• Title/Summary/Keyword: Chromosomal aberration assay

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Evaluation of the Genetic Toxicity of Synthetic Chemicals (XI) - a Synthetic Sulfonylurea Herbicide, Pyrazosulfuron-ethyl-

  • Ryu, Jae-Chun;Kim, Eun-Young;Kim, Young-Seok;Yun, Hye-Jung
    • Environmental Mutagens and Carcinogens
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    • v.24 no.1
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    • pp.33-39
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    • 2004
  • To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals including agrochemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. Pyrazosulfuron-ethyl [Ethyl-5-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)-1-methylpyrazole-4-carboxylate, $C_{14}H_{18}N{6}O_{7}S,$ M.W. =414.39, CAS No. 93697-74-6], is one of well known rice herbicide belong in the sulfonyl urea group. To clarify the genotoxicity of this agrochemical, Ames bacterial reversion assay, in vitro chromosomal aberration assay with Chinese hamster lung (CHL) fibroblast and bone marrow micronucleus assay in mice were subjected. In Ames assay, although pyrazosulfuron-ethyl revealed cytotoxic at 5,000-140 $\mug/plate$ in Salmonella typhimurium TA100, no dose-dependent mutagenic potential in 4.4~70 $\mug/plate$ of S. typhimurium TA 98, TA 100, TA1535 and TA 1537 both in the absence and presence of S-9 metabolic activation system was observed. Using CHL fibroblasts, the 50% cell growth inhibition concentration $(IC_{50})$ of pyrazosulfuron-ethyl was determined as 1,243 $\mug/mL,$ and no chromosomal aberration was observed both in the absence and presence of S-9 mixture in the concentration range of 311-1,243 $\mug/mL.$ And also, in vivo micronucleus assay using mouse bone marrow, pyrazosulfuron-ethyl revealed no remarkable induction of MNPCE (micronucleated polychromatic erythrocytes/1000 polychromatic erythrocytes) in the dose range of 625-2,500 mg/kg body weight when administered orally. Consequently, Ames bacterial gene mutation with Salmonella typhimurium, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pyrazosulfuron-ethyl in this study.

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Genotoxicity Study of Sophoricoside in Bacterial and Mammalian Cell System

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.183-184
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    • 2003
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.

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Comparison of L5178Y tk+/- Mouse Lymphoma Assay and In vitro Chromosome Aberration Test

  • Lee, Michael;Jung Kwon;Cho, Ji-Hee;Hong, Mi-Young;Kim, Eun-Joo;Junghee Han;Chung, Moon-Koo;Han, Sang-Seop
    • Toxicological Research
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    • v.18 no.2
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    • pp.129-138
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    • 2002
  • The mouse lymphoma assay (MLA) has been recently validated as a sensitive and specific test system to determine the genotoxic potential for a chemical. The objective of this study is to evaluate the utility of MLA for detecting mutagens. Especially, to compare MLA with the in vitro chromosomal aberration test (CA), we performed MLA using the microwell method with three chemicals (hydroxyurea, theophylline and amino acid copper complex), which were reportedly positive in the CA. In cell treated with hydroxyurea, anti-neoplastic agent that blocks DNA replication, evidence of a positive response was obtained without S9 mix for 4 h and 24 h. In addition, analysis of colony size distribution at concentration that gave an elevated mutant fraction showed that hydroxyurea induced a high proportion of small type colonies, indicating that hydroxyurea-induced mutation is associated with large chromosomal deletion. Conversely, negative MLA result was obtained for theophylline, which was wed as central nervous system stimulator. Although theophylline increased the mutant frequency at concentration of 1250 $\mu\textrm{g}$/$\textrm{m}{\ell}$ with S9 mix for 4 h, a concentration-related increase in mutant frequency was not observed. The MLA result of amino acid copper complex was considered equivocal because the positive result was obtained at concentration showing 10% or less RS or RTG. Thus, among 3 CA-positive chemicals, positive MLA result was obtained for one. The other two chemicals were negative and equivocal. However MLA, which evaluates mutagenic potential of chemicals through colony formation by cell grouth, may provide a higher predictivity of carcinogenesis than CA.

