Effect of the pasteurization process on the contents of ascorbigen, indole-3-carbinol, indole-3-acetonitrile, and 3,3'-diindolylmethane in fermented cabbage

The aim of the study was to investigate the effect of the pasteurization process on the content of ascorbigen, indole-3-carbinol, indole-3-acetonitrile, and 3,3'-diindolylmethane in fermented cabbage. Pasteurization was run at a temperature of 80 °C for 5-30 min. Significant changes were only observed in contents of ascorbigen and 3,3'-diindolylmethane. The total content of the compounds analyzed in cabbage pasteurized for 10-30 min was found to be decreased by ca. 20%, and the losses were due to thermal degradation of the predominating ascorbigen. Pasteurization was found not to exert any considerable effect on contents of indole-3-acetonitrile and indole-3-carbinol in cabbage nor did it affect contents of the compounds analyzed in juice.     

In vitro antiproliferative activity of isothiocyanates and nitriles generated by myrosinase-mediated hydrolysis of glucosinolates from seeds of cruciferous vegetables

A comparison of the effect of isothiocyanates and nitriles derived from some glucosinolates, namely, epi-progoitrin, sinalbin, glucotropaeolin, glucocheirolin, and glucoraphenin, on human erythroleukemic in vitro cultured cells was studied. Many studies have in fact evidenced that a consumption of vegetable containing glucosinolates could reduce the development of colorectal cancer. In the experimental conditions used, the production of isothiocyanates and nitriles from glucosinolates is almost quantitative as confirmed by HPLC or GC-MS analysis. The obtained results demonstrated that in general nitriles are considerably less potent than the corresponding isothiocyanates in inhibiting cancer cell growth. Particularly, the isothiocyanates inhibitory activity on K562 cells growth is higher in the case of products derived from epi-progoitrin, glucotropaeolin, glucoraphenin, and glucocheirolin; while for nitriles the higher activity in inhibiting K562 cells growth is showed by sinalbin-derived product. Considering the antiproliferative activity found for isothiocyanates and nitriles, further studies will be aimed to the possible application of glucosinolate-derived products as chemopreventive cancer agents for the reduction of colorectal cancer.     

Inhibition of cancer cell proliferation in vitro by fruit and berry extracts and correlations with antioxidant levels

The effects of 10 different extracts of fruits and berries on cell proliferation of colon cancer cells HT29 and breast cancer cells MCF-7 were investigated. The fruits and berries used were rosehips, blueberries, black currant, black chokeberries, apple, sea buckthorn, plum, lingonberries, cherries, and raspberries. The extracts decreased the proliferation of both colon cancer cells HT29 and breast cancer cells MCF-7, and the effect was concentration dependent. The inhibition effect for the highest concentration of the extracts varied 2-3-fold among the species, and it was in the ranges of 46-74% (average = 62%) for the HT29 cells and 24-68% (average = 52%) for the MCF-7 cells. There were great differences in the content of the analyzed antioxidants in the extracts. The level of the vitamin C content varied almost 100-fold, and the content of total carotenoids varied almost 150-fold among the species. Also in the composition and content of flavonols, hydroxycinnamic acids, anthocyanins, and phenolics were found great differences among the 10 species. The inhibition of cancer cell proliferation seen in these experiments correlated with levels of some carotenoids and with vitamin C levels, present at levels that can be found in human tissues. The same inhibition of cell proliferation could not be found by ascorbate standard alone. This correlation might indicate a synergistic effect of vitamin C and other substances. In MCF-7 cells, the anthocyanins may contribute to the inhibition of proliferation.     

Development and validation of a spectrophotometric method for quantification of total glucosinolates in cruciferous vegetables

Given their putative role in chemoprevention, validated methods are needed for quantification of total glucosinolates. Based on the colorimetric reaction of ferricyanide with 1-thioglucose, released by alkaline hydrolysis of glucosinolates, we developed a simple and sensitive method for spectrophotometric quantification of total glucosinolates in cruciferous vegetables. Lyophilized and ground vegetables are extracted with 80% boiling methanol. Extracted glucosinolates are isolated using a strong anion exchange column and then hydrolyzed with 2 N NaOH to release 1-thioglucose. Ferricyanide is added, and the decrease in absorbance is measured at 420 nm, with final values adjusted for background. Recovery of internal standard (sinigrin) was 107%. Intra- and interassay coefficients of variation were 5.4% and 15.8%, respectively. Dose response was linear with sinigrin and amount of plant material extracted (R(2) ≥ 0.99). Using sinigrin, the lower limit of quantification was 0.6 mg. This straightforward method may be an alternative to time-consuming and costly chromatographic methods.     

