Potentiation of the bioavailability of daidzin by an extract of Radix puerariae.

The dose effect of pure daidzin on the suppression of ethanol intake in Syrian golden hamsters was compared with that of crude daidzin contained in a methanol extract of Radix puerariae (RP). EC50 values estimated from the graded dose-response curves for pure daidzin and RP extract daidzin are 23 and 2.3 mg per hamster per day, respectively. Apparently the antidipsotropic activity of the RP extract cannot be accounted for solely by its daidzin content (22 mg/g). In addition to daidzin, six other isoflavones were identified in the RP extract and quantified--namely, puerarin (160 mg per g of extract), genistin (3.7 mg/g), daidzein (2.6 mg/g), daidzein-4',7-diglucoside (1.2 mg/g), genistein (0.2 mg/g), and formononetin (0.16 mg/g). None of these, administered either alone or combined, contributes in any significant way to the antidipsotropic activity of the extract. Plasma daidzin concentration-time curves determined in hamsters administered various doses of pure daidzin or RP extract by i.p.injection indicate that the crude extract daidzin has approximately 10 times greater bioavailability than the pure compound. Reconstruction of the dose-response effects for pure and crude daidzin using bioavailable daidzin rather than administered dose gives a single curve. Synthetic daidzin added to the RP extract acquires the bioavailability of the endogenous daidzin that exists naturally in the extract. These results show that (i) daidzin is the major active principle in methanol extracts of RP, and (ii) additional constituents in the methanol extract of RP assist uptake of daidzin in golden hamsters.

Daidzin suppresses ethanol consumption by Syrian golden hamsters without blocking acetaldehyde metabolism.

Daidzin is a potent, selective, and reversible inhibitor of human mitochondrial aldehyde dehydrogenase (ALDH) that suppresses free-choice ethanol intake by Syrian golden hamsters. Other ALDH inhibitors, such as disulfiram (Antabuse) and calcium citrate carbimide (Temposil), have also been shown to suppress ethanol intake of laboratory animals and are thought to act by inhibiting the metabolism of acetaldehyde produced from ingested ethanol. To determine whether or not daidzin inhibits acetaldehyde metabolism in vivo, plasma acetaldehyde in daidzin-treated hamsters was measured after the administration of a test dose of ethanol. Daidzin treatment (150 mg/kg per day i.p. for 6 days) significantly suppresses (> 70%) hamster ethanol intake but does not affect overall acetaldehyde metabolism. In contrast, after administration of the same ethanol dose, plasma acetaldehyde concentration in disulfiram-treated hamsters reaches 0.9 mM, 70 times higher than that of the control. In vitro, daidzin suppresses hamster liver mitochondria-catalyzed acetaldehyde oxidation very potently with an IC50 value of 0.4 microM, which is substantially lower than the daidzin concentration (70 microM) found in the liver mitochondria of daidzin-treated hamsters. These results indicate that (i) the action of daidzin differs from that proposed for the classic, broad-acting ALDH inhibitors (e.g., disulfiram), and (ii) the daidzin-sensitive mitochondrial ALDH is not the one and only enzyme that is essential for acetaldehyde metabolism in golden hamsters.

Isoflavones in food supplements: chemical profile, label accordance and permeability study in Caco-2 cells

Consumers nowadays are playing an active role in their health-care. A special case is the increasing number of women, who are reluctant to use exogenous hormone therapy for the treatment of menopausal symptoms and are looking for complementary therapies. However, food supplements are not clearly regulated in Europe. The EFSA has only recently begun to address the issues of botanical safety and purity regulation, leading to a variability of content, standardization, dosage, and purity of available products. In this study, isoflavones (puerarin, daidzin, genistin, daidzein, glycitein, genistein, formononetin, prunetin, and biochanin A) from food supplements (n = 15) for menopausal symptoms relief are evaluated and compared with the labelled information. Only four supplements complied with the recommendations made by the EC on the tolerable thresholds. The intestinal bioavailability of these compounds was investigated using Caco-2 cells. The apparent permeability coefficients of the selected isoflavonoids across the Caco-2 cells were affected by the isoflavone concentration and product matrix.

Enhanced aglycone production of fermented soybean products by Bacillus species

This study evaluated the effect of starter culture and fermentation period on the isoflavone content of protein-rich soybeans variety TG145. Initially, soybeans were washed, soaked in water for 16 h and autoclaved at 121°C for 40min. Three different bacterial starter cultures (~104 CFU/g) namely Bacillus subtilis BEST195, B. subtilis Asaichiban and B. subtilis TN51 were then added and the fermentation was allowed to proceed at 42°C for 24 h (natto-style) and 72 h (thua nao-style). The quantities of six major isoflavones (daidzin, genistin, glycitin, daidzein, genistein, and glycitein) were then determined in these fermented soybean products using reverse phase HPLC technique. Generally, our results clearly showed that the content of total isoflavones in the fermented products prepared by Bacillus starter cultures greatly increased ranging from 43 to 99% compared to that of the unfermented autoclaved soybeans. In addition, a dramatic increase of aglycones was also observed (> 400%) in the soybean products fermented by Bacillus subtilis strain TN51. This present study suggests a promising use of Bacillus starter cultures in improving isoflavone compounds especially the aglycones which would benefit for novel functional food development.

