• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제37권5호
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• pp.415-420
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• 2011

Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.

• 한국어병학회지
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• 제22권3호
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• pp.375-382
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• 2009

목적 유전자를 숙주 세포의 게놈에 삽입하거나 발현하는데 있어서, retrovirus 매개의 유전자 전달 시스템을 사용하게 되면, 복잡하고 힘든 절차를 거치게 된다. 본 연구에서는 목적 유전자의 BF-2 세포 게놈 내 삽입을 하기 위해, UV로 불활화한 어류 레트로바이러스인 SnRV를 사용한 간단한 방법에 대해 조사하였다. 우선, BF-2 세포를 사용한 transfection을 위해 Lipofectamine 2000과 Transome을 사용하여 최적 조건을 결정하였다. 0.5 $\mu\ell$ Lipofectamine 2000을 사용한 경우 0.5, 1 그리고 2 $\mu{g}$ DNA 사용에 대해 33.8, 40.6 그리고 40.2%의 transfection 효율을 보인 동시에 최소 80 % 이상의 높은 세포 생존율을 나타낸 반면, Transome을 사용한 transfection 효율은 모두 5% 이하였다. UV 처리 시간에 있어서는 5분간의 UV 처리로 SnRV의 감염성이 불활성화되는 것을 확인하였다. 다음으로 GFP 유전자의 양측에 SnRV에서 유래된 LTR 서열을 접하고 있는 cassette를 구축한 뒤 BF-2 세포에 transfection 하고, cassette 유전자의 삽입과 발현을 위해 UV로 불활화한 SnRV를 처리하였다. 그 결과 UV로 불활화한 SnRV를 1회 처리 또는 SnRV 무처리 BF-2 세포에서는 형광이 관찰되지 않았던 반면, 3회와 5회 처리한 BF-2 세포에서 형광발현이 확인되었다. 이러한 결과로, GFP 유전자가 불활화한 SnRV를 이용하여 BF-2 세포 게놈에 삽입되는 것을 확인하였다.

• Journal of Animal Science and Technology
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• 제45권1호
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• pp.1-12
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• 2003

본 연구에서는 국내 고유 재래 가축들의 유전정보와 유전학적 개량의 기초자료 제공을 목적으로 고분염 분석(high-resolution banding) 방법에 의한 한국재래계의 염색체 분염 표지를 제시하였다. 본 연구에 공시된 한국재래계로서는 축산기술연구소에서 계통화 시킨 황갈색 및 적갈색 계통으로 이들이 생산한 수정란의 초기 배자를 이용하여 염색체 분석을 수행하였으며 닭의 초기배자에 EtBr 및 colchicine을 처리함으로써 보다 양호한 고정도 염색체를 획득하였다. 한국재래계의 GTG-banding 결과 모든 상동염색체간에 뚜렷하고 특징적인 band 양상을 얻을 수 있었으며, Leghorn 및 국제표준핵형(ISSAK)과 비교시 염색체의 형태적 양상에서는 거의 차이가 없는 것으로 분석되었고 대표적 landmark간에도 거의 일치되는 양상을 보였다 그러나 대부분의 한국재래계의 대형염색체에서 더 많은 G-band의 분리 양상을 보이고 특히 1번 및 Z 염색체에서 특징적 분리 양상의 차이를 보였다. 한국재래계의 C-banding 분석에서는 세포별 heterochromatin의 다형성을 보이기는 하나 대부분의 염색체의 동원체와 말단부위에서 C- band가 나타났으며, Z 염색체 장완 말단부와 W 염색체 전체에서는 거의 모든 세포에서 C-band가 출현하였다. 또한 3번 염색체 동원체와 Z 염색체 장완 말단부에서 특징적 다형성을 나타내어 이들 염색체들에서는 상동염색체간 heterochromatin의 이형적 양상(heteromorphic)이 존재함을 밝혔다.

• Journal of Preventive Medicine and Public Health
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• 제19권2호
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• pp.244-251
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• 1986

항암제 mitomycin C가 정상인 배양 임파구의 SCE에 미치는 영향을 알아보기 위해 세포분열지수, 자매염색분체빈도, 세포분열주기 변동 및 염색체군별 SCE 빈도를 조사한 결과는 다음과 같이 요약할 수 있다. 1) 세포당 SCE빈도는 항암제 mitomycin C의 최저농도인 $6.25{\pm}{\times}10^{-9}$ M에서는 $13.1{\pm}2.8$이었고, 최고 농도인 $1.00{\times}10^{-7}$에서는 $75.8{\pm}8.2$로써 농도 증가에 따라 대수증가를 보였다. 그리고 세포분열지수는 완만한 감소경향을 나타냈다. 2) 세포분열주기 변동은 항암제 mitomycin C의 농도 $2.50{\times}10^{-8}M$에서 대조군에 비해 유의한 세포분열지연 현상을 보였으며 (p<0.05), $5{\times}10^{-8}M$ 이상에서는 세포분열지연이 매우 유의하게 나타났다(p<0.01). 3) 염색체군별 단위염색체당 SCE빈도는 항암제의 농도 증가에 따라 절대 빈도는 증가되었다 할지라도 A군에서 G군으로 갈수록 감소되어 특정염색체군과 SCE와의 관계는 보이지 않았다.

