Increased Butyrate Production During Long-Term Fermentation of In Vitro-Digested High Amylose Cornstarch Residues with Human Feces.

An in vitro semi-continuous long-term (3 wk) anaerobic incubation system simulating lower gut fermentation was used to determine variability in gut microbial metabolism between 4 predigested high amylose-resistant starch residues (SR): SRV, SRVI, SRVII, and SRGEMS in human fecal samples. Subjects participated twice, 5 mo apart: 30 in Phase I (15 lean, 9 overweight and 6 obese), 29 in Phase II (15 lean, 9 overweight, 5 obese); 13 of 15 lean subjects participated in both phases. Of the 4 SRs, SRV displayed the highest gelatinization temperature, peak temperature, enthalpy changes, and the least digestibility compared with the other SRs. In both phases, compared with blank controls, all SRs increased butyrate ∼2-fold which stabilized at week 2 and only SRV caused greater propionate concentration (∼30%) after 3 wk which might have been partly mediated by its lesser digestibility. Fecal samples from lean and overweight/obese subjects incubated with SRs showed similar short-chain fatty acid production across both time points, which suggests that resistant starch may benefit individuals across BMIs.

Soluble dietary fiber (Fibersol-2) decreased hunger and increased satiety hormones in humans when ingested with a meal.

We hypothesized that a digestion-resistant maltodextrin, Fibersol-2 (Archer Daniels Midland/Matsutani LLC, Decatur, IL, USA) may impact satiety by decreasing hunger, prolonging satiation, and/or increasing peripheral satiety signals. In a randomized, double-blind, placebo-controlled crossover study, healthy subjects (9 men and 10 women) underwent 3 treatments in which they consumed a standardized meal with a tea containing 0, 5, or 10 g of Fibersol-2. A visual analog scale questionnaire was given in 30-minute intervals to measure subjective appetite and satiety. Blood was drawn just before the meal (time 0) and at 30, 60, 90, 120, 180, and 240 minutes after meal for measurements of plasma ghrelin, cholecystokinin, gastrin, peptide YY, gastric inhibitory polypeptide, and glucagon-like peptide-1, all by enzyme-linked immunosorbent assay. There were significant delays in hunger and increased satiety for 1.5 to 2 hours after treatment with 10 g of Fibersol-2. These delays did not occur after ingesting 0 or 5 g Fibersol-2 at any time. Control and 5 g Fibersol-2 treatments did not suppress increases in hunger postmeal; hunger scores increased and satiety scores decreased significantly (P < .05) at all time points relative to the first postmeal assessment. Plasma peptide YY and glucagon-like peptide-1 were significantly increased by the ingestion of meal with tea containing 10 g Fibersol-2 compared with 0 or 5 g Fibersol-2 (P < .05). This study demonstrated that 10 g Fibersol-2 with a meal stimulated production of satiety hormones and enhanced satiety.

Resistant starch: promise for improving human health.

Ongoing research to develop digestion-resistant starch for human health promotion integrates the disciplines of starch chemistry, agronomy, analytical chemistry, food science, nutrition, pathology, and microbiology. The objectives of this research include identifying components of starch structure that confer digestion resistance, developing novel plants and starches, and modifying foods to incorporate these starches. Furthermore, recent and ongoing studies address the impact of digestion-resistant starches on the prevention and control of chronic human diseases, including diabetes, colon cancer, and obesity. This review provides a transdisciplinary overview of this field, including a description of types of resistant starches; factors in plants that affect digestion resistance; methods for starch analysis; challenges in developing food products with resistant starches; mammalian intestinal and gut bacterial metabolism; potential effects on gut microbiota; and impacts and mechanisms for the prevention and control of colon cancer, diabetes, and obesity. Although this has been an active area of research and considerable progress has been made, many questions regarding how to best use digestion-resistant starches in human diets for disease prevention must be answered before the full potential of resistant starches can be realized.

Echinacea sanguinea and Echinacea pallida extracts stimulate glucuronidation and basolateral transfer of Bauer alkamides 8 and 10 and ketone 24 and inhibit P-glycoprotein transporter in Caco-2 cells.