LC-MS/MS quantification of sulforaphane and indole-3-carbinol metabolites in human plasma and urine after dietary intake of selenium-fortified broccoli

This study aimed at developing a sensitive LC-MS/MS method for the quantification of sulforaphane (SFN) and indole-3-carbinol metabolites in plasma and urine after dietary intake of regular and selenium-fertilized broccoli using stable isotope dilution analysis. In a three-armed, placebo-controlled, randomized human intervention study with 76 healthy volunteers, 200 g of regular (485 μg of total glucosinolates and <0.01 μg of selenium per gram fresh weight) or selenium-fertilized broccoli (589 μg of total glucosinolates and 0.25 μg of selenium per gram fresh weight) was administered daily for 4 weeks. Glucoraphanin and glucobrassicin metabolites quantified in plasma and urine were SFN-glutathione, SFN-cysteine, SFN-cysteinylglycine, SFN-acetylcysteine, and indole-3-carboxaldehyde, indole-3-carboxylic acid, and ascorbigen, respectively. Dietary intake of selenium-fertilized broccoli increased serum selenium concentration analyzed by means of atomic absorption spectroscopy by up to 25% (p < 0.001), but affected neither glucosinolate concentrations in broccoli nor their metabolite concentrations in plasma and urine compared to regular broccoli.     

Determination of ciprofloxacin, the major metabolite of enrofloxacin, in milk by isopotential fluorimetry

A new method for the determination of ciprofloxacin, the major metabolite of enrofloxacin, for concentrations between 20 and 200 ng/mL by means of matrix isopotential synchronous fluorescence spectrometry and derivative techniques is proposed. This new method is useful for the determination of compounds in samples with unknown background fluorescence, such as ciprofloxacin in whey, without the need of tedious preseparation. The determination was performed in an ethanol/water medium (20% v/v) at pH 4.8, provided by adding a sodium acetate/acetic acid buffer solution. Since enrofloxacin is widely used as an antibacterial agent in veterinary medicine, the method was successfully applied to the determination of its main metabolite in milk. An exhaustive statistical analysis has been developed to all calibration graphs. This treatment includes robust regression such as least median of squares, which also detects outliers and leverage points. The overall least-squares regression has been applied to find the more exact straight line that fits the experimental data. The error propagation has been considered to calculate the detection limit and the repeatability of the method.     

Determination of major carotenoids in a few Indian leafy vegetables by high-performance liquid chromatography

Leafy vegetables [Basella rubra L., Peucedanum sowa Roxb., Moringa oleifera Lam., Trigonella foenum-graecum L., Spinacia oleracea L., Sesbania grandiflora (L.) Poir., and Raphanus sativus L.] that are commonly used by the rural population in India were evaluated in terms of their main carotenoid pattern. The extracted carotenoids were purified by open column chromatography (OCC) on a neutral alumina column to verify their identity by their characteristic UV-visible absorption spectra. Reverse-phase high-performance liquid chromatography (HPLC) on a C18 column with UV-visible photodiode array detection under isocratic conditions was used for quantification of isolated carotenoids. Acetonitrile/methanol/dichloromethane (60:20:20 v/v/v) containing 0.1% ammonium acetate was used as a mobile phase. The major carotenoids identified by both methods were lutein, beta-carotene, violaxanthin, neoxanthin, and zeaxanthin. Among the carotenoids identified, lutein and beta-carotene levels were found to be higher in these leafy vegetables. Results show that P. sowa and S. oleracea are rich sources of lutein (77-92 mg/100 g of dry wt) and beta-carotene (36-44 mg/100 g of dry wt) compared with other leafy vegetables. The purity of carotenoids eluted by OCC was clarified by HPLC, and they were found to be 92% +/- 3% for neoxanthin, 94% +/- 2% for violaxanthin, 97% +/-2% for lutein and zeaxanthin, and 90% +/- 3% for beta-carotene. It could be recommended to use P. sowa and S. oleracea as rich sources of lutein and beta-carotene for health benefits. The OCC method proposed is relatively simple and provides purified carotenoids for feeding trials.     