Flavonoid increase in soybean as a response to Nezara viridula injury and its effect on insect-feeding preference

The ability of the stink bug (Nezara viridula) to induce and/or increase production of chemical defenses, i.e., flavonoids, in immature seeds of five genotypes of soybean (BR-16, IAC-100, PI 227687, PI 229358, and PI 274454) was investigated under greenhouse and laboratory conditions. Samples from pods of each genotype damaged by stink bug were analyzed for flavonoid content with high performance liquid chromatography. A dual-choice test was conducted to evaluate the feeding preference of N. viridula comparing BR-16 pods treated with extracts of PI 227687 seeds (with and without stink-bug injury), with water-treated pods. Seeds of PI 227687 damaged by N. viridula presented the highest concentration (352 mug/g) of daidzin (4'-hydroxyisoflavone-7-glucoside). The same trend was observed with genistin (4',5,7-trihydroxyisoflavone-7-glucoside): PI 227687 contained 142.4 mug/g, PI 274454, 31.6 mug/g, and PI 229358, 38.9 mug/g. Seeds damaged by stink bugs had higher isoflavone contents (daidzin and genistin), compared to controls. However, after being damaged, PI 274454 and PI 229358 produced less genistin than the other genotypes and no differences in concentration between damaged and nondamaged plants of this genotypes were observed. The numbers of observations of the insect feeding and the numbers of stylet sheaths left in water-treated BR-16 pods were greater than in those treated with PI 227687 extracts. The insects fed for longer periods on BR-16 pods treated with extract of PI 227687 without injury compared to those that were treated with extract of PI 227687 previously injured by stink bugs. Extracts of PI 227687 pods ( damaged or not) were deterrent to adults of N. viridula, and insect injury increased concentrations of daidzin and genistin in PI 227687 seeds. The deterrence seemed to be more pronounced after pods had suffered stink-bug injury.

cis-Jasmone e Tween 20 na indução de metabólitos secundários em soja e seus efeitos em insetos-praga

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de isoflavonas de soja no ambiente e correlação com atividades estrogênica : estudo de caso da região de Dourados (MS)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Release of Flavonoids by the Soybean Cultivars McCall and Peking and Their Perception as Signals by the Nitrogen-Fixing Symbiont Sinorhizobium fredii1

Sinorhizobium fredii strain USDA191 forms N-fixing nodules on the soybean (Glycine max L. Merr.) cultivars (cvs) McCall and Peking, but S. fredii strain USDA257 nodulates only cv Peking. We wondered whether specificity in this system is conditioned by the release of unique flavonoid signals from one of the cultivars or by differential perception of signals by the strains. We isolated flavonoids and used nodC and nolX, which are nod-box-dependent and -independent nod genes, respectively, to determine how signals activate genes in the microsymbionts. Seeds of cv McCall and cv Peking contain the isoflavones daidzein, genistein, and glycitein, as well as their glucosyl and malonylglucosyl glycosides. Roots exude picomolar concentrations of daidzein, genistein, glycitein, and coumestrol. Amounts are generally higher in cv Peking than in cv McCall, and the presence of rhizobia markedly influences the level of specific signals. Nanomolar concentrations of daidzein, genistein, and coumestrol induce expression of nodC and nolX in strain USDA257, but the relative nolX-inducing activities of these signals differ in strain USDA191. Glycitein and the conjugates are inactive. Strain USDA257 deglycosylates daidzin and genistin into daidzein and genistein, respectively, thereby converting inactive precursors into active inducers. Although neither soybean cultivar contains unique nod-gene-inducing flavonoids, strain- and cultivar-specific interactions are characterized by distinct patterns of signal release and response.

The natural plant stress elicitor cis-jasmone causes cultivar-dependent reduction in growth of the stink bug, Euschistus heros and associated changes in flavonoid concentrations in soybean, Glycine max.

To test the hypothesis that the plant stress related elicitor cis-jasmone (cJ) provides protection in soybean pods against the seed-sucking stink bug pest, Euschistus heros, the growth of E. heros on cJ-treated pods was investigated using three soybean cultivars differing in insect susceptibility, i.e. BRS 134 (susceptible), IAC 100 (resistant) and Dowling (resistant). E. heros showed reduced weight gain when fed cJ-treated Dowling, whereas no effect on weight gain was observed when fed other treated cultivars. Using analysis of variance, a three factor (cultivar x treatment x time) interaction was observed with concentrations of the flavonoid glycosides daidzin and genistin, and their corresponding aglycones, daidzein and genistein. There were increases in genistein and genistin concentrations in cJ-treated Dowling at 144 and 120 h post treatment, respectively. Higher concentrations of malonyldaidzin and malonylgenistin in Dowling, compared to BRS 134 and IAC 100, were observed independently of time, the highest concentrations being observed in cJ-treated seeds. Levels of glycitin and malonylglycitin were higher in BRS 134 and IAC 100 compared to Dowling. Canonical variate analysis indicated daidzein (in the first two canonical variates) and genistein (in the first only) as important discriminatory variables. These results suggest that cJ treatment leads to an increase in the levels of potentially defensive isoflavonoids in immature soybean seeds, but the negative effect upon E. heros performance is cultivar-dependent.