The use of Echinacea as a medicinal herb is prominent in the United States, and many studies have assessed the effectiveness of Echinacea as an immunomodulator. We hypothesized that Bauer alkamides 8, 10, and 11 and ketone 24 were absorbed similarly either as pure compounds or from Echinacea sanguinea and Echinacea pallida ethanol extracts, and that these Echinacea extracts could inhibit the P-glycoprotein transporter in Caco-2 human intestinal epithelial cells. Using HPLC analysis, the permeation rate of Bauer alkamides by passive diffusion across Caco-2 cells corresponded with compound hydrophilicity (alkamide 8 > 10 > 11), independent of the plant extract matrix. Both Echinacea ethanol extracts stimulated apparent glucuronidation and basolateral efflux of glucuronides of alkamides 8 and 10 but not alkamide 11. Bauer ketone 24 was totally metabolized to more hydrophilic metabolites when administered as a single compound, but was also glucuronidated when present in Echinacea extracts. Bauer alkamides 8, 10, and 11 (175-230 µM) and ethanol extracts of E. sanguinea (1 mg/mL, containing ~ 90 µM total alkamides) and E. pallida (5 mg/mL, containing 285 µM total alkamides) decreased the efflux of the P-glycoprotein transporter probe calcein-AM from Caco-2 cells. These results suggest that other constituents in these Echinacea extracts facilitated the metabolism and efflux of alkamides and ketones, which might improve therapeutic benefits. Alkamides and Echinacea extracts might be useful in potentiating some chemotherapeutics, which are substrates for the P-glycoprotein transporter.

Artichoke extract lowered plasma cholesterol and increased fecal bile acids in Golden Syrian hamsters.

A study was conducted in hamsters to determine if artichoke leaf extract (ALE) could lower plasma total and non-HDL cholesterol by increasing fecal excretion of neutral bile acids and sterols. Sixty-four Golden Syrian hamsters (8 week old) were fed control diet or a similar diet containing ALE (4.5 g/kg diet) for 6 weeks. No significant changes for total cholesterol, HDL, non-HDL cholesterol triglycerides or fecal neutral sterols and bile acids were found after 21 days for ALE-fed animals compared with controls. But after 42 days, ALE-fed male hamsters had significantly lower total cholesterol (15%), non-HDL cholesterol (30%) and triglycerides (22%) and female hamsters fed ALE showed reductions of 15% for total cholesterol, 29% for non-HDL cholesterol and 29% for triglycerides compared with controls. Total neutral sterol and bile acids concentrations increased significantly by 50% and 53% in fecal samples of ALE fed males, and 82.4% and 25% in ALE fed females compared with controls. The ALE lowered hamster plasma cholesterol levels by a mechanism involving the greater excretion of fecal bile acids and neutral sterols after feeding for 42 days.

Glycemic index, insulinemic index, and satiety index of kefir.

OBJECTIVES: To determine glycemic, insulinemic, and satiety indices of 3 types of kefir. METHODS: This study was divided into 3 phases. In phase 1, 50 g of available carbohydrate from low-fat strawberry kefir or orange kefir was tested, and in phase 2, low-fat plain kefir containing 25 g of available carbohydrates was tested for glycemic index (GI), in both cases compared with an equivalent amount of glucose. In phase 3, 1000-kJ portions of all 3 types of kefirs were compared with white bread with the same energy content to determine the insulinemic index (II) and satiety index (SI) of all 3 kefirs. In all phases, a single-meal, randomized crossover design was performed in which the test meals were given to healthy adults, 5 men and 5 women. RESULTS: The total incremental plasma glucose area under the curve (iAUC) for strawberry, orange, and plain kefirs was significantly lower compared with the respective high-GI control food, which was glucose solution. However, the IIs and SIs of kefir did not differ significantly from the white bread. CONCLUSION: Kefir is a low- to moderate-GI food; however, its II was high. Although kefir had higher water content, the SI of kefir was not significantly different from white bread.

Plasma caffeic acid is associated with statistical clustering of the anticolitic efficacy of caffeic acid in dextran sulfate sodium-treated mice.

