• Title/Summary/Keyword: Ames test

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Evaluation of Genotoxicity of Water and Ethanol Extracts from Rhus verniciflua Stokes(RVS)

  • Kim, Ji-Young;Oh, Se-Wook;Han, Dae-Seok;Lee, Michael
    • Toxicological Research
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    • v.24 no.2
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    • pp.151-159
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    • 2008
  • Rhus verniciflua Stokes(RVS), one of traditional medicinal plants in Asia, was found to have pharmacological activities such as antioxidative and antiapoptotic effects, raising the possibility for the development of a novel class of anti-cancer drugs. Thus, potential genotoxic effects of RVS in three short-term mutagenicity assays were investigated, which included the Ames assay, in vitro Chromosomal aberration test, and the in vivo Micronucleus assay. In Ames test, the addition of RVS water extracts at doses from 313 up to 5000 mg/plate induced an increase more than 2-fold over vehicle control in the number of revertant colonies in TA98 and TA1537 strains for detecting the frame-shift mutagens. The similar increase in reversion frequency was observed after the addition of RVS ethanol extracts. To assess clastogenic effect, in vitro chromosomal aberration test and in vivo micronucleus assay were performed using Chinese hamster lung cells and male ICR mice, respectively. Both water and ethanol extracts from RVS induced significant increases in the number of metaphases with structural aberrations mostly at concentrations showing the cell survival less than 60% as assessed by in vitro CA test. Also, there was a weak but statistically significant increase in number of micronucleated polychromatic erythrocytes(MNPCEs) in mice treated with water extract at 2000 mg/kg while ethanol extracts of RVS at doses of up to 2000 mg/kg did not induce any statistically significant changes in the incidence of MNPCEs. Therefore, our results lead to conclusion that RVS acts as a genotoxic material based on the available in vitro and in vivo results.

Bacterial Reverse Mutation Test of Clean Natural using Salmonella typhimurium (천연소독제 Clean Natural의 Salmonella typhimurium에 대한 복귀돌연변이시험)

  • Chun Myung-Sun;Han Sang-Wook;Cho Yoon-Hee;Lim Yeong-Yun;Kim Eui-Gyung;Lee Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.20 no.3
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    • pp.175-178
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    • 2005
  • Clean Natural is a new disinfectant of which main components are propolis and wood vinegar from Quercus mongolica. To evauate the bacterial reverse mutation of Clean Watural, the in vitro Ames test using Salmonella typhimurium TA98, TA100, TA102, TA1535 and TA1537 were performed with clean natural at the concentrations 0, 5, 2.5, and 1.25 mg/ml/plate. Clean Natural was negative in Ames test with Salmonella typhymuyium with and without rat liver microsomal enzyme (S-9 fraction). These results indicate that Clean Watural does not cause bacterial reverse mutation.

Bacterial Reverse Mutation Test of Wild Ginseng Culture Extract (산삼배양추출물의 세균을 이용한 복귀돌연변이시험)

  • Song Si-Whan;Yang Deok Chun;Choung Se Young
    • Journal of Food Hygiene and Safety
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    • v.19 no.4
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    • pp.193-197
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    • 2004
  • To evaluate the bacterial reverse mutation of wild ginseng culture extract, the in vitro Ames test using Salmonella typhimurium (TA100, TA1,535, TA98, TA1,537) and Escherichia coli (WP2 uvrA) were performed with wild ginseng extract at the concentrations 0, 1.6, 8, 40, 200, 1,000, 2,500 and $5,000{\mu}g/ml/plate$. Wild ginseng culture extract was negative in Ames test with both Salmonella typhimurium or Escherichia coli with and without rat liver microsomal enzyme (S-9 fraction). According to these results, we concluded that wild ginseng culture extract did not cause bacterial reverse mutation.

