• Title/Summary/Keyword: Ames Test

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Genetic Toxicity Test of o-Nitrotoluene by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Eun-Mi;Lee, So-Youn;Lee, Woo-Sun;Kang, Jin-Seok;Han, Eui-Sik;Go, Seo-Youn;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-Nie
    • Molecular & Cellular Toxicology
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    • v.3 no.2
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    • pp.107-112
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    • 2007
  • o-Nitrotoluene is used to synthesize artificial dyes and raw materials of urethane resin. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of onitrotoluene. TA1535 and TA98 cells were treated with o-nitrotoluene to test its toxicity by basic genetic toxicity test. Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to o-nitrotoluene was analyzed using Affymatrix genechip. The result of Ames test was that o-nitrotoluene treatment did not increase the mutations both in base substitution strain TA1535 and in frame shift TA98. o-Nitrotoluene has not increased micronuclei in CHO cells. But onitrotoluene increased DNA damage in L5178Y cell. Two-hundred two genes were initially selected as differentially expressed genes in response to o-nitrotoluene by microarray analysis and forty four genes among them were over 2 times of log fold changed. These forty four genes could be candidate biomarkers of genetic toxic action of o-nitrotoluene related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to the DNA damage will be useful to understand the detailed mechanism of action of o-nitrotoluene.

Genetic Toxicity Test of 8-Hydroxyquinoline by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Woo-Sun;Kim, Hyun-Joo;Lee, Eun-Mi;Kim, Joo-Hwan;Suh, Soo-Kyung;Kwon, Kyung-Jin;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-N.
    • Molecular & Cellular Toxicology
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    • v.3 no.2
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    • pp.90-97
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    • 2007
  • 8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.

In Vitro Studies on the Genotoxic Effects of Wood Smoke Flavors

  • Chung, Young-Shin;Ahn, Jun-Ho; Eum, Ki-Hwan;Choi, Seon-A;Oh, Se-Wook;Kim, Yun-Ji;Park, Sue-Nie;Yum, Young-Na;Kim, Joo-Hwan;Lee, Michael
    • Toxicological Research
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    • v.24 no.4
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    • pp.321-328
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    • 2008
  • Smoke flavors based on the thermal decomposition of wood have been applied to a variety of food products as an alternative for traditional smoking. Despite its increasing use, the available genotoxicity data on wood smoke flavors (WSF) are still controversial. Thus, potential genotoxic effects of WSF in four short-term in vitro genotoxicity assays were investigated, which included the Ames assay, chromosomal aberration assay, micronucleus test and the alkaline comet assay. WSF did not cause any mutation in the Ames assay using five tester strains at six concentrations of 0.16, 0.31, 0.63, 1.25, 2.5 and 5 ${\mu}l/plate$. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. No statistically significant increase in the number of metaphases with structural aberrations was observed at the concentrations of 1.25, 2.5, and 5 ${\mu}l/ml$. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that WSF has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, based on the results obtained from these four in vitro studies, it is concluded that WSF is not a mutagenic agent in bacterial cells and causes no chromosomal and DNA damage in mammalian cells in vitro.

Mutagenic and Antimutagenic Effects of Hemp Seed Oil Evaluated by Ames Salmonella Testing (삼종실유의 항돌연변이 효과)

  • Jeun, Jung-Ae;Cho, Hee-Joon;Jun, Hee-Jin;Lee, Ji-Hae;Jia, Yao-Yao;Cho, Kyoung-Sang;Kim, Eun-Soo;Lee, Sung-Joon
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.396-400
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    • 2011
  • We examined the in vitro mutagenic and antimutagenic effects of hexane-extracted hemp (Cannabis sativa L. subsp. sativa var. sativa) seed oil (HO) with and without S9-mediated metabolic activation, using the TA98 and TA100 Salmonella Typhimurium strains. The MTT assay revealed no cytotoxicity in HepG2 cells for HO quantities $\leq400g$/mL. In the mutagenicity test, revertant colonies did not exceed spontaneous colonies in number. Colony numbers did not increase in either strain after HO treatment, with or without metabolic activation. HO showed no mutagenic effects and did not induce a mutation in either strain. In the antimutagenicity test, HO reduced the number of mutated colonies induced by 4NQO in both strains. The inhibition rates of HO (TA98, 21-91%; TA100, 21-85%) indicated a potent reduction in mutagenicity induced by 4NQO. HO showed no significant mutagenicity and may have antimutagenic effects, as assessed by Ames testing.

A Study on the Mutagenicity of Thermally Oxidized Safflower Oil (가열산화 홍화유의 돌연변이원성에 관한 연구)

  • 안명수;이진영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.120-127
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    • 2000
  • Deep-fat frying is a common cooking practice. There has been considerable concern regarding the mutagenic and carcinogenic potential of thermally oxidized oils. Studies on deep-fried foods so far have revealed not much on the mutagenicity of the oils in the foods. Therefore, in the present study, it was attempted to investigate the mutagenicity ofthe thermally oxidized safflower oil. Oil was heated in a home-fryer at a temperature of 180$\pm$3$^{\circ}C$ for 48 hours. Oil samples were taken at 0, 8, 16, 24, 32, 40 and 48 hours of heating, respectively. Each sample was used to study the changes in peroxide value (POV), acid value (AV), iodine value (IV), conjugated dienoic acid (CDA) content, %, and fatty acid composition. Another series of samples were fractionated into non-polar and polar fractions by column chromatography. The mutagenicity of the samples taken from the thermally oxidized oils, as well as the non-polar and polar fractions of the thermally oxidized oils, was investigated with the Ames test. The Ames test was carried out with and without metabolic activation. Bacterial tester strains used in the present study were the histidine auxotrophic strains of Salmonella typhimurium TA100, TA1535 and TA102 were used for the detection of base pair mutations, and TA98 and TA1537 for frame shift mutations. Each series of samples was dissolved in tetraphydrofuran (inhibitor-free) and tested at doses ranging from 0.05 to 5 mg/plate. None of the oil samples taken during the 48 hour oxidation period showed any mugagenic activity. This was the case, even after the activaton with 59 mix. Also, none of the polar and non-polar fractions showed any mutagenic activity on all the strains tested.

