• Title/Summary/Keyword: reverse mutation test

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Mutagenecity Test of Intralipidos (Intralipidos에 대한 변이원성시험)

  • Jung, Ji-Youn;Lee, Won-Woo;Ihm, Jong-Hee;Nam, Jeong-Seok;Che, Jeong-Hwan;Li, Guang-Xun;Kang, Byeong-Cheol;Yi, Beoung-Hi;Park, Jae-Hak;Lee, Yong-Soon
    • Toxicological Research
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    • v.14 no.3
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    • pp.453-457
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    • 1998
  • In order to evaluate the mutagenic potential of Intralipidos produced by Greenmate cooperation. We performed Salmonella typhimurium reversion assay, chromosomal aberration test on chinese hamster ovarian cells and in vivo micronucleus assay using mouse bone marrow cells. In the reverse mutation test using Salmonella typhimurium TA98 and TA100, Intralipidos did not increase the number of revertant at any of the concentration tested in this study. Intralipidos did not increase the number of cells having structural or numberical chromosome aberration in cytogenetic test. In mouse micronucleus test, no significant increase were observed in the occurrence of micornucleated polychromatic erythrocytes in ICR male mice intraperitoneally administered with Intralipidos. These results indicate that Intralipidos has no genetic toxicity under these experimental conditions.

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Genotoxicity Studies on Corn Silk Extract Containing High Maysin (고메이신 함유 옥수수수염 추출물의 유전독성학적 안전성 연구)

  • Ha, Ae Wha;Kang, Hyeon Jung;Kim, Sun Lim;Kim, Myung Hwan;Kim, Woo Kyoung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1045-1052
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    • 2017
  • In this study, a battery of genetic-toxicity studies on corn silk extract with high maysin content were performed according to internationally accepted protocols. In a mutation test using Salmonella Typhimurium TA1535, TA1537, TA98, and TA100, the number of mutant colonies did not significantly increase up to a maximum concentration of $5,000{\mu}g/plate$ in the presence or absence of the S9 metabolic activation system. In the chromosome aberration test using Chinese hamster lung fibroblasts, negative results were observed in the concentration up to $1,250{\mu}g/mL$ of corn silk extract. In the micronucleus test using ICR mice, incidence of polymorphonuclear erythrocytes with a maximum concentration of 2,000 mg/kg corn silk extract did not show any significant difference compared to the negative control group. Based on these results, the test substance, con silk extract, did not influence genotoxicity.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay (염기서열결정과 Line Probe 분석법에 의한 Rifampin내성 결핵균의 rpoB 유전자 분석)

  • Lee, Min-Ki;Kim, Yun-Seong;Lee, Hyo-Jin;Cheon, Du-Su;Yun, Sang-Myung;Park, Sam-Seok;Kim, Cheol-Min;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.251-263
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    • 1997
  • Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

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Evaluation of Reverse Hybridization Assay for Detecting Fluoroquinolone and Kanamycin Resistance in Multidrug-Resistance Mycobacterium tuberculosis Clinical Isolates (다제내성결핵 균주에서 Reverse Hybridization Assay를 이용한 Fluoroquinolone, Kanamycin 신속 내성 검사의 유용성)

  • Park, Chin-Su;Sung, Nack-Moon;Hwang, Soo-Hee;Jeon, Jae-Hyun;Won, Young-Sub;Min, Jin-Hong;Kim, Cheon-Tae;Kang, Hyung-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.1
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    • pp.44-49
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    • 2012
  • Background: Multidrug-resistant tuberculosis (MDR-TB) is an increasing public health problem and poses a serious threat to global TB control. Fluoroquinolone (FQ) and aminoglycoside (AG) are essential anti-TB drugs for MDR-TB treatment. REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$ (M&D, Wonju, Korea) were evaluated for rapid detection of FQ and kanamycin (KM) resistance in MDR-TB clinical isolates. Methods: M. tuberculosis (n=67) were isolated and cultured from the sputum samples of MDR-TB patients for extracting DNA of the bacilli. Mutations in genes, gyrA and rrs, that have been known to be associated with resistance to FQ and KM were analyzed using both REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$, respectively. The isolates were also utilized for a conventional phenotypic drug susceptibility test (DST) as the gold standard of FQ and KM resistance. The molecular and phenotypic DST results were compared. Results: Sensitivity and specificity of REBA MTB-FQ$^{(R)}$ were 77 and 100%, respectively. Positive predictive value and negative predictive value of the assay were 100 and 95%, respectively, for FQ resistance. Sensitivity, specificity, positive predictive value and negative predictive value of REBA MTB-KM$^{(R)}$ for detecting KM resistance were 66%, 94%, 70%, and 95%, respectively. Conclusion: REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$ evaluated in this study showed excellent specificities as 100 and 94%, respectively. However, sensitivities of the assays were low. It is essential to increase sensitivity of the rapid drug resistance assays for appropriate MDR-TB treatment, suggesting further investigation to detect new or other mutation sites of the associated genes in M. tuberculosis is required.

