• Title/Summary/Keyword: Chromosomal aberration (CA)

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Antimutagenic Effects of Ginsenoside Rb$_1$, Rg$_1$ in the CHO-K1 Cells by Benzo[a]pyrene with Chromosomal Aberration Test and Comet Assay

  • Kim, Jong-Kyu;Kim, Soo-Jin;Rim, Kyung-Taek;Cho, Hae-Won;Kim, Hyeon-Yeong;Yang, Jeong-Sun
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.126-132
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    • 2009
  • The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.

In Vitro Mammalian Chromosomal Aberration Test of Allyl Chloride for Workers' Health (근로자의 건강보호를 위한 알릴 염화물의 포유류 배양세포 염색체이상시험)

  • Rim, Kyung-Taek;Kim, Soo-Jin
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.24 no.2
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    • pp.160-168
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    • 2014
  • Objectives: Chemical hazard evaluations are important for workers' health and working environments. Allyl chloride (CAS No. 107-05-1) is used in many industries, leading to concerns about the possibility of threats to the health of workers. Since only insufficient or controversial information is available about potential related hazards, an in vitro mammalian chromosomal aberration (CA) assay was conducted in order to gain additional information concerning any such hazards. Moreover, toxicological information from this study could be applied for workers' rights to know, and to prepare or update the Materials Safety Data Sheet (MSDS) for a number of industries. Methods and Results: The assay was performed using the Chinese hamster lung fibroblast cell (ATCC, CRL-1935), by the direct method (-S9) and by the metabolic activated method (+S9 mix). Using the direct method, the seven dosages in the 48-hour treatment group did not show that the frequency of CA is proportionate to the dosage. The frequency of CA is not proportionate to the dosage addition for a six-hour treatment using the metabolic activated method. Conclusions: From these findings, it was decided that this chemical does not induce chromosomal aberrations under the tested conditions.

Evaluation of the Genetic Toxicity of Cyclopentane and Ammonium Nitrate - In vitro Mammalian Chromosomal Aberration Assay in Chinese Hamster Ovary Cells

  • Kim, Soo-Jin;Rim, Kyung-Taek;Kim, Jong-Kyu;Kim, Hyeon-Yeong;Yang, Jeong-Sun
    • Safety and Health at Work
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    • v.2 no.1
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    • pp.17-25
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    • 2011
  • Objectives: In this study, the in vitro mammalian chromosomal aberration (CA) assay was conducted to gain additional information concerning the hazards associated with the use of cyclopentane and ammonium nitrate. While these two chemicals had already been tested by many methods, they had not been studied in the CA test. Methods: The assay was performed using the ovarian infantile cell (CHO-K1 cell), by the direct method (-S9) and by the metabolic activated method (+S9 mix). Results: Using the direct method, the 7 dosages in a 48 hour treatment group did not show that the frequency of CA is proportion to the dosage addition. The frequency of CA is not proportion to the dosage addition for a 6 hour treatment using the metabolic activated method. Conclusion: From these findings, it was decided that the 2 chemicals do not induce chromosomal aberrations under the tested conditions.

Mutagenicity Study of (R)-JG-381, A New Antidiabetic Agent (항당뇨물질 (R)-JG-381의 변이원성 시험)

  • 오우용;주상섭;박형근;함광수;조장섭;이선미
    • Biomolecules & Therapeutics
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    • v.8 no.3
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    • pp.248-254
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    • 2000
  • (R)-JG-381, a R form of alkylglycidic acid derivative, was examined for mutagenicity in the reverse mutation test on bacteria, chromosomal aberration test on cultured mammalian cells and micronucleus test in mice. In the reverse mutation test on bacteria using Salmonella typhimurium strain TA98, TA100, TA102, TA1535, TA1537 with or without a metabolic activation system (S9 mix), (R)-JG-381 did not affect the revertant colonies but significantly increased revertant colonies in one test strain, TA98, compared with the vehicle control. In the chromosomal aberration (CA) test using cultured Chinese Hamster Lung fibroblast(CHL) cells, the number of aberrant cells was clot increased in the presence or absence of 59 mix at concentration of the (R)-JG-381 0.025 $\mu$l/m1 to 0.1 $\mu$l/m1, compared with vehicle control. In the micronucleus (MN) test, micronucleated polychromatic erythrocytes in the (R)-JG-381-treated mice were not different from those of the vehicle-treated mice.

