• Title/Summary/Keyword: Micronuclei

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Genetic Toxicity Test of 1,2-Dibromoethane by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ki-Y.;Kim, Ji-H.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Shee, Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.14 no.4
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    • pp.246-252
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    • 2006
  • 1,2-Dibromoethane(DBE) has been widely used as a soil fumigant, an additive to leaded gasoline and an industrial solvent. In this study, we have carried out in vitro genetic toxicity test of 1,2-dibromoethane and microarray analysis of differentially expressed genes in response to 1,2-dibromoethane. 1,2-Dibromoethane showed mutations in base substitution strain TA1535 both with and without exogenous metabolic activation. 1,2-Dibromoethane showed mutations in frame shift TA98 both with and without exogenous metabolic activation. 1,2-Dibromoethane showed DNA damage based on single cell gel/comet assay in L5178Y cells both with and without exogenous metabolic activation. 1,2-Dibromoethane increased micronuclei in CRO cells both with and without exogenous metabolic activation. Microarray analysis of gene expression profiles in L5178Y cells in response to 1,2-dibromoethane selected differentially expressed 241 genes that would be candidate biomarkers of genetic toxic action of 1,2-dibromoethane.

Phytomonitoring of the Genotoxicity of Environmental Pollutants: An Application to Armenian Nuclear Power Plant

  • Kim, Jin Kyu;Aroutiounian, Rouben M.;Nebish, Anna A.;Kim, Jin-Hong
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.181-185
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    • 2015
  • Today the biosafety evaluation, a common problem of vital importance, is based on internationally proved test-systems, standards and techniques. The paradigm of biosafety includes multidisciplinary approach, a combination of physical, chemical and biological tests to monitor the environmental level of pollutants and needs to be improved by modern approaches. The genetic risk of environmental pollutions has long been studied by many researchers. In this study, used was the known sensitive plant test-system, clones of plant Tradescantia (spiderwort) able to detect gene mutations (frequency of mutational events and formation of micronuclei) in combination with chemical and, in some instances, with radiological measurements. In addition, male gametophyte generation of fruit trees was applied as bioindicators of genotoxicity. The obtained results did not show any significant increase along with wind direction. As for the male gametophyte assay, the fertility of the investigated fruit-trees near to NPP did not significantly differ from that of the control point. The influence of the NPP on the male generative system of the investigated taxa of fruit trees for the investigated year was not revealed. The system described needs to be expanded by species of interest (human) as there is a difficulty to transfer the revealed dose correlations to humans. The development of this idea includes various levels: population (epidemiological studies), individual, cellular, molecular (DNA), etc.

Toxicity Evaluation of TA, a Pharmacopuncture Medicine, in an in Vivo Micronucleus Test (ICR 마우스를 이용한 소핵시험을 통한 TA 약침의 안전성 평가)

  • Hwang, Ji Hye;Jung, Hyo Won;Jung, Chul
    • Korean Journal of Acupuncture
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    • v.36 no.1
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    • pp.74-80
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    • 2019
  • Objectives : TA, a polyherbal extract, typically is used for pharmacopuncture therapy on patients with traffic accident-related injuries and musculoskeletal diseases. This study was performed to evaluate the safety of the TA extract, using a micronucleus test. Methods : The dose range and sampling time were first established. An in vivo micronucleus test was then performed to determine the induction of micronuclei in mouse bone marrow cells after a single intramuscular administration of TA to 7-week-old ICR mice (0.2 ml/animal, at 24 hours post-dosing). Results : The incidence of micro-nucleated polychromatic erythrocytes (PCEs) in PCEs in the TA group was similar to that in the negative-control group, while that in the positive-control group was significantly greater. The positive- and negative-control groups did not differ in the ratio of PCEs to total erythrocytes. Conclusions : Our toxicity study indicates that the TA extract does not induce micronucleus formation in mouse bone marrow cells.

