• Title/Summary/Keyword: Chromosome aberration assay

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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.

Reduction of Radiation-induced Chromosome Aberration and Apoptosis by Dithiothreitol

  • Kim, Jeong Hee;Lee, Eun Ju;Hyun, Jin Won;Kim, Sung Ho;Mar, Woongchon;Kim, Jin Kyu
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.683-687
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    • 1998
  • We have examined in vitro and in vivo radioprotective effects of a well-known thiol-containing compound, dithiothreitol (DTT). The treatment of both 0.5 and 1mM of DTT significantly increased clonogenic survival of ${\gamma}$-ray irradiated Chinese hamster (V79-4) cells. In order to investigate the possible radioprotective mechanism of DTT, we measured gamma-ray induced chromosome aberration by micronucleus assay. In the presence of 0.5mM or 1mM DTT, the frequencies of micronuclei were greatly reduced in all dose range examined (1.5-8 GY). Slightly higher reduction in micronucleus formation was observed in 1mM DTT-treated cells than in 0.5mM DTT-treated cells. In addition, incubation with both 0.5 and 1mM of DTT prior to gamma-ray irradiation reduced nucleosomal DNA fragmentation at about same extent, this result suggests that treatment of DTT at concentrations of 0.5 and 1mM reduced radiation-induced apoptosis. In vivo experiments, we also observed that DTT treatment reduced the incidence of apoptotic cells in mouse small intestine crypts. In irradiated control group 4.4${\pm}$0.5 apoptotic cells per crypt were observed. In DTT-administered and irradiated mice, only 2.1${\pm}$0.4 apoptotic cells per crypt was observed. In vitro and in vivo data obtained in this study showed that DTT reduced radiation-induced damages and it seems that the possible radioprotective mechanisms of action of DTT are prevention of chromosome aberration.

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Genotoxicity Study of Water Extract of Cordyceps militaris Grown Upon Protuetja dreujtarsis (굼벵이 유래 밀리타리스 동충하초 열수 추출물의 유전독성평가)

  • Jo, Wol-Soon;Nam, Byung-Hyouk;Choi, Yoo-Jin;Oh, Su-Jung;Kang, Eun-Young;Lee, Sang-Ho;Jeong, Min-Ho
    • Toxicological Research
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    • v.23 no.3
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    • pp.245-251
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    • 2007
  • Water extract of Cordyceps militaris grown upon Protuetja dreujtarsis (CMPD) was examined for the genetic toxicity-bacterial mutagenicity, chromosome aberration, and micronucleus formation. For mutagenicity assay, bacterial reversion test with Salmonella typhimurium TA98, TA100, TA1535, TA 1537, and E. coli WP2uvrA were performed. The extract at the concentrations of $50{\sim}5,000{\mu}g/plate$ did not induce mutagenicity at all. Chromosome aberration test was performed by using Chinese lung (CHL) cells. There was no significant chromosome aberration in CHL cells with S-9 mixture at the concentrations of $312.5{\sim}1,250{\mu}g/ml$ of the extract and without S-9 mixture at the concentrations of $1.2{\sim}19.5{\mu}g/ml$ of the extract. For micronucleus test, ICR mice were treated with the extract at the dose of 0.5, 1, and 2g/Kg. The frequencies of the micronucleated polychromatic erythrocytes (MNPCE) in bone marrow preparations of the extract-treated group were not increased compared to the untreated control group. Taken together, our results show that water extract of CMPD did not induce any harmful genotoxicity.

Study on Mutagenicity of DehydroevodiamineㆍHCl(DHED) (치료제 DehydroevodiamineㆍHCl(DHED)의 변이원성 연구)

  • 성이숙;정성윤;정주연;채규영;진미령;최봉웅;장병모;김대경
    • YAKHAK HOEJI
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    • v.46 no.3
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    • pp.208-212
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    • 2002
  • Dehydroevodiamine HCl (DHED), which is a component separated from Evodia rutaecarpa Bentham, has novel anticholinesterase and antiamnesic activities in the scopolamine-induced amnesia model. Several studies suggest that DHED might be an effective drug for the Alzheimer's disease and the vascular type of dementia. In order to evaluate the mutagenic potential of DHED, Salmonella typhimurium reversion assay, chromosomal aberration test on Chinese hamster lung cells, in vivo micronucleus assay using mouse bone marrow cells, and comet assay were performed. DHED did not increase the number of revertant in the reverse mutation test using Salmonella typhimurium TA1535, TA1537, TA98, TA100. DHED HCl, at concentration of 5 and 10 $\mu\textrm{g}$/mι, increased the number of chromosome aberrated Chinese hamster lung cells with 5 and 10%, respectively. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocyte was observed in ICR mice orally administered with DHED. DHED was tested for ability to induce genotoxic effect in L5178Y cells (mouse lymphoma cells) using the single cell gel electrophoresis assay (comet assay). In comet assay, tail moment did not increase in L5178Y cells treated with 10, 100, 300 $\mu$M DHED.

