• Title/Summary/Keyword: micronucleus assay

검색결과 191건 처리시간 0.029초

Response of Odontoblast to the Bio-Calcium Phosphate Cement

  • Kim, Jin-Woo;Kim, Sung-Won;Kim, Gyoo-Cheon;Kim, Yong-Deok;Kim, Cheol-Hun;Kim, Bok-Joo;Kim, Uk-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제33권4호
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    • pp.301-307
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    • 2011
  • Purpose: If the tooth structure is damaged, then it is impossible to regenerate the tooth. The materials used to restore the tooth structure are not related to the composition of the tooth. The materials used to restore the structure can't replace the natural tooth because they just fill the defective structure. Calcium phosphate cement remineralizes the dentin and almost replaces the natural tooth, but there are some disadvantages. We conducted basic tests with Biomimetic CPC (Bio-CPC) to make sure of the possibility of the biomaterial to remineralize the defective tooth structure. Methods: In this study, the bioactivity and biocompatibility of Bio-CPC were evaluated for its potential value as the bio-material for regeneration of damaged tooth structure by conducting a cell toxicity assay (WST-1 assay), a cytokinesis-block micronucleus assay, a chromosomal aberration test, total RNA extraction and RT-PCR on MDPC-23 mouse odontoblast-like cells. Results: The in vitro cytotoxicity test showed that the Bio-CPC was fairly cytocompatible for the MDPC-23 mouse odontoblast-like cells. Conclusion: Bio-CPC has a possibility to be a new biomaterial and further study of Bio-CPC is needed.

Genotoxicity of Taxol and 10-Deacetyl Baccatin III Using Single Cell Gel Electrophoresis (Comet Assay) in Chinese Hamster Lung Fibroblast

  • Kim, Hyun-Joo;Kim, Kyung-Ran;Youn, Ji-Youn;Kim, Min-Hee;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
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    • pp.61-61
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    • 1996
  • Taxol is used as cancer therapeutic agent. It has been known as weak posotive of chromosome aberration assay in vitro in our previous results (Ryu et al., 1996) and potent clastogens in the mouse bone marrow micronucleus (Tinwell and Ashby, 1994). We performed microgel electrophoresis to determine the effect of taxol and it's precursor 10-deacetyl baccatin III(DAB) on DNA. Microgel electrophoresis is useful, rapid, simple, visual, and sensitive technique for measuring DNA breakage and repair mechanisms in mammalian 근ells. The range of concentration used for taxol were 854, 427, 213.5, 106.8, 53.4 Ug/ml, for DAB 910 ,455, 227.5 U9/ml, Cell viability always exceed 85%. We analyzed the results by using the special software of image analyzer for this comet assay (Komet 3.0). By using this image analyzer software , we can get the result as the tail moment ((mean of tail length - mean of head lengh) x tail%DNA/100). A slight increase in DNA migration was observed for taxol at the concentration of 854 Ug/m4 in the absence of S9 mixture. No increased DNA migration was observed after treatment with DAB.

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Genetic Toxicity Test of Glycidol by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ji-H.;Kim, Ki-Y.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Sheen, Yhun-Y.
    • Biomolecules & Therapeutics
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    • 제14권4호
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    • pp.240-245
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    • 2006
  • The primary use for glycidol is as a stabilizer in the manufacture of vinylpolymers, however, it is also used as an intermediate in the production of pharmaceuticals, as an additives for oil and synthetic hydraulic fluids, and as a diluting agent is same epoxy resins. In this study, we have carried out in vitro genetic toxicity test of glycidol and microarray analysis of differentially expressed genes in response to glycidol. The result of Ames test showed mutations with glycidol treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, glycidol showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with glycidol treatment showed DNA damage both with and without exogenous metabolic activation. Glycidol increased micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to glycidol by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of glycidol.