We hypothesized that interindividual variability in the bioavailability of caffeic acid (CA) would influence its anticolitic efficacy and that mice may be appropriate for modeling human gut microbial metabolism of CA, which is thought to influence CA bioavailability. Anaerobic human fecal and mouse cecal sample mixtures were incubated with CA derivatives from Echinacea purpurea and compound disappearance rates were measured, which were similar in both sample types. CA metabolism, including formation of its main metabolite, m-hydroxyphenylpropionate, in the mouse cecum may usefully model human gut metabolism of this compound. Ten-week-old CD-1/IGS female mice were fed 120 mg CA/kg (n = 36) or control diet for 7 d (n = 12); one-half of each group then drank 1.25% dextran sulfate sodium (DSS) in water for 5 d. DSS-treated mice fed CA showed lessened colitic damage than did mice given DSS alone, with longer colons, greater body weight, and colonic Cyp4b1 expression. Cluster analysis of the cecal histopathological score showed that mice with severe cecal damage (mean cecal score = 8.5; n = 11) also had greater myeloperoxidase (MPO) activity and lower plasma CA compared with mice showing mild cecal damage (mean cecal score = 4.5; n = 4) (P < 0.05). Cecal score was positively correlated with colonic MPO activity (r = 0.72; P < 0.05) and negatively correlated with plasma CA (r = -0.57; P < 0.05). These studies indicated that the anticolitic efficacy of CA was related to variability in CA bioavailability, which may be influenced by gut microbial metabolism of this compound.

Lesser in vitro anaerobic cecal isoflavone disappearance was associated with greater apparent absorption of daidzein and genistein in Golden Syrian hamsters.

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.

Inhibition of azoxymethane-induced preneoplastic lesions in the rat colon by a cooked stearic acid complexed high-amylose cornstarch.

This study evaluated a novel stearic acid complexed high-amylose cornstarch (SAC) for the prevention of preneoplastic lesions in the colon of azoxymethane (AOM)-treated Fisher 344 rats fed resistant starches at 50-55% of the diet for 8 weeks. Uncooked SAC (r-SAC) diet was compared with raw normal-cornstarch diet (r-CS) or raw high-amylose cornstarch diet (r-HA), and water-boiled CS (w-CS) was compared with w-HA and w-SAC, respectively. w-SAC markedly reduced mucin-depleted foci (MDF) numbers compared with w-HA or w-CS. r-HA significantly decreased aberrant crypt foci (ACF) numbers compared with r-CS or r-SAC. Increased cecum weight and decreased cecum pH were observed in the SAC or HA groups. The highest amounts of total or individual short-chain fatty acids (SCFAs) in cecum and of butyrate or propionate in feces were observed in the AOM-treated w-SAC group. This study revealed the effectiveness of a novel resistant starch in inhibiting colonic preneoplastic lesions and the importance of high-moisture cooking on the suppression of colon carcinogenesis by this resistant starch.

Permeability of rosmarinic acid in Prunella vulgaris and ursolic acid in Salvia officinalis extracts across Caco-2 cell monolayers.

ETHNOPHARMACOLOGICAL RELEVANCE: Rosmarinic acid (RA), a caffeic acid-related compound found in high concentrations in Prunella vulgaris (self-heal), and ursolic acid (UA), a pentacyclic triterpene acid concentrated in Salvia officinalis (sage), have been traditionally used to treat inflammation in the mouth, and may also be beneficial for gastrointestinal health in general. AIM OF THE STUDY: To investigate the permeabilities of RA and UA as pure compounds and in Prunella vulgaris and Salvia officinalis ethanol extracts across human intestinal epithelial Caco-2 cell monolayers. MATERIALS AND METHODS: The permeabilities and phase II biotransformation of RA and UA as pure compounds and in herbal extracts were compared using Caco-2 cells with HPLC detection. RESULTS: The apparent permeability coefficient (P(app)) for RA and RA in Prunella vulgaris extracts was 0.2 ± 0.05 × 10(-6)cm/s, significantly increased to 0.9 ± 0.2 × 10(-6)cm/s after ß-glucuronidase/sulfatase treatment. P(app) for UA and UA in Salvia officinalis extract was 2.7 ± 0.3 × 10(-6)cm/s and 2.3 ± 0.5 × 10(-6)cm/s before and after ß-glucuronidase/sulfatase treatment, respectively. Neither compound was affected in permeability by the herbal extract matrix. CONCLUSION: RA and UA in herbal extracts had similar uptake as that found using the pure compounds, which may simplify the prediction of compound efficacy, but the apparent lack of intestinal glucuronidation/sulfation of UA is likely to further enhance the bioavailability of that compound compared with RA.

