phytoestrogens; Plant Extracts

Isoflavones intake is associated with health benefits. The metabolism of isoflavones by bacteria plays a key role in their biotransformation. Therefore, commercial soy drink was fermented by 11 lactic acid bacteria (LAB) and 9 bifidobacteria strains. The majority of the strains showed deglycosylation of the isoflavone glycosides present in soy drink and appearance of the aglycones daidzein, genistein and glycitein. Moreover, we observed the further transformation of daidzein into O-desmethylangolensin (O-DMA) and tetrahydrodaidzein, alongside with dihydrodaidzein (DHD) and a putative isomer of DHD. On the other hand, genistein was transformed by nearly all strains into 6-hydroxy-O-desmethylangolensin (6-hydroxy-O-DMA), but no dihydrogenistein production was registered. A high concentration of 2-(4-hydroxyphenyl)-propionic acid was observed, suggesting the degradation of O-DMA and 6-hydroxy-O-DMA. The potential of LAB and Bifidobacterium strains to produce functional soy drink enriched with bioactive isoflavones is demonstrated in this work.

Topics: Bifidobacterium; Fermented Foods; Genistein; Humans; Isoflavones; Lactobacillales; Propionates; Soy Milk

Efficient aquaculture is depending on sustainable protein sources. The shortage in marine raw materials has initiated a shift to "green aquafeeds" based on staple ingredients such as soy and wheat. Plant-based diets entail new challenges regarding fish health, product quality and consumer risks due to the possible presence of chemical contaminants, natural toxins and bioactive compounds like phytoestrogens. Daidzein (DAI), genistein (GEN) and glycitein (GLY) are major soy isoflavones with considerable estrogenic activities, potentially interfering with the piscine endocrine system and affecting consumers after carry-over. In this context, information on isoflavone biotransformation in fish is crucial for risk evaluation. We have therefore isolated hepatic fractions of Atlantic salmon (Salmo salar), the most important species in Norwegian aquaculture, and used them to study isoflavone elimination and metabolite formation. The salmon liver microsomes and primary hepatocytes were characterized with respect to phase I cytochrome P450 (CYP) and phase II uridine-diphosphate-glucuronosyltransferase (UGT) enzyme activities using specific probe substrates, which allowed comparison to results in other species. DAI, GEN and GLY were effectively cleared by UGT. Based on the measurement of exact masses, fragmentation patterns, and retention times in liquid chromatography high-resolution mass spectrometry, we preliminarily identified the 7-O-glucuronides as the main metabolites in salmon, possibly produced by UGT1A1 and UGT1A9-like activities. In contrast, the production of oxidative metabolites by CYP was insignificant. Under optimized assay conditions, only small amounts of mono-hydroxylated DAI were detectable. These findings suggested that bioaccumulation of phytoestrogens in farmed salmon and consumer risks from soy-containing aquafeeds are unlikely.

Topics: Animals; Aquaculture; Biotransformation; Chromatography, Liquid; Genistein; Glucuronides; Glucuronosyltransferase; Glycine max; Hepatocytes; Isoflavones; Microsomes, Liver; Phytoestrogens; Salmo salar; UDP-Glucuronosyltransferase 1A9

Soy isoflavones are popular ingredients with anti-adipogenic and anti-lipogenic properties. The anti-adipogenic and anti-lipogenic properties of genistein are well-known, but those of genistin and glycitein remain unknown, and those of daidzein are characterized by contrasting data. Therefore, the purpose of our study was to investigate the effects of daidzein, glycitein, genistein, and genistin on adipogenesis and lipogenesis in 3T3-L1 cells. Proliferation of 3T3-L1 preadipocytes was unaffected by genistin and glycitein, but it was affected by 50 and 100 µM genistein and 100 µM daidzein for 48 h. Among the four isoflavones, only 50 and 100 µM genistin and genistein markedly suppressed lipid accumulation during adipogenesis in 3T3-L1 cells through a similar signaling pathway in a dose-dependent manner. Genistin and genistein suppress adipocyte-specific proteins and genes, such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT-enhancer-binding protein α (C/EBPα), and adipocyte binding protein 2 (aP2)/fatty acid-binding protein 4 (FABP4), and lipogenic enzymes such as ATP citrate lyase (ACL), acetyl-CoA carboxylase 1 (ACC1), and fatty acid synthase (FAS). Both isoflavones also activate AMP-activated protein kinase α (AMPKα), an essential factor in adipocyte differentiation, and inhibited sterol regulatory element-binding transcription factor 1c (SREBP-1c). These results indicate that genistin is a potent anti-adipogenic and anti-lipogenic agent.

Topics: 3T3-L1 Cells; Acetyl-CoA Carboxylase; Adipocytes; Adipogenesis; AMP-Activated Protein Kinases; Animals; Anti-Obesity Agents; ATP Citrate (pro-S)-Lyase; CCAAT-Enhancer-Binding Protein-alpha; Cell Survival; Fatty Acid Synthases; Fatty Acid-Binding Proteins; Gene Expression Regulation; Glycine max; Isoflavones; Lipogenesis; Mice; PPAR gamma; Sterol Regulatory Element Binding Protein 1

Soybean (Glycine max L.) is a good source of natural antioxidants and commonly consumed as fermented products such as cheonggukjang, miso, tempeh, and sufu in Asian countries. The aim of the current study was to examine the influence of novel endophytic bacterial strain,

Topics: Amino Acids; Antioxidants; Bacillus amyloliquefaciens; Fermentation; Food Hypersensitivity; Genistein; Glycine max; Isoflavones; Nutritive Value; Phenols

Soy isoflavones are bioactive phytoestrogens with known health benefits. Soybean embryo extract (SEE) has been consumed as a source of isoflavones, mainly daidzein, glycitein, and genistein. While previous studies have reported the anti-obesity effects of SEE, this study investigates their molecular mechanisms and the synergistic effects of co-treatment with SEE and enzymatically modified isoquercitrin (EMIQ). SEE upregulated genes involved in lipolysis and brown adipocyte markers and increased mitochondrial content in differentiated C3H10T1/2 adipocytes in vitro. Next, we use a high-fat diet-induced obesity mouse model to determine the anti-obesity effect of SEE. Two weeks of single or combined treatment with SEE and EMIQ significantly reduced body weight gain and improved glucose tolerance. Mechanistically, SEE treatment increased mitochondrial content and upregulated genes involved in lipolysis in adipose tissue through the cAMP/PKA-dependent signaling pathway. These effects required a cytosolic lipase adipose triglyceride lipase (ATGL) expression, confirmed by an adipocyte-specific ATGL knockout mouse study. Collectively, this study demonstrates that SEE exerts anti-obesity effects through the activation of adipose tissue metabolism and exhibits a synergistic effect of co-treatment with EMIQ. These results improve our understanding of the mechanisms underlying the anti-obesity effects of SEE related to adipose tissue metabolism.

Topics: Adipocytes; Adipose Tissue; Animals; Cell Differentiation; Diet, High-Fat; Genistein; Glycine max; Humans; Isoflavones; Lipolysis; Mice; Obesity; Phytoestrogens; Plant Extracts; Quercetin; Seeds

Topics: Animals; Estradiol; Estrogens; Female; Genistein; Glycine max; Inflammation; Isoflavones; Ovary; Oxidative Stress; Phytoestrogens; Polycystic Ovary Syndrome; Rats

Lactobacillus rhamnosus strain ASCC 1520 with high soy isoflavone transformation ability was used to ferment soymilk and added to the diet of mice. The impact of L. rhamnosus fermentation on soy isoflavone metabolites and intestinal bacterial community, in conjunction with fecal enzyme activity and short-chain fatty acids (SCFA) excretion was evaluated. Antibiotics intervention resulted in a decrease in fecal enzyme activities and SCFA. Although long-term intake of soymilk or L. rhamnosus-fermented soymilk did not affect the fecal β-glucuronidase and β-galactosidase activities, it improved the β-glucosidase activity when antibiotics were concomitantly administered. Soymilk or fermented soymilk administration increased the isoflavone metabolites (O-DMA and equol) excreted in urine. Antibiotics decreased the daidzein excretion and its metabolites but showed little effect on glycitein and genistein excretion. Principal coordinates analysis (PCoA) of the 16s rRNA gene sequencing data found a remarkable shift in gut microbiota after soymilk administration and antibiotics treatment. Matastats test of the relative abundance of bacterial taxa revealed Odoribacter (Bacteroidales family), Lactobacillus (Lactobacillales order), and Alistipes (Rikenellaceae family) were enriched in soymilk while bacterial taxa from Bacteroides and Lactobacillus were enriched in L. rhamnosus-fermented soymilk. Furthermore, there was less decrease in bacterial taxa with fermented soymilk group even when antibiotics were concomitantly administered. Overall, this study revealed that the gut microbiota of a healthy host is enough for the whole isoflavone metabolism under normal conditions. Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used. PRACTICAL APPLICATION: Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used.

Topics: Animals; Bacteria; beta-Glucosidase; Fatty Acids, Volatile; Feces; Fermentation; Food Microbiology; Gastrointestinal Microbiome; Genistein; Isoflavones; Lactobacillales; Male; Mice; Mice, Inbred BALB C; Soy Milk

Isoflavones have been reported to stimulate muscle growth. The aim of this

Topics: Anabolic Agents; Animals; Cell Line; Drug Therapy, Combination; Genistein; Glycine max; In Vitro Techniques; Insulin-Like Growth Factor I; Isoflavones; Mice; Muscle Fibers, Skeletal; Myoblasts; Myosin Heavy Chains; Plant Extracts; Receptor, IGF Type 1

S-Equol is a metabolite of daidzein, a type of soy isoflavone, and three reductases are involved in the conversion of daidzein by specific intestinal bacteria. S-Equol is thought to prevent hormone-dependent diseases. We previously identified the equol producing gene cluster (eqlABC) of Eggerthella sp. YY7918. Daidzein reductase (DZNR), encoded by eqlA, catalyzes the reduction of daidzein to dihydrodaidzein (the first step of equol synthesis), which was confirmed using a recombinant enzyme produced in Escherichia coli. Here, we purified recombinant DZNR to homogeneity and analyzed its enzymological properties. DZNR contained FMN, FAD, and one 4Fe-4S cluster per 70-kDa subunit as enzymatic cofactors. DZNR reduced the CC bond between the C-2 and C-3 positions of daidzein, genistein, glycitein, and formononetin in the presence of NADPH. R-Dihydrodaidzein and R-dihydrogenistein were highly stereo-selectively produced from daidzein and genistein. The K

Topics: Actinobacteria; Coenzymes; Equol; Escherichia coli; Flavin Mononucleotide; Flavin-Adenine Dinucleotide; Genistein; Humans; Iron; Isoflavones; NADP; Oxidoreductases; Oxidoreductases Acting on CH-CH Group Donors; Protein Multimerization; Protein Structure, Quaternary; Protein Subunits; Stereoisomerism; Sulfur

Topics: Anticarcinogenic Agents; Apoptosis; Breast Neoplasms; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Female; Glycine max; Humans; Isoflavones; MCF-7 Cells

Doenjang (DJ), a fermented soybean product used in soups, stews, and sauces, contains high quality proteins, fats, vitamins, minerals, and other functional ingredients, including isoflavones and saponins. This study investigated whether DJ improves the bioavailability of isoflavones compared to boiled soybean (BS) in sham-operated or ovariectomized (OVX) rats. We also examined the effects of ovariectomy on the differences in bioavailability of isoflavones. BS and DJ were administered in sham-operated and OVX rats, and blood samples were collected. Twenty-six isoflavone-derived metabolites were identified. Pharmacokinetic analysis revealed that T

Topics: Animals; Biological Availability; Estrogens; Female; Fermentation; Fermented Foods; Genistein; Glycine max; Isoflavones; Ovariectomy; Rats; Rats, Sprague-Dawley; Soy Foods

Black soybeans are rich in isoflavones, which have several beneficial health effects. In this study, a validated method based on UHPLC-MS/MS was developed to screen black-soybean metabolites in rat urine, bile, and plasma and to quantify the compounds (daidzein, genistein, glycitein, and daidzin) and their metabolites (daidzein-4'-β-d-glucuronide, genistein-7-β-d-glucuronide, and genistein-4'-β-d-glucuronide) in plasma. Thirty-seven compounds were tentatively detected in the biological samples. The method was fully validated in quantitative experiments, including in assessments of linearity (2.5-100 ng/mL for daidzein, genistein, and glycitein; 10-100 ng/mL for daidzin; 5-3125 ng/mL for genistein-7-β-d-glucuronide; and 5-1562.5 ng/mL for daidzein-4'-β-d-glucuronide and genistein-4'-β-d-glucuronide), matrix effects (85-115%), recovery (80-105%), precision (<10%), and accuracy (<10%). The compounds were stable throughout sample storage, treatment, and analysis. The method was first applied to detect IFs and metabolites in rats after oral administration of black-soybean extract. These results support the potential of this method for successful application in pharmacokinetic studies.

Topics: Animals; Bile; Chromatography, High Pressure Liquid; Genistein; Glucuronides; Glycine max; Isoflavones; Male; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity; Tandem Mass Spectrometry

The effect of supplementing a basal diet for dairy cows with "Soybean extract 40" (Biomedica, Prague, Czech Republic), containing 40% soybean isoflavones, on the contents of daidzein, glycitein, genistein, and equol in milk as well as fresh and mature yogurts was estimated. To determine the contents of these isoflavonoids, an efficient analytical LC-MS (TOF) technique was used. The "Soybean extract 40" used in our study contained an especially high proportion of daidzein (307gkg

Topics: Animals; Cattle; Chromatography, Liquid; Equol; Glycine max; Isoflavones; Mass Spectrometry; Milk; Yogurt

1. Soybean is a common source of protein in many pet foods. Slow glucuronidation of soy-derived isoflavones in cats has been hypothesized to result in accumulation with adverse health consequences. Here, we evaluated species' differences in soy isoflavone glucuronidation using urine samples from cats and dogs fed a soy-based diet and liver microsomes from cats compared with microsomes from 12 other species. 2. Significant concentrations of conjugated (but not unconjugated) genistein, daidzein and glycitein, and the gut microbiome metabolites, dihydrogenistein and dihydrodaidzein, were found in cat and dog urine samples. Substantial amounts of conjugated equol were also found in cat urine but not in dog urine. 3. β-Glucuronidase treatment showed that all these compounds were significantly glucuronidated in dog urine while only daidzein (11%) and glycitein (37%) showed any glucuronidation in cat urine suggesting that alternate metabolic pathways including sulfation predominate in cats. 4. Glucuronidation rates of genistein, daidzein and equol by cat livers were consistently ranked within the lowest 3 out of 13 species' livers evaluated. Ferret and mongoose livers were also ranked in the lowest four species. 5. Our results demonstrate that glucuronidation is a minor pathway for soy isoflavone metabolism in cats compared with most other species.

