• Title/Summary/Keyword: Heat extracted antigen

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Studies on the Enhancing Effect of Polymyxin B on the Antibodies Response of Enterobacterial Antigens (Pomyxin B의 장계세균항원(腸系細菌抗原)에 대(對)한 항체산생(抗體産生) 증강작용(增强作用)에 관(關)한 연구(硏究))

  • Lee, Jae-Koo
    • The Journal of the Korean Society for Microbiology
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    • v.6 no.1
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    • pp.29-40
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    • 1971
  • Various kinds of antibiotics are generally believed to have inhibitory effects on the antibody response. However, as polymyxin B which belongs to the cyclic polypeptide group of antibiotic was found to have some enhancing effects on the antibody response of rabbits to enterobacterial common antigen(CA) under specified conditions, experiments were carried out on this problem with the following results. 1. When mixture of polymyxin B and CA derived from Salmonella typhimurium(STM) was treated 30 minutes at $37^{\circ}C$ and injected three times into rabbits by intravenous route, the antibody response to CA was weaker than rabbits injected CA only. 2. Mixture of polymyxin B and CA showed a marked antibody production when injected into rabbits primed with small amounts of heat-extracted antigen of STM, while the injection of CA alone showed low titers of response. 3. Mixture of polymyxin B and heat-extracted CA-containing antigen of Escherichia coli 014 also showed a increased antibody production than CA alone in rabbits primed with antigen of STM. 4. The effect of polymyxin B appeared in different ways. This antibiotic did not enhance the CA antibody response in rabbits primed with small amounts of E. coli 0111 and 055, but enhance in rabbits primed with Shigella flexneri. 5. No enhancing effect on the antibody response was observed by polymyxin B in rabbits primed with CA. 6. No enhancing effect on the antibody response was also noted in rabbits primed with STM antigen in case polymyxin B and CA were administered simultaneously but in veins of different places. 7. Bacitracin did not enhance the CA antibody response in primed rabbits with STM antigen, but neomycin slightly enhance the response. 8. Lipopolysaccharide showed no priming effect on the CA antibody response, and no enhancement of the CA antibody response in rabbits printed with STM. 9. The priming effect of STM antigen against CA antibody response was very weak as compared with the effect of CA derived from STM antigen.

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Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.51-57
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    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

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Study on the Production and Management of Aquatic Animal : Rapid and Optimized Diagnosis of Edwardsiellosis by Coagglutination Test with Antibody Sensitized Staphylococcus aureus (수산 생물의 생산과 관리에 관한 기초연구 : 항체 감작 Staphylococcus aureus에 의한 Coagglutination Test기법을 이용한 Edwardsiellosis의 신속 진단)

  • HA Jae Yi;SOHN Sang Gyu;HUH Min-Do;JEONG Hyun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.5
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    • pp.620-628
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    • 1996
  • To avoid the self-agglutination of Staphylococcus aureus sensitized with rabbit antibody in the absence of antigen, we determined the optimum concentration of rabbit antibody for sensitization. It was analyzed by using three different kinds of S. aureus strains at various concentraions of antibody. The optimized coagglutination test using the S. aureus sensitized with rabbit antibody was applied to the diagnosis of edwardsiellosis in field and in laboratory. The presence of E. tarda as low as $10\;{\mu}g/ml$ was detected by this method. Moreover, it showed good coagglutination results against several different forms of antigens such as FKC, EDTA or heat extracted antigen of E. tarda. E. tarda strains, isolated from the flounders suffering from edwardsiellosis in fields, showed some cross-reactions to the E. tarda 219 analyzed by both agglutination and coagglutination test with rabbit anti-E, tarda 219 antibody. The degree of cross-reactions analyzed was enough to apply the coagglutination test for the diagnosis of edwardsiellosis in field. Thus, even 1,000 fold diluted tissue homogenate of infected flounder naturally contained enough E. tarda as an antigen to show good coagglutination with S. aueus sensitized with rabbit anti-E, tarda 219 antibody. The successful application of this method to the homogenate and heat extract of tissues from naturally or artificially infected flounder or tilapia preyed that coagglutination test was a simple and rapid reliable dignostic technique suitable for using in laboratory and field without any special equipments.

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Qualitative Enzyme-Linked Immunosorbent Assay (ELISA) for the Diagnosis of Edwardsiellosis (Edwardsiellosis의 진단을 위한 정성적 ELISA법)

  • Kim, Myoung-Sug;Hwang, Eun-A;Huh, Min-Do;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.24-31
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    • 1999
  • Optimization and standardization of solid phase enzyme immunoassay were done for the diagnosis of edwardsiellosis in fish. The analyzed degree of immobilized antibody on surface of solid phase with peroxidase saturation method showed the optimized result by using partially purified $50{\mu}g/ml$ of rabbit anti-E. tarda Edk-2 antibody in sodium bicarbonate buffer for overnight incubation to cover the surface of polystyrene beads. Optimized immunoreaction was observed in the treatment of $50{\mu}g/ml$ of biotin conjugated antibody followed extravidin-peroxidase diluted 1 : 2,000 in PBS. The detectable concentrations of the this method were $1{\times}10^5$ cells/ml and $1{\times}10^5$ cells/ml expressed as the source of antigen amount for EDTA extraction and heat extraction, respectively. High cross-reaction of solid phase ELISA with the prepared rabbit and-E. tarda Edk-2 was observed against E. tarda strains isolated from flounder suffering from edwardsiellosis in aquatic farms of Korea. It suggested that the potential of this solid phase of ELISA technique is very powerful for the application to different strains of E. tarda isolated in farms of many different areas.