Genotoxicity Study on Khal, a Halocidin Derivative, in Bacterial and Mammalian Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Jeon, Hee-Kyoung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.3
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    • pp.151-158
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    • 2006
  • Khal was a synthetic congener of halocidin, a heterodimeric peptide consisting of 19 and 15 amino acid residues detected in Halocynthia aurantium. This compound was considered a candidate for the development of a novel peptide antibiotic. The genotoxicity of Khal was subjected to high throughput toxicity screening (HTTS) because they revealed strong antibacterial effects. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay and chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of Khal was determined the concentration of $25.51\;{\mu}/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, Khal was not induced DNA damage in mouse lymphoma cell line. Also, the mutation frequencies in the Khal-treated cultures were similar to the vehicle controls. It is suggests that Khal is non-mutagenic in MOLY assay. And no clastogenicity was observed in Khal-treated Chinese hamster lung cells. The results of this battery of assays indicate that Khal has no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that Khal, as the optimal candidates with both no genotoxic potential and antibacterial effects must be chosen.

Genotoxicological Safety Evaluation of X-ray Irradiated Four Foods (X-선 조사식품 4종의 유전독성학적 안전성 평가)

  • Jung, Da-Woon;Huang, Yu-Hua;Song, Beom-Seok;Byun, Myung-Woo;Kang, Il-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1588-1593
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    • 2014
  • This study evaluated the genotoxic effects of 30 kGy of X-ray irradiation to four foods (chicken, egg powder, dried green onion, and black pepper). In bacterial reversion assay with Salmonella Typhimurium TA98, TA100, TA1535, and TA1537, the X-ray irradiated foods did not show a significantly increased number of revertant colonies in the presence or absence of the S9 metabolic activation system. In chromosomal aberration tests with Chinese hamster ovary (CHO) cells, the X-ray irradiated foods showed no increase in the frequency of chromosomal aberrations. In in vivo mouse micronucleus assay, the X-ray irradiated foods did not show any increase in the frequency of polychromatic erythrocytes with micronuclei. These results indicate that 30 kGy of X-ray irradiation to four foods (chicken, egg powder, dried green onion, and black pepper) showed no genotoxic effects under these experimental conditions.

The Evaluation of Antifungal Activities and Safeties of 6-(3,4-Dichlorophenyl)amino-7chloro-5,8-quinolinedione (6-(3,4-디클로로페닐)아미노-7-클로로-5,8퀴놀린디온의 항진균작용 및 안전성 평가)

  • Yun, Yeo-Pyo;Kim, Dong-Hyun;Lee, Byung-Mu;Heo, Moon-Young;Chung, Hae-Moon;Kang, Hye-Young;Choi, Jung-Ah;Kim, Do-Hee;Ryu, Chung-Kyu
    • YAKHAK HOEJI
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    • v.42 no.5
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    • pp.527-533
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    • 1998
  • 6-(3,4-Dichlorophenyl)amino-7-chloro-5,8-quinolinedione (RCK50) was tested for antifungal activities in mice systemically infected with Candida albicans. The therapeutic potential of RCK50 was also assessed in comparison with ketoconazole. CK50 had $ED_{50}$ 0.22${\pm}$0.01mg/kg. Ketoconazole as a positive control had $ED_{50}$ 6.00${\pm}$1.70mg/kg. Intraperitoneally administered RCK50 at the $ED_{50}$ for 7 days and 14 days reduced Candida albicans colony count in the kidneys and liver. And administered RCK50 at the $ED_{50}$ for 14 days improved survival rates. The genotoxicities of RCK50 had been evaluated. RCK50 was negative in Ames test with Salmonella typhimurium and chromosomal aberration test in CHL cells. RCK50 did not show any clastogenic effect in mouse peripheral blood and was negative in mouse micronucleus assay. These results indicate that RCK50 has no genotoxic potential under these experimental conditions. Acute oral toxicity studies of RCK50 were carried out in ICR mice of both sexes. RCK50 did not show acute oral toxicities and $LD_{50}$ values were over 2,850mg/kg in ICR mice.