A Study on the Mutagenicity of Autoxidized Soybean Oil (자동산화 대두유의 돌연변이원성에 관환 연구)

  • Lee, Jin-Young;Ahn, Myung-Soo
    • Korean journal of food and cookery science
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    • v.15 no.6
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    • pp.611-617
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    • 1999
  • Commercial soybean oil, which did not contain any antioxidant, were autoxidized at $60{\pm}2^{\circ}C$ for 79 days, and the changes of peroxide value(POV), thiobarbituric acid value(TBAV), conjugated dienoic acid(CDA) content, and fatty acid composition of the oil were studied during the 79 day-storage period. The samples with POV S of 0, 150, 300, 500, 450, 400 and 300 meq./kg oil were used for the test of mutagenic activity. The Ames test was carried out with and without metabolic activation. Bacterial strains used in this study were the histidine auxotrophic strains of S. typhimurium TA100, TA1535, and TA102 for the detection of base pair, and TA98 and TA1537 for frame shift mutations. Each series of samples was disso1ved in tetrahydrofuran(inhibitor free) and tested at doses ranging from 0.05 to 5 mg/plate. The autoxidized soybean oil increased significantly(p<0.05) the number of $His^+$ revertant colonies in cases of TA 98 with S9 mix, TA100 without S9 mix, 1535 and 1537 with and without S9, respectively. The samples having the highest peroxide values showed the strongest mutagenicity. It seemed that the amount of hydroperoxides in the oils was closely related to the mutagenic activity of the respective oils.

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Development of Fecal Microbial Enzyme Mix for Mutagenicity Assay of Natural Products

  • Yeo, Hee-Kyung;Hyun, Yang-Jin;Jang, Se-Eun;Han, Myung-Joo;Lee, Yong-Sup;Kim, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.838-848
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    • 2012
  • Orally administered herbal glycosides are metabolized to their hydrophobic compounds by intestinal microflora in the intestine of animals and human, not liver enzymes, and absorbed from the intestine to the blood. Of these metabolites, some, such as quercetin and kaempherol, are mutagenic. The fecal bacterial enzyme fraction (fecalase) of human or animals has been used for measuring the mutagenicity of dietary glycosides. However, the fecalase activity between individuals is significantly different and its preparation is laborious and odious. Therefore, we developed a fecal microbial enzyme mix (FM) usable in the Ames test to remediate the fluctuated reaction system activating natural glycosides to mutagens. We selected, cultured, and mixed 4 bacteria highly producing glycosidase activities based on a cell-free extract of feces (fecalase) from 100 healthy Korean volunteers. When the mutagenicities of rutin and methanol extract of the flos of Sophora japonica L. (SFME), of which the major constituent is rutin, towards Salmonella typhimurium strains TA 98, 100, 102, 1,535, and 1,537 were tested using FM and/or S9 mix, these agents were potently mutagenic. These mutagenicities using FM were not significantly different compared with those using Korean fecalase. SFME and rutin were potently mutagenic in the test when these were treated with fecalase or FM in the presence of S9 mix, followed by those treated with S9 mix alone and those with fecalase or FM. Freeze-dried FM was more stable in storage than fecalase. Based on these findings, FM could be usable instead of human fecalase in the Ames test.

Genetic Toxicity Test of Glycidol by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ji-H.;Kim, Ki-Y.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Sheen, Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.14 no.4
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    • pp.240-245
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    • 2006
  • The primary use for glycidol is as a stabilizer in the manufacture of vinylpolymers, however, it is also used as an intermediate in the production of pharmaceuticals, as an additives for oil and synthetic hydraulic fluids, and as a diluting agent is same epoxy resins. In this study, we have carried out in vitro genetic toxicity test of glycidol and microarray analysis of differentially expressed genes in response to glycidol. The result of Ames test showed mutations with glycidol treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, glycidol showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with glycidol treatment showed DNA damage both with and without exogenous metabolic activation. Glycidol increased micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to glycidol by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of glycidol.

Genotoxicity Assessment of Erythritol by Using Short-term Assay

  • Chung, Young-Shin;Lee, Michael
    • Toxicological Research
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    • v.29 no.4
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    • pp.249-255
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    • 2013
  • Erythritol is a sugar alcohol that is widely used as a natural sugar substitute. Thus, the safety of its usage is very important. In the present study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of erythritol. According to the OECD test guidelines, the maximum test dose was 5,000 ${\mu}g$/plate in bacterial reverse mutation tests, 5,000 ${\mu}g/ml$ in cell-based assays, and 5,000 mg/kg for in vivo testing. An Ames test did not reveal any positive results. No clastogenicity was observed in a chromosomal aberration test with CHL cells or an in vitro micronucleus test with L5178Y $tk^{+/-}$ cells. Erythritol induced a marginal increase of DNA damage at two high doses by 24 hr of exposure in a comet assay using L5178Y $tk^{+/-}$ cells. Additionally, in vivo micronucleus tests clearly demonstrated that oral administration of erythritol did not induce micronuclei formation of the bone marrow cells of male ICR mice. Taken together, our results indicate that erythritol is not mutagenic to bacterial cells and does not cause chromosomal damage in mammalian cells either in vitro or in vivo.

