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Genotoxicity Tests on 3-Methoxypropanoic acid methyl ester, Methanesulfonic acid and Quinoline
Jung Kwon,Ji-Young Kim,Seng-Min Back,Ji-Yeon Kim,Ok-Hee Kim,Wook-Joon Yu,Sun-kyoung Yoo,Hee-Young Ro,Eunju Kim,Hye-jin Han,Ilseob Shim,Hyun-Mi Kim,Pil Je Kim,Kyunghee Choi 한국실험동물학회 2013 한국실험동물학회 학술발표대회 논문집 Vol.2013 No.2
마이크로플레이트를 이용한 8종 화학물질의 복귀돌연변이 평가
김명준 ( Myoung Jun Kim ),허성무 ( Seongmoo Heo ),남수현 ( Suhyun Nam ),백승민 ( Seng-min Back ),박성훈 ( Seong Hoon Park ),오정자 ( Jeong Ja Oh ),정영신 ( Young-shin Chung ) 한국동물실험대체법학회 2021 동물실험대체법학회지 Vol.15 No.1
This study was conducted to establish the technology for bacterial reverse mutation test using 384-well microplates and liquid media instead of solid agar in order to evaluate the mutagenicity of test articles with limited amounts. The important advantage of this test method was considered that the usage of S9 fraction prepared from rat liver was smaller in comparison with the Ames test using solid agar. Five bacterial strains (S. typhimurium TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA) recommended in OECD TG 471 were used to evaluate 8 chemicals including 6 mutagens (Allyl chloride, 4-amino-2-nitrophenol, ethylene glycol dimethyl ether, benzo[α]pyrene, 7,12-dimethylbenzanthracene, 8-hydroxyquinoline) and 2 non-mutagens (dioctyl phthalate, sulfisoxazole). All experiments were performed using Ames MPFTM Penta II kits (Xenometrix, Switzerland). Three independent laboratories participated in the intra-laboratory and inter-laboratory studies. Results from this study were compared with reference mutagenicity data (NTP or published data). In results of this study, five out of six mutagens were evaluated to be positive, and the one mutagen (ethylene glycol dimethyl ether) to be negative. Two non-mutagens were determined to be negative in microplate tests. Both the intra-laboratory and the inter-laboratory study showed the 87.5% concordance in the overall results. It was concluded that a microplated reverse mutation test may be a compatible test method for the animal welfare as well as for the limited amount of substances.
생체 내 Pig-a 유전자 돌연변이시험에 대한 세 시험실의 전수가능성 평가
정영신 ( Young-shin Chung ),박범수 ( Bumsoo Pak ),한세희 ( Sehee Han ),이지연 ( Jiyeon Lee ),김지영 ( Jiyoung Kim ),백승민 ( Seng-min Back ),박초롱 ( Cho-rong Park ),김수환 ( Su-hwan Kim ),이종권 ( Jong-kwon Lee ) 한국동물실험대체법학회 2017 동물실험대체법학회지 Vol.11 No.1
In order to establish a pig-a gene mutation assay as a regulatory test for evaluating in vivo genotoxicity in Korea, the transferability study as a stage II validation was performed using the test substance, N-ethyl-N-nitrosourea (ENU) across three laboratories. The lead laboratory (KIT) standardized the protocol using Rat MutaFlow® Kit and transferred the methodology to two participating laboratories (Hoseo University and Biotoxtech Co. Ltd.). Sprague Dawley rats were treated with vehicle or three dose levels of ENU (10, 20, 40mg/kg) by oral gavages on the 3 consecutive days. On the day 15 after the first treatment, bloods were collected and analyzed to estimate the percentage of reticulocytes (%RETs), mutant type of RETs and erythrocytes (RBC) using flowcytometer. In results, the exposure of ENU induced the dose-dependent increases in the mutant frequencies of RBCCD59- and RETCD59-, while %RETs were not affected. Data comparisons among three participating laboratories indicated the good transferability with the high correlation coefficients (0.97-0.99) between the lead and participating laboratories. In conclusion, the pig-a gene mutation assay may be the effective test for the estimation of in vivo gene mutation and the validation studies may be needed using more test substances in Korea.