Antioxidant levels and inhibition of cancer cell proliferation in vitro by extracts from organically and conventionally cultivated strawberries

The effects of extracts from five cultivars of strawberries on the proliferation of colon cancer cells HT29 and breast cancer cells MCF-7 were investigated, and possible correlations with the levels of several antioxidants were analyzed. In addition, the effects of organic cultivation compared to conventional cultivation on the content of antioxidants in the strawberries and strawberry extracts on the cancer cell proliferation were investigated. The ratio of ascorbate to dehydroascorbate was significantly higher in the organically cultivated strawberries. The strawberry extracts decreased the proliferation of both HT29 cells and MCF-7 cells in a dose-dependent way. The inhibitory effect for the highest concentration of the extracts was in the range of 41-63% (average 53%) inhibition compared to controls for the HT29 cells and 26-56% (average 43%) for MCF-7 cells. The extracts from organically grown strawberries had a higher antiproliferative activity for both cell types at the highest concentration than the conventionally grown, and this might indicate a higher content of secondary metabolites with anticarcinogenic properties in the organically grown strawberries. For HT29 cells, there was a negative correlation at the highest extract concentration between the content of ascorbate or vitamin C and cancer cell proliferation, whereas for MCF-7 cells, a high ratio of ascorbate to dehydroascorbate correlated with a higher inhibition of cell proliferation at the second highest concentration. The significance of the effect of ascorbate on cancer cell proliferation might lie in a synergistic action with other compounds.     

In Vitro Suppression of Growth of Murine WEHI-3 Leukemia Cells and in Vivo Promotion of Phagocytosis in a Leukemia Mice Model by Indole-3-carbinol

Indole-3-carbinol (I3C), a potential anticancer substance, can be found in cruciferous (cabbage family) vegetables, mainly cauliflower and Chinese cabbage. However, the bioactivity of I3C on the apoptotic effects of murine leukemia WEHI-3 cells and promotion of immune responses in leukemia mice model are unclear. In this study, we investigated the effect of I3C on cell-cycle arrest and apoptosis in vitro and immunomodulation in vivo. I3C decreased the viable WEHI-3 cells and caused morphological changes in a concentration- and time-dependent manner. I3C also led to G0/G1 phase arrest, decreased the levels of cyclin A, cyclin D, and CDK2, and increased the level of p21(WAF1/CIP1). Flow cytometric analyses further proved that I3C promoted ROS and intracellular Ca(2+) production and decreased the levels of ΔΨ(m) in WEHI-3 cells. Cells after exposure to I3C for 24 h showed DNA fragmentation and chromatin condensation. Comet assay also indicated that I3C induced DNA damage in examined cells. I3C increased the levels of cytochrome c, FADD, GADD153, GRP78, and caspase-12 as well as induced activities of caspase-3, -8, and -9. Moreover, I3C attenuated NF-κB DNA binding activity in I3C-treated WEHI-3 cells as shown by EMSA and Western blotting analyses. In the in vivo study, we examined the effects of I3C on WEHI-3 leukemia mice. Results showed that I3C increased the level of T cells and decreased the level of macrophages. I3C also reduced the weights of liver and spleen, and it promoted phagocytosis by macrophages as compared to the nontreated leukemia mice group. On the basis of our results, I3C affects murine leukemia WEHI-3 cells both in vitro and in vivo.     

Urinary excretion of 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a ring-fission metabolite of (-)-epicatechin, in rats and its in vitro antioxidant activity

There is great interest in the nutritional potential of (-)-epicatechin, a common polyphenolic constituent of many foods and beverages, because of its potent antioxidant capacity. To better evaluate the biological role of (-)-epicatechin, we studied the urinary excretion of 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a ring-fission metabolite of (-)-epicatechin by intestinal microflora, in rats as well as its antioxidant activity in vitro. The method for measuring the urinary levels of (-)-epicatechin and 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone was based on the enzymatic hydrolysis of beta-glucuronidase and sulfatase, and was subsequently determined by HPLC coupled to an electrochemical detector. Following administration of (-)-epicatechin at doses of 0, 20, 40, and 80 mumol per rat, (-)-epicatechin and 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone were excreted into the urine within 24 h in a dose-dependent manner. Urinary 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone was mostly in the conjugated form, with a higher ratio of conjugation than (-)-epicatechin. We assessed the relative antioxidant potentials for scavenging radicals in the aqueous phase as expressed in the Trolox equivalent antioxidant capacity (TEAC). The results demonstrated that the degradation of (-)-epicatechin into 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone attenuated the antioxidant ability of the former. However, 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone showed stronger antioxidant activity than l-ascorbic acid. These results led us to suppose that 5-(3',4'-dihydroxyphenyl)-gamma-valerolactone, a microbial metabolite of (-)-epicatechin, circulating in the body may also at least be biologically active in terms of contributing to its combined antioxidant effect.     