Efficacy of a mycotoxin binder against dietary fumonisin, deoxynivalenol, and zearalenone in rats.

It was hypothesized that a mycotoxin binder, Grainsure E, would inhibit adverse effects of a mixture of fumonisin B1, deoxynivalenol, and zearalenone in rats. For 14 and 28 days, 8-10 Sprague-Dawley rats were fed control diet, Grainsure E (0.5%), toxins (7 µg fumonisin B1/g, 8 µg of deoxynivalenol/g and 0.2 µg of zearalenone/g), toxins (12 µg of fumonisin B1/g, 9 µg of deoxynivalenol/g, and 0.2 µg of zearalenone/g + Grainsure E), or pair-fed to control for food intake of toxin-fed rats. After 28 days, decreased body weight gain was prevented by Grainsure E in toxin-fed female rats, indicating partial protection against deoxynivalenol and fumonisin B1. Two effects of fumonisin B1 were partly prevented by Grainsure E in toxin-fed rats, increased plasma alanine transaminase (ALT) and urinary sphinganine/sphingosine, but sphinganine/sphingosine increase was not prevented in females at the latter time point. Grainsure E prevented some effects of fumonisin B1 and deoxynivalenol in rats.

Bacteroides uniformis is a putative bacterial species associated with the degradation of the isoflavone genistein in human feces.

Inter-individual variation in isoflavone absorption depends on gut microbial degradation and affects the efficacy of these compounds. We hypothesized that inter-individual variation in fecal isoflavone disappearance coincided with variation in bacterial species. In vitro anaerobic fecal disappearance of isoflavones was measured from 33 participants by HPLC. Fecal microbial 16S rRNA variable region PCR products were obtained from 4 participants with the greatest and least genistein or glycitein degradation and were subjected to denaturing gradient gel electrophoresis. DNA bands with a homology of 90-95% to Bacteroides uniformis and Faecalibacterium prausnitzii were present in greater intensities in fecal samples showing a genistein disappearance rate constant of 1.47 ± 0.14 h(-1) compared with those with a genistein disappearance rate constant of 0.15 ± 0.03 h(-1) (P < 0.05). Human fecal bacterial species with DNA sequences 90-100% homologous to Tannerella forsythensis and 4 other species were present in greater intensities in fecal samples showing a glycitein disappearance rate constant of 0.57 ± 0.30 h(-1) compared with fecal samples with a glycitein disappearance rate constant of 0.08 ± 0.03 h(-1) (P < 0.05). In high degraders, B. uniformis may be a candidate for genistein degradation and T. forsythensis for glycitein degradation, based on fecal isoflavone degradation in the presence of these species. Bacteroides acidifaciens increased isoflavone disappearance in anaerobic human fecal incubations under nutrient-rich and -depleted conditions, suggesting this species as one responsible for the generally high degradation of isoflavones by humans. These fecal microbes are candidate biomarkers for interindividual variation in isoflavone uptake and efficacy.

(n-3) Fatty Acids: Clinical Trials in People with Type 2 Diabetes.

Recent human clinical trials of the effects of (n-3) fatty acids on participants with type 2 diabetes (T2D) were reviewed, focusing on 11 clinical trials conducted within the past 4 y, and subsequent to a Cochrane Database meta-analysis of this topic. Doses of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in these studies were mostly in the range of ∼2 g/d provided for 6 wk to 6 mo. Summarizing across these studies, there were no changes in fasting glucose or insulin compared with baseline or placebo. (n-3) Fatty acids generally decreased serum triglycerides but had varying effects on serum cholesterol, LDL cholesterol, and HDL cholesterol. A few studies indicated beneficial effects of (n-3) fatty acids on arterial blood flow. The effects of EPA and/or DHA have not yet been studied in clinical trials in participants at risk for T2D; the prevention or exacerbation of T2D by fish oil or EPA and DHA supplements of amounts >0.5 g/d deserves study. The prevention of adverse vascular effects of T2D by (n-3) fatty acids may be a promising direction for further study.

Greater apparent absorption of flavonoids is associated with lesser human fecal flavonoid disappearance rates.