발효 탐라오가피 추출물의 유전독성 연구 (Genotoxicity Study from the Extracts of Fermented Acanthopanax koreanum)

  • 조명래;김영현;김재민;이상종;신현무;이옥환
    • 한국식품위생안전성학회지
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    • 제31권2호
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    • pp.107-112
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    • 2016
  • 본 연구는 발효 탐라오가피(fermented A. koreanum) 추출물의 유전독성을 연구하기 위하여, 미생물복귀돌연변이 시험, 마우스 골수세포를 이용한 소핵시험, 염색체 이상시험을 연구하였다. 미생물복귀돌연변이 연구에서 발효 탐라오가피 추출물은 Salmonella typhimurium TA98, TA100, TA1535, TA1537와 Escherichia coli WP2uvrA에 대하여 대사활성계의 존재(+S-9 Mix) 및 부재(-S-9 Mix) 하에서 돌연변이 유도를 보이지 않았다. 또한, ICR 마우스를 이용한 소핵실험에서 발효 탐라오가피 추출물은 500, 1,000, 2,000 mg/kg 농도에서 MNPCE/2,000 PCE 와 PCE/200 RBC의 소핵형성을 유발하지 않았다. 한편, CHO-K1 세포를 이용한 염색체 이상실험에서 발효 탐라오가피 추출물은 대사활성계의 존재 6시간 처리군, 대사활성계 부재 6시간 처리군 및 대사활성계 부재 24시간 처리군에서 염색체 이상을 보이지 않았다. 따라서, 본 연구결과 발효 탐라 오가피 추출물은 유전독성을 나타내지 않음을 알 수 있었다.

Inhalation Toxicity of Particulate Matters Doped with Arsenic Induced Genotoxicity and Altered Akt Signaling Pathway in Lungs of Mice

  • Park, Jin-Hong;Kwon, Jung-Taek;Minai-Teherani, Arassh;Hwang, Soon-Kyung;Chang, Seung-Hee;Lim, Hwang-Tae;Cho, Hyun-Seon;Cho, Myung-Haing
    • Toxicological Research
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    • 제26권4호
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    • pp.261-266
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    • 2010
  • In the workplace, the arsenic is used in the semiconductor production and the manufacturing of pigments, glass, pesticides and fungicides. Therefore, workers may be exposed to airborne arsenic during its use in manufacturing. The purpose of this study was to evaluate the potential toxicity of particulate matters (PMs) doped with arsenic (PMs-Arsenic) using a rodent model and to compare the genotoxicity in various concentrations and to examine the role of PMs-Arsenic in the induction of signaling pathway in the lung. Mice were exposed to PMs $124.4{\pm}24.5\;{\mu}g/m^3$ (low concentration), $220.2{\pm}34.5\;{\mu}g/m^3$ (middle concentration), $426.4{\pm}40.3\;{\mu}g/m^3$ (high concentration) doped with arsenic $1.4\;{\mu}g/m^3$ (Low concentration), $2.5\;{\mu}g/m^3$ (middle concentration), $5.7\;{\mu}g/m^3$ (high concentration) for 4 wks (6 h/d, 5 d/wk), respectively in the whole-body inhalation exposure chambers. To determine the level of genotoxicity, Chromosomal aberration (CA) assay in splenic lymphocytes and Supravital micronucleus (SMN) assay were performed. Then, signal pathway in the lung was analyzed. In the genotoxicity experiments, the increases of aberrant cells were concentration-dependent. Also, PMs-arsenic caused peripheral blood micronucleus frequency at high concentration. The inhalation of PMs-Arsenic increased an expression of phosphorylated Akt (p-Akt: protein kinase B) and phpsphorylated mammalian target of rapamycin (p-mTOR) at high concentration group. Taken together, inhaled PMs-Arsenic caused genotoxicity and altered Akt signaling pathway in the lung. Therefore, the inhalation of PMs-Arsenic needs for a careful risk assessment in the workplace.

