• 대한수의학회지
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• 제38권3호
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• pp.606-613
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• 1998

Cadmium is one of the well-known environmental toxicants and induces cancer in rodents and human, but its carcinogenic mechanism has not been well demonstrated until now. Genotoxic effects of cadmium in Salmonella typhimurium TA98, TA100 and TA1535/pSK1002 or in WB-F344 rat liver epithelial cells were investigated to elucidate the tumor initiating effects of cadmium. TA98, TA100 and TA1535/pSK1002 tester strains were used to detect frameshift mutation, base-pair mutation and SOS repair response, respectively, in Salmonella mutation test. Reverse mutations from histidine to $histidin^+$ of Salmonella typhimurium TA98 and TA100 by $CdCl_2$ were not significantly different from control up to the maximum doses ($100{\mu}M$ and $200{\mu}M$ in TA98 and TA100, respectively) at which non-cytotoxicity was observed. DNA SOS repair responses(${\beta}$-galactosidase activity) generally did not show significant increases compared to control in both of the conditions with or without metabolic activation in Salmonella typhimurium TA1535/pSK1002 by $CdCl_2$. But the activities of ${\beta}$-galactosidase by $400{\mu}M$ of $CdCl_2$ in metabolic activation condition and by 130 and $400{\mu}M$ of $CdCl_2$ in non-metabolic activation condition were more decreased than those of control. DNA single strand breaks for 4hrs were observed only in WB-F344 rat liver epithelial cells treated with $200{\mu}M$ of $CdCl_2$. As a conclusion, $CdCl_2$ did not induce gene mutation in microbials but induce DNA single strand breaks in rat liver epithelial cells.

• Molecular & Cellular Toxicology
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• 제1권4호
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• pp.230-236
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• 2005

Formaldehyde is a common environmental contaminant found in tobacco smoke, paint, garments, diesel and exhaust, and medical and industrial products. Formaldehyde has been considered to be potentially carcinogenic, making it a subject of major environmental concern. However, only a little information on the mechanism of immunological sensitization and asthma by this compound has been known. So, we performed with Jurkat cell line, a human T lymphocyte, to assess the induction of DNA damage and to identify the DEGs related to immune response or toxicity by formaldehyde. In this study, we investigated the induction of DNA single strand breaks by formaldehyde using single cell gel electrophoresis assay (comet assay). And we compared gene expression between control and formaldehyde treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity ($IC_{30}$) of formaldehyde was determined above the 0.65 mM in Jurkat cell in 48 h treatment. Based on the $IC_{30}$ value from cytotoxicity test, we performed the comet assay in this concentration. From these results, 0.65 mM of formaldehyde was not revealed significant DNA damages in the absence of S-9 metabolic activation system. And the one differentially expressed gene (DEG) of formaldehyde was identified to zinc finger protein 292 using $GeneFishing^{TM}$ method. Through further investigation, we will identify more meaningful and useful DEGs on formaldehyde, and then can get the information on the associated mechanism and pathway with immune response or other toxicity by formaldehyde exposure.

• Journal of Periodontal and Implant Science
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• 제42권5호
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• pp.151-157
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• 2012

Purpose: Cyclooxygenase (COX) enzyme catalyzes the production of prostaglandins, which are important mediators of tissue destruction in periodontitis. Single nucleotide polymorphisms of $COX_2$ enzyme have been associated with increasing susceptibility to inflammatory diseases. The present study evaluates the association of two single nucleotide polymorphisms in $COX_2$ gene (-1195G>A and $8_{473}$C>T) with chronic periodontitis in North Indians. Methods: Both SNPs and their haplotypes were used to explore the associations between $COX_2$ polymorphisms and chronic periodontitis in 56 patients and 60 controls. Genotyping was done by polymerase chain reaction followed by restriction fragment length polymorphism. Chi-square test and logistic regression analysis were performed for association analysis. Results: By the individual genotype analysis, mutant genotypes (GA and AA) of $COX_2$-1195 showed more than a two fold risk (odds ratio [OR]>2) and $COX_2$ $8_{473}$ (TC and CC) showed a reduced risk for the disease, but the findings were not statistically significant. Haplotype analysis showed that the frequency of the haplotype AT was higher in the case group and a significant association was found for haplotype AT (OR, 1.79; 95% confidence interval, 1.03 to 3.11; P=0.0370) indicating an association between the AT haplotype of $COX_2$ gene SNPs and chronic periodontitis. Conclusions: Individual genotypes of both the SNPs were not associated while haplotype AT was found to be associated with chronic periodontitis in North Indians.