Genotoxicity Study of ChondroT (ChondroT의 유전독성 연구)

  • Kim, Sun-Gil;Kim, Joo Il;Kim, Ji-Hoon;Yoon, Chan Suk;Jeong, Ji-Won;Na, Chang-Su;Kim, Seon-Jong
    • Journal of Korean Medicine Rehabilitation
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    • v.31 no.1
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    • pp.59-79
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    • 2021
  • Objectives This study was performed to observe the genotoxic effect of the ChondroT. Methods To evaluate the genotoxicity of ChondroT, an experiment of bacterial reverse mutation test, in vitro mammalian chromosomal aberration test and mammalian erythrocyte micronucleus test in mouse was conducted. Results TA98, TA100 and TA1537 strains in the absence of metabolic activation system (S9 mix), the number of revertant colonies being greater than 2-fold of the respective negative control value. Both in -S9 mix and +S9 mix, the frequencies of aberration cells with structural aberration and numerical aberrations of chromosome were less than 5%. There was no increase of polychromatic erythrocyte with one or more micronuclei at any dose of test substance compared to the negative control group (p<0.05). Conclusions In TA98, TA100 and TA1537 strains in the absence of metabolic activation system (S9 mix), the number of revertant colonies was greater than 2-fold of the respective negative control value, showing positive results. ChondroT was considered to be non-clastogenic to Chinese hamster lung (CHL/IU) cells under the present experimental condition. and ChondroT was determined not to induce an increased frequency of micronuclei in the bone marrow cells of male ICR mice under the present experimental condition.

Evaluation of Genotoxicity and 28-day Oral Dose Toxicity on Freeze-dried Powder of Tenebrio molitor Larvae (Yellow Mealworm)

  • Han, So-Ri;Yun, Eun-Young;Kim, Ji-Young;Hwang, Jae Sam;Jeong, Eun Ju;Moon, Kyoung-Sik
    • Toxicological Research
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    • v.30 no.2
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    • pp.121-130
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    • 2014
  • The larval form of Tenebrio molitor (T. molitor) has been eaten in many countries and provides benefits as a new food source of protein for humans. However, no information exists regarding its safety for humans. The objective of the present study was to evaluate the genotoxicity and repeated dose oral toxicity of the freeze-dried powder of T. molitor larvae. The genotoxic potential was evaluated by a standard battery testing: bacterial reverse mutation test, in vitro chromosome aberration test, and in vivo micronucleus test. To assess the repeated dose toxicity, the powder was administered once daily by oral gavage to Sprague-Dawley (SD) rats at dose levels of 0, 300, 1000 and 3000 mg/kg/day for 28 days. The parameters which were applied to the study were mortality, clinical signs, body and organ weights, food consumption, ophthalmology, urinalysis, hematology, serum chemistry, gross findings and histopathologic examination. The freezedried powder of T. molitor larvae was not mutagenic or clastogenic based on results of in vitro and in vivo genotoxicity assays. Furthermore, no treatment-related changes or findings were observed in any parameters in rats after 28 days oral administration. In conclusion, the freeze-dried powder of T. molitor larvae was considered to be non-genotoxic and the NOAEL (No Observed Adverse Effect Level) was determined to be 3000 mg/kg/day in both sexes of SD rats under our experimental conditions.