Peripheral Blood Lymphocytes as In Vitro Model to Evaluate Genomic Instability Caused by Low Dose Radiation

  • Tewari, Shikha;Khan, Kainat;Husain, Nuzhat;Rastogi, Madhup;Mishra, Surendra P;Srivastav, Anoop K
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권4호
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    • pp.1773-1777
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    • 2016
  • Diagnostic and therapeutic radiation fields are planned so as to reduce side-effects while maximising the dose to site but effects on healthy tissues are inevitable. Radiation causes strand breaks in DNA of exposed cells which can lead to chromosomal aberrations and cause malfunction and cell death. Several researchers have highlighted the damaging effects of high dose radiation but still there is a lacuna in identifying damage due to low dose radiation used for diagnostic purposes. Blood is an easy resource to study genotoxicity and to estimate the effects of radiation. The micronucleus assay and chromosomal aberration can indicate genetic damage and our present aim was to establish these with lymphocytes in an in vitro model to predict the immediate effects low dose radiation. Blood was collected from healthy individuals and divided into 6 groups with increasing radiation dose i.e., 0Gy, 0.10Gy, 0.25Gy, 0.50Gy, 1Gy and 2Gy. The samples were irradiated in duplicates using a LINAC in the radiation oncology department. Standard protocols were applied for chromosomal aberration and micronucleus assays. Metaphases were stained in Giemsa and 200 were scored per sample for the detection of dicentric or acentric forms. For micronuclei detection, 200 metaphases. Giemsa stained binucleate cells per sample were analysed for any abnormality. The micronuclei (MN) frequency was increased in cells exposed to the entire range of doses (0.1-2Gy) delivered. Controls showed minimal MN formation ($2.0%{\pm}0.05$) with triple MN ($5.6%{\pm}2.0$) frequency at the lowest dose. MN formation increased exponentially with the radiation dose thereafter with a maximum at 2Gy. Significantly elevated numbers of dicentric chromosomes were also observed, even at doses of 0.1-0.5Gy, compared to controls, and acentric chromosomes were apparent at 2Gy. In conclusion we can state that lymphocytes can be effectively used to study direct effect of low dose radiation.

자주달개비 분석법을 이용한 카펫 방출 휘발성 유기화합물의 생물학적 영향 평가 (Biological Effects of Volatile Organic Compounds from Carpet Materials as Assessed by the Tradescantia Assay)

  • 김진규;신해식;이영엽;이진홍
    • 환경생물
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    • 제25권3호
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    • pp.191-196
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    • 2007
  • 실내 공기는 대기와는 달리 실내 건축 자재에서 유래된 물질로 오염될 수 있다. 본 연구는 실내자재인 카펫에서 방출되는 휘발성 유기화합물의 생물학적 영향을 평가하기 위하여 수행되었다. 카펫과 자주달개비 BNL 4430 꽃차례를 환경노출시험용기에 넣고 일정시간 노출을 실시하였고 흡착관의 VOCs에 대한 화학분석을 실시하였다. 화학분석결과 카펫에서는 12종의 VOCs가 방출되는 것이 확인되었으며 이중 스틸렌$(71.9{\mu}g\;m^{-3})$과 톨루엔$(49.6{\mu}g\;m^{-3})$의 농도가 높았다. 환경노출시험용기에서 카펫에서 방출되는 VOCs에 24시간 노출된 자주달개비 실험군의 미세핵 빈도는 100사분자 당 $7.73{\pm}0.75MCN$으로서 TO-14 표준혼합기체 1ppm에 4시간 노출된 실험군의 미세핵 빈도인 $7.31{\pm}0.70$와 유사한 높은 값을 나타내었다. 반면 표준혼합기체 1ppm에 2시간 노출된 실험군의 경우 미세핵 자연발생 빈도와 유사한 수준을 보였다. 이 같은 결과로부터 카펫에서 방출된 휘발성 유기화합물이 함유되어 있는 실내공기에 장시간 노출될 경우 생물유전독성이 유발된다는 것이 확인되었다. 본 연구에 적용한 생물-화학 병용분석 기법은 실내 공기오염의 생물학적 감시에 매우 효율적임이 입증되었다.