• 한국식품위생안전성학회지
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• 제29권4호
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• pp.383-390
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• 2014

The safety of a new natural plant composition (ADP) was assessed on the genotoxicity study and 14-day repeat dose toxicity study. ADP contains a mixed water extract obtained from the mixture of Phellodendron cortex (Phellodendron amurense) and Anemarrhena rhizoma (Anemarrhena asphodeloides), and poses the contractile properties mediated by alpha-adrenoceptor of the prostate and urethra as well as antioxidant and anti-inflammatory properties. In order to evaluate genetic safety, in vivo micronucleus test was performed in ICR mice orally administered with three dose levels of 1250, 2500, 5000 mg/kg body weight, and vehicle and positive control. In the 14 days study, Sprague-Dawley rats were treated with ADP at the dose levels of 500, 1000, 2000 mg/kg once a day, and clinical signs, body weights, hematology, serum biochemistry, necropsy findings and organ weights were monitored and examined. In experimental results, ADP treatment, compared with vehicle control, did not induce the micronucleated erythrocytes from mouse bone marrow. In the 14 days study, any significant and toxicological differences in all measurements of parameters were not observed in ADP treatment groups of animals, compared with vehicle treatment. The No-Observed-Adverse-Effect-Level (NOAEL) of ADP in the 14 days study was determined to be greater than 2000 mg/kg/day in both sexes.

• Restorative Dentistry and Endodontics
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• 제31권1호
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• pp.36-49
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• 2006

본 연구의 목적은 기존 상용화된 근관 봉함재인 레진계 봉함재 (AH 26, AH Plus), 산화 아연 유지놀계 봉함재(Tubliseal EWT Pulp Canal Sealer EWT), 수산화 칼슘계 봉함재 (Sealapex), 기존의 인산삼칼슘계 봉함재(Sankin Apatite type I, II, III)와 새로이 개발된 인산칼슘계 근관 봉함재 (CAPSEAL I, CAPSEAL II)의 세포독성과 유전독성을 비교 평가하고자 하였다. MTT test를 통해 세포독성을 평가하였으며 미생물을 이용한 복귀돌연변이 시험 (Ames test)으로 유전독성을 평가하였다. 이 연구의 결과는 아래와 같다. 1. 즉시군이 24시간군에 비해 MTT assay에서 세포독성이 높게 나타났다 (p<0.001). 2. 즉시군에서 CAPSEAL I과 CAPSEAL II는 AH 26, AH Plus, Tubliseal EWT Pulp Canal Sealer EWT, Sealapex와 SARCS II 보다 낮은 세포독성을 보였다 (p<0.01). 3. 즉시군에서 AH 26은 TA98과 TA100에 각각 S9 fraction을 처리한 경우와 그렇지 않은 경우 모두 유전독성을 나타냈으며, AH Plus 또한 TA100에 S9 fraction을 처리한 경우와 그렇지 않은 경우 유전독성을 나타냈다. 4. 즉시군에서 Tubliseal EWT, Pulp Canal Sealer EWT Sealapex가 TA100균주에 S9 fraction을 처리하였을 때 유전독성 양성반응이 나타났으며, 그 외의 경우는 모두 음성반응을 나타냈다. 5. 24시간군에서는 SARCS II가 TA98균주에서 S9 fraction처리했을 때와 처리하지 않았을 때 모두 유전독성이 나타났고, AH 26은 TA98에 S9 fraction을 처리하였을 때 유전독성이 나타났다 그 외의 경우는 모두 음성반응을 보였다. 6. CAPSEAL I과 CAPSEAL II는 유전독성에서 모두 음성반응을 나타냈다. 7. CAPSEAL I과 CAPSEAL II 두 근관봉함재 간에는 세포독성실험과 유전독성실험에서 즉시군과 24시간군 모두에서 통계학적으로 유의할 만한 차이를 보이지 않았다.

