• Title/Summary/Keyword: Micronuclei assay

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Genotoxicological Safety Evaluation of Crude Antifungal Compounds Produced by Bacillus subtilis SN7 (Bacillus subtilis SN7이 생성한 조항균 물질의 유전독성학적 안정성평가)

  • Chang, Hae-Choon;Koh, Sang-Bum;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.28 no.1
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    • pp.131-141
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    • 2017
  • This study was carried out to perform genotoxicological safety evaluation of crude antifungal compounds produced by Bacillus subtilis SN7 (B. subtilis SN7) isolated from meju. Bacterial reverse mutation assay with Salmonella typhimurium TA98, TA100, TA1535, and TA1537 or Escherichia coli WP2uvrA in the presence and absence of the S9 metabolic activation system was carried out, and the crude antifungal compounds produced by B. subtilis SN7 showed no significant increase in the number of revertant colonies. In the chromosomal aberration tests using Chinese hamster lung (CHL) cells, sample treatment groups showed no increase in the frequency of chromosome aberrations compared to the negative control group. Furthermore, in the micronucleus formation test, the crude antifungal compounds showed no significance increase in the frequency of polychromatic erythrocytes with micronuclei. These results suggest that the crude antifungal compounds produced by B. subtilis SN7 isolated from meju showed no harmful genotoxic effects.

Genetic Toxicity Test of Emodin by Ames, Micronucleus, Comet Assays and Microarray Analysis Showing Differential Result

  • Go, Seo-Y.;Kwon, Kyoung-J.;Park, Sue-N.;Sheen, Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.15 no.3
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    • pp.192-198
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    • 2007
  • Emodin (1,3,8-trihydroxy-6-methylanthraquinone) is a major constituent of rhubarb. Although it has been claimed to have a wild spectrum of therapeutic value, its side effects, especially in human kidney cells have not been well characterized. In this study, we have carried out in vitro genetic toxicity test of emodin and microarray analysis of differentially expressed genes in response to emodin. The result of Ames test showed mutations with emodin treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, emodin showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with emodin treatment showed DNA damage both with and without exogenous metabolic activation. Emodin did not increase micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to emodin by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of emodin.

Study on the Biological Effects of TSP Collected from the Subway Station with Tradescantia Bioassay (지하철 시설내 부유먼지에 함유된 돌연변이원의 생물학적 영향 평가)

  • Kim, Jin-Kyu;Shin, Hae-Shik;Lee, Jeong-Joo;Kim, Kyun;Lee, Jin-Hong
    • Environmental Analysis Health and Toxicology
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    • v.17 no.3
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    • pp.245-252
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    • 2002
  • Airborne pollutants in the subway facilities can be potentially harmful to the health of passengers. This study was designed to examine whether the suspended particulates have mutagenic or carcinogenic effect on the plant cell systems. Total suspended particulates were collected with a high volume air sampler, in the entrance, the waiting room, and the platform of each subway station. The biological end -points in this experiment were the pink mutations in stamen hairs and micronuclei in the pollen mother cells of Tradescantia. The exudates were collected by shaking the filter papers from the sampler in distilled water for 24 hours. All the plant cuttings exposed to the exudates resulted in positive responses. The micronucleus assay proved more reliable and sensitive to the test than the stamen hair assay. The results indicate that the air particulates can give an adverse effect on the health of subway passengers.

Genotoxicity Studies on Carrageenan: Short-term In Vitro Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Choi, Seon-A;Oh, Se-Wook;Park, Sue-Nie;Yum, Young-Na;Kim, Joo-Hwan;Seo, Young-Rok;Lee, Michael
    • Toxicological Research
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    • v.25 no.1
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    • pp.51-58
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    • 2009
  • Carrageenan is a naturally-occurring sulfated polygalactan which has been widely used in the dairy industry and a gelling agent in non-dairy products. In this study, four short-term in vitro genotoxicity assays were investigated to evaluate the potential genotoxic effects of carrageenan. The mutagenicity of carrageenan was evaluated up to a maximum dose of 5 mg/plate in Ames test. There was no increase in the number of revertant colonies compared to its negative control at any dose in all of strains tested. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. Carrageenan was not considered to be clastogenic in this assay at up to the highest feasible concentration which could be evaluated. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that carrageenan 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, our results indicate that carrageenan was not genotoxic based on four in vitro genotoxicity results.

Genotoxicological Safety of High-Dose Irradiated Porridges (고선량 조사된 시판 분말죽의 유전독성학적 안전성평가)

  • Kang, Il-Jun;Kang, Young-Hee;Chung, Cha-Kwon;Oh, Sung-Hoon;Lee, Ju-Woon;Byun, Myung-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.261-266
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    • 2005
  • Gamma irradiation at 30 kGy was applied to porridge to evaluate its possible genotoxicity. The genotoxicity of irradiated porridge was evaluated by Salmonella Typhimurium reversion assay, chromosomal aberration test and in vivo micronucleus assay. The results were negative in the bacterial reversion assay with S. Typhimurium TA98, TA100, TA1535 and TA1537. No mutagenicity was detected in the assay both with and without metabolic activation. In chromosomal aberration tests with CHL cells and in vivo mouse micronucleus assay, no significant difference in the incidences of chromosomal aberration and micronuclei was observed between nonirradiated and 30 kGy-irradiated porridge. These results indicate that porridge irradiated at 30 kGy did not show any genotoxic effects under these experimental conditions.