Topics: Animals; Cats; Dogs; Equol; Estradiol; Ferrets; Genistein; Glucuronidase; Glycine max; Herpestidae; Isoflavones; Liver; Microsomes, Liver; Species Specificity

Isoflavone profile is greatly affected by heating process. However, kinetic analyses of isoflavone conversion and degradation using a continuous industry processing method have never been characterized. In this study, Proto soybean was soaked and blanched at 80 °C for 2 min and then processed into soymilk, which underwent UHT (ultra-high temperature) at 135 to 150 °C for 10 to 50 s with a pilot plant-scale Microthermics processor. The isoflavone profile was determined at different time/temperature combinations. The results showed that all isoflavone forms exhibited distinct changing patterns over time. In the soymilk under UHT conditions, the degradation (disappearance) of malonyldaizin and malonylgenistin exhibited first-order kinetics with activation energies of 59 and 84 kj/mole, respectively. At all UHT temperatures, malonylgenistin showed higher rate constants than malonyldaidzin. However, malonylglycitin changed irregularly under these UHT temperatures. The increase of genistin, daidzin, glycitein and acetlydaidzin during heating demonstrated zero-order kinetics and the rate constants increased with temperature except for the conditions of 145 to 150 °C for 50 s. Overall, genistein series exhibited higher stability than daidzein series. Under all UHT conditions, total isoflavone decreased from 12% to 24%.

Topics: Food Handling; Glucosides; Glycine max; Hot Temperature; Humans; Isoflavones; Kinetics; Soy Milk

Isoflavones may play a role in the prevention of hormone-related cancers. Equol is an isoflavone metabolized from daidzein in the presence of certain intestinal bacteria. Slackia sp. strain NATTS, a newly identified equol-producing bacterium, was recently isolated from human feces in Japan. We investigated the association of serum levels and dietary intake of isoflavones and Slackia sp. strain NATTS with the risk of prostate cancer in a case-control study among Japanese men.. Fifty-six patients with newly diagnosed prostate cancer and 56 hospital controls were enrolled in this study. Isoflavones were assessed by measurement of serum levels and administration of a food frequency questionnaire. Slackia sp. strain NATTS in feces was also measured. The odds ratios (ORs) and 95 % confidence intervals (CIs) for prostate cancer were then determined using a logistic regression model.. The adjusted ORs for prostate cancer in comparison with the highest to lowest categories were 0.06 (95 % CI 0.02-0.24) for serum genistein, 0.18 (95 % CI 0.06-0.52) for daidzein, 0.16 (95 % CI 0.06-0.46) for glycitein, 0.52 (95 % CI 0.22-1.22) for equol, 0.86 (95 % CI 0.30-2.48) for dietary genistein, and 0.80 (95 % CI 0.28-2.28) for dietary daidzein. The adjusted OR for prostate cancer in comparison with values above versus below the median was 0.95 (95 % CI 0.42-2.16) for Slackia sp. strain NATTS.. Our study findings suggest that high serum levels of genistein, daidzein, and glycitein are significantly associated with a decreased risk of prostate cancer among Japanese men.

Topics: Actinobacteria; Aged; Case-Control Studies; Diet; Equol; Feces; Genistein; Humans; Isoflavones; Japan; Male; Middle Aged; Odds Ratio; Prostatic Neoplasms; Surveys and Questionnaires

In this research, a microbial endophytic strain obtained from the rhizosphere of the conifer Taxus baccata and designated as Streptomyces sp. AC35 (FJ001754.1 Streptomyces, GenBank) was investigated. High 16S rDNA gene sequence similarity suggests that this strain is closely related to S. odorifer. The major fatty acid profile of intracellular lipids was also carried out to further identify this strain. Atomic force microscopy and scanning acoustic microscopy were used to image our strain. Its major excreted substances were extracted, evaluated for antimicrobial activity, purified, and identified by ultraviolet-visible spectroscopy (UV-vis), liquid chromatography-mass spectrometry (LC-MS/MS) and nuclear magnetic resonance as the bioactive isoflavone aglycones-daidzein, glycitein and genistein. Batch cultivation, performed under different pH conditions, revealed enhanced production of antimycin components when the pH was stable at 7.0. Antimycins were detected by HPLC and identified by UV-vis and LC-MS/MS combined with the multiple reaction monitoring. Our results demonstrate that Streptomyces sp. AC35 might be used as a potential source of effective, pharmaceutically active compounds.

Topics: Antimycin A; Genistein; Isoflavones; Streptomyces

To ascertain the association between soya consumption, isoflavone intakes and oesophageal cancer risk in remote north-west China, where the incidence of oesophageal cancer is known to be high.. Case-control study. Information on habitual consumption of soya foods and soya milk was obtained by personal interview. The intakes of isoflavones were then estimated using the US Department of Agriculture nutrient database. Logistic regression analyses were performed to assess the association between soya consumption, isoflavone intakes and oesophageal cancer risk.. Urumqi and Shihezi, Xinjiang Uyghur Autonomous Region, China.. Participants were 359 incident oesophageal cancer patients and 380 hospital-based controls.. The oesophageal cancer patients consumed significantly less (P < 0·001) total soya foods (mean 57·2 (sd 119·0) g/d) and soya milk (mean 18·8 (sd 51·7) ml/d) than the controls (mean 93·3 (sd 121·5) g/d and mean 35·7 (sd 73·0) ml/d). Logistic regression analyses showed an inverse association between intake of soya products and the risk of oesophageal cancer. The adjusted odds were OR = 0·33 (95 % CI 0·22, 0·49) and OR = 0·48 (95 % CI 0·31, 0·74) for consuming at least 97 g of soya foods and 60 ml of soya milk daily (the highest tertiles of consumption), respectively, relative to the lowest tertiles of consumption. Similarly, inverse associations with apparent dose-response relationships were found between isoflavone intakes and oesophageal cancer risk.. Habitual consumption of soya products appears to be associated with reduced risk of oesophageal cancer in north-west China.

Topics: Aged; Anticarcinogenic Agents; Antioxidants; Case-Control Studies; China; Diet; Esophageal Neoplasms; Female; Genistein; Humans; Incidence; Isoflavones; Logistic Models; Male; Middle Aged; Risk; Soy Foods; Soy Milk

To examine the association between soybean products and risk of type 2 diabetes, we measured four isoflavone biological markers--genistein, daidzein, glycitein and equol--in a nested case-control study.. The study population was composed of 693 cases (316 women and 377 men) and 698 matched controls (317 women and 381 men) within the Korean Genome and Epidemiology Study. The concentrations of isoflavone biomarkers were measured using HPLC-MS/MS on plasma samples that were collected at baseline. A stratified analysis was undertaken to examine the association between plasma isoflavone concentrations and risk of type 2 diabetes according to sex and equol production. Logistic regression models were used to compute ORs and 95% CIs adjusted for confounders.. In women, compared with the lowest quartile of plasma concentration of genistein, the highest quartile exhibited a significantly decreased risk of diabetes (OR 0.58, 95% CI 0.35, 0.95). When stratified by equol-producing status in women, the OR for diabetes in the highest vs the lowest quartile of genistein concentration was 0.31 (95% CI 0.16, 0.60) in equol producers, but genistein concentration was not associated with risk of diabetes in equol non-producers (p for interaction = 0.013). In men, isoflavone concentrations were not associated with risk of diabetes, regardless of equol-producing status.. High plasma concentrations of genistein were associated with a decreased risk of type 2 diabetes in women. This inverse association was prominent in equol-producing participants. These results suggest a beneficial effect of a high intake of soybean products on risk of type 2 diabetes in women.

Topics: Asian People; Case-Control Studies; Chi-Square Distribution; Chromatography, High Pressure Liquid; Diabetes Mellitus, Type 2; Diet; Equol; Female; Genistein; Genome, Human; Humans; Isoflavones; Logistic Models; Male; Middle Aged; Odds Ratio; Prospective Studies; Protective Factors; Republic of Korea; Risk Assessment; Risk Factors; Sex Factors; Soy Foods; Tandem Mass Spectrometry

There is a growing interest in the pharmaceutical field concerning isoflavones topical delivery systems, especially with regard to their skin care properties and antiherpetic activity. In this context, the present work describes an ultra-fast liquid chromatography method (UFLC) for determining daidzein, glycitein, and genistein in different matrices during the development of topical systems containing isoflavone aglycones (IA) obtained from soybeans. The method showed to be specific, precise, accurate, and linear (0.1 to 5 µg mL(-1)) for IA determination in soybean acid extract, IA-rich fraction obtained after the purification process, IA loaded-nanoemulsions, and topical hydrogel, as well as for permeation/retention assays in porcine skin and porcine esophageal mucosa. The matrix effect was determined for all complex matrices, demonstrating low effect during the analysis. The stability indicating UFLC method was verified by submitting IA to acidic, alkaline, oxidative, and thermal stress conditions, and no interference of degradation products was detected during analysis. Mass spectrometry was performed to show the main compounds produced after acid hydrolysis of soybeans, as well as suggest the main degradation products formed after stress conditions. Besides the IA, hydroxymethylfurfural and ethoxymethylfurfural were produced and identified after acid hydrolysis of the soybean extract and well separated by the UFLC method. The method's robustness was confirmed using the Plackett-Burman experimental design. Therefore, the new method affords fast IA analysis during routine processes, extract purification, products development, and bioanalytical assays.

Topics: Administration, Topical; Animals; Biological Transport; Chromatography, High Pressure Liquid; Esophagus; Furaldehyde; Genistein; Glycine max; Hydrogels; Hydrolysis; Isoflavones; Mucous Membrane; Permeability; Plant Extracts; Skin; Swine

This study examined the macronutrients, phytochemicals, and antioxidant activities of yellow soybean sprout (YSS) and green soybean sprout (GSS) with different germination days. YSS and GSS were obtained by sprouting soybean in darkness or with light exposure at 21 °C. Lipid, protein, carbohydrate, and ash contents were analyzed before and after soybean germination. Phytochemicals (total phenolic compounds, saponin, and isoflavone) were also determined. DPPH, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC) were determined to examine the antioxidant activities of soybean sprout. Results showed YSS had a higher yield than GSS. Based on dry mass composition, 7-d germination of GSS decreased 14% protein, 37% lipid, 22% carbohydrate, and 16% ash, whereas 7-d germination of YSS decreased 6% protein and 47% lipid. Carbohydrate did not change and ash significantly increased for the 7-d germinated YSS. Lipid was greatly metabolized in germination, which explained why the protein relative percentage in dried soybean sprout was higher than that in the corresponding soybean. Total phenolic compounds and saponin (mg/g soybean sprout, dry basis) had the same accumulation trend in soybean sprout with the increases in germination days. Aglycone isoflavones (genistein, glycitein, and daidzein) and daidzin showed an increased trend, whereas malonylgenistin and malonylglycitin showed a decreased trend with germination days for both GSS and YSS. The change in other isoflavones did not show definite trends. GSS had 20% more antioxidant activities than YSS (7-d germinated soybean sprout). The increases in ORAC antioxidant activity suggest eating GSS may be more beneficial than GSS for promoting human health.

Topics: Antioxidants; Biphenyl Compounds; Carbohydrates; Color; Germination; Glucosides; Glycine max; Isoflavones; Light; Lipids; Phenol; Phenols; Phytochemicals; Picrates; Proteins; Saponins

The aim of this study was to examine the association of serum isoflavones, adiponectin, and insulin levels with ovarian cancer risk.. We gathered cases with histologically confirmed epithelial ovarian cancer at Sapporo Medical University Hospital from October 2010 to September 2012. Potential controls were recruited from female inpatients without any history of cancer or diabetes mellitus in different wards of the same hospital over the same period of time. Serum isoflavones, adiponectin, and insulin levels were measured in order to estimate associations with ovarian cancer risk in a case-control study. Data from 71 cases and 80 controls were analyzed with a logistic regression model adjusting for known risk factors.. A significant reduction in ovarian cancer risk was observed for the high tertile of serum daidzein level versus the low (Ptrend<0.001). A significant reduction in ovarian cancer risk was also observed for the high tertile of serum glycitein level versus the low (Ptrend=0.005). Furthermore, a significant reduction in ovarian cancer risk was observed for the high tertile of serum adiponectin level versus the low (Ptrend=0.004). Conversely, serum insulin level showed significantly elevated risk for ovarian cancer with the high tertile versus the low Ptrend<0.001).. Decreased serum isoflavones levels, such as those for daidzein and glycitein, decreased serum adiponectin levels, and increased serum insulin levels could be shown to be associated with elevated risk of ovarian cancer.

Topics: Adiponectin; Carcinoma, Ovarian Epithelial; Case-Control Studies; Female; Humans; Insulins; Isoflavones; Logistic Models; Middle Aged; Neoplasms, Glandular and Epithelial; Ovarian Neoplasms; Risk Factors

Isoflavones from soybeans [ Glycine max (L.) Merr.] have a significant impact on human health to reduce the risk of several major diseases. Breeding soybean for high isoflavone content in the seed is possible through marker-assisted selection (MAS) which can be based on quantitative trait loci (QTL). The objective of this study was to identify QTL controlling isoflavone content in a set of 'MD96-5722' by 'Spencer' recombinant inbred line (RIL) populations of soybean. Wide variations were found for seed concentrations of daidzein, glycitein, genistein, and total isoflavones among the RIL populations. Three QTL were identified on three different linkage groups (LG) represented by three different chromosomes (Chr). One QTL that controlled daidzein content was identified on LG A1 (Chr 5), and two QTL that underlay glycitein content were identified on LG K (Chr 9) and LG B2 (Chr 14). Identified QTL could be functional in developing soybean with preferable isoflavone concentrations in the seeds through MAS.

Topics: Glycine max; Inbreeding; Isoflavones; Polymorphism, Single Nucleotide; Quantitative Trait Loci; Seeds

Malonyl isoflavone glucosides are water-soluble derivatives of soybean hypocotyls. This study compared the hydrolysis and absorption of malonyl isoflavone glucosides and nonmalonyl isoflavone glucosides in rats. Plasma concentrations of isoflavones were measured after oral administration of malonyl isoflavone glucosides or isoflavone glucosides. After fasting, the duodenum, jejunum, ileum, and colon were excised, and homogenates were prepared. The extent of hydrolysis of each glucoside by each intestinal homogenate was measured. Plasma levels of isoflavone aglycones, such as daidzein and glycitein, were higher in rats administered malonyl isoflavone glucosides than in those administered isoflavone glucosides. The area under the curve of daidzein in plasma of rats administered malonyl isoflavone glucosides was also significantly greater than that in those administered isoflavone glucosides. A transport experiment using Caco-2 cells suggested that degradation of malonyl glucosides to aglycones is necessary for intestinal absorption. Malonyl isoflavone glucosides were hydrolyzed only in the colon, whereas hydrolysis of isoflavone glucosides occurred in the jejunum, ileum, and colon. These results indicated more effective absorption of malonyl isoflavone glucosides than of nonmalonyl isoflavone glucosides. Moreover, effective absorption of malonyl isoflavone aglycones in the colon contributed to the significant increase in plasma isoflavone levels.