Inhibition of iron uptake by phytic acid, tannic acid, and ZnCl2: studies using an in vitro digestion/Caco-2 cell model.

The objective of this study was to document the effects of phytic acid, tannic acid, and zinc on iron uptake in an in vitro digestion/Caco-2 cell culture model. The effects of phytic acid and tannic acid on iron uptake were measured at increasing molar ratios of FeCl3 to phytic acid or tannic acid. Maximal inhibition of iron uptake by phytic acid occurred at a 1:10 ratio of Fe to phytic acid. Dialyzable Fe decreased with a low Fe to phytic acid ratio but increased with Fe:phytic acid ratios greater than 1:3 indicating that more iron was soluble at higher phytic acid levels but less available. As in human studies, heme iron was less inhibited by phytic acid than nonheme iron. Tannic acid was a more potent inhibitor of nonheme iron uptake, as maximal inhibition (97.5%) of iron uptake occurred at a ratio of 1:1 or less. The addition of ZnCl2 to the digest at ratios of 1:0.5 and 1:1 decreased iron uptake by 57 and 80%, respectively. Overall, the results agree qualitatively with studies in humans and demonstrate the relative effects of these compounds on iron uptake in this model system. This study provides key information for determining iron availability under more complex meal conditions.     

Analysis of lidocaine and its major metabolite, monoethylglycinexylidide, in elk velvet antler by liquid chromatography with UV detection and confirmation by electrospray ionization tandem mass spectrometry

A sensitive liquid chromatographic (LC) method with UV detection was developed for the determination of residues of lidocaine (LID) and its major metabolite, monoethylglycinexylidide (MEGX), in elk velvet antler. The drugs were extracted from alkaline velvet antler homogenates, cleaned up on a C(18) solid-phase extraction cartridge, and separated on an Inertsil ODS-3 (3.0 x 250 mm, 5 microm) column using an isocratic mobile phase made up of 0.05 M phosphate buffer (pH 4.0)/acetonitrile (88:12, v/v) at a flow rate of 1.0 mL/min. The limits of quantification for LID and its major metabolite, MEGX, were 10 and 20 ng/g, respectively. The method was validated and used to measure the concentration of residues of LID and MEGX in elk velvet antlers harvested after either LID anesthesia or application of a drug-free control method (electro-anesthesia, EA). No LID or MEGX residues were detected in any of the antlers harvested after EA application. No MEGX residues were detected in any of the velvet antlers harvested after LID application, but residues of LID ranging in concentration from 68 to 4300 ng/g were detected in the three sections of the velvet antlers harvested after LID administration. LC-tandem mass spectrometry was used to confirm the presence of lidocaine detected in the velvet antlers.     

Inhibitory effect of a quercetin metabolite, quercetin 3-O-beta-D-glucuronide, on lipid peroxidation in liposomal membranes.

To study the antioxidant activity of quercetin 3-O-beta-D-glucuronide (Q3GA), which is one of the quercetin metabolites in the blood after intake of quercetin-rich food, the inhibitory effect of Q3GA on lipid peroxidation was estimated using phosphatidylcholine large unilamellar vesicles (PC LUV) as a biomembrane model. Iron ion, an aqueous peroxyl radical generator, a peroxynitrite generator, or lipoxygenase was used as the inducer of lipid peroxidation. In all cases, Q3GA inhibited lipid peroxidation significantly, although its inhibitory effect was lower than that of quercetin aglycon. The ultrafiltration of PC LUV containing Q3GA revealed that Q3GA has low but significant affinity with the membranes of phospholipid bilayers. It is therefore likely that Q3GA acts as an efficient antioxidant in membranous lipid peroxidation through its localization in the phospholipid bilayer. This conjugated quercetin metabolite seems to retain the ability to protect cellular and subcellular membranes from peroxidative attack by reactive oxygen species and peroxidative enzymes.     