It was hypothesized that 5,7,4'-OH-flavonoids disappeared more rapidly from human fecal incubations and were less absorbable by humans than flavonoids without 5-OH moieties. Anaerobic fecal disappearance rates over 24 h were determined for 15 flavonoids in samples from 20 men and 13 women. In these anaerobic fecal mixtures, flavonoids with 5,7,4'-OH groups, genistein, apigenin, naringenin, luteolin, kaempferol, and quercetin (disappearance rate, k=0.46+/-0.10 h(-1)), and methoxylated flavonoids, hesperetin and glycitein (k=0.24+/-0.21 h(-1)), disappeared rapidly compared with flavonoids lacking 5-OH (e.g., daidzein, k=0.07+/-0.03 h(-1)). Apparent absorption of flavonoids that disappeared rapidly from in vitro fecal incubations, genistein, naringenin, quercetin, and hesperetin, was compared with that of daidzein, a slowly disappearing flavonoid, in 5 men and 5 women. Subjects ingested 104 micromol of genistein and 62 micromol of daidzein (soy milk), 1549 micromol of naringenin and 26 micromol of hesperetin (grapefruit juice), and 381 micromol of quercetin (onions) in three test meals, each separated by 1 week. Blood and urine samples were collected over 24 h after each test meal. Plasma flavonoid concentrations ranged from 0.01 to 1 microM. The apparent absorption, expressed as percentage of ingested dose excreted in urine, was significantly less for naringenin (3.2+/-1.7%), genistein (7.2+/-4.6%), hesperetin (7.3+/-3.2%), and quercetin (5.6+/-3.7%) compared with daidzein (43.4+/-15.5%, p=0.02). These data affirmed the hypothesis that the 5,7,4'-OH of flavonoids limited apparent absorption of these compounds in humans.

Deoxynivalenol suppresses circulating and splenic leukocyte subpopulations in BALB/c mice: dose response, time course and sex differences.

It was hypothesized that suppression of peripheral blood leukocyte subsets were markers of exposure to dietary deoxynivalenol (DON), a Fusarium graminearum mycotoxin in grain, at 1.0 mg kg(-1) but not at lesser doses in BALB/c mice. Groups of 10 female and 10 male BALB/c mice were fed 0, 0.25, 0.5, 1.0, and 2.0 mg kg(-1) DON for 14 and 28 days. Using flow cytometry with staining for leukocyte surface markers, the percentage of CD19(+) leukocytes (B cells) in peripheral blood was decreased in both sexes of BALB/c mice after 14 days of exposure to 1.0 or 2.0 mg kg(-1) DON, whereas exposure to DON over 28 days did not inhibit B cells compared to the control diet. The percentage of mononuclear cells in peripheral blood was decreased in female BALB/c mice fed 1 and 2 mg kg(-1) DON after 14 days compared with control diet. The percentage of CD11b(+) leukocytes (monocytes) in peripheral blood and total CD11b(+) splenic leukocytes were decreased only in female mice fed 1.0 and 2.0 mg kg(-1) DON after 28 days compared with control diet, which shows the greater sensitivity to DON in females compared to males. It was concluded that BALB/c mice adapted to DON exposure because peripheral blood cellular effects of DON at 14 days disappeared by 28 days with the exception of monocyte changes in females. This suggests that female sex hormones potentiate one potential marker of DON immunotoxicity in BALB/c mice.

Increased CYP4B1 mRNA is associated with the inhibition of dextran sulfate sodium-induced colitis by caffeic acid in mice.