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Mutagenicity Studies on Nonspecific Immunostimulator BARODON® (비특이 면역증강제 BARODON®에 대한 유전독성시험)

  • 서민수;조성대;안남식;정지원;양세란;박준석;박기수;홍인선;조은혜
    • Toxicological Research
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    • v.19 no.2
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    • pp.141-146
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    • 2003
  • A nonspecific immunostimulator $BARODON^{\circledR}$ was tested for mutagenicity using Ames Salmonella tester strains TA98, TA1 00, TA 102, TA 1535 and TA 1537 with or without metabolic activation (59 mix). None of the fresh species showed mutagenicity. In the reverse mutation test using Salmonella phimurium TA98, TA100, TA102, TA1535 and TA1537 did not increase the number of revertants at all doses tested (5, 2.5 or 1.25 mg/ml). Chromosome aberration test was carried out in Chinese hamster lung (CHL) cell line. The cells were treated with $BARODON^{\circledR}$ (1, 0.5 or 0.25 mg/ml), while positive control group was treated with Mitomycin C (0.1 mg/ml). The results show that there is no statistically significant difference between positive control and treatment groups. In mouse micronucleus test, there was significant increase in the ratio of micronucleated polychromatic erythrocyte (MNPCE) in the high dose group (10% $BARODON^{\circledR}$), while there is no significance between control and low (2.5% $BARODON^{\circledR}$) or middle (5% $BARODON^{\circledR}$ dose groups. Taken together, this results suggest that below 5% $BARODON^{\circledR}$ might not have mutagenic potential in vitro and vivo systems.

Evaluation of Antifungal Activities and Safeties of 6-[(N-2,4-Dibromophenyl) amino]-7-Chloro-5,8-Quinolinedione (6-[(N-2,4-디브로모페닐)아미노]-7-클로로-5,8-퀴놀린디온의 항진균작용 및 안전성 평가)

  • 유충규;김동현;윤여표;허문영;권상미;정성희
    • Journal of Food Hygiene and Safety
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    • v.11 no.4
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    • pp.299-306
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    • 1996
  • 6-[(N-3,4-Dibromophenyl)amino]-7-chloro-5,8-quinolinedione(FCK13) was tested for antifungal activities. The MIC values were determined by the two-fold dilution method. The therapeutic potential of RCK13 had been assessed in comparison with ketoconazole and fluconazole against systemic infections with candida albicans in normal mice. RCK13 had ED50,0.80$\pm$0.21 mg/kg but ketoconazole had ED50, 8.00$\pm$0.73 mg/kg respectively. And administered RCK13 at the ED50 for 14 days improved survival rates as well as ketoconazole. Acute oral toxicity studies of RCK13 were carried out in ICR mice of both sexes. These acute oral toxicities of RCK13 were low and LD50 values were over 2,850 mg/kg in ICR mice. The genotoxicities of RCK13 had been evaluated. RCK13 was negative in Ames test with Salmonella typhimurium and chromosomal aberration test in CHL cells. The clastogenicity was tested on the RCK13 with in vivo mouse micronucleus assay. RCK13 did not show any clastogenic effect in mouse peripheral blood and was negative in mouse micronucleus assay. These results indicate that RCK13 has no genotoxic potential under these experimental conditions.

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The Evaluation of Antifungal Activities and Safeties of 6-(4-Iodophenyl)amino-7-chloro-5,8-quinolinedione (6-(4-요오도페닐)아미노-7클로로-5,8-퀴놀린디온의 항진균작용 및 안전성 평가)

  • 유충규;윤여표;허문영;이병무;강혜영;이유진
    • Journal of Food Hygiene and Safety
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    • v.14 no.1
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    • pp.55-59
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    • 1999
  • 6-(4-Iodophenyl)amino-7-chloro-5,8-quinolinedione (RCK9) was evaluated for antifungal activities. The MIC values of RCK9 were determined against A. flavus, c. albicans, C. neoformans and F. oxysporium. The RCK9 showed generally potent antifungal activities against the tested fungi. Acute oral toxicity studies of RCK9 were carried out in ICR mice of both sexes. These acute oral toxicities of RCK9 had been evaluated. RCK9 were low and LD50 values were over 2,850 mg/kg in ICR mice. The genotoxicities of RCK9 had been evaluated. RCK9 was negative in Ames test with Salmonella typhimurium and chromosomal aberration test in CHL cells. The clastogenicity was tested on the RCK9 with in vivo mouse micronucleus assay. RCK9 did not show any clastogenic effect in mouse peripheral blood and was negative in mouse micronucleus assay. The results indicate that RCK9 has no genotoxic potential under these experimental conditions.

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