Determination of free and conjugated indole-3-acetic acid, tryptophan, and tryptophan metabolites in grape must and wine.

Tryptophan (Trp) and its metabolites, especially indole-3-acetic acid (IAA), are considered to be potential precursors of 2-aminoacetophenone (AAP), an aroma compound that causes an "untypical aging off-flavor" (UTA) in Vitis vinifera wines. In this study, RP-HPLC with fluorescence detection was used for the qualitative and quantitative analysis of Trp and Trp metabolites in grape musts and wines to which different viticultural measures had been applied (time of harvest, soil treatment, leaf plucking, vine prune). An alkaline hydrolysis was developed to release bound IAA and Trp. A sensitive and selective determination of different Trp metabolites was achieved after solid phase extraction using a strong anion exchange material. In the examined grape musts, more than 95% of the total IAA was bound either as ester conjugate or as amide conjugate. Free IAA and other Trp metabolites were below the detection limit (<3 microg/L) or could be determined only in traces. Their amounts increased significantly during fermentation, whereas the amount of Trp decreased. It could be shown that the different viticultural measures applied (except the vine prune) as well as the climatic conditions of the vintage exhibited significant influences on the amounts of Trp and Trp metabolites in grape musts or wines.     

In vitro metabolism of trans-permethrin and its major metabolites, PBalc and PBacid, in humans

To estimate the metabolic profile of trans-permethrin in humans, a comparison of the in vitro metabolism of trans-permethrin in humans and rats was conducted using hepatic microsomes, and cytochrome P450 and UDP-glucuronyltransferase isoforms, which catalyze the metabolism of 3-phenoxybenzyl alcohol (PBalc) and 3-phenoxybenzoic acid (PBacid), respectively. In humans and rats, the major metabolic reaction of trans-permethrin in microsomal incubations was the cleavage of ester linkage to give PBalc, followed by oxidation to 4'-OH-PBalc, 4'-OH-PBacid, and PBacid. As to 4'-hydroxylation of PBalc, several CYPs were able to catalyze the reaction, and CYP2E1 was identified as a predominant isoform. PBacid and its conjugates (glucuronide and glycine) are major urinary metabolites of trans-permethrin in mammals. PBacid is also a metabolite of several pyrethroids, and has been used as a biomarker of human exposure to pyrethroids. Our study indicated that there was no difference in glucuronyltransferase activity of PBacid between humans and rats, and that only UGT1A9 can catalyze the glucuronidation of PBacid among human UGTs. Some UGT1A9 variants are known to have poor glucuronidation activity. From these results, it was assumed that deficiency or polymorphism of UGT1A9 might affect the profile of PBacid and its conjugates in urine collected from persons exposed to trans-permethrin or other pyrethroids. These results are helpful for understanding the metabolism of trans-permethrin in humans and determining methods for quantification of target analytes for assessment of human exposure to trans-permethrin and other pyrethroids that give PBacid and its conjugates as urinary metabolites.     

Determination of beta-carotene and lutein available from green leafy vegetables by an in vitro digestion and colonic fermentation method

Green leafy vegetables (Spinacea oleracea, Cnidoscolus aconitifolius, and Solanum americanum) contain a high amount of beta-carotene (27-52 mg/100 g of dry sample) and lutein (140-193 mg/100 g of dry sample). The amount of beta-carotene and lutein released from the food matrix by the action of digestive enzymes ranged from 22 to 67% and from 27 to 77%, respectively. There was a significant correlation between the enzymatic release of carotenoids (lutein + beta-carotene) and the content of Klason lignin, nonstarch polysaccharides, and resistant protein. The carotenoids released by the in vitro colonic fermentation ranged from 2 to 11%, and part of them (0.251-4.03 mg/100 g of original dry sample) remained intact in the fermentation media and could be potentially absorbed in the colon. A significant part of carotenoids seems to be unavailable in the intestinal tract (16% in S. oleracea to 58% in C. aconitifolius).     