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Evaluation of the Genetic Toxicity of Synthetic Chemicals (Ⅵ) -In vitro Chromosomal Aberration Assay with 17 Chemicals in Chinese Hamster Lung Cells - (합성화학물질들의 유전독성평가(Ⅵ) -Chinese hamster lung세포를 이용한 17종 합성화학물질들의 염색체 이 상 시험 -)

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Youn-Jung;Jeon, Hee-Kyung
    • Environmental Analysis Health and Toxicology
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    • v.18 no.2
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    • pp.111-120
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    • 2003
  • The validation of many synthetic chemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, the regulation and evaluation of the chemical hazard playa very important role to environment and human health. The clastogenicity of 17 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 2-Nitroaniline (CAS No. 88-74-4) induced chromosomal aberrations with statistical significance at the concentration of 86.3 ${\mu}$g/ml in the absence of metabolic activation system. 1-Chloroanthraquinone (CAS No. 82-44-0) which is one of the most cytotoxic chemical among 17 chemicals tested revealed no clastogenicity in the range of 0.8 ∼ 3.0 ${\mu}$g/ml both in the presence and absence of S-9 metabolic activation system. From the results of chromosomal aberration assay with 17 synthetic chemicals in Chinese hamster lung cells in vitro, 2-Nitroaniline (CAS No. 88-74-4) revealed weak positive clastogenic results in this study.

Genotoxicity Studies of Chrysin (Chrysin의 유전독성에 관한 연구)

  • Jee Seungwan;Kim Changhwan;Park Misun;Eom Miok;Ryeom Taikyung;Kim Okhee;Kang Hoil
    • Toxicological Research
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    • v.21 no.1
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    • pp.71-75
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    • 2005
  • Chrysin (5,7-dihydroxyflavone) is a flavonoid compound contained in many fruits, vegetables and honey. In our experiment, we investigated genotoxicity of chrysin using bacterial reverse mutation assay, chromosomal aberration test, in vivo micronucleus test. In bacterial reverse mutation assay, chrysin did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA102 with and without metabolic activation. In chromosome aberration test, chrysin did not also induce structural and numerical abberations regardless of metabolic activation in Chinese hamster lung fibroblast cells. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocytes (MNPCE) was observed in ICR male mice orally administered with chrysin at the dose of 0.5, 1.0, 2.0 g/kg body weight. Taken together these results, chrysin has no mutagenic potential in our experiment.

Modification of Survival and Blood-forming Stem Cells in Cobalt-60 Gamma Irradiated Mice by Aloe vera (Aloe vera투여가 Cobalt-60 감마선 조사를 받은 마우스의 생존율과 조혈간세포에 미치는 영향)

  • Choi Min-cheol;Sung Jai-Ki
    • Journal of Veterinary Clinics
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    • v.7 no.2
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    • pp.451-469
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    • 1990
  • The present study was carried out to investigate whether the aloe had a radioprotective effect in mice exposed to cobalt-60 gamma radiation or not. The survival ratio of mice for 30 days, hematopoiesis of blood-forming stem cells by spleen colony assay, chromosomal aberration frequency of bone marrow cells and histopathological findings of bone marrow were investigated. The survival ratios of aloe administered groups with concentration of 250, 500, 1,000 and 1,500mg for 3 days before irradiation and control group in cobalt-60 gamma irradiated mice(700rads whole body irradiation, dose rate of 50rads/min.) were 77.4, 79.3, 80.6, 90.0 and 53.1%, respectively. The survival ratios of pre-irradiation aloe administered groups were superior to those of post-irradiation aloe groups and control group. In spleen colony assay, Aloe vera administration before irradiation enhanced the recoveries of numbers of blood-forming stem cells of bone marrow of irradiated mice. There were decreased chromosomal aberrations of bone marrow cells at the first day after irradiation in aloe administered groups compared to that of control group. Histopathological findings in the bone marrow of irradiated mice were hypocellularity due to the depletion of myelocytes, abundant of fat vacuoles and these changes were weakened in aloe administered groups compared to that of control group.

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