• 한국해양바이오학회지
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• 제1권4호
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• pp.298-304
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• 2006

In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

• International Journal of Oral Biology
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• 제40권4호
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2001년도 추계학술대회(제116회) 및 13회 Workshop 제3회 한ㆍ일 치과보존학회 공동학술대회 초록집
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• pp.561.1-561
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• 2001

The aim of this study was to examine the localization of bacteria in the apices of roots with refractory apical periodontitis by a scanning electron micrscopy(SEM), and to identify the extra-radical bacteria using a immunohistological method. Eleven teeth were prepare for immunostaining and 4 for SEM. Immunostaining was performed with LSAB method and specific antisera against 18 bacteria selected for this study. Specimens for SEM observations were dried using a t-butyl alcohol freeze dryer and coated with gold-palladium.(omitted)

• 한국가축번식학회지
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• 제25권3호
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• pp.287-292
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• 2001

난자의 동결보존 방법은 보다 효과적 난자은행을 개발하기 위한 필수불가결한 요소이다. 본 연구에서는 보다 효과적인 유리화 동결법을 개발하기 위해 유리화 동결 응해 후 염색체와 방추사의 손상이 일어나는지를 알아보고자 본 실험을 수행하였다. 난구세포에 둘러싸인 난자를 5.5 M etylene glycol과 1.0 M sucrose 용액에 노출 후 electron microscope grid에 부착시킨 후 액체질소에서 냉동 보존하였다. 동결 융해 후 생존한 난자들은 방추사의 이상성을 보기 위해 immunostaining 방법을, 염색체의 이상성을 보기 위해 karyotyping을 시행하였다. 실험결과 염색체 이상빈도는 대조군의 경우 19.6%, 동결 융해군은 32.8%로 관찰되었고, 방추사의 이상빈도는 대조군의 경우 20.2%, 동결 융해군의 경우 32.3%로 대조군에 비해 염색체와 방추사의 이상성이 증가되었다. 결론적으로 마우스의 성숙난자를 유리화 동결시킨 후 염색체와 방추사의 이상성이 증가됨을 관찰하였다.

• Journal of Chest Surgery
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• 제31권12호
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• pp.1200-1205
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• 1998

연구목적: 폐암은 protooncogene의 활성화와 종양억제유전인자(tumor suppressor gene)의 비활성화 등 다단 과정에 의하여 발생한다. 치료목적의 완전 절제가 가능하였던 제 IIIA기 비소세포폐암 환자에서 cyclin D1, p53, bcl-2 gene의 변이가 폐암에 미치는 영향을 조사하고자 하였다. 대상 및 방법: 1990년부터 1995년까지 연세의료원에서 치료목적의 완전절제가 가능하였던 stageIIIA 비소세포폐암 환자 100명의 paraffin block과 임상기록을 이용하였다. 각 환자의 조직절편을 labelled streptavidin-biotin method로 immunohistochemical 염색하였고 cyclin D1, p53, Bcl-2 immunostaining을 위한 조직절편들은 immunostaining하기 전 citrate buffer 내에서 10분에서 20분간 microwave oven으로 전처치한 후 cyclin D1은 NCL-cyclin D1-GM으로 p53는 lone DO-7으로 bcl-2는 clone 124로 overnight incubation하였다. 수술 후 평균 추적조사기간은 24.1 개월(range; 2∼84 개월)이었다. 결과: 100예의 폐암 중 56예가 편평상피세포암, 37예가 선암, 5예가 adenosquamous cell carcinoma, 2예가 대세포암이었고 수술 후 5년 생존율은 32.1%이었다. cyclin D1의 양성율은 35 %, p53는 56 %, bcl-2는 17 %였으나 cyclin D1, p53, Bcl-2 단백질 양성 발현과 생존율과의 상관관계는 없었다. 결론: 연구결과 cyclin D1, p53, Bcl-2 단백질 양성 발현이 비소세포폐암 발생기전과 연관되어 있으나 수술 후 예후인자로서는 부적당한 것으로 판단되었다.

• 대한세포병리학회지
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• 제12권2호
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• pp.81-87
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• 2001

The differentiation between reactive mesothelial and carcinoma cells in serous effusion cytology can be a diagnostic challenge based on morphology alone. The expression of some cell adhesion molecules may be helpful in the differential diagnosis. This study evaluated the usefulness of E-cadherin Immunocytochemistry for discrimination of carcinoma cells from reactive mesothelial cells. Alcohol fixed, paraffin embedded cell blocks taken from 42 reactive and 102 malignant serous effusions with histologically confirmed diagnoses were immunostained with monoclonal antibody to E-cadherin by LSAB method. E-cadherin expression was identified in only 2 benign reactive serous effusions(5%) whereas 91 malignant serous effusions(89%) expressed E-cadherin The differences in immunostaining for E-cadherin between reactive and malignant serous effusions were statistically significant(p < 0.001). The sensitivity and specificity of the E-cadherin immunostaining for carcinoma cells were 89% and 95%, respectively. In conclusion, E-cadherin is a useful diagnostic adjunct for differentiation between reactive mesothelial and carcinoma cells in serous effusions.