Analysis of Chromosome Aberrations and Micronuclei in Peripheral Lymphocytes of Nurses in a Hospital (병원 근무 간호사들의 염색체 이상 및 소핵 빈도에 미치는 위험요인)

  • Jeon Hye Jeong;Kim Tae Hwan;Choi Soo Yong;Chung Hai won
    • Environmental Mutagens and Carcinogens
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    • v.25 no.4
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    • pp.150-156
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    • 2005
  • Analysis of chromosome aberration (CA) and cytokinesis-block micronucleus (CBMN) assay in peripheral lymphocytes of nurses exposed to low levels of anticancer drug and Ethylene Oxide(EO) gas in a hospital were performed. The frequency of CA was increased in the exposed compared to the controls whereas no increase of the frequency of MN was found. The frequencies of chromatid type CA were 1.2, 3.91 and 9.67 per 500 cells in the controls, workers exposed to anticancer drug and workers exposed to EO, respectively. Lower frequency of CA in nurses handling anticancer drugs with safety covers compared to those without safety covers was observed, but it was not statistically significant. The frequency of CA in nurses handling anticancer drugs increased by the frequency of mixing anticancer drugs. Poisson regression analysis showed a significant association of the frequency of chromatid type CA with age, duration of wort exposure to anticancer drug and EO gas exposure, but no association of the frequency of chromosome type CA with any variables. The results suggested that there were associations between CA and the occupational exposure to low levels of anticancer drug and EO gas.

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Studies on the Genotoxicity of the Gamma-irradiated Panax Ginseng Radix In Vitro and In Vivo (방사선조사 인삼의 유전독성에 관한 연구)

  • 하광원;정해관;오혜영;허옥순;손수정;한의식;정성철;최부영;김영미
    • Journal of Food Hygiene and Safety
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    • v.9 no.2
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    • pp.67-74
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    • 1994
  • This study was aimed to find out the comparative effects between non-irradiated, and 5kGy-10kGy of gamma-irradiated Panax Ginseng Radix powder on the genotoxicity for identification of possibility of DNA damage causing cancer. Four different short-term mutagenicity tests were used: (1) Salmonella typhimurium reversion assay (Ames test) (2) Chromosome aberration test in cultured Chinese hamster lung (CHL) fibroblast cells. (3) Micronucleus test in ddY mouse (4) Somatic mutation and recombination test in the wing cells of Drosophila melanogaster.Gamma-irradiated Panax Ginseng Radix powder revealed negative results in these four mutagenicity tests. This means gamma-irradiated ginseng could be safe on the genotoxic point of view.

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In vitro Genotoxicological Safety of Fresh Vegetable-Extract Juice by Gamma Irradiation (감마선 조사된 녹즙의 In vitro 유전독성학적 안전성 평가)

  • 이현자;강근옥;육홍선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1227-1236
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    • 2001
  • Genotoxicological safety on 10 kGy-gamma irradiated vegetable juices such as Oenanthstolonifera DC., Daucus carota L., Brassica oleracea var. acephala and Angelica keiskei was determined by the Salmonella typhmurium reversion assay, the SOS Chromotest using in Escherichia cloi PQ37 and chromosome aberration test in cultured Chinese hamster lung (CHL) fibroblast cells. Vegetable juices exposed to 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests.

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Genotoxic evaluation of Gyllus bimaculatus in 3 sets of mutagenicity tests

  • Ahn, Mi-Young;Pae, Hey-Jin;Lee, Byung-Mu;Ryu, Kang-Sun
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.120.1-120.1
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    • 2003
  • The mutagenic potential of Gryllus bimaculatus was evaluated using the short-term genotoxicity tests including Ames, chromosome aberration and micronuclei tests. In salmonella typhimurium assay, G. bimaculatus did not show any mutagenic response in the absence or presence of S9 mix with TA98, TAl00, TA1535, and TA1537. In chromosome aberration test, G. bimaculatus did not show any significant effect on Chinese Hamster Ovary (CHO) cells compared with control. (omitted)

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Genotoxicity Studies of the Complex of Acriflavine and Guanosine (Acriflavine과 Guanosine 복합체(AG60)의 유전독성시험)

  • 정영신;홍은경;김상건;안의태;이경영;강종구
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.106-111
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    • 2002
  • AG6O, the complex of acriflavine and guanosine, has been shown to possess the synergistic antitumorigenic activity in the previous paper (J. Pharm. Pharmacol. 1997, 49:216). In this study, we have investigated the genotoxic properties of AG60 using in vitro and in vivo system such as Ames bacterial reversion test, chromosomal aberration assay and micronucleus assay. In Ames reverse mutation test, AG60 treatment at the dose range up to 250 $\mu\textrm{g}$/plate caused the dose-independent random induction of the mutagenic colony formation in S. typhimurium TA98, TA100, TA1537, and E. coli WP2uvrA, while any mutagenic effect of AG60 wasn't observed in S. typhimurium TA1535. Any significant chromosomal aberration wasn't observed in chinese hamster lung (CHL) fibroblast cells incubated with PBS or AG60 at the concentrations of 2.5, 5, 10 $\mu\textrm{g}$/$m\ell$ for 24 hours without but even with 59 metabolic activation system for 6 hours. In vivo ICR mice, the intramuscular injection of AG60 at the doses of 7.15, 14.3, and 28.6 mg/kg did not induce the frequency of micronucleus formation. However, mitomycin C, as one of the positive controls at the dose of 2 mg/kg caused the 8.4% induction in the frequency of micronucleus and 24% increase in the chromosomal aberration.

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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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