• Title/Summary/Keyword: micronucleus assay

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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.

Study on the genotoxicity of soi1 leachate from two polluted sites in Cheongju with Tradescantia-micronuclus assay (자주달개비 미세핵 분석법을 이용한 청주공단주변 토양침출수의 유전독성 평가)

  • Kim Jin Gyu;Lee Byeong Heon;Sin Hae Sik;Lee Jin Hong
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.119-122
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    • 2002
  • Soil contaminants are common in industrialized sites, They can affect directly soil and indirectly ground water and food. Soil mutagens and carcinogens are of great interest due to their potentially hazardous effects on human health. The aim of this study was to monitor the genotoxicity of contaminated soils, Soil leachates were collected from two polluted sites and one control site in Cheongju. Tradescantia BNL 4430 clone was used as experimental matierials. Chromosomal damages induced by soil leachates were detected by the Tradescantia-micronucleus assay. It is known from the result that Tradescantia-micronucleus assay is an excellent botanical tool for detection of biological risk due to environmental toxicants.

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In vitro and In vivo Evaluation of Genotoxicity of Stevioside and Steviol, Natural Sweetner (천연감미료 스테비오사이드와 스테비올의 생체내, 시험관내 유전독성평가)

  • 오혜영;한의식;최돈웅;김종원;손수정;엄미옥;강일현;강혁준;하광원
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.614-622
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    • 1999
  • The standard operation procedure of mouse lymphoma L5178Y $tk^{+/-}-3.7.2C$ gene mutation assay (MOLY) has been regarded as a sensitive in vitro mammalian cell gene mutation assay that is capable of detecting clastogens as well as mutagens. Using MOLY, one of natural sweetner, stevioside (5mg/ml) and its aglycon, steviol ($340{\;}\mu\textrm{g}/ml$) were evaluated the mutagenicity. Stevioside and steviol did not induce mutagenicity in MOLY. On the other hand, stevioside (250mg/kg, B.W.) and steviol (200mg/kg, B.W.) were also evaluated their ability to induce micronuclei in regenerating hepatocytes and bone marrow cells of ddY mice. From these results, stevioside and steviol did not induce any mutagenic effect both MOLY and in vivo micronucleus test.

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Evaluation of Genotoxicity of Water and Ethanol Extracts from Rhus verniciflua Stokes(RVS)

  • Kim, Ji-Young;Oh, Se-Wook;Han, Dae-Seok;Lee, Michael
    • Toxicological Research
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    • v.24 no.2
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    • pp.151-159
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    • 2008
  • Rhus verniciflua Stokes(RVS), one of traditional medicinal plants in Asia, was found to have pharmacological activities such as antioxidative and antiapoptotic effects, raising the possibility for the development of a novel class of anti-cancer drugs. Thus, potential genotoxic effects of RVS in three short-term mutagenicity assays were investigated, which included the Ames assay, in vitro Chromosomal aberration test, and the in vivo Micronucleus assay. In Ames test, the addition of RVS water extracts at doses from 313 up to 5000 mg/plate induced an increase more than 2-fold over vehicle control in the number of revertant colonies in TA98 and TA1537 strains for detecting the frame-shift mutagens. The similar increase in reversion frequency was observed after the addition of RVS ethanol extracts. To assess clastogenic effect, in vitro chromosomal aberration test and in vivo micronucleus assay were performed using Chinese hamster lung cells and male ICR mice, respectively. Both water and ethanol extracts from RVS induced significant increases in the number of metaphases with structural aberrations mostly at concentrations showing the cell survival less than 60% as assessed by in vitro CA test. Also, there was a weak but statistically significant increase in number of micronucleated polychromatic erythrocytes(MNPCEs) in mice treated with water extract at 2000 mg/kg while ethanol extracts of RVS at doses of up to 2000 mg/kg did not induce any statistically significant changes in the incidence of MNPCEs. Therefore, our results lead to conclusion that RVS acts as a genotoxic material based on the available in vitro and in vivo results.

Genotoxicity Study of GST Extract (GST 추출물의 유전독성평가)

  • Lee, Chul Wha;Han, Jong Min;Lee, Mi Young;Jung, In Chul;Jin, Mirim;Kim, Seung Hyung;Park, Yang Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.6
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    • pp.621-629
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    • 2014
  • This study aimed to evaluate the genotoxicity of GST (Gamisasangja-tang). For examining genotoxicity, we carried out bacterial reverse mutation assay, chromosome aberration assay, micronucleus induction test according to OECD guidelines. Bacterial reverse mutation assay: In GST treating group, regardless of existence S9 mix, revertant colonies counts appeared to be less than twice of negative control group and dose dependent increase. In positive control group, revertant colonies counts were shown to be more than twice of negative control croup. Chromosome aberration assay: All cell line showed repetition rate of abnormal chromosome aberration less than 5%, regardless of treating time, existence of S9 mix, and no significant change ($p{\succeq}0.05$) compared with negative control group. Micronucleus induction test: Micronucleated polychromatic erythrocytes (MNPCE) repetition rate of Polychromatic erythrocytes (PCE) showed no significant changes compared with negative control group ($p{\succeq}0.05$). PCE portion of total erythrocytes also showed no significant changes ($p{\succeq}0.05$). Our results showed that GST didn't induce any genotoxicity.