한국산 겨우살이 추출물의 안전성 평가 (Safety Evaluation of Korean Mistletoe Extract)

  • 김인보;정주성;윤택준;김종배
    • 한국식품영양학회지
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    • 제26권3호
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    • pp.383-390
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    • 2013
  • 본 연구에서는 겨우살이 열수 추출물인 미슬로 C의 안전성을 검토하고자 유전 독성 및 실험동물을 이용한 안전성 검사를 실시하였다. 미슬로 C의 미생물 돌연변이 실험을 S. typhimurium의 히스티딘 요구성 균주와 E. coli의 트립토판 요구성 균주를 이용하여 대사 활성계 적용 및 비적용 하에서 복귀돌연변이 시험을 실시한 바, $5,000{\mu}g/plate$의 처리 농도까지 복귀돌연변이 집락은 나타나지 않았다. ICR 마우스에게 500, 1,000 및 2,000 mg/kg를 경구 투여하고, 골수세포를 수집하여 소핵을 측정한 결과, 정상마우스의 경우와 비교하여 유의한 소핵은 관찰되지 않았기에 미슬로 C는 유전독성을 유발하지 않는 것으로 판단되었다. 식품의약안전청의 의약품 등의 독성시험기준에 따라 암 수 SD 계열의 랫드에 시험물질을 0, 500, 1,000 및 2,000 mg/kg/day의 용량으로 1회 경구 투여한 후, 14일간의 체중 변화 및 사망률을 조사한 결과, 대조군과 비교하여 유의한 체중 변화는 없었으며, $LD_{50}$은 2,000 mg/kg 이상인 것으로 사료된다. 또한 0, 250, 500 및 1,000 mg/kg/day의 용량으로 13주간 반복 투여하면서 실험동물의 일반증상, 체중변화, 혈액 및 혈액생화학적 변화, 부검소견, 조직학적인 변화를 관찰하였다. 시험기간 중 암 수 모든 군에서 시험물질 투여에 기인한 일반적인 증상 변화는 관찰되지 않았고, 시험물질의 반복 투여로 인한 사망 마우스 역시 관찰되지 않았다. 따라서 미슬로 C를 13주간의 랫드에 대한 13주 반복 경구 투여 결과, 무독성량은 최소한 1,000 mg/kg 이하인 결과를 나타냈으며, 이 농도에서 독성을 유발하는 표적장기는 관찰되지 않았다.

Genetic Toxicity Test of 8-Hydroxyquinoline by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Woo-Sun;Kim, Hyun-Joo;Lee, Eun-Mi;Kim, Joo-Hwan;Suh, Soo-Kyung;Kwon, Kyung-Jin;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-N.
    • Molecular & Cellular Toxicology
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    • 제3권2호
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    • pp.90-97
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    • 2007
  • 8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.

인체 말초 혈액 림프구에서 저선량 방사선 조사에 의해 유도되는 적응 반응 (Adaptive Response Induced by Low Dose Ionizing Radiation in Human Lymphocytes)

  • 김정희;이경종;조철구;류성렬;김태환;김성호
    • Journal of Radiation Protection and Research
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    • 제20권2호
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    • pp.97-102
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    • 1995
  • 인체 말초 혈액 림프구에서 저선량의 감마선에 의해 유도되는 적응 반응을 관찰하였다. 인체 림프구를 저선량의 감마선(priming dose 0.01Gy) 을 조사한 후 여러 시간 간격 후 고선량 (challenging dose, 1.5Gy) 을 조사하였다. 저선량을 미리 조사한 림프구와 조사하지 않은 림프구에서 발생된 미세핵의 빈도를 계수하였다. 비세핵 발생 빈도는 저선량 조사 4시간 후 고선량을 조사하였을 때 최대 감소치를 보였다. 저선량과 고선량 조사 시간차가 7시간 또는 20시간이었을 때 미세핵 발생 빈도는 약간 감소하였으나 유의성은 없었다. 본 연구에서는 $G_0$상태의 세포주기에서 저선량을 조사하였을 경우 적응 반응이 나타나는 것이 관찰 되었다. 미세핵 분석법은 실험 방법이 비교적 간단하고, 기타 염색체 분석법에 비하여 빠른 시간내 결과를 도출 할 수 있어 추후 방사선에 대한 적응 반응의 연구에 유용한 지표가 될 수 있을 것이다.

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