Susceptibility to inflammatory bowel diseases depends upon interactions between the genetics of the individual and induction of chronic mucosal inflammation. We hypothesized that administration of dietary phenolics, caffeic acid and rutin, would suppress upregulation of inflammatory markers and intestinal damage in a mouse model of colitis. Colitis was induced in C3H/ HeOuJ mice (8 weeks old, 6 male/6 female per treatment) with 1.25% dextran sulfate sodium (DSS) for 6 d in their drinking water. Rutin (1.0 mmol (524 mg)/kg in diet), caffeic acid (1.0 mmol (179 mg)/kg in diet), and hypoxoside extract (15 mg/d, an anticolitic phenolic control) were fed to the mice for 7 d before and during DSS treatment, as well as without DSS treatment. Body weight loss was prevented by rutin and caffeic acid during DSS treatment. Colon lengths in mice fed caffeic acid and hypoxoside during DSS treatment were similar to DSS-negative control. Food intake was improved and myeloperoxidase (MPO) was decreased with each phenolic treatment in DSS-treated mice compared with DSS treatment alone. Colonic mRNA expression of IL-17 and iNOS were inhibited when IL-4 was increased by each phenolic treatment combined with DSS, whereas CYP4B1 mRNA was increased only by caffeic acid in DSS-treated mice, compared with DSS treatment alone. Colonic and cecal histopathology scores of DSS-treated mice were significantly more severe (P < 0.01) than in mice fed caffeic acid before and during DSS treatment, based on mucosal height, necrosis, edema, erosion, and inflammatory cell infiltration. Although both rutin and caffeic acid suppressed the expression of selected inflammatory markers, only caffeic acid protected against DSS-induced colitis, in association with normalization of CYP4B1 expression. The inhibition of DSS-induced colitic pathology by caffeic acid was mediated by mechanisms in addition to anti-inflammatory effects that deserve further study.

Isoflavone glycitein diminished plasma cholesterol in female golden Syrian hamsters.

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.

Synthesis and characterization of deoxynivalenol glucuronide: its comparative immunotoxicity with deoxynivalenol.

Deoxynivalenol (DON) is a mycotoxin commonly contaminating wheat, barley and corn. DON glucuronide (DONGLU) is a major DON metabolite. We synthesized and purified DONGLU and tested its immunotoxicity, hypothesizing that DONGLU would be much less toxic to K562 cells compared with DON. DONGLU was synthesized using rat liver microsomes, uridine-5'-diphosphoglucuronic acid and DON, and purified with a Sephadex LH-20 column and reverse phase HPLC. beta-Glucuronidase hydrolysis formed a product with retention time and UV spectrum identical with DON. Using atmospheric pressure chemical ionization in negative mode, the molecular mass (M-1) of purified DONGLU was 471 g/mol; in agreement with an expected molecular weight of 472 g/mol. MS and NMR indicated that the glucuronide moiety was conjugated with the carbon-3-hydroxyl group of DON. The cytotoxicity of DON and DONGLU were compared in cell culture using human erythroleukemia cell line K562. Fifty percent inhibition of cell number was observed with a DON concentration of 1.31 microM using a methylthaizol tetrazolium (MTS) cell viability assay whereas no significant cytotoxicity was observed for DONGLU at up to 270 microM. DONGLU did not influence DON toxicity at 0.5 microM, 1.3 microM and 8.4 microM concentration combinations of each compound. These data verified that DONGLU is a detoxification product of DON.

High urinary isoflavone excretion phenotype decreases plasma cholesterol in golden Syrian hamsters fed soy protein.

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.

Metabolism of glycitein (7,4'-dihydroxy-6-methoxy-isoflavone) by human gut microflora.

Gut microbial disappearance and metabolism of the soy isoflavone glycitein, 7,4'-dihydroxy-6-methoxyisoflavone, were investigated by incubating glycitein anaerobically with feces from 12 human subjects. The subjects' ages ranged from 24 to 53 years with a body mass index (BMI) of 20.9-25.8 kg/m(2) (mean BMI = 24.0 +/- 1.1 kg/m(2)). Glycitein disappearance followed an apparent first-order rate loss. Fecal glycitein disappearance rates for the subjects segregated into three different groups described as high (k = 0.67 +/- 0.14/h), moderate (k = 0.34 +/- 0.04/h), and low (k = 0.15 +/- 0.07/h) glycitein degraders (p < 0.0001). There was no dose effect on the disappearance rates for each subject from 10 to 250 microM glycitein (average k = 0.32 +/- 0.03/h, p > 0.05). Four putative glycitein metabolites, characterized by liquid chromatography-mass spectrometry (electrospray ionization using positive ionization mode), were dihydroglycitein, dihydro-6,7,4'-trihydroxyisoflavone, and 5'-O-methyl-O-desmethylangolensin. Two subjects produced a metabolite tentatively identified as 6-O-methyl-equol, and one subject produced daidzein as an additional metabolite of glycitein. These results show that glycitein is metabolized by human gut microorganisms and may follow metabolic pathways similar to other soy isoflavones.