Topics: Animals; Area Under Curve; Biological Transport; Caco-2 Cells; Colon; Female; Glucosides; Glycine max; Humans; Hydrolysis; Hypocotyl; Intestinal Absorption; Intestinal Mucosa; Isoflavones; Malonates; Rats; Rats, Wistar; Tissue Distribution

Soybeans and other legumes investigated as fishmeal replacements in aquafeeds contain phytoestrogens capable of binding to and activating estrogen receptors. Estradiol has catabolic effects in salmonid white muscle, partially through increases in protein turnover. The current study determines whether phytoestrogens promote similar effects. In rainbow trout (Oncorhynchus mykiss) primary myocyte cultures, the phytoestrogens genistein, daidzein, glycitein, and R- and S-equol reduced rates of protein synthesis and genistein, the phytoestrogen of greatest abundance in soy, also increased rates of protein degradation. Increased expression of the ubiquitin ligase fbxo32 and autophagy-related genes was observed with high concentrations of genistein (100 μM), and R- and S-equol (100 μM) also up-regulated autophagy-related genes. In contrast, low genistein concentrations in vitro (0.01-0.10 μM) and in vivo (5 μg/g body mass) decreased fbxo32 expression, suggesting a potential metabolic benefit for low levels of genistein exposure. Phytoestrogens reduced cell proliferation, indicating that effects of phytoestrogens extend from metabolic to mitogenic processes. Co-incubation of genistein with the estrogen receptor (ER) antagonist, ICI 182,780, ameliorated effects of genistein on protein degradation, but not protein synthesis or cell proliferation, indicating that effects of genistein are mediated through ER-dependent and ER-independent mechanisms. Collectively, these data warrant additional studies to determine the extent to which dietary phytoestrogens, especially genistein, affect physiological processes that impact growth and nutrient retention.

Topics: Animals; Cell Proliferation; Cells, Cultured; Equol; Female; Gene Expression; Genistein; Isoflavones; Muscle Cells; Muscle Proteins; Muscles; Oncorhynchus mykiss; Phytoestrogens; Protein Biosynthesis; Proteolysis; Receptors, Estrogen

To assess dietary isoflavone intake between population and hospital outpatient controls and examine if cancer risks estimated for isoflavone using hospital outpatient controls would be different from those using population controls. Three parallel case-control studies on leukemia, breast, and colorectal cancers in China in 2009-2010 were conducted, using population and hospital outpatient controls to separately match 560 incident cases at a 1:1 ratio. A validated food frequency questionnaire was administered by face-to-face interview. Conditional logistic regression analysis was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). The 2 control groups had closely similar distributions of dietary isoflavone intake. Risk estimates for breast cancers were adjusted ORs (95% CI) of 0.39 (0.23-0.66) and 0.31 (0.18-0.55) for daidzein, 0.35 (0.20-0.61) and 0.28 (0.16-0.52) for genistein, 0.66 (0.41-1.08) and 0.53 (0.32-0.88) for glycitein, and 0.53 (0.33-0.85) and 0.43 (0.26-0.71) for total isoflavone using hospital outpatient and population controls respectively. The study found that hospital outpatient controls were comparable to population controls in measured dietary intake of isoflavone in the Chinese hospital setting.

Topics: Adult; Aged; Breast Neoplasms; Case-Control Studies; China; Colorectal Neoplasms; Diet; Female; Genistein; Humans; Isoflavones; Leukemia; Male; Middle Aged; Neoplasms; Odds Ratio; Outpatient Clinics, Hospital; Risk Factors

To evaluate whether soybean extracts and estrogens present additive effects on adult rat uterus.. Fifty ovariectomized rats were randomly divided into five equal groups of ten animals: Control, treated with vehicle; SE46 and SE120, treated with 46 and 120 mg/kg soybean concentrated extract (SE), respectively; EE, treated with conjugated equine estrogens (CE) 50 μg/kg; SE120 + EE, treated with 50 μg/kg (CE) plus 120 mg/kg SE. The substances were administered daily by gavage for 21 consecutive days. Thereafter the animals were weighed and killed by decapitation; trunk blood was collected for hormone determinations. Uteri were removed immediately and fixed in 10% formaldehyde, followed by dehydration, embedding in paraffin and 6-m sections staining with hematoxylin and eosin for histomorphometric analyses of myometrium and endometrium. After ANOVA analysis of the data, the study was complemented with the Tukey-Kramer test for multiple comparisons.. The concentrated extract of soybean at high concentration (SE 120 kg/mg) and estrogens proved to have a trophic effect on the uterus (endometrium and myometrium) of castrated rats. In groups SE120, EE and SE120 + EE, all morphometric parameters examined (number of glands, eosinophils, blood vessels and the glandular area) were increased. No significant addictive effects of soybean extract plus estrogens were detected in the SE120 + EE group.. Our results indicate that soy extract has a trophic effect on rat uterine structures. Treatment of ovariectomized rats with a concentrated soy extract in combination with conjugated estrogens had no addictive effect on the uterine response.

Topics: Analysis of Variance; Animals; Endometrium; Estradiol; Estrogens; Estrogens, Conjugated (USP); Female; Genistein; Glycine max; Isoflavones; Myometrium; Organ Size; Ovariectomy; Phytoestrogens; Plant Extracts; Progesterone; Rats; Uterus

The antioxidant activity and contents of various polyphenol classes in the seeds of seven soybean varieties of different seed color and one yellow seed cultivar, representing a reference genotype, were evaluated. Total polyphenols and tannins were determined after extraction of plant material with 70% aqueous acetone, and total flavonoids were extracted with methanol and acetic acid, whereas anthocyanins were extracted with 20% aqueous ethanol. In addition, isoflavone content and composition were determined using high-performance liquid chromatography analysis. Antioxidant activity of seed extracts was evaluated by the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity assay. A positive linear correlation between antioxidant activity and contents of total polyphenols and anthocyanins was established. The highest antioxidant activity was observed in the extracts of black and brown varieties, which also showed high levels of all polyphenol classes examined. Yellow seed had the highest total isoflavone content (3.62 mg/g of dry material). The highest concentration of total daidzein was determined in black seeds (>2.0 mg/g of dry material), and the highest total glycitein and genistein contents occurred in the yellow cultivar (0.53 and 1.49 mg/g of dry material, respectively). According to our results, varieties of black and brown seeds could be of special interest not only for their large content of total polyphenols, ranging from 4.94 to 6.22 mg of gallic acid equivalents/g of dry material, but also for their high content of natural antioxidants such as anthocyanins.

Topics: Anthocyanins; Antioxidants; Biphenyl Compounds; Chromatography, High Pressure Liquid; Color; Genistein; Glycine max; Isoflavones; Picrates; Plant Extracts; Polyphenols; Seeds; Species Specificity; Tannins

Phytochemicals found in soy and other legumes have been speculated to reduce the risk of endometrial cancer; however, inconsistent findings have been reported in the few epidemiological studies conducted to date.. We conducted a prospective analysis of 46 027 nonhysterectomized postmenopausal women who were recruited into the Multiethnic Cohort (MEC) Study between August 1993 and August 1996 and provided detailed baseline information on diet and other endometrial cancer risk factors. A total of 489 women diagnosed with incident endometrial cancer were identified through the Surveillance, Epidemiology, and End Results tumor registry linkages during a median follow-up period of 13.6 years. Cox proportional hazards models were used to estimate multivariable-adjusted relative risks (RRs) and 95% confidence intervals (CIs) for endometrial cancer associated with dietary intake of legumes, soy, and tofu, and for total isoflavones and specific isoflavones (daidzein, genistein, or glycitein). Truncated (age 50-89 years) age-adjusted incidence rates were calculated by applying age-specific rates within isoflavone quintiles to the overall MEC population eligible for endometrial cancer. To estimate the percentage of endometrial cancers that may have been prevented by consuming the highest quintile of total isoflavones, the partial population attributable risk percent was calculated.. A reduced risk of endometrial cancer was associated with total isoflavone intake (highest vs lowest quintile, ≥7.82 vs <1.59 mg per 1000 kcal/d, RR = 0.66, 95% CI = 0.47 to 0.91), daidzein intake (highest vs lowest quintile, ≥3.54 vs <0.70 mg per 1000 kcal/d, RR = 0.64, 95% CI = 0.46 to 0.90), and genistein intake (highest vs lowest quintile, ≥3.40 vs <0.69 mg per 1000 kcal/d, RR = 0.66, 95% CI = 0.47 to 0.91). No statistically significant association with endometrial cancer risk was observed for increasing intake of legumes, soy, tofu, or glycitein. Truncated age-adjusted incidence rates of endometrial cancer for the highest vs lowest quintile of total isoflavone intake were 55 vs 107 per 100 000 women per year, respectively. The partial population attributable risk percent for total isoflavone intake lower than the highest quintile was 26.7% (95% CI = 5.3% to 45.8%).. This study suggests that greater consumption of isoflavone-containing foods is associated with a reduced risk of endometrial cancer in this population of nonhysterectomized postmenopausal women.

Topics: Aged; Aged, 80 and over; Cohort Studies; Endometrial Neoplasms; Fabaceae; Feeding Behavior; Female; Genetic Predisposition to Disease; Genistein; Glycine max; Growth Inhibitors; Humans; Incidence; Isoflavones; Life Style; Middle Aged; Phytoestrogens; Postmenopause; Proportional Hazards Models; Prospective Studies; Risk Assessment; Risk Factors; SEER Program; Soy Foods; Surveys and Questionnaires; United States

The effectiveness of soy isoflavones to prevent bone loss in postmenopausal women is controversial. While consumption of soy in Vietnam is very high, we recently reported a prevalence of osteoporosis comparable to that of many Western populations. In the present study, we analyzed the isoflavone content of soy drink products commercially available in Vietnam and Sweden, and we also compared these products to "home-made" soy drink from beans of different origin. The amounts of the bioactive aglycones (daidzein, glycitein and genistein) and their glycoside isomers were quantified by high-pressure liquid chromatography. We found that the total isoflavone content was low in all preparations, around 70-100 mg/L and of this only 10% were bioactive aglycones. Of these, the Vietnamese products contained significantly lower levels of glycitein than the products from Sweden and "home-made" soy drink preparations. The results show that consumption of several liters of soy drink per day would be needed to achieve threshold levels for a protective effect on bone. There was no significant association between total protein and isoflavone content in different products. Accurate labeling of soy drink and other products eg of aglycone and glycoside content would allow health professionals and researchers to better explore the possible benefits of soy in dietary intervention studies.

Topics: Analysis of Variance; Chromatography, High Pressure Liquid; Genistein; Isoflavones; Soy Milk; Soybean Proteins; Sweden; Vietnam

To understand the relationship between the role of soy isoflavones and estrogen receptor (ER)-β in colon tumorigenesis, we investigated the cellular effects of soy isoflavones (composed of genistein, daidzein, and glycitein) in DLD-1 human colon adenocarcinoma cells with or without ER-β gene silencing by RNA interference (RNAi). Soy isoflavones decreased the expression of proliferating cell nuclear antigen (PCNA), extracellular signal-regulated kinase (ERK)-1/2, AKT, and nuclear factor (NF)-κB. Soy isoflavones dose-dependently caused G2/M cell cycle arrest and downregulated the expression of cyclin A. This was associated with inhibition of cyclin dependent kinase (CDK)-4 and up-regulation of its inhibitor p21(cip1) expressions. ER-β gene silencing lowered soy isoflavone-mediated suppression of cell viability and proliferation. ERK-1/2 and AKT expressions were unaltered and NF-κB was modestly upregulated by soy isoflavones after transient knockdown of ER-β expression. Soy isoflavone-mediated arrest of cells at G2/M phase and upregulation of p21(cip1) expression were not observed when ER-β gene was silenced. These findings suggest that maintaining the expression of ER-β is crucial in mediating the growth-suppressive effects of soy isoflavones against colon tumors. Thus upregulation of ER-β status by specific food-borne ER-ligands such as soy isoflavones could potentially be a dietary prevention or therapeutic strategy for colon cancer.

Topics: Adenocarcinoma; Anticarcinogenic Agents; Cell Cycle; Cell Proliferation; Colonic Neoplasms; Cyclin-Dependent Kinase Inhibitor p21; Estrogen Receptor beta; Genistein; Humans; Isoflavones; NF-kappa B

Isoflavones are among the major bioactive compounds found in a wide variety of plant-derived foods, especially in soybeans and soy-based foods. In this study, the effect of a soy-derived isoflavone mixture (designated as SI-I, containing 71% daidzein, 14.3% genistein and 14.7% glycitein) on HeLa cells and its mechanism were investigated. SI-I in concentration range 5-80 μg/ml significantly reduced the survival rate of HeLa cells by MTT assay, whereas showed no side effect on that of L929 cells. After HeLa cells were exposed to 10, 20 and 40 μg/ml SI-I for 4 days, typical apoptotic morphological changes, including nuclear fragmentation, cytoplasm shrinkage and decrease of cell volume, were observed by fluorescence microscope and CLSM, respectively. FCM analysis revealed that the percentages of early apoptotic cells with lost Δψm increased by 2.27, 2.74 and 4.05 folds respectively, compared with control. The results showed that SI-I inhibited HeLa cell growth through inducing apoptosis via the mitochondrial pathway and comparisons with reported data indicated that synergistic effect existed between the isoflavone species contained in SI-I. It is proposed that natural soy-derived isoflavones are potential candidates as chemotherapeutic agents against human cervical cancer.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Proliferation; Female; Genistein; Glycine max; HeLa Cells; Humans; Isoflavones; Membrane Potential, Mitochondrial; Microscopy, Fluorescence; Mitochondria; Uterine Cervical Neoplasms

There has been great interest in the potential beneficial and adverse health effects of dietary isoflavones. Determination of tissue concentrations of isoflavone metabolites provides an insight into the potential bioactivity of dietary isoflavones. However, data on the distribution of isoflavones in animal models fed dietary isoflavones are limited. In this study, additional data on the distribution of isoflavones in serum and/or tissues of rats and pigs fed dietary isoflavones were generated.. Rats (male and female) were fed a casein control diet (containing no isoflavones) and an isoflavone-supplemented diet (containing an alcohol-washed soy protein isolate plus NOVASOY, providing a total of 1,047 mg/kg of total isoflavones). Female pigs were fed a control diet (without soy) containing 17.5 mg/kg of isoflavones, a soy diet containing 582.8 mg/kg of isoflavones or a soy diet supplemented with a daily dose of 2.3 g (equivalent to 42.0 and 14.5 mg/kg of body weight at the onset and end of treatment, respectively) of crystalline genistein. The concentrations of isoflavones in serum and tissues (liver and mammary gland) and in tissues (liver and mammary gland) of pigs were determined via a sensitive and rapid method using liquid chromatography/mass spectrometry.. Rats fed the control diet containing no isoflavones had nondetectable levels of isoflavone metabolites in serum, liver and mammary gland samples. Rats fed the isoflavone-supplemented diet had the greatest levels of equol, followed by genistein, daidzein and glycitein, respectively, in their serum, livers and mammary glands. The concentrations of total isoflavones (daidzein, equol and genistein plus glycitein) in serum were significantly (p < 0.05) greater in male rats vs. female rats, but the reverse was true in the case of livers. Concentrations of daidzein, equol, genistein and glycitein were lowest (p < 0.05) in the livers of pigs fed the control diet, and in the mammary glands of female pigs there was only an effect of feeding soy plus genistein on the concentrations of daidzein and equol (p <0.05).. The current data therefore show gender as well as species differences in the tissue distribution of isoflavones.

