• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1995.04a
• /
• pp.95-95
• /
• 1995

뇌신경 변성 / 퇴행과 관련된 중요한 병인론 중의 하나는 변성 과정에서 형성된 유리기(free radical)로 인한 항산화계의 평형 소실로 알려져 있다. Aspalatone (APT)의 예상되는 항산화 효능을 검정하기 위하여 본 실험에서는 Kainic acid (KA) 유발 뇌변성 모델을 적용하였다. KA 모델은 변연계의 간질성 경련과 신경세포 변성에 대하여 재현성 있는 병변 모델을 제공해 주며, 이와 같은 신경세포의 병독 기전에 산소 유리기가 관여함이 강력히 시사되고 있기 때문이다. KA 투여로 인하여 지속적이고도 전형적인 간질성 경련이 관찰되고 1일 이내에 높은 치사율을 보였으나 APT으로 인하여 그 간질성 경련 행위와 비율이 억제되고 KA 유발 치사율도 억제되었다. 최종 KA 투여 3일 후에 얻어진 흰쥐 해마 및 대뇌 피질에서 항산화 효소인 Superoxide dismutase (SOD), Catalase (Cat.), Glutathione peroxidase (GSH-PX) 및 과산화지질의 지표인 Malondialdehyde (MDA)를 검정하였다. 대조군에 비하여 KA는 뇌조직의 SOD-1을 유도하였으나, Cat.와 GSH-PX의 활성은 현저히 유도되지 않았고, 반면에 MDA 치는 현저히 증가하였다. 즉, Cat., GSH-PX와 같은 $H_2O$$_2$중화제가 동반 유도하지 않는 SOD의 유도는 세포내 축적되는 $H_2O$$_2$로 인하여 Fenton/Haber-Weiss 반응을 가속화하여 과산화지질화를 촉진함을 시사한다. APT 병용 투여로 SOD는 현저히 유도되지 않았으나 특히 Cat.가 현저히 유도되어지고 MDA는 억제되었다. 이와 같은 생화학적인 결과는 다음의 형태학적인 소견과 일치한다. Fos 관련 항원 (FRA)와 SOD-1을 면역세포화학 (Immunocytochemistry)적 방법으로 이중 표식 (double-labelling) 하였다. FRA는 KA로 인한 신경세포의 자극에 대한 지표로 응용하였고, SOD-1은 퇴행성 뇌질환에서 산화적 손상의 지표로 사용하였다. KA 투여로 해마의 dentate gyrus (DG) 내에 강한 면역환성 (immunoreactivity)이 나타났고 pyramidal cell layer (PCL)와 glia에 SOD-1이 강하게 염색되었다. APT 병용 투여로 상당수의 경련이 일어나지 않은 흰쥐는 해마의 DG에 FRA가 경미하게 염색되었고, PCL에 SOD-1도 경미하게 나타났으나, 경련이 나타난 쥐에서는 KA만을 투여한 흰쥐와 구별되지 않았다. 이상의 APT의 항산화 효과는 KA로 인한 뇌세포 변성 개선에 중요한 인자로 작용할 것으로 사료되나, 보다 명확한 APT의 기전을 검색하고 직접 임상에 응응하기 위하여는 보다 다양한 실험 조건이 보완되어야 찰 것으로 생각된다.

• Parasites, Hosts and Diseases
• /
• v.35 no.3
• /
• pp.203-210
• /
• 1997

The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

• Korean Journal of Veterinary Research
• /
• v.40 no.1
• /
• pp.130-137
• /
• 2000

In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

• Korean Journal of Veterinary Research
• /
• v.35 no.1
• /
• pp.143-148
• /
• 1995

Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.

• Korean Journal of Veterinary Research
• /
• v.34 no.2
• /
• pp.321-326
• /
• 1994

Korean isolate, porcine epidemic diarrhea virus (KPEDV-9) was adapted through serial passages in vero cell. The viral yield reached up to $10^{5-6}$ $TCID_{50}/ml$ at the passage level of 90th. The cell adapted virus was characterized through genetic and morphological examinations. The RNA extracted from virus infected cell revealed the presence of RNA band with molecular size of >20Kb. The electron microscopic examination on purified virus showed the pleomorphic appearance of enveloped particles with 5-10nm surface projections, which fit with the shape of coronavirus. The etiological survey on swine diarrhea by immunofluorescence test(FA) indicated 17.5% positive rate on the PEDV infection. In addition, the incidence were detected both in piglets within two weeks old as well as fattening pigs. Serological survey by ELISA revealed the overall 45% positive result, thus, indicating the PEDV infection are widespread throughout this country.