• 대한임상검사과학회지
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• 제47권3호
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• pp.132-139
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• 2015

최근 $p16^{INK4a}$/Ki-67 이중면역염색은 액상세포검사에서 자궁경부 이형성증과 암을 조기에 발견하기 위한 새로운 생체 표지자로 대두되고 있다. 저자들은 자궁경부질도말의 액상세포검체 총 109례에서 CINtec$^{(R)}$ PLUS kit를 사용하여 $p16^{INK4a}$/Ki-67 이중면역염색을 시행하였고, 그 결과를 액상세포검사, HPV hybrid capture II (HC II) 검사 그리고 조직학적 진단과 서로 비교하였다. $p16^{INK4a}$/Ki-67 양성 발현은 세포학적 진단에서 저등급 편평상피내병변 이상의 증례 그리고 조직학적 진단에서 자궁경부 상피내종양 1등급 이상의 증례에서 유의미하게 높았다. 액상세포검사상 비정형 편평 상피세포 소견을 보이는 46례 중, 31례 (67.4%)가 $p16^{INK4a}$과 Ki-67 양성 소견을 보였고, 이러한 양성 증례들은 조직검사에서도 대부분 자궁경부 상피내종양 1등급 이상의 병변에 해당하였다. 자궁경부 상피내종양 1등급 이상의 병변을 발견하기 위한 $p16^{INK4a}$/Ki-67 이중면역염색의 민감도는 액상세포검사보다 높은 89.0%였고, 특이도는 73.5%로 HC II 검사보다 높게 분석되었다. 따라서, $p16^{INK4a}$/Ki-67 이중면역염색 방법은 액상세포검사법의 민감도와 HC II 검사법의 특이도를 보완하기 위한 진단적 검사로 유용하다고 할 수 있다.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.35-53
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• 1996

The development and repair requires the formation of new tissues comprised of various extracellular matrix components. The present study investigated the formation and distribution of the major ECM components such as type I collagen, type III collagen, fibronection, bone sialoprotein, and osteonection during development and repair. For developing observation. Sprague-Dawley rats weighing $27{\pm}1gm$ were sacrificed. For repair observation, Sprague-Dawley rats weighing $110{\pm}5gm$ were used. The pulp perforation were prepared on mesial surface of the maxillary first molar by using 1/2round bur. At 5 days after perforation, rats were sacrificed by perfusion with 3 % paroformaldehyde. The maxillary first molar region were cut, demineralized, dehydrated and embedded in paraffin. Immunostaining the ECM components was achieved by the avidin-biotin complex method. The results as follows : 1. Bright immunoreaction for fibronectin was present in the basement membrane at the inner epithelial-mesenchymal interface, especially concentrated in the blood vessel walls, cell membrane of odontoblasts, and initial predentin. 2. Type I and III collagen was observed in the newly formed pulp tissue, predentin, and its intensity increased as more of these components during repair. 3. Strong immunostaining for bone sialoprotein and osteonectin was found in dentin while no or weaker staining was observed loose connective tissue of the pulp. 4. These results suggest that develpment and repair is achieved through a series of cell differentiation and attachment by the specific ECM components.

• Asian-Australasian Journal of Animal Sciences
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• 제23권11호
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• pp.1412-1420
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• 2010

In this study, changes in testicular tissues of rats subjected to xylene and formaldehyde inhalation were evaluated. Three experimental groups were included in the study. Each group of rats was exposed to formaldehyde (6 ppm), technical xylene (300 ppm) or a combination of these two agents (150 ppm+3 ppm) for 8 weeks (8 h/d). Control groups were maintained for a period of eight weeks under the same conditions. Staining methods (triple staining, strep ABC method) were applied to examine histometric changes and relaxin like factor (RLF) expression in the testicular tissue. Immunostaining for RLF showed that density of staining for RLF decreased in rats exposed to formaldehyde. Formaldehyde or a combination of formaldehyde and xylene led to a decrease in seminiferous epithelial height. In conclusion, exposure of rats to formaldehyde and xylene-formaldehyde combinations adversely affects Leydig cells (RLF) and seminiferous epithelium of testicular tissue.

• 대한수의학회지
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• 제35권1호
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• pp.67-73
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• 1995

아무르장지뱀(Takydromus amurensis)의 위장관 점막에 분포하는 내분비세포의 분포와 출현빈도 및 세포의 종류를 알아보기 위하여 몇가지 도은법 및 면역조직화학적 방법으로 관찰하였다. Grimelius법에만 염색된 은호성세포(argyrophil cell)는 유문부에 다수로 십이지장에 중등도로 국한되어 출현하였다. 한편 bovine CG면역반응세포는 유문부에서 최고의 빈도로 전장관에 걸쳐 분포하였다. BPP면역반응세포는 소장부위에서만 동정되었다. 따라서 아무르장지뱀의 위장관 내분비세포에서 은호성세포와 bovine CG면역반응세포는 분포와 출현빈도에서 일치하지 않았다.