Topics: Animals; Body Weight; Chromatography, Liquid; Diet; Dietary Supplements; Equol; Female; Genistein; Isoflavones; Liver; Male; Mammary Glands, Animal; Mass Spectrometry; Rats; Rats, Sprague-Dawley; Soybean Proteins; Swine

Our hypothesis in this study was that in vitro disappearance of isoflavones from fecal or cecal contents of Golden Syrian hamsters paralleled the apparent absorption of these compounds, comparable with previous findings from in vitro human fecal incubations. Two studies were conducted to test this idea: one on in vitro fecal (study 1, n = 20/sex) and the other on in vitro cecal contents (study 2, n = 10/sex) ability to degrade isoflavones. According to HPLC analysis, urinary isoflavone excretion was significantly less by 2-4 fold in males compared with females in both studies. Fecal isoflavone excretion was not significantly different between sexes or isoflavones (study 1) and was <0.5% of ingested dose. In vitro anaerobic fecal isoflavone degradation rate constants from study 1 were minimal with no significant correlation between urinary and fecal isoflavone excretion. However, in vitro anaerobic cecal isoflavone degradation rate constants (study 2) were greater and significantly correlated with urinary excretion of daidzein (R = 0.90; p = 0.01) and genistein (R = 0.93; p = 0.004), but not glycitein (R = 0.50; p = 0.3). Both male and female hamsters showed a pattern of urinary isoflavone excretion similar to that found in humans (daidzein > genistein). Hamster in vitro cecal isoflavone degradation rate constants seemed to be analogous to human in vitro fecal isoflavone degradation rate constants for genistein and daidzein. The sex difference in isoflavone excretion in hamsters and the instability in glycitein excretion across studies coupled with the paucity of human data on this isoflavone deserve further investigation.

Topics: Absorption; Anaerobiosis; Animals; Biological Availability; Cecum; Cricetinae; Feces; Female; Genistein; Isoflavones; Male; Mesocricetus; Regression Analysis

Seeds of soybean [Glycine max (L.) Merr.] accumulate more isoflavones than any tissue of any plant species. In other plant parts, isoflavones are usually released to counteract the effects of various biotic and abiotic stresses. Because of the benefits to the plant and positive implications that consumption may have on human health, increasing isoflavones is a goal of many soybean breeding programs. However, altering isoflavone levels through marker-assisted selection (MAS) has been impractical due to the small and often environmentally variable contributions that each individual quantitative trait locus (QTL) has on total isoflavones. In this study, we developed a Magellan × PI 437654 F(7)-RIL population to construct a highly saturated non-redundant linkage map that encompassed 451 SNP and SSR molecular markers and used it to locate genomic regions that govern accumulation of isoflavones in the seeds of soybean. Five QTLs were found that contribute to the concentration of isoflavones, having single or multiple additive effects on isoflavone component traits. We also validated a major locus which alone accounted for up to 10% of the phenotypic variance for glycitein, and 35-37% for genistein, daidzein and the sum of all three soybean isoflavones. This QTL was consistently associated with increased concentration of isoflavones across different locations, years and crosses. It was the most important QTL in terms of net increased amounts of all isoflavone forms. Our results suggest that this locus would be an excellent candidate to target for MAS. Also, several minor QTLs were identified that interacted in an additive-by-additive epistatic manner, to increase isoflavone concentration.

Topics: Analysis of Variance; Chromosome Mapping; Crosses, Genetic; Epistasis, Genetic; Genetic Linkage; Genetic Loci; Genetic Variation; Genistein; Glycine max; Inbreeding; Isoflavones; Phenotype; Quantitative Trait Loci; Recombination, Genetic; Seeds

Phytoestrogens have attracted attention as being safer alternatives to hormone replacement therapy (HRT) and as chemopreventive reagents for breast cancer because dietary soy isoflavone intake has been correlated with reduction in risk. To identify safe and effective phytoestrogen candidates for HRT and breast cancer prevention, we investigated the effects of daidzein, genistein, coumestrol, resveratrol and glycitein on cell growth, cell cycle, cyclin D1 expression, apoptosis, Bcl-2/Bax expression ratio and p53-dependent or NF-kappaB-dependent transcriptional activity in MCF-7 breast cancer cells. Phytoestrogens, except for glycitein, significantly enhanced estrogen-response-element-dependent transcriptional activity up to a level similar to that of 17beta-estradiol (E(2)). E(2) increased cell growth significantly, coumestrol increased cell growth moderately, and resveratrol and glycitein reduced cell growth. Phytoestrogens, except for glycitein, stimulated the promotion of cells to G(1)/S transition in cell cycle analysis, similar to E(2). This stimulation was accompanied by transient up-regulation of cyclin D1. While genistein, resveratrol and glycitein all increased apoptosis and reduced the Bcl-2/Bax ratio, resveratrol reduced this ratio more than either genistein or glycitein. Moreover, resveratrol significantly enhanced p53-dependent transcriptional activity, but slightly reduced NF-kappaB-dependent transcriptional activity. On knockdown analysis, genistein, resveratrol and glycitein all reduced the Bcl-2/Bax ratio in the presence of apoptosis-inducing stimuli, and estrogen receptor (ER) alpha silencing had no effect on these reductions. In contrast, in the absence of apoptosis-inducing stimuli, only resveratrol reduced the ratio, and ERalpha silencing abolished this reduction. Thus, resveratrol might be the most promising candidate for HRT and chemoprevention of breast cancer due to its estrogenic activity and high antitumor activity.

Topics: Apoptosis; bcl-2-Associated X Protein; Breast Neoplasms; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Coumestrol; Estradiol; Estrogen Receptor alpha; Female; Genistein; Humans; Isoflavones; Phytoestrogens; Proto-Oncogene Proteins c-bcl-2; Resveratrol; Stilbenes

A reliable chromatographic method for the determination of soy isoflavones (genistein, daidzein and glycitein) using a coulometric detection has been developed and applied to analyse plasma of postmenopausal women. The chromatographic separation was performed on a C18 reversed phase column with a mobile phase composed of acetonitrile-phosphate buffer mixture. Coulometric detection was carried out at +0.500 V. A careful and rapid solid phase extraction procedure on hydrophilic/lipophilic cartridges was chosen for plasma sample purification with and without hydrolysis obtaining good extraction yield values for all the analytes (>90.0%). The enzymatic hydrolysis step was necessary for the determination of the total amount of soy isoflavones. The limit of quantitation was 0.5 ng mL(-1) for genistein and 0.25 ng mL(-1) for daidzein and glycitein. The method was found to be precise and accurate. Thus, the proposed method is suitable for the analysis of soy isoflavones (free and total amounts) in plasma of postmenopausal women under treatment with the SoymenGN dietary supplement.

Topics: Chromatography, High Pressure Liquid; Electrochemistry; Female; Genistein; Humans; Isoflavones; Middle Aged; Postmenopause; Solid Phase Extraction

The production of beta-maltooligosaccharides of glycitein and daidzein using Lactobacillus delbrueckii and cyclodextrin glucanotransferase (CGTase) as biocatalysts was investigated. The cells of L. delbrueckii glucosylated glycitein and daidzein to give their corresponding 4'- and 7-O-beta-glucosides. The beta-glucosides of glycitein and daidzein were converted into the corresponding beta-maltooligosides by CGTase. The 7-O-beta-glucosides of glycitein and daidzein and 7-O-beta-maltoside of glycitein showed inhibitory effects on IgE antibody production. On the other hand, beta-glucosides of glycitein and daidzein exerted 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging activity and supeoxide-radical scavenging activity.

Topics: Animals; Anti-Allergic Agents; Antibody Formation; Antioxidants; Glycosides; Immunoglobulin E; Isoflavones; Oligosaccharides; Rats

Several dietary intervention studies examining the health effect of soy isoflavones allude to the importance of equol in establishing the cardiovascular response to soy protein. Although, the specific mechanism by which this action occurs has not been established. The aim of this study was to investigate the inhibitory effect of soy-isoflavones and the metabolite of daidzein, equol, on agonist-induced platelet responses dependent on thromboxane A(2) (TxA(2)) receptor.. Competitive radioligand binding assay was used to screen for affinity of these compounds to the TxA(2) receptor. The effect of equol on platelet activation, evaluate through of release of the ATP, by analogs of TxA(2) was analyzed. The effect of equol on platelet aggregation was investigated with ADP, U46619 (a TxA(2) mimic) and the calcium ionophore A23187.. The data showed that aglycone isoflavones and equol bind to TxA(2) receptor in the micromol/L range, whereas their glucoside derivates had very low binding activity for this receptor. Under equilibrium conditions, the following order of the relative affinity in inhibiting [(3)H]-SQ29585 binding was: equol>genistein>daidzein>glycitein>>genistin, daidzin, glycitin. Equol interaction was reversible and competitive for labeled-SQ29548 with not apparent decrease in the number of TxA(2) binding sites. In addition, from platelet activation studies, equol effectively inhibited ATP secretion elicited by the TxA(2) analog U46619. On the other hand, equol inhibited the platelet aggregation induced by U46619 and A23187, while it failed to inhibit that induced by ADP.. The aglycone isoflavones from soy, and particularly equol, have been found to have biological effects attributable to thromboxane A(2) receptor antagonism. These findings may help elucidate how dietary isoflavone modulate platelet function and explain why soy-rich foods are claimed to have beneficial effects in the prevention of thrombotic events.

Topics: Binding, Competitive; Bridged Bicyclo Compounds, Heterocyclic; Calcium; Equol; Fatty Acids, Unsaturated; Genistein; Humans; Hydrazines; Isoflavones; Platelet Activation; Platelet Aggregation; Platelet Aggregation Inhibitors; Receptors, Thromboxane A2, Prostaglandin H2

The relative contribution of the small intestine to absorption and microbial metabolism of ingested isoflavonoids (IFN) was investigated in swine with canulae in distal ilea to facilitate collection of chyme (canula open). Weaned swine were fed a single meal containing ground roasted soybean and corn with canulae open followed by a second test soy diet at 48 h with canulae closed to allow passage of chyme into the large intestine. All remaining feedings were soy-free (corn-casein diet). Ileal effluent and urine were collected for 16 and 48 h, respectively, and analyzed for IFN and microbial metabolites of IFN. IFN in ileal effluent were present entirely as aglycones. IFN equivalents excreted for 24 h after ingesting the soy diet were not significantly different when canulae were open or closed. Urinary IFN aglycone equivalents on day 2 were similar to those on day 1 when canulae remained closed, but less than 10% of that on day 1 when canulae were open for 16 h postfeeding. Urinary concentrations of dihydrodaidzein, dihydrogenistein, O-desmethylangolensin, and equol exceeded IFN aglycone equivalents. These findings suggest extensive preabsorptive conversion of IFN glucosides to aglycones in the small intestine and relatively efficient microbial metabolism of IFN in weaned swine.

Topics: Animals; Chromatography, High Pressure Liquid; Genistein; Glucosides; Glycine max; Ileum; Intestinal Absorption; Isoflavones; Swine

Isoflavones are derived from dietary sources and considered to promote health by preventing the onset of such chronic diseases as cardiovascular disease, cancer, and osteoporosis. Valid and reliable estimation of isoflavone intake is a prerequisite to establishing biological functions of isoflavones on health risks.. This study aimed to validate the approach of estimating dietary isoflavone intake with respective urinary isoflavone concentrations in US adults.. Data from the US Department of Agriculture isoflavone database and dietary recalls of 2,908 US adults with urinary isoflavone data in the 1999-2002 National Health and Nutrition Examination Survey were used.. Dietary isoflavone was consumed by only 35% of adults in a day with an average intake of 3.1 mg/day, which resulted in a mean intake of 1.0 mg/day for all US adults. The isoflavone intakes were from genistein (55%), diadzein (35%), glycitein (7%), biochanin A (2%), and formononetin (2%). Both daily total and energy adjusted isoflavone intake differed by race/ethnicity subgroups (P<0.05) and was associated positively with income (P<0.01) and inversely with body mass index (P<0.05). Geometric mean urinary isoflavone concentration was 5.0 ng/mL among isoflavone consumers and the urinary genistein and daidzein excretion correlated with their isoflavone intake levels (P<0.01).. In large population-based studies, estimated dietary isoflavone intake can be validated by urinary isoflavones. Further studies are needed at an individual level to validate dietary isoflavone intake by urinary isoflavone concentration.

Topics: Adult; Aged; Alcohol Drinking; Biomarkers; Black or African American; Cross-Sectional Studies; Diet; Exercise; Female; Genistein; Hispanic or Latino; Humans; Isoflavones; Male; Middle Aged; Nutrition Surveys; Poverty; Smoking; Soy Foods; United States; White People; Young Adult

Increased consumption of soy and soy isoflavones is associated with a reduced risk for prostate cancer (PCa). PCa progression is characterized, in part, by a loss of luminal/basal epithelial differentiation; however, the effects of soy isoflavones on cellular differentiation in the prostate are unknown. The present study examined the effects of the soy isoflavone glycitein on cellular differentiation in prostate epithelial cells (RWPE-1, WPE1-NB14, and RWPE-2). Glycitein significantly inhibited RWPE-1 cellular proliferation at concentrations ranging from 0.4 to 50 microM. Expression of the luminal epithelial cell marker cytokeratin 18 was not affected by glycitein treatment in the WPE1-NB14 and RWPE-2 cell lines. However, expression of cytokeratin 18 and prostate specific antigen (PSA) was decreased in the RWPE-1 cell line in response to glycitein treatment, whereas the expression of the basal epithelial cell markers p63 and cytokeratin 5 remained unchanged. These data suggest that glycitein may induce basal cell differentiation in the RWPE-1 cell line.