Protective Effect of Buplueri Radix (BR) Against 1,2,4-benzentriol Induced DNA Damage in Human Lymphocytes (Buplueri Radix 의 1,2,4-benzentriol에 의해 유발된 DNA Damage에 대한 보호효과에 대한 연구)

  • Lee, Young-Joon;Kang, Su-Jin
    • Journal of Society of Preventive Korean Medicine
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    • v.12 no.2
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    • pp.51-59
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    • 2008
  • Objectives : Buplueri Radix (BR), used medical plant in Korea traditional medicine, contains various compounds, including a series of triterpene saponins known as saikosaponins. We performed this study for the protective effect of BR against oxidative damage induced by 1,2,4-benzentriol(BT) in human lymphocytes. Methods : In order to investigate the protective effect of BR against carcinogens, genotoxicity induced by benzene metabolite, BT were performed using cytokinesis-block micronucleus(CBMN) assay and comet assay. Results : The frequency of micronucleus at 25, 50 and $100{\mu}M$ concentration of BT were $8{\pm}2.36$, $23{\pm}2.31$, $35{\pm}4.17$ respectively. In addition of BR with concentration of 25 and $50{\mu}g/mL$, MN frequencies were significantly decreased. According to comet assay, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 while BT with BR treatment decreased DNA breakage. No genotoxicity was observed by BR($25{\sim}50{\mu}g/mL$) treatment alone on DNA breakage. Since BT can induce DNA damage through the generation of reactive oxygen species(ROS), we examined the level of ROS in human lymphocytes treated with BT and/or BR using DCF-DA, ROS-sensitive probe. The generation of ROS in BT-treated cells was also observed, and BR addition inhibited the level of BT-induced DNA damage. Conclusions : From above results it is suggested that BR could protect the cell and DNA from pro-oxidant effect by ROS by BT

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The induction of Micronucleus and Aneuploidy in human lymphocytes by Hydroquinone and its association with Genetic Polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene

  • Chung, Eun-Jung;Kim, Tae-Yon;Kim, Jin-Sik;Kim, Yang-Ji;Cho, Yoon-Hee;Chung, Hai-Won
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.176-176
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    • 2002
  • In order to investigate whether the induction of micronucleus and aneuploidy in human lymphocytes by Hydroquinone (HQ) is associated with genetic polymorphisms of CYP1A1, GSTM1, GSTT1, NQO1 gene, the cytokinesis-block micronucleus (CBMN) assay in combination with fluorescence in situ hybridization (FISH) technique using specific centromeric probes for chromosome 7 and 8 and PCR-RFLP based genotyping for 30 healthy people were performed.(omitted)

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Computerized Image Analysis of Micronucleated Reticulocytes in Mouse Bone Marrow (컴퓨터 이미지 분석법을 이용한 마우스 골수세포에서 소핵의 계수)

  • 권정;홍미영;고우석;정문구;이미가엘
    • Toxicological Research
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    • v.18 no.4
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    • pp.369-374
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    • 2002
  • The present study was performed to validate an automated image analysis system (Loats Automated Micronucleus Scoring System) for the mouse bone marrow micronucleus assay, comparing with conventional microscopic scoring. Two studies were conducted to provide slides for a comparison of micro-nucleated polychromatic erythrocytes (MNPCEs) values collected manually to those collected by the auto-mated system. Test article A was used as an example of a compound negative for the induction of micronuclei and test article B was wed as a micronucleus-inducing agent to elicit a positive response. Cyclophosphamide was included to provide an positive control in two studies. Bone marrow samples were collected 24 h after administration of test article A and B in male ICR mice. The cells were fixed with absolute methanol and stained with May-Grunwald and Giemsa. The number of MNPCEs was determined by the analysis of 1000 total PCEs per bone marrow sample. In addition to micronucleus scoring, an index of bone marrow toxicity based on PCE ratio (% of PCEs to total erythrocytes) was determined for each sample. The automated and manual scoring was similar when the MNPCEs incidence induced by each test article was less than 10. However manual scoring was able to effectively enumerate micronucleated PCEs in mouse bone marrow when MNPCEs incidence was more than 10, such as cyclophosphamide treatment. Conversely, PCE ratio was superior in computer-assisted image analysis. Taken together, it is suggested that improvement of the automated image analysis may be necessary to render the automatic scoring as sensitive as manual scoring for routine counting of micronuclei, especially because it is superior in objectivity and high throughput scoring.

Radiation exposure dose in human blood lymphocytes as assessed by the CBMN assay

  • Ryu, Tae Ho;Kim, Jin-Hong;Kim, Jin Kyu
    • Journal of Ecology and Environment
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    • v.37 no.4
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    • pp.195-200
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    • 2014
  • The chances of accidental exposure are augmented as the application of ionizing radiation increases in various fields. Such accidental exposures may occur at nuclear power plants, laboratories, and hospitals. Cytogenetic assays have been used for estimating radiation dose in the situation of the accidents. The micronucleus assay has several advantages over the other cytogenetic methods as it is simple and fast. The present study aimed at investigation of the micronuclei frequencies in cytokinesis-block cells in human blood lymphocytes after ${\gamma}$-irradiation and at establishment of a standard dose response relationship. The samples of peripheral blood were obtained from 6 different donors aged between 24 and 30 years old. The bloods were irradiated in vitro with 0-5 Gy. A linear quadratic dose-response equation was obtained by scoring the micronuclei in binucleated cells; $y=27.87x^2+46.13x+2.08$ ($r^2=0.99$). Irradiation caused a significant decrease in the nuclear division index. Necrotic and apoptotic cells increased in number after irradiation in a dose-dependent manner. In conclusion, the conventional cytokinesis-block micronucleus assay has proven to be the great technique in biological dosimetry. Dose-response calibration curve derived from CMBN assay could be used to estimate the exposure dose during a radiological emergency.