• Korean Journal of Veterinary Research
• /
• v.34 no.1
• /
• pp.135-139
• /
• 1994

In the year 1992/93 leucocytozoonosis could be first diagnbosed in 87 chickens of 4 chicken farms in Honam districts. The diagnosis was confirmed by detection of the blood merozoites or gametocytes and histological finding of the schizonts from various organs with some clinical signs. Cases of leucocytozoonosis only occurred from the end of June to the middle of September. Artificial infection could be observed by means of inoculation of infected blood merozoites. The schizonts were found in the liver and cardiac muscle of the different chickens recovered from the natural infection, respectively, in September and next February. Thus the relapse or long-term infection in cold seasons might be possible. The unique gametocyte antigen polypeptide was 50.1 kD.

• Korean Journal of Veterinary Research
• /
• v.57 no.4
• /
• pp.223-226
• /
• 2017

This study was undertaken to develop new analytical methods for assessment of anticoccidials. High-performance liquid chromatography (HPLC) was found to be a fast, reliable, and practical method. The anticoccidials used in this experiment were toltrazuril and diclazuril, and the analysis factors were specificity, linearity, accuracy, repeatability, and intermediate precision. The linearity of each anticoccidial was better than 0.99, and the accuracies were 99.5% and 99.1% with relative SD of 0.5 and 0.4, respectively. To assess whether the developed HPLC method could be effectively applied, toltrazuril and diclazuril post-market veterinary products (five products) that are currently sold were tested. The results revealed no non-compliant items and the method was applied successfully. Therefore, the newly developed HPLC method for anticoccidial assessment described in this study may be useful as a reference method in the Korean Standards of Veterinary Pharmaceuticals for the analysis of toltrazuril and diclazuril.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.4 no.1
• /
• pp.34-40
• /
• 2001

Purpose: The purpose of this study was to evaluate the clinical efficacy of reverse transcription-polymerase chain reaction (RT-PCR) in detecting rotaviral gastroenteritis in children comparing with that of commercial immunoassays. Methods: Stools from 79 children admitted Korea University Hospital due to diarrhea were collected from December 1999 to February 2000. Immunoassays were done using commercial rotavirus Latex kit and Rotatec (ELISA) kit. RT-PCR was performed to amplify group A rotavirus, most commonly pathogenic to human, using VP4- and VP7-specific primers. The detection rates of immunoassays and RT-PCR were compared. Results: ELISA assay was superior to LA assay and moderately concordant with RT-PCR in detecting rotaviral gastroenteritis. Conclusion: Although RT-PCR is known very sensitive, it does not have significant advantage over immunoassay in detecting rotaviral gastroenteritis.

• Korean Journal of Microbiology
• /
• v.37 no.1
• /
• pp.49-55
• /
• 2001

Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. The anthrax toxin contains three components, including the protective antigen (PA), which binds to eucaryotic cell surface receptors and mediates the transport of toxins into the cell. In this study, the entire 2,294-nucleotide protective antigen gene (pag) was sequenced from 4 of B. anthracis strains to identify potential variation in the toxin and to further our understanding of B. anthracis evolution in Korea. Sequence alignment of the entire PA gene from 30 strains representative of the four B. anthracis diversity groups revealed mutations. The mutation of B. anthracis BAK are located adjacent to a highly antigenic region crossing the junction between PA domains 3 and 4 shown to be critical to LF binding. The different mutational combinations observed in this study give rise to 11 PA genotypes and 4PA phenotypes. Three-dimensional analysis of all the amino acid changes (Ala to Val) observed in BAK indicated that these changes are not only close sequentially but also very close in three-dimensional space to the antigenic region importan tfor LF binding. Phylogenetic (cladistic) analysis of the pag corresponded with previous strain grouping based on chromosomal variation, suggesting that plasmid evolution in B. anthracis has occurred with little or no horizontal transfer between the different strains.

• Analytical Science and Technology
• /
• v.17 no.3
• /
• pp.211-216
• /
• 2004

Commercial one-step rapid fecal occult blood (FOB) kit which was used as a screening test to detect traces of blood in stool samples was evaluated for the feasibility of the forensic identification of human blood. The sensitivity was determined and compared with the conventional Leucomalichite green (LMG) method. In addition, the specificity of the kit and the effects of various chemicals and environmental factors were examined. FOB kit was specific for human hemoglobin and more sensitive than LMG test (approximately 100 times). FOB kit showed positive band using at least 1,000,000-fold diluted human blood. The antigen was very stable regardless of storage temperature and boiling. The positive reaction was not affected by LMG and Luminol, the traditional tests for identification of bloodstain. As a results, FOB test kit could be effectively applied to identification of human blood at crime scene and crime laboratories.