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An anti-clastogenic Role of Selenium in Arsenic- and Chromium-induced Oxidative Stress Causing Chromosomal Damages (비소와 크롬에 의한 산화적 스트레스와 염색체 상해에 대한 셀레늄의 방어 효과)

  • 기혜성;손은희;박영철;맹승희;정해원
    • Journal of Environmental Health Sciences
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    • v.23 no.4
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    • pp.9-15
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    • 1997
  • This experiment was carried out to examine the roles of selenium in arsenic- and chromium-induced oxidative stress, which results in chromosomal damage, such as sister chromatid exchange (SCE) and chromosomal aberration (CA). For this purpose, the frequency of CA and SCE related to the level of 0xidative stress were analyzed. Selenium decreased the frequency of CA induced by As. In order to evaluate the effect of selenium on clastogenic factors, media from As- and Cr-treated cells were ultrafiltered and added again to cells in the presence or absence of selenium. Selenium decreased the frequency of SCE by As and Cr. This observation indicates the possibility of presence of clastogenic factor. In addition, the clastogenic factor would be involed in oxidative stress since selenium decreased the level of oxidative stress. Thus, it is suggested that selenium may play a role as an anti-clastogenic effector by preventing the oxidative stress, thereby decreasing the frequency of Asand Cr-induced chromosomal damage.

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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.

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.

Evaluation of Chromosomal Alteration in Electrical Workers Occupationally Exposed to Low Frequency of Electro Magnetic Field (EMFs) in Coimbatore Population, India

  • Balamuralikrishnan, Balasubramanian;Balachandar, Vellingiri;Kumar, Shanmugam Suresh;Stalin, Nattan;Varsha, Prakash;Devi, Subramaniam Mohana;Arun, Meyyazhagan;Manikantan, Pappuswamy;Venkatesan, Chinnakulandhai;Sasikala, Keshavarao;Dharwadkar, Shahnaz N.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2961-2966
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    • 2012
  • Extremely low frequency electro magnetic fields (EMFs) have been classified as possibly carcinogenic to humans by the International Agency for Research on Cancer. An increased number of chromosomal alterations in peripheral lymphocytes are correlated with elevated incidence of cancer. The aim of the present study was to assess occupationally induced chromosomal damage in EMF workers exposed to low levels of radiation. We used conventional metaphase chromosome aberration (CA) analysis and the micronucleus (MN) assay as biological indicators of non ionizing radiation exposure. In the present study totally 70 subjects were selected including 50 exposed and 20 controls. Informed written consent was obtained from all participants and the study was performed in accordance with the Declaration of Helsinki and the approval of the local ethical committee. A higher degree of CA and MN was observed in exposed subjects compared to controls, the frequency of CA being significantly enhanced with long years of exposure (P<0.05). Moreover increase in CA and MN with age was noted in both exposed subjects and controls, but was significantly greater in the former. The results of this study demonstrated that a significant induction of cytogenetic damage in peripheral lymphocytes of workers occupationally exposed to EMFs in electric transformer and distribution stations. In conclusion, our findings suggest that EMFs possess genotoxic capability, as measured by CA and MN assays; CA analysis appeared more sensitive than other cytogenetic end-points. It can be concluded that chronic occupational exposure to EMFs may lead to an increased risk of genetic damage among electrical workers.

Assessment of Sensitivity of Photo-Chromosomal Assay in the Prediction of Photo-carcinogenicity (광염색체이상시험의 광발암성 예측능력에 대한 평가)

  • Hong Mi-Young;Kim Ji-Young;Lee Young Mi;Lee Michael
    • Toxicological Research
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    • v.21 no.2
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    • pp.99-105
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    • 2005
  • Photo-mutagenic compounds have been known to alter skin cancer rates by acting as initiators or by affecting subsequent steps in carcinogenesis. The objectives of this study are to investigate the utility of photo-chromosomal aberration (photo-CA) assay for detecting photo-clastogens, and to evaluate its ability to predict rodent photocarcinogenicity. Photo-CA assay was performed with five test substances that demonstrated positive results in photo-carcinogenicity tests: 8-Methoxypsoralen (photoactive substance that forms DNA adducts in the presence of ultraviolet A irradiation), chlorpromazine (an aliphatic phenothiazine an alpha-adrenergic blocking agent), lomefloxacin (an antibiotic in a class of drugs called fluoroquinolones), anthracene (a tricyclic aromatic hydrocarbon a basic substance for production of anthraquinone, dyes, pigments, insecticides, wood preservatives and coating materials) and Retinoic acid (a retinoid compound closely related to vitamin A). For the best discrimination between the test substance-mediated genotoxicity and the undesirable genotoxicity caused by direct DNA absorption, a UV dose-response of the cells in the absence of the test substances was firstly analyzed. All 5 test substances showed a positive outcome in photo-CA assay, indicating that the photo-CA test is very sensitive to the photo-genotoxic effect of UV irradiation. With this limited data-set, an investigation into the predictive value of this photo-CA test for determining the photo-carcinogenicity showed that photo-CA assay has the high ability of a test to predict carcinogenicity. Therefore, the photo-CA test using mammalian cells seems to be a sensitive method to evaluate the photo-carcinogenic potential of new compounds.