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석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),한주희 ( Ju Hee Han ),고애선 ( Ae Sun Koh ),박재학 ( Jae Hak Park ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
Zebrafish is proposed as an ideal vertebrate cancer model. Because zebrafish is easy to maintain and breed, and her fecundity is high enough (200~300 eggs per fish). Powerful genetics and cellular biology tools are also available in zebrafish. Even, histologically many organs are very similar with humans. We have raised over 1,000 fish for the transgenesis study through injections of various genes in a closed recirculating water system. In the end of the study, before euthanasia, we biopsied the negative control fish at 24 months of age. The histological analysis of one case of zebrafish tumorous change revealed biliary carcinoma, based upon the H&E staining. The local invasion, fibrosis, and increased mitotic index were observed in Laboratory maintained wild type zebrafish. This is the first documentation of the spontaneous biliary carcinoma in wild type zebrafish in Korea.
제브라피쉬 장내에서 Vancomycin-resistant enterococci 분리
석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),한주희 ( Ju Hee Han ),고애선 ( Ae Sun Kohv ),정우경 ( Woo Kyung Jung ),박재학 ( Jae Hak Park ) 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
The use of zebrafish embryos is not regulated by current Animal Protection law (full amendment, January 26, 2007) in Korea and therefore considered as a refinement, if not replacement of animal experiments because the complicated physiological environment is absent in vitro system. The skin ulcers and abdominal swelling were observed in Laboratory maintained zebrafish. Contents of abdominal swelling found at necropsy are peritoneal exudates. Vancomycin resistant enterococci (VRE) were detected for VanC-1 (Enterococcus gallinarum) by PCR on cultured colonies from the affected fish gut, exudates. In addition, unhatched eggs were observed in the abdominal exudates, based upon the H&E staining. This is the first documentation of the precise identification of VRE with VanC-1 gene in laboratory-maintained zebrafish. The pathogenesis to VRE infection in zebrafish was required in the following study.
Bovine Corneal Opacity Assay법을 이용한 안점막 자극 물질의 검증에 관한 연구
이현경 ( Hyun Kyoung Lee ),노경진 ( Kyoung Jin Noh ),석승혁 ( Seoung Hyeok Seok ),백민원 ( Min Won Baek ),이희영 ( Hui Young Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),김배환 ( Bae Hwan Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.2
Bovine corneal opacity and permeability assay have been proposed as reliable alternatives to the Draize test. In this study, we measured the opacity of the bovine cornea by quantifying the ability of light to pass through it using opacitometer. We obtained the regular value of basal opacity using bovine cornea collected within three hours after killing. For opacity alone, values obtained for 3 chemicals that are already known as irritants showed a correlation of r=0.994 (p=0.001) with Draize score about same materials. Comparing to reference Draize data from ECVAM (European Centre for the Validation of Alternative Methods), a correlation coefficient with our Draize score for the 3 chemicals was 0.982 (p=0.003). Bovine cornea test data of additional 3 chemicals already known as irritants showed a correlation of r=0.968 (p<0.001) with reference Draize data from ECVAM. Then the bovine cornea opacity test and Draize test with 3 cosmetics were performed. In both tests, we obtained similar results that showed all cosmetics were non-irritant. These results indicate that bovine corneal opacity assay has the potentiality to be developed as alternatives to rabbit eye irritation test.
사람피부와 돼지피부를 이용한 OECD Guideline 428에 따른 피부흡수시험법 검증
한주희 ( Ju Hee Han ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박귀례 ( Kui Lea Park ),손경희 ( Kyung Hee Sohn ),주정흔 ( Jung Hun Ju ),임채형 ( Chae Hyung Lim ),박재학 ( Ja 한국동물실험대체법학회 2009 동물실험대체법학회지 Vol.3 No.1
In vitro skin absorption test has been proposed as a reliable alternative method to in vivo skin absorption test. In many other countries, in vitro skin absorption has been conducted according to OECD guideline which was adopted in 2000. However, in vitro skin absorption test hasn`t been validated yet in Korea. In this study, experimental conditions were set and validated according to a detailed protocol of OECD guideline 428. We compared penetration properties of caffeine (log P -0.01) and benzoic acid (log P 1.83) through human split-thickness skin and pig ear full-thickness skin using Franz cell. The permeation of caffeine through the human skin (10.16%) was higher than through pig ear full-thickness skin (1.68%). The permeation of benzoic acid through human skin and pig ear skin were similar. Hydrophilic caffeine penetrated the human split-thickness skin more efficiently than pig ear full-thickness skin. Our data showed a similar pattern with compared other reports that regarded skin absorption of caffeine and benzoic acid through split-thickness skin and full-thickness skin. In conclusion, we validated in vitro skin absorption test in Korea
연구논문 : Bovine Corneal Opacity Assay법을 이용한 안점막 자극 물질의 검증에 관한 연구
이현경 ( Hyun Kyoung Lee ),노경진 ( Kyoung Jin Noh ),석승혁 ( Seoung Hyeok Seok ),백민원 ( Min Won Baek ),이희영 ( Hui Young Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),김배환 ( Bae Hwan Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.2
Bovine corneal opacity and permeability assay have been proposed as reliable alternatives to the Draize test. In this study, we measured the opacity of the bovine cornea by quantifying the ability of light to pass through it using opacitometer. We obtained the regular value of basal opacity using bovine cornea collected within three hours after killing. For opacity alone, values obtained for 3 chemicals that are already known as irritants showed a correlation of r=0.994 (p=0.001) with Draize score about same materials. Comparing to reference Draize data from ECVAM (European Centre for the Validation of Alternative Methods), a correlation coefficient with our Draize score for the 3 chemicals was 0.982 (p=0.003). Bovine cornea test data of additional 3 chemicals already known as irritants showed a correlation of r=0.968 (p<0.001) with reference Draize data from ECVAM. Then the bovine cornea opacity test and Draize test with 3 cosmetics were performed. In both tests, we obtained similar results that showed all cosmetics were non-irritant. These results indicate that bovine corneal opacity assay has the potentiality to be developed as alternatives to rabbit eye irritation test.