Topics: Cell Cycle; Cell Differentiation; Cell Proliferation; Cells, Cultured; Epithelial Cells; Genistein; Humans; Isoflavones; Keratin-18; Keratin-5; Male; Prostate; Prostate-Specific Antigen

There are limited reports on the bioavailability and pharmacokinetics of isoflavones in elderly humans and aged animals. The present study was conducted to assess the effect of glycosidation of isoflavones on their bioavailability and pharmacokinetics in aged (20 month old) male Fischer-344 (F-344) rats. The F-344 rat, developed by the National Institute on Aging, is an inbred rat model that is commonly used for aging studies and resembles many features of aging humans. Three sources of isoflavones; Novasoy (a commercial supplement), a mixture of synthetic aglycons (daidzein, genistein and glycitein), and a mixture of synthetic glucosides (daidzin, genistin, and glycitin) were tested. Following administration, blood samples were collected at different times (0-48 h post-oral gavage and 0-8 h post-IV dosing). Plasma isoflavones and 7-hydroxy-3-(4'-hydroxyphenyl)-chroman (a metabolite of daidzein) were measured by LC/MS. The extent of absorption was determined by comparing the area under the curve (AUC) of the plasma-concentration time curve after intravenous (IV) administration with that following oral administration. The extent of bioavailability was then calculated as: %bioabailability = (AUC(or)/AUC(IV))x(Dose(IV)/Dose(or))x100. Bioavailabilities for genistein were significantly (p = 0.013) higher for the aglycon (35 +/- 9%) compared with the glucoside forms (11 +/- 3%). In contrast, the bioavailabilities for glycitein were significantly (p = 0.011) higher in Novasoy (27 +/- 13%) and the glucoside form (21 +/- 10%) compared with the aglycon (8 +/- 3%). No significant differences in the bioavailability of daidzein were observed in aged rats dosed with aglycon, glucoside or Novasoy. However, aged rats were able to produce equol as early as 8 h post-dosing. In summary, the source of isoflavones had significant effects on genistein and glycitein bioavailability in aged male rats.

Topics: Aging; Animals; Biological Availability; Diet; Equol; Genistein; Glucosides; Glycine max; Glycosylation; Injections, Intravenous; Isoflavones; Male; Phytoestrogens; Rats; Rats, Inbred F344

Soybean is an important Brazilian agricultural commodity that contains a high concentration of isoflavones. Many studies showed that isoflavones are active in the prevention of many human diseases. However, the correct processing techniques used to prepare the soy foodstuffs are important to maintain the active forms. The objective of this study was to evaluate the effect of gamma irradiation on the isoflavone contents of the defatted soybean flour when compared with soybean molasses, a derivative from the soybean food production. After extracting phenolic compounds with methanol aqueous solution (80%), isoflavones were detected by reverse-phase high-performance liquid chromatography/diode-array detector. The radiation doses of 2 and 5 kGy presented a small effect on the isoflavones content of defatted soy flour. Samples irradiated at 50 kGy showed lower isoflavone contents. The observed reduction in the concentration of isoflavones-daidzein, glycitein and genistein-induced by gamma radiation in soy molasses was not significant in defatted soy flour, thus suggesting that isoflavones in defatted soy flour were not eliminated by gamma radiation at rates up to 50 kGy.

Topics: Chromatography, High Pressure Liquid; Diet; Flour; Food Handling; Gamma Rays; Genistein; Glycine max; Humans; Isoflavones; Seeds; Soy Foods

The goal of this study was to evaluate how oral antibiotics (OABX) change the appearance of isoflavones (IFLs) in adults and children after soy consumption. The urinary IFL excretion rate (UIER) known to reflect circulating IFLs was hypothesized to change due to intestinal microflora changes by OABX. Subjects provided urine collections in pairs of a baseline urine and an overnight urine before and after consuming soy nuts first during OABX treatment and then again when healthy. During OABX versus when healthy, UIER (nmol/h/kg) in adults (n = 12) was increased (P < 0.05) for daidzein (35.2 +/- 7.2 vs. 18.9 +/- 2.4), daidzein + genistein + glycitein [nonmetabolites (NM); 42.6 +/- 8.0 vs. 23.6 +/- 2.9), and total isoflavonoids (Total IFLs; daidzein + genistein + glycitein + dihydrodaidzein + dihydrogenistein + equol + O-desmethylangolensin) (51.5 +/- 10.3 versus 29.6 +/- 4.7). In contrast, children (n = 7) showed reduced UIER (P < 0.05) when on OABX versus when healthy for daidzein (36.3 +/- 6.4 vs. 46.8 +/- 4.7), dihydrodaidzein (1.2 +/- 0.6 vs. 3.0 +/-1.1), NM (46.3 +/- 8.2 vs. 59.5 +/- 6.0), dihydrodaidzein + dihydrogenistein + equol + O-desmethylangolensin (1.0 +/- 0.8 vs. 4.3 +/- 1.3), and Total IFLs (48.2 +/- 8.5 vs. 63.8 +/- 6.4).

Topics: Administration, Oral; Adolescent; Adult; Age Factors; Anti-Bacterial Agents; Biomarkers; Child; Child, Preschool; Female; Follow-Up Studies; Genistein; Glycine max; Humans; Isoflavones; Male; Phytoestrogens; Soy Foods

Complete separation of aglycones and glucosides of selected isoflavones (genistin, genistein, daidzin, daidzein, glycitin, glycitein, ononin, sissotrin, formononetin, and biochanin A) was possible in 1.5 min using an ultrahigh-pressure liquid chromatography (U-HPLC) on a different particular chemically modified stationary phases with a particle size under 2 microm. In addition, selected separation conditions for simultaneous determination of isoflavones together with a group of phenolic acids (gallic, protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, and sinapic acid) allowed separation of all 19 compounds in 1.9 min. Separations were conducted on a non-polar reversed phase (C(18)) and also on more polar phases with cyanopropyl or phenyl groups using a gradient elution with a mobile phase consisting of 0.3% aqueous acetic acid and methanol. Chromatographic peaks were characterised using parameters such as resolution, symmetry, selectivity, etc. Individual substances were identified and quantified using UV-vis diode array detector at wavelength 270 nm. Limits of detection (3S/N) were in the range 200-400 pg ml(-1). Proposed U-HPLC technique was used for separation of isoflavones and phenolic acids in samples of plant materials (Trifolium pratense, Glycine max, Pisum sativum and Ononis spinosa) after acid hydrolysis of the samples and modified Soxhlet extraction.

Topics: Caffeic Acids; Chromatography, High Pressure Liquid; Coumaric Acids; Gallic Acid; Genistein; Glycine max; Hydroxybenzoates; Isoflavones; Molecular Structure; Pisum sativum; Plant Extracts; Propionates; Trifolium; Vanillic Acid

We evaluated the reproducibility and validity of the FFQ used in the Shanghai Men's Health Study (SMHS) for assessing dietary isoflavone intake, using multiple 24-h dietary recalls (24-HDR) and urinary isoflavones as the reference criteria, with data from the dietary validation study of the SMHS. A total of 196 study subjects completed the 24-HDR and 2 FFQ and donated a quarterly spot urine sample during the 1-y study period. Levels of urinary isoflavones were measured in a random sample of 48 study participants. The correlation coefficient between the 2 FFQ administered 1 y apart was 0.50 for soy protein intake and ranged from 0.50 to 0.51 for isoflavone intake. The correlations of isoflavone intake from the second FFQ with those from the multiple 24-HDR ranged from 0.38 (genistein) to 0.44 (glycitein), and the correlations with urinary isoflavone levels were 0.48 for total isoflavones, 0.44 for daidzein, 0.42 for genistein, and 0.54 for glycitein. The intraclass correlation coefficients for the 4 spot urine samples were 0.36, 0.42, and 0.40 for daidzein, genistein, and glycitein, respectively, and 0.62, 0.68, and 0.55 for their metabolic products equol, dihydrodaidzein, and O-desmethylangolensin, respectively. These results suggest that the SMHS FFQ can reliably and accurately measure usual intake of isoflavones, and that the levels of isoflavones in urine samples are relatively stable among men in Shanghai.

Topics: Asian People; Diet; Genistein; Humans; Isoflavones; Male; Middle Aged; Reproducibility of Results; Surveys and Questionnaires

Traditionally, food is associated with energetic and nutritious characteristics such as sources of carbohydrates, proteins and lipids. Recently, however, foods with bioactive substances such as isoflavones have received great attention. The objective of this study was therefore to evaluate the presence of the isoflavones daidzein, glycitein, genistein and their conjugated forms in grains and leaves of several leguminous plants utilized largely in Brazilian cuisine. Grains used in Peruvian cuisine were also analyzed. After extracting phenolic compounds with methanol (80%), isoflavones as detected by reversed-phase high-performance liquid chromatography/ diode-array detector were only found in chickpeas and soybean. Chickpea extracts showed only the isoflavone genistein at 31 microg/g defatted flour. Detection of these isoflavones was confirmed by electrospray ionization mass and tandem mass spectrometric experiments. For soybean, a distinct distribution of isoflavones was found in hypocotyls and cotyledon. The highest concentration of isoflavones found was approximately 5.9 mg/g for hypocotyls, whereas the total concentration of isoflavone was around 0.4 mg/g for the cotyledons (dry matter). These results indicate that isoflavone concentrations vary within the different tissues of the leguminous species tested.

Topics: Chromatography, High Pressure Liquid; Fabaceae; Food Analysis; Genistein; Humans; Isoflavones; Spectrometry, Mass, Electrospray Ionization

The purpose of the present study was to determine whether children experience a higher systemic exposure to isoflavonoids when consuming a body weight-adjusted dose of soya compared with adults. Forty study participants were recruited from a local Waldorf school, including twenty-one children and nineteen adults. Participants collected a baseline urine sample and ate immediately thereafter a body weight-adjusted dose of soya nuts (15 g/54.4 kg equivalent to 0.615 (SD 0.036) mg total isoflavones/kg) followed by a 12 h urine collection. Nineteen children and eighteen adults completed the protocol correctly (fourteen child-parent pairs). Children, compared with adults, showed a statistically significant (P < 0.05 by unpaired t test) higher urinary isoflavone excretion rate for daidzein (+39%), genistein (+44%), all non-metabolites (daidzein + genistein + glycitein; +41%) and total isoflavonoids (+32%). Isoflavones are more bioavailable in children v. adults. Urine is an excellent medium to determine systemic isoflavone exposure in children due to its non-invasiveness and high compliance, in particular when collected overnight; it also allows evaluation of completeness of specimen collection.

Topics: Adolescent; Adult; Aging; Biological Availability; Body Weight; Child; Child Nutritional Physiological Phenomena; Child, Preschool; Female; Genistein; Humans; Isoflavones; Male; Soy Foods

The objective of the present study is to explore the association between lower urinary tract symptoms (LUTS) and dietary isoflavone in elderly men. In a large prospective cohort of 2000 Chinese men, the association between dietary isoflavone and LUTS were studied using standardized structured questionnaires. Dietary intake was assessed by a modified version of the Block FFQ. LUTS were assessed by the Chinese version of the International Prostatic Symptoms Score. Demographic, lifestyle and other medical information were also collected and were adjusted for in the analysis. After excluding 299 men with history of bladder surgery, bladder or prostate cancer or who were current users of medication for urinary problems, the association between LUTS and dietary isoflavone was explored. A total of 96.2 % of subjects reported some consumption of genistein, glycitein or daidzein. In ordinal multinominal logistic regression, subjects with dietary total isoflavone of more than 5.1 mg were significantly less likely to suffer from more severe LUTS (dietary intake from 5.1 to 9.5 mg: OR 0.59; 95 % CI 0.44, 0.80; from 9.6 to 14.3 mg: OR 0.81; 95 % CI 0.61, 1.09; from 14.4 to 21.7 mg: OR 0.68; 95 % CI 0.51, 0.92; 21.8 mg and above: OR 0.73; 95 % CI 0.54, 0.98) after adjustment for covariates. In this cross-sectional study, we showed a strong inverse association between dietary intake of isoflavone and the risk of LUTS.

Topics: Aged; Alcohol Drinking; China; Cross-Sectional Studies; Diet; Feeding Behavior; Genistein; Glycine max; Humans; Isoflavones; Logistic Models; Male; Prospective Studies; Prostatic Hyperplasia; Prostatic Neoplasms; Risk Factors; Smoking; Surveys and Questionnaires; Urination Disorders

To compare endogenous and exogenous beta-glucosidases for the hydrolysis of the predominant isoflavone glucosides in soymilk in order to improve the biological activity.. beta-glucosidase activity of probiotic organisms, including Bifidobacterium animalis ssp. lactis Bb12, Lactobacillus acidophilus ATCC 4461 and Lactobacillus casei 2607 in soymilk, was evaluated and was related to the increase in the concentration of isoflavone aglycones during fermentation. The concentrations of isoflavone compounds in soymilk were monitored using a Varian model HPLC with an Amperometric electrochemical detector. The aglycone composition, also known as aglycone equivalent ratio, has been considered to be important for the delivery of health benefits of isoflavones, and was monitored during the fermentation of soymilk. Comparison of the hydrolytic effectiveness of both exogenous and endogenous enzyme during 4-h incubation in soymilk was conducted using the Otieno-Shah (O-S) index. Results showed that exogenous enzyme exhibited faster rate of isoflavone glucoside hydrolysis than that by endogenous enzyme. Highest O-S indices were obtained after 4, 3 and 2 h of incubation with enzyme solution having beta-glucosidase activity of 0.288 U ml(-1), 0.359 U ml(-1) and 0.575 U ml(-1), resulting into aglycone concentration increments of 5.87-, 6.07- and 5.94-fold, respectively. Conversely, aglycone concentration in the soymilk with B. animalis ssp. lactis Bb12, L. casei 2607 and L. acidophilus 4461 increased by 3.43-, 2.72- and 3.03-fold, respectively, after 4 h of fermentation at 37 degrees C. In addition, the O-S index of endogenous enzyme was much lower than that of the exogenous enzyme over the same 4-h incubation period. Optimum aglycone equivalent ratios coincided with highest O-S indices and highest aglycone concentrations in soymilk hydrolysed with exogenous enzyme. The same correlation of O-S indices and highest aglycone concentrations occurred for endogenous enzyme during the 24 h of fermentation.. Obtaining highest aglycone concentration and optimum aglycone equivalent ratio could provide a critical beginning point in clinical trials for the realization of unique health benefits of soy isoflavones.. Screening for beta-glucosidase activities of probiotics in soymilk and comparing their hydrolytic potentials with that of exogenous beta-glucosidase could find wide applications in the development of different aglycone-rich functional soy beverages.

Topics: Bifidobacterium; Cellulases; Chromatography, High Pressure Liquid; Culture Media; Fermentation; Food Microbiology; Genistein; Hydrolysis; Isoflavones; Lacticaseibacillus casei; Lactobacillus acidophilus; Phytoestrogens; Probiotics; Soy Milk

Adult T-cell leukemia occurs in human T-lymphotropic virus type I-infected individuals and is endemic to the south-western area of Kyushu in Japan. In this communication, we examined the effect of soy isoflavones on the growth of adult T-cell leukemia cells in vitro and in vivo. In the in vitro study, daidzein and genistein but not glycitein significantly inhibited the proliferation of ED-40515 and Hut102 cells in a dose-dependent manner. Among the isoflavones studied, genistein had the highest growth-inhibitory effect; however, genistein did not exert an apparent growth-inhibitory effect on Jurkat and Molt-4 cells, which were non-adult T-cell leukemia cells. Genistein prevented the G(1)/S or G(2)/M transition at 3 and 10 or 30 microM, respectively. Genistein upregulated p21 protein expression together with p53 accumulation. In addition, treatment with 30 microM genistein strongly induced phosphorylation of checkpoint kinase (CHK) 2 and p53 at serines 15, 20 and 37. Caffeine, an inhibitor of ataxia-telangiectasia mutated protein kinase, alleviated the genistein-induced p53 and CHK2 phosphorylation, suggesting the involvement of DNA damage at 30 microM. However, marked phosphorylation of CHK2 and p53 could not be detected at 3 and 10 microM genistein. These data indicate that genistein has biphasic growth-inhibitory properties. The in vivo studies demonstrated that soy-derived isoflavones significantly inhibit ED-40515 cell growth and infiltration into various organs in non-obese diabetic severe combined-immunodeficiency common gamma-chain knockout mice. Taken together, it is evident that soy isoflavones might serve as a promising compound for the treatment of adult T-cell leukemia.