Cytokinesis-blocked micronuclei in the human peripheral lymphocytes following low dose γ-rays irradiation (저선량의 감마선 피폭된 사람 말초 임파구의 미소핵을 이용한 방사선 생물학적 피폭선량 측정법 연구)

  • Kim, Tae-hwan
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.99-104
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    • 2001
  • To determine if micronucleus (MN) assay could be used to predict the absorbed dose of victims after accidental radiation exposure, we carried out to assess the absorbed dose depending on the numerical changes of MN in human peripheral blood lymphocytes after $^{60}Co\;{\gamma}-rays$ exposure in the range of 0.25 to 1 Gy, respectively. The MNs were observed at very low doses, and the numerical changes according to doses. Satisfactory dose-effect calibration curve is observed after low dose irradiation of human lymphocytes in vitro. When plotting on a linear scale against radiation dose, the line of best fit was $Y=(0.02{\pm}0.0009)+(0.033{\pm}0.010)D+(0.012{\pm}0.012)D^2$. The dose-response curve for MN induction immediately after irradiation was linear-quadratic and has a significant relationship between the frequencies of MN and dose. These data show a trend towards increase of the numbers of MN with increasing dose. The number of MN in lymphocytes that were observed in the control group is $0.1610{\pm}0.0093/cell$. Accordingly, MN assay in human peripheral lymphocytes could be a useful in viva model for studying radio-protective drug sensitivity or screening test, microdosimertic indicator and radiation-induced target organ injury. Since MN assay is simple, rapid and reproducible, it will also be a biodosimetric indicator for individual dose assessment after accidental exposure.

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Evaluating the Micronucleus Induction Potential for the Genotoxicity Assay Using the Human Skin Model, KeraSkinTM (인공피부모델 KeraSkinTM을 이용한 유전독성 평가)

  • Lee, Su-Hyon;Jung, Haeng-Sun;Kim, Seol-Yeong;Kim, Hye Soo;Lim, Kyung-Min;Chung, Young-Shin;Choe, Tae-Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.211-216
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    • 2016
  • Micronucleus test is genotoxicity assay for detection of micronuclei in the cytoplasm of interphase cells. The reduction and replacement of in vivo toxicity testing on animals require the development of in vitro models to predict the genotoxicity or other tests for cosmetic products. In this study, we evaluated a genotoxicity assay for topically applied chemicals using a three-dimensional human reconstructed skin model, KeraSkin$^{TM}$. Two genotoxins, mitomycin C (MMC) and methyl methanesulfonate (MMS), induced significant dose-related increases in cytotoxicity and micronuclei induction in the skin model. In contrast, two non-genotoxins, 4-nitrophenol (4-NP) and trichloroethylene (TCE), induced cytotoxicity but not micronucleus formation. In conclusion, micronucleus test using human skin model may be useful for predicting in vitro genotoxic potentials of cosmetic products.

Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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Cytogenetic Radiation Adaptive Response Assessed by Metaphase Analysis and Micronuclei Test in Human Lymphocytes and Mouse Bone Marrow Cells (인체말초혈액 림프구와 마우스골수세포에서 중기염색체 분석법과 미소핵검사법을 이용한 방사선적응반응 평가)

  • Min, Jung-Jun;Bom, Hee-Seung;Lee, Seung-Yeon;Choi, Keun-Hee;Jeong, Hwan-Jeong;Song, Ho-Cheon;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.6
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    • pp.525-533
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    • 1998
  • Purpose: Radiation adaptive response in human peripheral lymphocytes and mouse bone marrow cells was investigated using both metaphase analysis and micronucleus assay. We assessed the correlation between both tests. Materials and Methods: Two groups of the human peripheral lymphocytes and mouse bone marrow cells were exposed to low dose (conditioning dose, 0,18 Gy) or high dose (challenging dose, 2 Gy) ${\gamma}$-rays. The other 4 groups were exposed to low dose followed by high dose after several time intervals (4, 7, 12, and 24 hours, respectively). The frequencies of chromosomal aberrations in metaphase analysis and micronuclei in micronucleus assay were counted. Results: Chromosomal aberrations and micronuclei of preexposed group were lower than those of the group only exposed to high dose radiation. Maximal reduction in frequencies of chromosomal aberrations were observed in the group to which challenging dose was given at 7 hour after a conditioning dose (p<0.001). Metaphase analysis and micronucleus assay revealed very good correlation in both human lymphocytes and mouse bone marrow cells (r=0.98, p<0.001 ; r=0.99, p=0.001, respectively). Conclusion: Radiation adaptive response could be induced by low dose irradiation in both human lymphocytes and mouse bone marrow cells. There was a significant correlation between metaphase analysis and micronucleus assay.

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Grapefruit Juice Suppresses Azoxymethane-induced Colon Aberrant Crypt Formation and Induces Antioxidant Capacity in Mice

  • Madrigal-Bujaidar, Eduardo;Roaro, Laura Martino;Garcia-Aguirre, Karol;Garcia-Medina, Sandra;Alvarez-Gonzalez, Isela
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6851-6856
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    • 2013
  • In the present report we determined the protective capacity of grapefruit juice (GJ) against molecular and cellular damage in azoxymethane (AOM) treated mice. Animals were daily administered GJ orally (0.8, 4.1, and 8.2 ${\mu}l/g$) for seven weeks, as well as intraperitoneally (ip) injected with AOM twice (weeks 2 and 3 of the assay). Control groups administered with water, with the high dose of GJ, and with AOM injected in weeks 2 and 3 were also included. The results showed a significant, dose-dependent protection of GJ on the number of colon aberrant crypts (AC) induced by AOM. The highest inhibitory effect was reached with the highest tested dose of GJ, decreasing ACF by 51% and 43% at weeks 4 and 7 of the assay. Regarding protein and lipid oxidation we also found a dose-dependent decrease caused with GJ in comparison with the increased levels produced by AOM. Therefore, our results established chemopreventive potential for GJ, and suggested effects related to its antioxidant capacity. Finally, we found that the tested agents induced neither micronuclei increase nor alteration in bone marrow cytotoxicity.