Evaluation on the Safety of ${\gamma}$-Irradiated Angelica gigas Nakai: Stability of Active Components and Safety in Genotoxicity Test (감마선 조사 당귀(Angelica gigas Nakai)의 유효성분 안정성 및 유전독성학적 안전성 연구)

  • 조성기;유영법
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.300-306
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    • 2000
  • In the present studies, we assessed the stability of active components and toxicological safety of irradiated Angelica gigas Nakai(Danggui). In order to confirm the stability of active components in the ${\gamma}$-irradiated roots of Danggui, the quantitative analysis of decursin and decursinol angelate of ${\gamma}$-irradiated sample was carried out by high performance liquid chromatographic (HPLC) methods using reverse phase columns and normal phase columns. From the root of Danggui, decursin and decursinol angelate were isolated by a silica gel column chromatography(toluene : ether (1 : 1), Hexane : EtOAc(15 : 1)). And then the structures were confirmed in the 1H and 13C-NMR analysis. The HPLC chromatograms of decursin and decursinol angelate in ${\gamma}$-irradiated Danggui were similar with those of non-irradiated sample. In the examination of in vitro genotoxicity of the water extract from ${\gamma}$-irradiated Danggui using Salmonella reversion assay(Ames test) and micronucleus test in Chinese hamster ovary (CHO) cells, mutagenicity was not exhibited in the two assays with or without metabolic activation. These resutls suggest that active components in the ${\gamma}$-irradiated Danggui should be stable and that the safety of ${\gamma}$-irradiated Danggui could be revealed in further test in vivo.

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Antimutagenic Effect of Rresveratrol on Trip P-1 in Salmonella typhimurium TA98 (Trp P-1 변이원성에 대한 Resveratrol의 항돌연변이 효과)

  • 장귀현;안병용;권용주;최동성
    • The Korean Journal of Food And Nutrition
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    • v.14 no.4
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    • pp.329-332
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    • 2001
  • The antimutagenic activity of resveratrol on the mutagenicity induced by Trp P-1 (3-amino-1,4-dime-thyl-5H-pyrido{4,3-b}indole) was studied using the Ames test with Salmonella typhimurium TA98 and 100. Trp P-1 showed strong mutagenecity in S. typhimurium TA98, but was higly decreased mutagenecity in S. typhimurium TA100. This result suggests that the mutagenecity of Trp P-1 can be mainly induced by the DNA lesions causing frame shift. Resveratrol itself did not show antibacterial effect upon 300 $\mu\textrm{g}$/assay. Resveratrol showed the strongest inhibitory effect with dose dependent manner on the mutagenicity induced by Trp P-1. The inhibition rates of resveratrol at concentration of 2, 10, 25, 50, 100, 300 $\mu\textrm{g}$/assay were 13%, 37%, 52%, 65%, 81%, 89%, respectively.

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Antimutagenic Effect of Tansen (Salvia miltriorrhiza Bunge) (단삼의 항산화적 항돌연변이 효과)

  • 안병용;김동길;최동성
    • Microbiology and Biotechnology Letters
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    • v.27 no.3
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    • pp.197-202
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    • 1999
  • To confirm the effects of binlang(Areca catechu L.) and tansen(Salvia miltriorrhiza bung) on the mutagenicity induced by hydrogen peroxide, SOS Chromotest with Escherichia coli PQ37 and Ames test with Salmonella typhimurium TA104 were perlormed. Methanol-soluble parts of their water extracts showed high inhibitory effect against the mutagenicity of hydrogen peroxide in two bacterial mutation assays. Step-wise fractionation of methanol-soluble part from tansen was done using ethyl acetate, butanol and water. Among these fraction was further partitioned by Sephadex LH-20 column chromtography, and 6 subfractions were obtained. The fraction III showed the strongest inhibitory effects against the mutagenic activities induced by hydrogen peroxide. The inhibition rates of fraction III at concentration of 500$\mu\textrm{g}$/assay were 28%, 30% and 15% against 4-NQO, MNNG and B(a)P, respectively. But the mutagenic potency of AFB1 was increased.

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