Topics: Animals; Cell Cycle; Cell Division; Cell Line, Tumor; Genistein; Glycine max; Humans; Isoflavones; Jurkat Cells; Leukemia-Lymphoma, Adult T-Cell; Mice; Mice, Knockout; Mice, SCID; Transplantation, Heterologous

The soybean isoflavones, daidzein, genistein, and glycitein, were hypothesized to act as cholesterol-lowering components, separate from soy protein. Pure synthetic daidzein, genistein, or glycitein (0.9 mmol/kg diet) or a casein-based control diet was fed to groups of 10 female Golden Syrian hamsters for 4 weeks. Hamsters fed glycitein had significantly lower plasma total (by 15%) and non-HDL (by 24%) cholesterol compared with those fed casein (P<0.05). Daidzein and genistein's effects on these lipids did not differ from the effects of either casein or glycitein. Plasma HDL cholesterol and triglyceride concentrations were not significantly affected by dietary treatments. The percentage of urinary recovery of the ingested dose of each isoflavone was glycitein>daidzein>genistein (33.2%, 4.6%, 2.2%, respectively), with the apparent absorption of glycitein significantly greater than that of the other isoflavones. These data suggest that glycitein's greater cholesterol-lowering effect was due to its greater bioavailability, as reflected in its urinary recovery compared with that of the other isoflavones.

Topics: Animals; Biological Availability; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Cricetinae; Diet; Female; Genistein; Isoflavones; Mesocricetus; Triglycerides

To investigate the long-term effect of soybean isoflavones on changes in parvalbumin (PV) immunoreactivity in the hippocampus in normal female, ovariectomized (OVX) female and normal male rats.. Ten-month-old rats were assigned to one of 9 groups (n = 7 in each group) based on body weight using a randomized complete-block design. The groups were: control diet-treated females, OVX females, and males; 0.3 g/kg isoflavone-treated females, OVX females, and males; and 1.2 g/kg isoflavone-treated females, OVX females, and males. The PV immunostaining was conducted by using the standard avidin-biotin complex method.. PV immunoreactivity and the number of PV-immunoreactive neurons in all the groups after isoflavone treatment were significantly changed in the hippocampal CA1 region and in the dentate gyrus, but not in the hippocampal CA2/3 region. PV immunoreactivity and the number of PV-immunoreactive neurons in the control diet OVX females were similar to those in the control diet, and were greater than those in the control diet normal females. PV immunoreactivity and the number of PV-immunoreactive neurons in all the isoflavone-treated groups decreased dose-dependently after isoflavone treatment.. Long-term administration of isoflavones may induce a reduction of PV in interneurons in the hippocampal CA1 region and in the dentate gyrus. The reduction of PV in these regions suggests that the long-term administration of isoflavones may cause a change in calcium homeostasis in the hippocampal CA1 region and in the dentate gyrus.

Topics: Animals; Dentate Gyrus; Female; Genistein; Glycine max; Hippocampus; Isoflavones; Male; Neurons; Ovariectomy; Parvalbumins; Random Allocation; Rats; Rats, Sprague-Dawley

This study was conducted to compare the transformation of both isoflavone derivatives (aglycones, beta-glucosides, and acetyl and malonyl gluclucosides) and beta-glucosidase activity in kojis fermented with various generally recognized as safe filamentous fungi including Aspergillus awamori, Aspergillus oryzae, Aspergillus sojae, Rhizopus azygosporus, and Rhizopus sp. no. 2. Solid fermentation was performed to prepare the kojis by inoculating the steamed black beans with starter organism and culturing at 30 degrees C for 3 days. Results revealed that fermentation caused a marked increase in the content of aglycone (daidzein, glycitein, and genistein), the bioactive isoflavone, and a significant reduction in the content of beta-glucoside isoflavone (daidzin, glycitin, and genistin), compared with the unfermented steamed black bean. The extent of increased aglycone and reduced beta-glucoside isoflavone content varied with the starter organism used. Among the various black bean kojis prepared, the Rhizopus sp. no. 2 koji showed the highest level of enhancement in aglycone content. In the Rhizopus sp. no. 2 koji, the percentage of aglycone to total isoflavone increased from an initial approximately 2.9 to approximately 58.9% after fermentation. In comparison, the percentages found in kojis prepared with other starter organisms ranged from 18.9 to 38.9% after fermentation. Further preparations of black bean kojis with A. awamori at different cultivation temperatures (25, 30, and 35 degrees C) and various fermentation periods (1-5 days) revealed that koji prepared at 30 degrees C for 4 days showed the highest content of aglycones, with 7.7-, 5.7-, and 4.8-fold increases in the content of daidzein, genistein, and glycitein, respectively. In addition, the increase of aglycone content and the increase of beta-glucosidase activity during the fermentation of this koji showed a similar trend.

Topics: Aspergillus; beta-Glucosidase; Fermentation; Food Handling; Genistein; Isoflavones; Phaseolus; Rhizopus; Temperature

The inhibitory effects of isoflavones (daidzein, genistein, and glycitein) on human cytochrome P450 (CYP) 2A6 activities were investigated. Daidzein, genistein, and glycitein uncompetitively inhibited nicotine C-oxidation catalyzed by recombinant CYP2A6 expressed in baculovirus-infected insect cells with Ki values of 1.3 +/- 0.3 microM, 0.7 +/- 0.2 microM, and 5.2 +/- 0.8 microM, respectively, but not coumarin 7-hydroxylation. Effects of the intake of soy isoflavones on in vivo nicotine metabolism were investigated with 7 healthy Japanese homozygotes of CYP2A6*1. The cotinine/nicotine ratio of the plasma concentrations 2 hours after chewing 1 piece of nicotine gum under the basal condition (after abstaining from soy foods for 1 week) was 8.8 +/- 2.6 (4.4-11.4). The ratio was significantly (P < .05) reduced to 6.7 +/- 1.6 (4.0-8.2) after consumption of a soy isoflavone supplement (60 mg of total isoflavones/d) for 5 days. The authors found that isoflavone contained in soy products significantly decreased nicotine metabolism.

Topics: Animals; Aryl Hydrocarbon Hydroxylases; Baculoviridae; Chewing Gum; Cotinine; Cytochrome P-450 CYP2A6; Dietary Supplements; Genistein; Genotype; Homozygote; Humans; Insecta; Isoflavones; Mixed Function Oxygenases; Nicotine; Oxidation-Reduction; Phenotype; Soy Foods

Consumption of soya-based nutrients is increasing in modern society because of their potentially protective effects against chronic diseases. Soya products are also heavily advertised as alternative drugs for relief from symptoms of the menopause and for hormone replacement therapy. However, because of their oestrogenic activity, negative effects of isoflavones have been postulated. Therefore, we analysed influences of soya isoflavones, major soya constituents with endocrine activity, on thyroxine (T4) binding to its distribution proteins. Serum binding of (125)I-labelled L-T4 was analysed in the absence or presence of increasing concentrations of soya isoflavones using non-denaturing PAGE for analysis. Complete displacement of [(125)I]T4 binding to transthyretin (TTR) was observed in human serum incubated with genistein at concentrations >10 microM; interference started at >0.1 microM. Glycitein showed decreased and daidzein the lowest displacement potency. [(125)I]T4 was displaced to albumin in rat and to T4-binding globulin in human serum. Soya isoflavones also obstruct [(125)I]T4 binding to TTR in human cerebrospinal fluid (CSF). The inhibitory effect was confirmed in direct binding assays using purified TTR with 50% inhibitory concentration values of 0.07 microM for genistein, 0.2 microM for glycitein and 1.8 microM for daidzein. The present study underlined a potent competition of soya isoflavones for T4 binding to TTR in serum and CSF. Isoflavones might alter free thyroid hormone concentrations resulting in altered tissue availability and metabolism. As a consequence of this interference, one could expect a disturbance in the feedback regulation of hormonal networks, including the pituitary-thyroid-periphery axis during development and in adult organisms.

Topics: Animals; Binding, Competitive; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Genistein; Growth Inhibitors; Humans; Iodine Radioisotopes; Isoflavones; Ligands; Prealbumin; Rats; Rats, Wistar; Thyroxine; Thyroxine-Binding Proteins

The soy isoflavones genistein and daidzein, as well as the daidzein metabolite equol, have structural similarities to mammalian estrogens and bind with varying affinity to both known subtypes of the estrogen receptor. Consequently, prospective studies in both humans and animals have begun to evaluate the potential effects of isoflavones on estrogen receptor-mediated phenomena. However, many diets of laboratory-housed animals derive their protein from soy and thus likely contain substantial quantities of isoflavones. Exposing experimental subjects to these isoflavones via such diets could confound studies, particularly those evaluating the effects of estrogen or estrogen-like ligands. The aim of this study was to compare the levels of circulating concentrations of isoflavones and their metabolites in monkeys fed either a soy-free diet, a soy-based diet providing 130 mg of isoflavone (daidzein, genistein, and glycitein aglycon equivalents) daily, or a commercially available 'chow' diet containing an unspecified amount of soybean meal. Animals consuming the commercial diet had serum concentrations of daidzein, genistein, and glycitein that were significantly higher than those of animals fed a soy-free diet but similar to those of monkeys fed a soybased diet formulated to be high in isoflavones. Notably, animals fed the commercial diet also had serum equol concentrations that were similar to or, in some cases, in excess of serum concentrations in the animals fed the soy diet. These data argue for the use of soy-free diets in studies investigating estrogenic effects on physiologic or behavioral endpoints.

Topics: Animal Nutritional Physiological Phenomena; Animals; Diet; Female; Genistein; Isoflavones; Laboratory Animal Science; Macaca fascicularis; Macaca mulatta; Male; Phytoestrogens; Soy Foods

Apparent absorption of isoflavones varies greatly among individuals but is relatively stable within an individual. We hypothesized that high urinary isoflavone excreters would show less plasma non-HDL cholesterol (non-HDL-C) than low isoflavone excreters after soy protein feeding. Fifty Golden Syrian hamsters were fed a high-fat/casein diet (n = 10) or a high-fat/soy protein diet (n = 40) for 4 wk. We identified 2 distinct urinary isoflavone excretion phenotypes based upon HPLC analysis of urinary glycitein using a pairwise correlation plots analysis, or based upon total urinary isoflavone using a hierarchical cluster test. High isoflavone excreters showed greater urinary isoflavones (P < 0.05) than did low isoflavone excreters at wk 1 and 4. The low urinary glycitein excretion phenotype was more stable than the high urinary glycitein excretion phenotype by McNemar's test. High urinary isoflavone excreters had significantly less non-HDL-C than did the low isoflavone excreters or casein-fed controls (P < 0.05). Plasma total and non-HDL-C were negatively correlated with urinary daidzein, glycitein, and total isoflavone excretion (r = -0.45 to -0.58, P < 0.05). Urinary isoflavone excretion phenotypes predicted the cholesterol-lowering efficacy of soy protein. Isoflavone absorbability, probably due to gut microbial ecology, is an important controllable variable in studies of effects of soy protein on blood lipids.

Topics: Animals; Cholesterol; Cholesterol, HDL; Cricetinae; Female; Genistein; Isoflavones; Mesocricetus; Phenotype; Soybean Proteins; Triglycerides

Topics: Female; Fungi; Genistein; Glycine max; Hot Flashes; Humans; Isoflavones; Phytoestrogens; Pilot Projects; Postmenopause

Soy isoflavones display estrogenic activity in humans and animals, and thus are referred to as phytoestrogens. This study was performed to observe the effects of the soy isoflavones genistein, daidzein, and glycitein on cell cultures of rat skeletal muscles. [3H]Thymidine incorporation was used to determine cell proliferation, while protein synthesis and degradation were determined by tracking radiolabeled leucine. For the proliferation studies, insulin, estradiol, genistein, daidzein, or glycitein was supplemented at 0, 0.04, 0.08, 0.16, 0.31, 0.63, 1.25, 2.5, 5, 10, or 20 microM, respectively, or in combinations with final concentrations of 0, 0.1, 1, or 10 microM. Genistein reacted most similarly to estradiol, inhibiting proliferation at > or = 1 microM (P < .001). A combination of phytoestrogens resulted in significant inhibition of cell proliferation, but not to the extent observed with genistein alone. For the protein synthesis and degradation experiments, treatments of 0.1 microM dexamethasone or 1 microM concentrations of insulin, genistein, daidzein, or glycitein were used. Phytoestrogens did not inhibit or stimulate protein degradation or synthesis (P > .05). A one-tailed univariate analysis of variance revealed a trend (P < or = .1) in protein stimulation with genistein and glycitein treatments. These results suggest that the tyrosine kinase inhibiting activity of genistein may be affecting phosphorylation of the mitosis-promoting factor, preventing the advancement of the mitotic cell cycle. In addition, at higher total combined concentrations, daidzein and glycitein may be able to outcompete genistein for receptor sites. These results suggest that soy isoflavones in the diet may potentially modulate normal growth and development in humans and animals that ingest soy-based products.

Topics: Animals; Cell Division; Cell Line; Dexamethasone; Estradiol; Genistein; Glycine max; Insulin; Isoflavones; Muscle Proteins; Muscle, Skeletal; Phosphorylation; Phytoestrogens; Rats

A method for simultaneous detection and quantification is presented to determine the presence of isoflavones and bisphenol A in a biological sample. A coulometric array detector was used with reversed-phase high-performance liquid chromatography (HPLC). Daidzein (1), glycitein (2), genistein (3) and their glucoside conjugates, daidzin (4), glycitin (5) and genistin (6), were measured as phytochemicals. Also assayed here was equol (7), a metabolite from compound 1, and bisphenol A (8), an industrial chemical that acts as an endocrine disrupter. All chemicals were simultaneously detected by using a 600-mV single detection voltage with high efficacy. A mixture of 1, 3 and 8 was orally administered to rats, and the levels of these three chemicals in the serum were clearly increased after a 4 kU beta-glucuronidase treatment. The levels of compounds 1 and 3 in the serum were detected at 1665 and 2040 ng/ml, while 8 was at a low level of 417 ng/ml. Compound 7 in the serum was not detected until after enzymatic hydrolysis (72 ng/ml). These results suggest that this analytical method would be useful for metabolic and pharmacokinetic studies on isoflavones and bisphenol A.