Analysis of pendimethalin residues in fruit, nuts, vegetables, grass, and mint by gas chromatography

Pendimethalin [N-(1-ethylpropyl)-3,4-dimethyl-2,6-dinitrobenzenamine], in the formulation of Prowl (a commercial herbicide), was applied to various crops. Analysis of pendimethalin and its metabolite [4(1-ethylpropyl)amino-2-methyl-3,5-dinitrobenzyl alcohol] was accomplished by utilizing liquid-liquid partitioning, gel permeation chromatography (GPC) for nuts and mint, solid-phase extraction (SPE) cleanup, and gas chromatography (GC) with a nitrogen--phosphorus detector (NPD). Method validation recoveries for fruits, nuts, vegetables, grass, and mint are given for both compounds. Pendimethalin average recoveries ranged from 71% to 126% over two levels of fortification. Pendimethalin metabolite average recoveries ranged from 69% to 123% over two levels of fortification. The quantitation limit for all crops except mint was 0.050 ppm. The quantitation limit for mint and mint oil was 0.10 ppm. Residues greater than the limit of quantitation were found for pendimethalin in apple pomace, fresh and dry fig, grass screenings, mint oil, almond hulls, green onion, and tomato pomace (wet and dry). Residues greater than the limit of quantitation were found for pendimethalin metabolite in grass screenings, grass straw, and almond hulls. All other crop analyses for pendimethalin and its metabolite were below the limit of quantitation.     

Relative inhibition of lipid peroxidation, cyclooxygenase enzymes, and human tumor cell proliferation by natural food colors

The most abundant water soluble natural food colors are betacyanins and anthocyanins. Similarly, lycopene, bixin, beta-carotene, and chlorophyll are water insoluble colors. Pure betanin, bixin, lycopene, chlorophyll, beta-carotene, and cyanidin-3-O-glucoside were isolated from Beta vulgaris, Bixa orellana,Lycopersicum esculentum, Spinacia oleracea, Daucus carrota, and Prunus cerasus, respectively. These natural pigments, alone and in combination, were evaluated for their relative potencies against cyclooxygenase enzymes and tumor cell growth inhibition by using MCF-7 (breast), HCT-116 (colon), AGS (stomach), CNS (central nervous system), and NCI-H460 (lung) tumor cell lines. Among the colors tested, betanin, cyanidin-3-O-glucoside, lycopene, and beta-carotene inhibited lipid peroxidation. However, all pigments tested gave COX-1 and COX-2 inhibition and showed a dose-dependent growth inhibition against breast, colon, stomach, central nervous system, and lung tumor cells, respectively. The mixtures of these pigments were also evaluated for their synergistic effects and chemical interactions at various concentrations. The mixture of anthocyanin and betanin negated their efficacy in the cell growth inhibitory assay and did not enhance the COX enzyme inhibitory activity. This is the first report of a comparative evaluation and the impact on biological activities of these pigments alone and in combination.     

Inhibition of reactive nitrogen species effects in vitro and in vivo by isoflavones and soy-based food extracts

Recent studies have shown that soy isoflavone inhibits inducible nitric oxide (NO) synthase activities and is reported to have peroxynitrite scavenging ability. Consequently, we investigated whether isoflavones (daidzein and genistein) and extracts from soy-based products (miso, soymilk, tofu, soy sprout, black soybean, soybean, and yuba) would inhibit the reactive nitrogen species (RNS) effect in vitro and in vivo. In the in vitro experiments [including the protection of cellular DNA from peroxynitrite or sodium nitroprusside damage, an inhibitory effect on nitric oxide production from lipopolysaccharide (LPS)-induced RAW 264.7 cells, and nitric oxide scavenging ability], extracts from soy-based foods showed a potent antioxidant activity and an inhibiting effect on RNS activity. These effects were correlated with total isoflavone content. In the in vivo experiments, rats were given isoflavones (4.0 mg/kg bw) or soy-based product extracts (1.0 g/kg bw) orally for 1 week and were injected with vehicle H(2)O (1 mL/kg bw) or LPS (10 mg/kg bw) on the day 7. Twelve hours after treatment, the rats were killed, and blood serum was collected for analysis. The intraperitoneal administration of LPS resulted in an increase in serum nitrite, nitrate, and nitrotyrosine concentrations. These are stable metabolite end products of nitric oxide, to 4-, 16-, and 5-fold levels, (4, 10 microM and 58 +/- 14 pmol/mL), of the placebo control, respectively. Results showed that oral administration of isoflavones and extracts from soy-based products significantly decreased serum nitrite, nitrate, and nitrotyrosine levels in LPS-induced rats. This study demonstrates that soy isoflavone supplementation may inhibit RNS-induced oxidation both in vitro and in vivo.