• Toxicological Research
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• 제21권2호
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• pp.99-105
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• 2005

Photo-mutagenic compounds have been known to alter skin cancer rates by acting as initiators or by affecting subsequent steps in carcinogenesis. The objectives of this study are to investigate the utility of photo-chromosomal aberration (photo-CA) assay for detecting photo-clastogens, and to evaluate its ability to predict rodent photocarcinogenicity. Photo-CA assay was performed with five test substances that demonstrated positive results in photo-carcinogenicity tests: 8-Methoxypsoralen (photoactive substance that forms DNA adducts in the presence of ultraviolet A irradiation), chlorpromazine (an aliphatic phenothiazine an alpha-adrenergic blocking agent), lomefloxacin (an antibiotic in a class of drugs called fluoroquinolones), anthracene (a tricyclic aromatic hydrocarbon a basic substance for production of anthraquinone, dyes, pigments, insecticides, wood preservatives and coating materials) and Retinoic acid (a retinoid compound closely related to vitamin A). For the best discrimination between the test substance-mediated genotoxicity and the undesirable genotoxicity caused by direct DNA absorption, a UV dose-response of the cells in the absence of the test substances was firstly analyzed. All 5 test substances showed a positive outcome in photo-CA assay, indicating that the photo-CA test is very sensitive to the photo-genotoxic effect of UV irradiation. With this limited data-set, an investigation into the predictive value of this photo-CA test for determining the photo-carcinogenicity showed that photo-CA assay has the high ability of a test to predict carcinogenicity. Therefore, the photo-CA test using mammalian cells seems to be a sensitive method to evaluate the photo-carcinogenic potential of new compounds.

• 한국발생생물학회지:발생과생식
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• 제18권2호
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• pp.117-125
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• 2014

Vitrification method is widely used in oocyte cryopreservation for IVF but the birth rates are lower than that of the fresh oocyte. One of the known main reasons is structural instability of meiotic spindle and chromosome systems of mature oocyte. To get the best way for keeping competence of matured oocytes, we studied the best conditions for vitrification focused on equilibration times. The mature oocytes were underwent vitrification with current popular method and analyzed the survival rates, microtubule stability and DNA integrity. The survival rates of recovered oocyte are almost same between groups and are more than 93%. The structural configuration of meiotic spindle was well kept in 10 min equilibration group and the stability rate was almost same with that of control. The chromosomal breakdown was observed in all experimental groups, but the chromosomal stability was higher in 10 min equilibration group than the other groups. The 10 min equilibration group showed best condition compared with the other groups. Based on these results, the equilibration time is one of the key factors in successful keeping for competence of mature oocyte. Although, more fine analysis about the effects of physical stress on oocyte during vitrification is needed to define the optimal condition, it is suggested that the optimal equilibration time to get competent oocyte in mouse is 10 min. Information acquired this study may provide insight into intracellular structural events occurring in human oocytes after vitrification and application for cryopreservation of human oocyte.

• 한국산림과학회지
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• 제74권1호
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• pp.56-60
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• 1986

GD 기본(基本) 배지(培地)에 서로 다른 생장조절물질(生長調節物質)을 첨가(添加)한 4 가지 배지(培地)에 잣나무 배(胚)에서 유래(由來)한 Callus를 1년(年) 6개월간(個月間) 배양(培養)시켜 염색체수(梁色體數) 변이(雙異)를 조사(調査)하였다. 그 결과(結果) $0.1mg/{\ell}$와 NAA $0.1mg/{\ell}$에 2, 4-D $0.1mg/{\ell}$를 첨가(添加)한 배지(培地) 즉(卽) Auun 단독(單獨) 및 배합(培合) 처리(處理)한 배지(培地)에서는 2n 비율(比率)이 각각(各各) 29%와 17%로 낮게 나타났다. 그러나 Auxin 과 Cytokinin 을 종합(複合) 처리(處理)한 배지(培地) 즉(卽) 2, 4-D $0.1mg/{\ell}$와 BAP $0.1mg/{\ell}$에서는 52 %, 2, 4-D $0.1mg/{\ell}$와 Kinetin $0.1mg/{\ell}$에서는 36 %로써 비교적(比較的) 높은 빈도(頻度)로 2n 세포(細胞)가 나타났다. 2n 세포(細胞)의 빈도(頻度)는 기관분화(器官分化)와 상관(相關)이 높은 것으로 생각 할 수 있었다.

• 한국환경성돌연변이발암원학회지
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• 제21권1호
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• pp.14-22
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• 2001

The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. In this respect, administrative authorities has great concern to regulate and to evaluate the chemical hazard to environment and human health. The clastogenicity of 28 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. Glycidylacrylate which is one of the most cytotoxic chemical among 28 chemicals tested revealed clastogenicity in the range of 0.31-1.25 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. Neopentyl glycol (340-1360 $\mu\textrm{g}$/$m\ell$) also revealed weak positive result both in the presence and absence of metabolic activation system. Cyanoguanidine (/$420.5-841 $\mu\textrm{g}$m\ell$) and N-butylchloride ($231.5-926 $\mu\textrm{g}$/m\ell$) revealed weak positive result only in the absence of S-9 metabolic activation system. Nevertheless total aberration percentages of N-butylchloride in the presence of metabolic activation system, and 3,4＇-dichlorobenztrifluoride in the absence of S-9 metabolic activation revealed above 5% aberration, there is no statistical significance. From the results of chromosomal aberration assay with 28 synthetic chemicals in Chinese hamster lung cells, glycidylacrylate (CAS No. 106-90-0), neopentyl glycol (CAS No. 126-30-7), N-butyl chloride (CAS No. 109-69-3) and cyanoguanidine (CAS No. 461-58-5) revealed positive clastogenic results in this study.

• Toxicological Research
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• 제14권3호
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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.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 춘계학술대회
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• pp.183-184
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• 2003

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.