Topics: Administration, Oral; Animals; Benzhydryl Compounds; Calibration; Chromatography, High Pressure Liquid; Colorimetry; Genistein; Glucuronidase; Isoflavones; Male; Phenols; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity

Isoflavones identified as inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in soybean paste were assayed using the catalytic portion of Syrian hamster HMG-CoA reductase, and the kinetic values were measured using HMG-CoA and NADPH. The inhibition of HMG-CoA reductase by these inhibitors was competitive with HMG-CoA and noncompetitive with NADPH. Ki values for genistein, daidzein, and glycitein were 27.7, 49.5, and 94.7 microM, respectively.

Topics: Algorithms; Animals; Chromatography, High Pressure Liquid; Cricetinae; Genistein; Glycine max; Hydroxymethylglutaryl CoA Reductases; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Isoflavones; Kinetics; Mass Spectrometry; Mesocricetus; NADP; Plasmids

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 4.8 g soya protein and on average 22 mg 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.

Topics: Aging; Animals; Bacteria; Child; Child Nutritional Physiological Phenomena; Child, Preschool; Equol; Feces; Genistein; Humans; Infant; Infant Formula; Infant Nutritional Physiological Phenomena; Isoflavones; Milk; Soy Milk

To examine a potential role for soybean phytoestrogens in postmenopausal bone loss, twenty-four 12-week-old Sprague-Dawley rats were divided randomly into 4 groups and given controlled diets for 16 weeks. The treatment groups were as followed: sham operated, ovariectomized (OVX) control, OVX + isoflavone extract (6.25 g/kg), and OVX + 17beta-estradiol (4 mg/kg). OVX treatments reduced femoral and fourth lumbar vertebral bone density and mineral content (p<0.01), decreased uterine weight (p<0.01), accelerated body weight increases (p<0.05), and increased the activities (p<0.01) of both serum alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP). Supplementation with isoflavone prevented the losses of bone density and mineral content caused by OVX (p<0.01). Although both isoflavone and 17beta-estradiol exhibited similar bone-sparing ability on the OVX-induced bone loss, the effect of isoflavone was not the same as that of 17beta-estradiol on the serum ALP and TRAP, body weight increase, and uterine weight change. We concluded that dietary supplementation with soybean isoflavone can prevent postmenopausal bone loss via a different mechanism of estrogen in OVX rats.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Density; Calcium; Eating; Estradiol; Female; Genistein; Glycine max; Humans; Isoenzymes; Isoflavones; Organ Size; Osteoporosis; Osteoporosis, Postmenopausal; Ovariectomy; Plant Extracts; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

3-Hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase is the rate-limiting enzyme in the biosynthesis of cholesterol in mammals. Some microbial metabolites have been found to be HMG-CoA reductase inhibitors. Korean soybean paste is a unique food fermented by many microorganisms. The enzymatic method using the catalytic domain of Syrian hamster HMG-CoA reductase was employed for the screening of HMG-CoA reductase inhibitors. Soybean paste extract was fractionated by vacuum liquid chromatography. Fractions showing relatively high HMG-CoA reductase inhibition were further purified through Sephadex LH-20 column chromatography and C18 preparative HPLC, and the inhibitory compounds were identified as genistein, daidzein, and glycitein.

Topics: Animals; Chromatography, High Pressure Liquid; Cricetinae; Genistein; Glycine max; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent; Isoflavones; Mass Spectrometry; Molecular Structure; Plant Extracts

The presence of plant lignans in 14 different soy-based health supplements is reported here for the first time together with the analysis of the isoflavone content, for which these products are commercialized. Six plant lignans, i.e., secoisolariciresinol, matairesinol, syringaresinol, lariciresinol, isolariciresinol, and pinoresinol, have been identified and quantified by gas chromatography-mass spectrometry, and a positive correlation has been found between the levels of plant lignans and the levels of isoflavones in the different products. Additional quantification of plant lignans and isoflavones in soybeans has been carried out, and results are provided to allow the comparison of the average levels in soybeans and soy-based supplements.

Topics: Dietary Supplements; Gas Chromatography-Mass Spectrometry; Genistein; Glycine max; Isoflavones; Lignans

We isolated from soybean miso 8-hydroxyglycitein and 6-hydroxydaidzein as DPPH-radical scavengers, and elucidated their chemical structures by mass spectrometric, and (1)H- and (13)C-NMR spectrosopic analyses. These compounds showed DPPH-radical scavenging activity as high as that of alpha-tocopherol, 8-hydroxygenistein and 8-hydroxydaidzein. This is the first report of the isolation of 8-hydroxyglycitein from a natural source.

Topics: Biphenyl Compounds; Cell Proliferation; Free Radical Scavengers; Glycine max; HL-60 Cells; Humans; Hydrazines; Isoflavones; Molecular Structure; Nuclear Magnetic Resonance, Biomolecular; Phytoestrogens; Picrates

To evaluate the labeling accuracy and product uniformity of available soy isoflavone products in the eastern Washington State region for 13 products and to perform a cost comparison based on the isoflavone content in milligrams for 11 products.. Thirteen (13) isoflavone products (7 tablet and 6 capsule formulations) were randomly obtained from health food, grocery, and pharmacy outlets in eastern Washington State. Four different samples of the same product were extracted using 75% ethanol. Each sample was analyzed for isoflavone content using a gradient high-performance liquid chromatography (HPLC) method. Pure isoflavonoid standards were used to quantify the content of genistein, daidzein, and glycitein and their respective glycosides. The amount of total isoflavonoids per purchased product was then divided into its purchase price in order to make cost comparisons between the products based on a 50 mg/d dose.. The weight variation within each product was generally small (<4%). However, there was significant variability in the composition of the products. Only 4 of the 13 products contained at least 90% of the isoflavone content claimed on the label and 2 of products contained impurities in the HPLC chromatogram that exceeded 40%. There was no difference in the total isoflavonoid contents of the 2 identical products made by the same manufacturer, but the percentages of the main components had changed significantly over time. Based on a 50-mg isoflavone per day dose, the cost of a 30-day supply ranged from $3.20 to $65.88.. There is a lack of uniformity among the products tested and the majority (67%) of the products contained less than 90% of labeled amounts. There is significant variability in the compositions between the products and in the composition of the same product over time. Consumers cannot trust isoflavonoid product labels to represent the product's content accurately or that product pricing is a reflection of isoflavone content.

Topics: Chromatography, High Pressure Liquid; Dietary Supplements; Food Labeling; Genistein; Humans; Isoflavones; Soybean Proteins; Washington

In addition to oil and soyfoods, soybean is also produced for vegetable use. The importance of consuming vegetable soybean for the prevention of chronic diseases is well documented. The objectives of this study were to determine the magnitude of genotype x year interactions for isoflavone concentration and pattern, estimate heritabilities, and identify genotypes with a stable isoflavone concentration and pattern. Thirty-one soybean genotypes from maturity groups (MGs) III to VI were grown at Randolph Research Farm of Virginia State University, Petersburg, Virginia, during 3 years. The genotypes were harvested at immature green pod stage (R6-R7) and analyzed for isoflavone contents. Significant (P < 0.05) differences among the genotypes were found for genistein, daidzein, glycitein, and total isoflavones. The genotype x year interactions were also significant (P < 0.05) for the seed traits analyzed, indicating that the performance of the genotype changes from year to year. However, genotypes Pella and Aoda consistently showed with higher means than the overall means for all the seed traits throughout the 3 years. MG differences were also observed for genistein, daidzein, and total isoflavone content. Low- to moderate-heritability estimates of 54, 45, 58, and 64% were observed for genistein, daidzein, glycitein, and total isoflavone content, respectively, suggesting that the seed traits are equally influenced by environments and genetic variations. In general, for all seed traits with the exception of daidzein, the percentage contribution of genotype to the total sum of square was higher than the genotype x year interaction. The seed traits were interdependent and the associations among them were positive and significant suggesting that simultaneous selection and improvements are possible.

Topics: Environment; Food Analysis; Genistein; Genotype; Glycine max; Isoflavones; Quantitative Trait, Heritable

The estrogenic isoflavones of soybeans and their glycosides are products of the shikimate pathway, the target pathway of glyphosate. This study tested the hypothesis that nonphytotoxic levels of glyphosate and other herbicides known to affect phenolic compound biosynthesis might influence levels of these nutraceutical compounds in glyphosate-resistant soybeans. The effects of glyphosate and other herbicides were determined on estrogenic isoflavones and shikimate in glyphosate-resistant soybeans from identical experiments conducted on different cultivars in Mississippi and Missouri. Four commonly used herbicide treatments were compared to a hand-weeded control. The herbicide treatments were (1) glyphosate at 1260 g/ha at 3 weeks after planting (WAP), followed by glyphosate at 840 g/ha at 6 WAP; (2) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied preemergence (PRE), followed by glyphosate at 1260 g/ha at 6 WAP; (3) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by glyphosate at 1260 g/ha at full bloom; and (4) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by acifluorfen at 280 g/ha plus bentazon at 560 g/ha plus clethodim at 140 g/ha at 6 WAP. Soybeans were harvested at maturity, and seeds were analyzed for daidzein, daidzin, genistein, genistin, glycitin, glycitein, shikimate, glyphosate, and the glyphosate degradation product, aminomethylphosphonic acid (AMPA). There were no remarkable effects of any treatment on the contents of any of the biosynthetic compounds in soybean seed from either test site, indicating that early and later season applications of glyphosate have no effects on phytoestrogen levels in glyphosate-resistant soybeans. Glyphosate and AMPA residues were higher in seeds from treatment 3 than from the other two treatments in which glyphosate was used earlier. Intermediate levels were found in treatments 1 and 2. Low levels of glyphosate and AMPA were found in treatment 4 and a hand-weeded control, apparently due to herbicide drift.

Topics: Drug Resistance; Genistein; Glycine; Glycine max; Glyphosate; Herbicides; Isoflavones; Isoxazoles; Organophosphonates; Seeds; Shikimic Acid; Tetrazoles

Phytoestrogens (isoflavones and lignans) are of increasing interest due to their potential to prevent certain types of complex diseases. However, epidemiological evidence is needed on the levels of phytoestrogens and their metabolites in foods and biological fluids in relation to risk of these diseases. We report an assay for phytoestrogens which is sensitive, accurate, and uses low volumes of sample. Suitable for epidemiological studies, the assay consists of a simple sample preparation procedure and has been developed for the analysis of five isoflavones (daidzein, O-desmethylangolensin, equol, genistein, and glycitein) and two lignans (enterodiol and enterolactone), which requires only 200 microl of urine and utilizes one solid-phase extraction stage for sample preparation prior to derivatization for GC/MS analysis. Limits of detection were in the region 1.2 ng/ml (enterodiol) to 5.3ng/ml (enterolactone) and the method performed well in the UK Government's Food Standards Agency-sponsored quality assurance scheme for phytoestrogens. For the first time, average levels of all the above phytoestrogens were measured in samples of urine collected from a free living population sample of women. Results show a large range in both the amount and the type of phytoestrogens excreted.

Topics: 4-Butyrolactone; Equol; Female; Gas Chromatography-Mass Spectrometry; Genistein; Humans; Isoflavones; Lignans; Molecular Structure; Phytoestrogens; Plant Preparations; Reproducibility of Results; Sensitivity and Specificity

Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-dependent diseases. Soy and soy foods are rich sources of isoflavones, which have been shown to possess several biological activities. In this study, the metabolism of soy isoflavones daidzein, genistein and glycitein was investigated in human subjects. The aim was to find and identify urinary phase I metabolites of isoflavones, which have an intact isoflavonoid skeleton, and which might possess some bioactivity. Six volunteers included three soy bars per day into their normal western diet for a 2-week period. Daily urine samples were collected before, and after the supplementation period. Urine samples were hydrolyzed with Helix pomatia, extracted with diethyl ether, purified with Sephadex LH-20 chromatography, and analyzed as trimethylsilyl derivatives using gas chromatography-mass spectrometry (GC-MS). The structures of the isoflavone metabolites were identified using authentic reference compounds. The metabolites, for which authentic reference compounds were not available, were identified by the interpretation of mass spectra. Several new isoflavone metabolites were identified, and the presence of previously reported metabolites confirmed. The metabolic pathways of daidzein, genistein and glycitein are presented on the basis of the identification of the metabolites in human urine after soy supplementation.

Topics: Adult; Chromatography, Gas; Dietary Supplements; Female; Genistein; Glycine max; Humans; Isoflavones; Male; Reference Standards; Spectrometry, Mass, Electrospray Ionization

The soy isoflavones daidzin, glycitin, and genistin were purified from defatted soy flour using preparative-scale reverse-phase HPLC. The stabilities of the three isoflavones at different heating temperatures were investigated. Daidzin, glycitin, and genistin were lost at a rate of 26, 27, and 27% of their original concentration, respectively, after 3 min at 185 degrees C. At 215 degrees C, decreases of daidzin, glycitin, and genistin were 65, 98, and 74% after 3 min and 91, 99, and 94% after 15 min, respectively. The order of the thermal stabilities, from lowest to highest, was glycitin, genistin, and daidzin. Acetyl daidzin and acetyl genistin, daidzein, glycitein, and genistein were produced during heating at temperatures above 135 degrees C. The rate of binding of an acetyl group to form acetyl daidzin and acetyl genistin from daidzin and genistin was higher than the rate of loss of a glucoside group to form daidzein and genistein. However, acetyl daidzin and acetyl genistin decreased sharply at temperatures above 200 degrees C, while daidzein, glycitein, and genistein were relatively stable over 30 min. The stability of daidzein was higher than that of glycitein or genistein.

Topics: Acetylation; Chromatography, High Pressure Liquid; Genistein; Glycine max; Hot Temperature; Isoflavones; Kinetics

Soybean isoflavone concentrations vary widely, but the contribution of soil fertility and nutrient management to this variability is unknown. Field experiments from 1998 to 2000 on soils with low to high exchangeable potassium (K) concentrations evaluated K application and placement effects on isoflavone concentrations and composition of soybean in various tillage and row-width systems. Soybean seed yield and concentrations of daidzein, genistein, glycitein, leaf K, and seed K were measured. Significant increases in daidzein, genistein, and total isoflavone were observed with direct deep-banded K or residual surface-applied K on low-K soils. Positive effects of K fertilization on isoflavones were less frequent on medium- to high-testing K soils. Both individual and total isoflavones were often positively correlated with seed yield, leaf K, and seed K on low-K soils. Appropriate K management could be an effective approach to increase isoflavone concentrations for soybeans produced on low- to medium-K soils.