Genotoxicity Study of Sophoricoside, a Constituent of Sophora japonica, in Bacterial and Mammalian Cell System

  • Kim, Youn-Jung;Park, Hyo-Joung;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.99-105
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    • 2001
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to has an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial reversion test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay. As results, in the range of 1,250~40 $\mu\textrm{g}$/plate sophoricoside concentrations was not shown significant mutagenic effects in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains in Ames test. The 80% cell growth inhibition concentration (IC/SUB 80/) of sophoricoside was determined as above 5,000 $\mu\textrm{g}$/$m\ell$ in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line for the chromosomal aberration and comet assay, respectively. Sophoricoside was not induced chromosomal aberration in CHL fibroblast cell at concentrations of 700, 350 and 175 $\mu\textrm{g}$/$m\ell$ or 600, 300 and 150 $\mu\textrm{g}$/$m\ell$ in the absence or presence of S-9 metabolic activation system, respectively. Also, in the comet assay, the induction of DNA damage was not observed in L5178Y mouse lymphoma cell line both in the absence or presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside were observed in bacterial and mammalian cell systems used in these experiments.

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Evaluation of the Genotoxicity of Decursin and Decursinol Angelate Produced by Angelica gigas Nakai

  • Kim, Kang-Min;Kim, Tae-Ho;Park, Yun-Jung;Kim, Ik-Hwan;Kang, Jae-Seon
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.83-87
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    • 2009
  • In this study, we assessed the stability and toxicological safety of Angelica gigas Nakai (A. gigas Nakai) extract, which is comprised of decursin and decursinol angelate (D/DA). D/DA was tested for mutagenicity using Ames Salmonella tester strains (TA102, TA1535, and TA1537) with or without metabolic activation (S9 mix). No increase in the number of revertants was observed in response to any of the doses tested (1.25, 12.5, 125, and $1,250{\mu}/mLg$). In addition, a chromosome aberration test was conducted in the Chinese hamster lung (CHL) cell line. To accomplish this, cells were treated with D/DA (3.28, 13.12, 52.46, and $209.84{\mu}g/mL$) or with Mitomycin C ($0.1{\mu}/mLg$) as a positive control in the case of no metabolic activation or benzo(a)pyrene ($20{\mu}g/mL$) in the case of metabolic activation. No significant increase in chromosome aberrations was observed in response to treatment with any of these concentrations, regardless of activation of the metabolic system. According to these results, we concluded that D/DA did not induce bacterial reverse mutation or clastogenicity in vitro in the range of concentrations evaluated in these experiments.

Mutagenecity evaluation of insecticidal 2-carbomethoxy-4-chlorodiethyl phosphate in short-term bioassays (살충성 물질 2-carbomethoxy-4-chlorodiethyl phosphate의 유전독성 평가)

  • Lee, Je-Bong;Sung, Ha-Jung;Jeong, Mi-Hye;Kwon, Oh-Kyung;Lee, Hae-Keun;Kim, Young-Koo
    • The Korean Journal of Pesticide Science
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    • v.2 no.2
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    • pp.53-58
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    • 1998
  • For evaluating the mutagenic potential of 2-carbomethoxy-4-chlorodiethyl phosphate, three different short-term mutagenicity tests were used; Salmonella typhimurium preincubation assay with and without rat liver microsomal activation, chromosome aberration test in cultured chinese hamster lung fibroblast cell and in vivo micronucleus test in male mice bone marrow. In Salmonella typhimurium reverse mutation assay using TA98, TA100, TAl535 and TAl537, 2-carbomethoxy-4-chlorodiethyl phosphate did not show any mutagenic response in the presence and absence of S9 mix. It did not induce any significant structural chromosome aberrations in the absence of metabolic activation. In micronucleus test using ICR mice, the frequency of micronucleated polychromatic erythrocytes (MNPCE) increased in bone marrow cells treated with positive control, mitomycin-C, but 2-carbomethoxy-4-chlorodiethyl phosphate did not increase micronucleated polychromatic erythrocytes. These results indicate that 2-carbomethoxy-4-chlorodiethyl phosphate does not show any positive responses in short-term genotoxicity assays.

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