In Vivo Genotoxicity Evaluation of a No-Pain Pharmacopuncture Extract Using the Micronucleus Test

  • Ji Hye Hwang;Chul Jung
    • Journal of Pharmacopuncture
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    • v.26 no.4
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    • pp.366-372
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    • 2023
  • Objectives: We aimed to evaluate the genotoxicity of a recently developed no-pain pharmacopuncture (NPP) targeting muscle relaxation and analgesia using the micronucleus test. Methods: To evaluate the potential of NPP extracts to induce micronuclei in rat bone marrow cells, a micronucleus test was performed using male Sprague-Dawley rats. The test substance NPP was administered intramuscularly at concentrations of 0.25, 0.5, and 1 mL/animal. Saline was used as the negative control and cyclophosphamide as the positive control. Results: No NPP treatment-related deaths or abnormal changes in general appearance were observed at any dose level during the experimental period. No statistically significant differences in body weight were observed in any of the NPP dose groups compared to the saline negative control group. NPP did not cause a significant increase in the incidence of micronucleated polychromatic erythrocytes (PCEs) and PCEs or in the ratio of PCE-to-total erythrocytes. Conclusion: The NPP extract did not exhibit genotoxic in Sprague-Dawley rat bone marrow cells under the conditions of this study. Further toxicity studies of the NPP extract are required.

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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Mutagenicity Study of DA-3030, A New Recombinant Human G-CSF(rhG-CSF) (새로운 재조합 인 과립구 콜로니 자극인자 DA-3030의 변이원성연구)

  • 강경구;최성학;김옥진;안병옥;백남기;김계원;김원배;양중익
    • Biomolecules & Therapeutics
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    • v.2 no.3
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    • pp.286-291
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    • 1994
  • The mutagenicity of DA-3030(rhG-CSF)was studied by reverse mutation test, chromosome aberration test and micronucleus test. The reverse mutatuon test in bacteria was performed using salmonella typhimurium strain TA100, TA98, TA1535 and TA1537 with rhG-CSF in any of the concentrations(150, 75, 37.5, 18.75, 9.375 and 4,6875 $\mu\textrm{g}$/plate), no increase in the number of revertant colonies in each strain was observed, irrespective of treatment with the metabolic activation system(S-9 mix) The chromosome aberration test was carried out using CHL cells, cell line from chinese hamster lung. With 4 doses(75, 37.5, 18.75 and 9.375 $\mu\textrm{g}$/ml) of rhG-/CSF the cells were treated for 24 or 48 hours in the direct method or for 6 hours followed by 18 hour-expression time in the metabolic activation method. Results of the study showed, by the direct method or metabolic activation method, no trend toward increase in the number of aberrant metaphase. The micronucleus test was carried out using ICR mice at the age of 8 weeks. Three doses(862.5, 1725 and 3450 $\mu\textrm{g}$/kg) of DA-3030 were admintstered intraperitoneally with single shot and bone marrow cells were sampled at 24 hours after administration. Neither the number of polychromatic erythrocytes with micronuclei nor the ratio of normochromatic erythrocytes to polychromatic erythrocytes increased singinficantly in each dose, compared with a vehicle control. These results indicate that rhG-CSF has not mutagenic potential under the condiions.

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Comparative evaluation of the mutagenicity and genotoxicity of smoke condensate derived from Korean cigarettes

  • Kim, Ha Ryong;Lee, Jeong Eun;Jeong, Mi Ho;Choi, Seong Jin;Lee, Kyuhong;Chung, Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.30
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    • pp.14.1-14.7
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    • 2015
  • Objectives Cigarette smoking is associated with carcinogenesis owing to the mutagenic and genotoxic effects of cigarette smoke. The aim of this study was to evaluate the mutagenic and genotoxic effects of Korean cigarettes using in vitro assays. Methods We selected 2 types of cigarettes (TL and TW) as benchmark Korean cigarettes for this study, because they represent the greatest level of nicotine and tar contents among Korean cigarettes. Mutagenic potency was expressed as the number of revertants per ${\mu}g$ of cigarette smoke condensate (CSC) total particulate matter whereas genotoxic potency was expressed as a concentration-dependent induction factor. The CSC was prepared by the International Organization for Standardization 3308 smoking method. CHO-K1 cells were used in vitro micronucleus (MNvit) and comet assays. Two strains of Salmonella typhimurium (Salmonella enterica subsp. enterica ; TA98 and TA1537) were employed in Ames tests. Results All CSCs showed mutagenicity in the TA98 and TA1537 strains. In addition, DNA damage and micronuclei formation were observed in the comet and MNvit assays owing to CSC exposure. The CSC from the 3R4F Kentucky reference (3R4F) cigarette produced the most severe mutagenic and genotoxic potencies, followed by the CSC from the TL cigarette, whereas the CSC from the TW cigarette produced the least severe mutagenic and genotoxic potencies. Conclusions The results of this study suggest that the mutagenic and genotoxic potencies of the TL and TW cigarettes were weaker than those of the 3R4F cigarette. Further study on standardized concepts of toxic equivalents for cigarettes needs to be conducted for more extensive use of in vitro tests.