Topics: Agriculture; Fertilizers; Genistein; Glycine max; Isoflavones; Plant Leaves; Potassium; Seeds; Soil

Recent studies have reported that estrogen replacement therapy (ERT) reduces the risk of cardiovascular diseases in postmenopausal women. However, mechanisms responsible for this effect are not yet completely understood, and ERT is associated with carcinogenic side effects in women and feminizing effects in men. Because soybean isoflavones, a group of natural phytoestrogens, have only weak estrogenic activity and are not known to have side effects such as carcinogenesis and feminization, we evaluated the effects of genistein, daidzein and glycitein on the growth and DNA synthesis of aortic smooth muscle cells (SMC) from stroke-prone spontaneously hypertensive rats (SHRSP). SMC were cultured in dishes and proliferated on 10% dextran-coated charcoal/fetal bovine serum, and then treated with 0.1-30 micromol/L of genistein, daidzein or glycitein to investigate cell proliferation (cell number) and DNA synthesis (cell proliferation ELISA system), respectively. We also studied their effects on platelet-derived growth factor (PDGF)-BB (20 microg/L)-induced SMC proliferation. Soybean isoflavones inhibited proliferation and DNA synthesis of SMC from SHRSP in a concentration-dependent manner. Inhibition was significant at 3 micromol/L of genistein and 10 micromol/L of both daidzein and glycitein. For significant inhibition of PDGF-BB-induced SMC proliferation, concentrations as low as 0.1 micromol/L of each isoflavone were effective. These isoflavones, with their inhibitory effects on natural and PDGF-BB-induced SMC proliferation, may be useful in attenuatating such proliferation, a basic mechanism involved in atherosclerotic vascular change, thereby preventing atherosclerotic cardiovascular diseases.

Topics: Animals; Aorta; Cell Division; Cells, Cultured; Disease Susceptibility; DNA; Genistein; Glycine max; Growth Inhibitors; Hypertension; Isoflavones; Muscle, Smooth; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Stroke

Daily intake of isoflavones (daidzin, glycitin, genistin, daidzein, glycitein, and genistein) was determined quantitatively, based on the market basket method. Acid hydrolysis during extraction of foods was chosen to convert phytoestrogenes into the respective aglycons, facilitating HPLC analysis and allowing quantitation of total isoflavones as aglycones including both originally present glycosides and "free" aglycones. The isoflavones were extracted from samples with methanol and determined by reversed-phase HPLC analysis using a linear gradient of methanol-water as the eluent. From the results of hydrolysis, the daily intake of total isoflavon was 38.1 mg/adult Japanese. The values obtained by the market basket method and the National Nutrition Survey method were similar.

Topics: Adult; Aged; Child; Chromatography, High Pressure Liquid; Diet; Eating; Edible Grain; Glycine max; Humans; Isoflavones; Vegetables

Soy isoflavones are becoming of increasing interest as nutritional agents which can be used to combat osteoporosis and hyperlipidemia, and are also being considered as potential cancer chemopreventive compounds. However, prior to their formulation and distribution as therapeutic agents, thorough pharmacokinetic and toxicological assessment needs to be completed in men and women in a variety of health conditions in order to ensure their therapeutic efficacy and safety. At this time, studies of purified soy isoflavones are possible, and are being designed to fully evaluate the pharmacological utility of these preparations. In support of these studies, quantitative analysis of soy isoflavones in biological fluids can be accomplished with a wide variety of methods and analytical instrumentation. However, the relatively ubiquitous presence of high-performance liquid chromatography with ultraviolet detection (HPLC-UV) in most analytical laboratories, the relative ease of its operation, and the lesser expense of this instrumentation as compared to more sophisticated techniques such as liquid chromatography-mass spectrometry, offers some distinct advantages for its use in pharmacokinetic studies. In this manuscript, the development and validation of an HPLC-UV method for the quantitation of the principal soy isoflavones, genistein, daidzein, and glycitein, and their primary metabolites, in human plasma and urine is described. This analytical approach allows for pharmacologically relevant concentrations of the analytes and their principle metabolites to be detected, and has been validated in close agreement with the US Food and Drug Administration's guidelines for the validation of methods to be used in support of pharmacokinetic studies.

Topics: Chromatography, High Pressure Liquid; Genistein; Humans; Isoflavones; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet

An in vitro model, designated the Simulator of the Human Intestinal Microbial Ecosystem (SHIME), was used to study the effect of a soygerm powder rich in beta-glycosidic phytoestrogenic isoflavones on the fermentation pattern of the colon microbiota and to determine to what extent the latter metabolize the conjugated phytoestrogens. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 3 wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota were monitored. The microbiota were then subjected to a 2-wk treatment period by adding 2.5 g/d soygerm powder to the culture medium. The addition resulted into an overall increase of bacterial marker populations (Enterobacteriaceae:, coliforms, Lactobacillus: sp., Staphylococcus: sp. and Clostridium: sp.), with a significant increase of the Lactobacillus: sp. population. The short-chain fatty acid (SCFA) concentration increased approximately 30% during the supplementation period; this was due mainly to a significant increase of acetic and propionic acids. Gas analysis revealed that the methane concentration increased significantly. Ammonium and sulfide concentrations were not influenced by soygerm supplementation. Use of an electronic nose apparatus indicated that odor concentrations decreased significantly during the treatment period. The beta-glycosidic bonds of the phytoestrogenic isoflavones were cleaved under the conditions prevailing in the large intestine. The increased bacterial fermentation after addition of the soygerm powder was paralleled by substantial metabolism of the free isoflavones (genistein, daidzein and glycitein), resulting in recovery of only 12-17% of the supplemented isoflavones.

Topics: Bioreactors; Clostridium; Colon; Ecosystem; Electric Impedance; Enterobacteriaceae; Escherichia coli; Estrogens, Non-Steroidal; Feces; Fermentation; Glycine max; Humans; Intestines; Isoflavones; Lactobacillus; Models, Biological; Phytoestrogens; Plant Preparations; Staphylococcus

Isoflavonoids are a group of biologically active phytochemicals that humans are exposed to mainly through soy food intake. Because of the similar chemical structure of these compounds and estradiol, it has been hypothesized that isoflavonoids may be related to the risk of breast cancer. Overnight urine samples from 60 incident breast cancer cases and their individually matched controls were assayed for urinary excretion rates of five major isoflavonoids (daidzein, genistein, glycitein, equol, and O-desmethylangolensin) and total phenols. These subjects were from a large population-based case-control study conducted in Shanghai, and urine samples from breast cancer cases were collected before any cancer therapy to minimize the potential influence of the disease and its sequelae on study results. Urinary excretion of total phenols and all individual isoflavonoids, particularly glycitein, was substantially lower in breast cancer cases than controls. For total isoflavonoids, the mean excretion was 13.95 nmol/mg creatinine (SD, 20.76 nmol/mg creatinine) for cases and 19.52 nmol/mg creatinine (SD, 25.36 nmol/mg creatinine) for controls (P for difference = 0.04). The case-control difference was more evident when median levels of these compounds were compared, with the median excretion of all major isoflavonoids being 50-65% lower in cases than in controls. Individuals in the highest tertile of daidzein, glycitein, and total isoflavonoids had about half the cancer risk of those in the lowest tertile. The adjusted odds ratio for breast cancer was 0.14 (95% confidence interval, 0.02-0.88) for women whose urinary excretion of both phenol and total isoflavonoids was in the upper 50% compared with those in the lower 50%. The results from this study support the hypothesis that a high intake of soy foods may reduce the risk of breast cancer.

Topics: Adult; Anticarcinogenic Agents; Breast Neoplasms; Case-Control Studies; China; Chromans; Confidence Intervals; Creatinine; Equol; Estradiol; Estrogens, Non-Steroidal; Feeding Behavior; Female; Genistein; Glycine max; Humans; Isoflavones; Middle Aged; Monoamine Oxidase Inhibitors; Odds Ratio; Phenols; Risk Factors

Glycitein metabolism was compared with other isoflavones to begin to understand the effect of this compound. Total isoflavones of 4.5 micromol/kg body weight from soymilk (high in genistein and daidzein) and soygerm (high in daidzein and glycitein) was fed to seven women and seven men. To minimize interindividual variation, only subjects with moderate fecal isoflavone degradation rates (half-lives of daidzein and genistein were 15.7 and 8.9 h, respectively) were included. The average 48-h urinary excretion of glycitein, daidzein and genistein was approximately 55, 46 and 29% of the dose ingested, respectively, which was significantly different from each other in men and women (P < 0.001). Plasma isoflavone concentrations at 6 and 24 h after soymilk feeding paralleled relative amounts of isoflavones in soymilk (genistein > daidzein > glycitein) (P < 0.05) in men and women, but plasma isoflavone concentrations after soygerm feeding did not parallel soygerm isoflavone concentrations in women because genistein and glycitein did not differ from each other at 6 h after feeding. Six hours after soygerm dosing, plasma isoflavone concentrations paralleled soygerm isoflavone levels in men. Based on plasma isoflavone concentrations at 6 h after dosing, the bioavailabilities of daidzein and genistein were similar in men and women. At the high glycitein dose (soygerm), plasma concentration at 24 h after dosing suggested a modest gender difference in glycitein bioavailability.

Topics: Adult; Feces; Female; Genistein; Glycine max; Half-Life; Humans; Isoflavones; Male

Soy products contain high amounts of isoflavonoids, which have been shown to exhibit possible cancer-protective properties. Chinese populations in Asia, in particular, have a high level of soy intake and a relatively low risk of hormone-dependent cancers. In this study, we assessed the distributions of dietary soy isoflavonoids (daidzein, genistein, and glycitein) and urinary soy isoflavonoids and their metabolites (daidzein, genistein, glycitein, equol, and O-desmethylangolensin) among 147 Singapore Chinese (76 men and 71 women) ages 45-74 years, who are participants of the Singapore Cohort Study on diet and cancer. Urinary values were measured from spot samples collected 10-20 months following recruitment, when usual dietary habits were assessed by a structured food frequency/portion size questionnaire administered in person. Dietary levels of daidzein and genistein were comparable within individuals and about seven times higher than the level of dietary glycitein. All three dietary isoflavonoids showed an approximately 3.5-fold difference between the 25th and 75th percentile values. Similarly, daidzein was the most abundant and glycitein the least abundant of the five isoflavonoid compounds in urine. There was a 4.9-fold difference between the 25th and 75th percentile values for the sum of the five urinary isoflavonoids. Among study subjects, there were statistically significant, dose-dependent associations between frequency of overall soy intake and levels of urinary daidzein (two-sided P = 0.03) and sum of urinary daidzein, genistein, and glycitein (two-sided P = 0.04). In contrast, there were no associations between frequency of overall soy intake and levels of the two daidzein metabolites (equol and O-desmethylangolensin) in urine (two-sided P = 0.85 and 0.34, respectively). We suggest that within the range of exposures experienced by Singapore Chinese, urinary level of daidzein or the sum of daidzein, genistein, and glycitein obtained from a spot sample can serve as a biomarker of current soy consumption in epidemiological studies of diet-disease associations.

Topics: Aged; Antineoplastic Agents; Biomarkers; Cohort Studies; Diet; Female; Genistein; Glycine max; Humans; Isoflavones; Male; Middle Aged; Neoplasms, Hormone-Dependent; Singapore

Isoflavones are present in soybeans and its products in concentrations up to 300 mg/100 g, have estrogenic and antiestrogenic properties, and may be protective against hormone-related cancers. The purpose of this cross-sectional study was to investigate the association between urinary isoflavone excretion and self-reported soy intake. A total of 102 women of Caucasian, Native Hawaiian, Chinese, Japanese, and Filipino ancestry completed a dietary questionnaire for soy products consumed during the last year and during the 24-h period before urine collection. Overnight urine samples were analyzed for coumestrol and the soy isoflavones genistein, daidzein, and glycitein and their main human metabolites by reverse-phase high-pressure liquid chromatography. Soy protein and isoflavone intake (predominantly from tofu) were estimated using published nutritional databases. Wilcoxon's rank-sum test scores and Spearman rank correlation coefficients were computed. Japanese women excreted more daidzein, genistein, and glycitein than did Caucasian women, whereas Caucasian women excreted slightly more coumestrol. Soy intake differed significantly among ethnic groups. Dietary soy protein and isoflavone intakes during the previous 24 h were positively related to urinary isoflavone excretion [rs = 0.61 (P < 0.0001) and 0.62 (P < 0.0001), respectively]. Urinary excretion of isoflavones was also related to annual dietary soy protein and isoflavone intake [rs = 0.32 (P < 0.0012) and 0.31 (P < 0.0016), respectively]. The strong correlation between urinary isoflavone excretion and self-reported soy intake validates the dietary history questionnaire that is now used in a study exploring dietary risk factors for breast cancer.

Topics: Adult; Aged; Aged, 80 and over; China; Coumestrol; Cross-Sectional Studies; Female; Genistein; Glycine max; Hawaii; Humans; Isoflavones; Japan; Middle Aged; Philippines; White People

Five tempe-derived bacterial strains identified as Micrococcus or Arthrobacter species were shown to transform the soybean isoflavones daidzein and glycitein to polyhydroxylated isoflavones by different hydroxylation reactions. All strains converted glycitein and daidzein to 6,7,4'-trihydroxyisoflavone (factor 2) and the latter substrate also to 7,8,4'-trihydroxyisoflavone. Three strains transformed daidzein to 7,8,3',4'-tetrahydroxyisoflavone and 6,7,3',4'-tetrahydroxyisoflavone. In addition, two strains formed 6,7,8,4'-tetrahydroxyisoflavone from daidzein. Conversion of glycitein by these two strains led to the formation of factor 2 and 6,7,3',4'-tetrahydroxyisoflavone. The structures of these transformation products were elucidated by spectroscopic techniques and chemical degradation.

Topics: Arthrobacter; Biotransformation; Fermentation; Glycine max; Hydroxylation; Isoflavones; Kinetics; Micrococcus; Seeds

This study was undertaken to better understand the metabolic fate of dietary isoflavones in humans. Twelve volunteers were challenged with soya flour and urinary diphenol levels were then determined by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The presence of previously described urinary diphenols was confirmed, i.e. the isoflavones, daidzein and genistein; the isoflavonoid metabolites, equol, dihydrodaidzein (Int-O-D), O-desmethyl-angolensin (O-Dma); the lignan, enterolactone. Diphenols detected for the first time were the isoflavone, glycitein and five novel isoflavonoid metabolites which are tentatively identified as 6'-hydroxy-O-desmethylangolensin (6' OH-O-Dma), dihydrogenistein (Int-O-G), dehydro-O-desmethylangolensin (dehydro-O-Dma) and two isomers of tetrahydrodaidzein. Urinary excretion rates of the three isoflavones (daidzein, genistein, glycitein) over a 3-day period following soya challenge showed moderate variation (4x, 6x and 12x, respectively) between the 12 individuals suggesting some individual variabilities in ability to deconjugate and to absorb dietary isoflavones. However, urinary excretion rates of each of three major isoflavonoid metabolites (equol, O-Dma, 6' OH-O-Dma) showed more marked variation (922x, 17x, 15x, respectively); while some of this variability may reflect varying individual ability to ferment dietary isoflavones per se, an inverse relationship was found between urinary levels of equol and both O-Dma and 6' OH-O-Dma suggesting individual variability in the preferred metabolic pathways of dietary isoflavones.

Topics: Adult; Chromans; Chromatography, Gas; Equol; Female; Gas Chromatography-Mass Spectrometry; Genistein; Glycine max; Humans; Isoflavones; Lignans; Male; Middle Aged