• Toxicological Research
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• 제21권1호
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• pp.57-62
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• 2005

To investigate the mutant induction of wild ginseng culture extract, we performed chromosomal aberration assay with chinese hamster lung cell in vitro. The test concentration of the extract was decided for the standard with the 50% suppression of cell propagation in the cell. The concentrations for the chromosome test were 1,250, 2,500 and 5,000 ㎍/ml with metabolic activation (+S, 6 hours treatment), 1,100, 2,200 and 4,400 ㎍/ml without metabolic activation (-S, 6 hours treatment) 800, 1,600 and 3,200 ㎍/ml without metabolic activation (-S, 24 hours treatment). No significant increase in chromosome aberrations was observed at any of these concentrations both in the absence and presence of metabolic activation system. Cyclophosphamide monohydrate (CPA) and ethylmethanesulfonate (EMS) caused a significant increase in chromosome aberration. These results may be concluded that wild ginseng culture extract is not capable of inducing chromosome aberration in cultured chinese hamster lung cell regardless of metabolic activation and genotoxicity of that is negative under the present experimental condition.

• 한국식품위생안전성학회지
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• 제26권3호
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• pp.242-247
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• 2011

밤, 고구마, 우엉 등의 일부 식물체를 오랜 시간 보관하거나 가공하는 과정에서 표면에 녹변화합물이 형성된다. 본 연구에서는 이때 형성되는 녹변화합물이 유전독성 가능성을 내포한 novel trihydroxy acridine 유도체로서, 녹변화합물에 대한 복귀돌연변이시험을 수행하였다. 시험은 Salmonella typhimurium TA98, TA100 균주를 이용하였고 대사활성계 적용 유무에 따른 시험을 모두 수행하였다. 시험에 사용된 녹변화합물은 알라닌, 아르기닌, 아스파트산, 글라이신, 라이신, 페닐알라닌과 클로로겐산을 사용한 합성 및 고구마를 매트릭스로 하여 녹변화합물을 형성한 후 추출하는 두가지 방법을 통하여 준비하였다. 합성 녹변화합물과 추출 녹변화합물의 최대 처리 용량은 생육저해시험과 용해도, 흡광도를 고려 하여 각각 2000 ${\mu}g$/lplate, 50 ${\mu}g$plate로 결정 하였다. 녹변화합물 합성 과정 후 잔존하는 아미노산들과 클로로겐산의 영향을 배제하고자 이들을 또 다른 시험군으로 설정하고 각각 250 ${\mu}g$/plate, 1750 ${\mu}g$/plate의 농도로 처리하여 복귀돌연변이시험을 수행하였다. 시험결과 두 균주에서 농도에 의존적으로 콜로니 수가 증가하는 경향은 관찰되지 않았으며, 음성대조군 수치와 비교하여 두 배 이상의 콜로니 수도 나타나지 않았다. 이상의 결과를 종합할 때, 합성 녹변화합물과 추출 녹변화합물은 본 시험 조건에서 복귀돌연변이를 유발하지 않는 것으로 확인되었다.

• 한국환경성돌연변이발암원학회지
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• 제17권2호
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• pp.71-77
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• 1997

The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

• 한국환경성돌연변이발암원학회지
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• 제18권2호
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• pp.129-134
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• 1998

To evaluate the genotoxicity of CKD-602, an anticancer agent the in viかo reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the reverse mutation assay, CKD-602 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, and TA 1537 strains with and without metabolic activation. In the chromosome aberration test using CHL cells, there was an increased incidence of structural aberrations induced by CKD-602 without metabolic activation during 24 and 48 hours, but CKD-602 did not induce chromosome aberration with metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice. At 24 hours after treatment with CED-602 by i.p. once, there was an increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice.

• 한국환경성돌연변이발암원학회지
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• 제18권2호
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• pp.123-128
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• 1998

To evaluate the genotoxicity of SKP-450, an antihypertensive agent the in vitro reverse mutation assay using Salmonella typhimurium, the Chromosome aberration assay using Chinese hamster lung (CHL) cells and the in vivo micronucleus assay using bone marrow cells of ddY mice were performed. In the Reverse mutation test, SKP-450 did not induced mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 with and without metabolic activation. In the chromosome aberration assay using CHL cells, there was no increased incidence of structural and numerical aberrations with and without metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of male ddY mice at 30 hours after treatment with SKP-450 by p.o once. The results showed no increased incidence of micronucleated polychromatic erythrocytes in bone marrow of ddY male mice treated with SKP-450.