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.

Single Dose Oral Toxicity and Genotoxicological Safety Study of Ssanghwa-tang Fermented with Lactobacillus acidophyllus (유산균 발효 쌍화탕에 대한 단회 투여 경구 독성 및 유전 독성 연구)

  • Chung, Tae-Ho;Shim, Ki-Shuk;Kim, Dong-Seon;Lee, Jae-Hoon;Ma, Jin-Yeul
    • The Journal of Korean Medicine
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    • v.32 no.1
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    • pp.67-83
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    • 2011
  • Objectives: The purpose of this study was to examine the single dose toxicity with oral administration and genotoxicities of Ssanghwa-tang fermented with Lactobacillus acidophyllus. Materials and Methods: Clinical signs, weight changes, lethal doses$(LD_{50})$, and postmortem evaluation were determined by Globally Harmonized Classification System(GHCS) in a single-dose oral toxicity study. In vitro mammalian chromosomal aberration test was conducted with Ames test by cell proliferation suppression assessment using the cultivated CHO-K1(Chinese hamster ovary fibroblast) origins. Bacterial reversion assay was performed using Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and Escherichia coli (WP2uvrA). In vivo micronucleus test was performed using ICR mouse bone marrow. Results: No clinical sign was observed and none of the groups with doses up to 2000 mg/kg showed significant acute oral toxicity in the single dose oral administration. None of the sample doses taken during the 6 to 18 hour groups showed significant aberrant metaphases comparing to the negative control group in the in vitro mammalian chromosomal aberration test. No evidence of mutagenicity was seen for Escherichia coli (WP2uvrA) or Salmonella typhimurium (TA98, TA100, TA1535, and TA1537). No significant increase in the frequency of micronuclei was seen in the micronucleus test. Conclusion: These results indicate that the $LD_{50}$ value of Ssanghwa-Tang fermented with Lactobacillus acidophyllus may be over 2000 mg/kg and it have no acute oral toxicity and genotoxicity.

Protective Effects of a Herb, Menthae Herba, against Radiation-induced Oxidative DNA Damage

  • Jo, Sung-Kee;H, Heon-O;Uhee Jung;Kim, Sung-Ho;Byun, Myung-Woo
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.152-152
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    • 2003
  • As utilization of radiation in medicine, industry and biochemical research increases, the protection against radiation damage has become an important issue. Natural products such as herbal medicines are beginning to receive attention as modifiers on the radiation response. In the present study, the protective effect of a herb, Menthae Herba, against radiation-induced DNA damage was evaluated using alkaline single-cell gel electrophoresis (SCGE; comet assay) in the mouse peripheral blood Iymphocytes and the micronucleus formation test in the Chinese hamster ovary (CHO) cells. The tail moment, which was a marker of DNA damage in the SCGE, and the frequency of micronuclei was decreased in groups treated with Mentae Herba extract before exposure to 200 cGy of gamma-ray. We also confirmed its activities to scavenge DPPH and hydroxyl radicals. These experiments demonstrated that Menthae Herba was effective at reducing the radiation-induced damage of DNA and scavenging free radicals. It is plausible that scavenging of free radicals by Menthae Herba may have played an important role in providing the protection against the radiation-induced damage to the DNA. These results indicated that Menthae Herba might be a useful radioprotector and that radical scavenging appears to be one of the mechanisms of radiation protection.

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