카스가 아사코(Asako Kasuga),박종환(Jong-Hwan Park),석승혁(Seung-Hyeok Seok),백민원(Min-Won Baek),이희영(Hui-Young Lee),김동재(Dong-Jae Kim),박재학(Jae-Hak Park) 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.2
This title of a recent perspective in Science reflects the growing importance of a new model organism for developmental research, the zebrafish Danio reno. Due to its small size, rapid generation time, powerful genetic systems, and genomic resources, the zebrafish has emerged as an important model of vertebrate development and human disease. In this study, we will focus on the histology of zebrafish and it might be helpful for future research in this model system.
안점막 자극시험 대체시험법으로서 neutral red uptake assay 확립
이현경 ( Hyun Kyoung Lee ),노경진 ( Kyoung Jin Noh ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),이희영 ( Hui Young Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),김배환 ( Bae Hwan Kim ),박 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.1
The necessity of using animals to test whether new chemicals and products are eye irritants has been questioned over the last 20 years. During this process, numerous non animal methods have been proposed as reliable alternatives to the Draize test. The neutral red uptake assay is the in vitro method to test the eye irritation of chemicals and products. We need to validate the neutral red uptake assay as alternatives for the international harmonization. Therefore, we studied for the establishment of the neutral red uptake assay in Korea. In this study, we evaluated the eye irritation of five chemicals and three cosmetics by the Draize test and the neutral red uptake assay. The results of the neutral red uptake assay showed a correlation of r=0.893(p=0.107) with our Draize score in two chemicals. And the results of the neutral red uptake assay showed correlation of r=0.892(p=0.017) with the Draize score obtained from the ECVAM (European Centre for the Validation of Alternative Methods) in additional three chemicals. All of three cosmetics were evaluated as non irritants by our Draize test and the neutral red uptake assay. As a result, the neutral red uptake assay was established in Korea. For the international harmonization, the validation study has to be performed with more chemicals and products in more countries.
안점막자극시험의 대체시험법으로서 bovine corneal opacity assay의 확립
노경진 ( Kyoung Jin Noh ),석승혁 ( Seoung Hyeok Seok ),백민원 ( Min Won Baek ),이영희 ( Young Hui Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),이현경 ( Hyun Kyoung Lee ),김배환 ( Bae Hwan Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.1
The objective of this study is establishing of an alternative method using bovine cornea to rabbit eye irritation test. The bovine corneal opacity and permeability assay is checking opacity and permeability of bovine cornea after exposure of chemical irritants. In this study, we measured the opacity of the bovine cornea by quantifying the ability of light to pass through it using opacitometer. In addition, we optimized the basal opacity level before treatment of irritants to bovine corneas. With these results, we established the standard operating procedure of bovine corneal opacity assay.
연구논문 : 안점막자극시험의 대체시험법으로서 bovine corneal opacity assay의 확립
노경진 ( Kyoung Jin Noh ),석승혁 ( Seoung Hyeok Seok ),백민원 ( Min Won Baek ),이영희 ( Young Hui Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),이현경 ( Hyun Kyoung Lee ),김배환 ( Bae Hwan Kim ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.1
The objective of this study is establishing of an alternative method using bovine cornea to rabbit eye irritation test. The bovine corneal opacity and permeability assay is checking opacity and permeability of bovine cornea after exposure of chemical irritants. In this study, we measured the opacity of the bovine cornea by quantifying the ability of light to pass through it using opacitometer. In addition, we optimized the basal opacity level before treatment of irritants to bovine corneas. With these results, we established the standard operating procedure of bovine corneal opacity assay.
안점막 자극시험의 대체법으로서의 porcine corneal opacity 및 permeability assay
한주희 ( Ju Hee Han ),이현경 ( Hyun Kyoung Lee ),석승혁 ( Seung Hyeok Seok ),백민원 ( Min Won Baek ),이희영 ( Hui Young Lee ),김동재 ( Dong Jae Kim ),나이랑 ( Yi Rang Na ),박성훈 ( Sung Hoon Park ),김배환 ( Bae Hwan Kim ),박귀례 ( 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.1
The Draize eye irritation test has been changed to alternative methods based on 3 Rs (Replacement, Reduction, Refinement) of animal experiments. The bovine corneal opacity and permeability (BCOP) assay is well established as one of the alternative methods to the Draize test but the supply of bovine eyes from a slaughterhouse is not sufficient in actuality. The porcine corneal opacity and permeability(PCOP) assay have been proposed as a reliable alternative to the BCOP assay and the Draize test because porcine corneas have numerous advantages including structural similarities with human corneas and the stable capacity for a corneal supply. In this study, three cosmetics and seven chemicals that are already known as irritants at Draize score and the bovine corneal opacity(BCO) assay were tested by the PCOP assay for the first time in Korea. The porcine corneal opacity was measured by quantifying the ability of light to pass through a cornea using an opacitometer. The permeability was measured at 10 and 30 min time points after chemical exposure. The PCOP assay showed the high correlation with the Draize score and BCO assay. These results showed that the PCOP assay can be used as an alternative method for the BCO assay and the Draize test although it needs further studies about insoluble chemicals and others to increase reliability.