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Difference of Gene Expression between Hypertrophic Scar Keratinocytes and Normal Keratinocytes (비후성 반흔 각질세포와 정상 각질세포의 유전자 비교분석)

  • Choi, Sung-Won;Chung, Ho-Yun;Lim, Young-Kook;Kim, Hoon-Nam;Oh, Ji-Won;Kim, Moon-Kyu;Jeon, Sae-Hwa;Hong, Yong-Taek
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.317-322
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    • 2010
  • Purpose: There is no clear evidence of the original cause of hypertrophic scar, and the effective method of treatment is not yet established. Recently the steps of searching in gene and molecular level are proceeding. we are trying to recognize the difference between keratinocytes of hypertrophic scar and normal skin. Then we do support the comprehension of the scar formation mechanism and scar management. Methods: Total RNAs were extracted from cultured keratinocytes from 4 hypertrophic scars and normal skins. The cDNA chips were prepared. A total of 3063 cDNAs from human cDNA library were arrayed. And the scanning data were analyzed. Results: On microarray, heat shock protein, pyruvate kinase, tumor rejection antigen were more than 2 fold intensity genes. Among them, heat shock 70 kd protein showed the strongest intensity difference. Conclusion: In this study, it can be concluded that heat shock proteins play an important role in the process of wound healing and scar formation. This study provides basic biologic information for scar research. The new way of the prevention and treatment of scar formation would be introduced with further investigations.

Analysis of X Irradiation Related Genes in HL60 Cells Using cDNA Microarray (cDNA Microarray를 이용한 HL60 세포주에서 방사선 조사 관련 유전자의 검색 및 분석)

  • Park, Keon-Uk;Hwang, Mi-Sun;Suh, Seong-Il;Suh, Min-Ho;Kwon, Taeg-Kyu;Park, Jong-Wook;Cho, Jae-We;Choi, Eun-Ju;Baek, Won-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.4
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    • pp.299-308
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    • 2000
  • Recently developed cDNA microarray or DNA chip technology allows expression monitoring of expression of hundreds and thousands of genes simultaneously and provides a format for identifying genes as well as changes in their activity. In order to search for changes in gene expression after X irradiation in HL60 cells, cDNA microarray technique was done. In this study, expression of 588 human genes (including oncogenes, tumor suppressor genes, cell cycle regulator genes, intracellular signal transduction modulator genes, apoptosis related genes, transcription factor genes, growth factors and receptor genes, cytokine genes, etc) were analyzed. For cDNA microarray analysis mRNAs were extracted from control and 8 Gy-irradiated HL60 cells. As a result the changes in expression of several genes were observed. This alteration of gene expression was confirmed by reverse transcription-polymerase chain reaction. The expression of heat shock 60 KD protein, c-jun, erythroid differentiation factor, CPP32, myeloid cell nuclear differentiation antigen, MAP kinase-activated protein kinase, interleukin-8, monocyte chemotactic peptide 1 and RANTES genes was increased, but the expression of p55CDC gene was decreased after X irradiation.

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Development of Immunoassay Systems for the Assay of Soy Protein in Meat Products; The Assay of Soy Protein in Meat Blends and Commercial Product by Enzyme-linked Immunosorbent Assay(ELISA) (육제품에 첨가된 대두단백 정량을 위한 면역분석법 개발에 관한 연구 ; ELISA에 의한 고기유화물 및 시판육제품에 첨가된 대두단백 정량)

  • Kim, Cheon-Jei;Kim, Jong-Bae;Kim, Byung-Cheol;Lee, Seoung-Bae;Jung, Sung-Won;Choe, Doo-Young;Ko, Won-Sick
    • Korean Journal of Food Science and Technology
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    • v.24 no.5
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    • pp.397-403
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    • 1992
  • This study was carried out for the development of assay method to quantify the soy protein content in meat homogenate, emulsion-type sausage and commercial meat products by Enzyme-linked Immunosorbent Assay(ELISA). The standard antigen was extracted before and after heat treatment. It was observed that the degree of reaction was not varied significantly according to the heating temperature. The recovery rate in meat homogenate and emulsion-type sausage was not varied significantly according to the heating temperature. The reaction was not interfered with fat and spices of the samples. Samples with 10% soy protein showed lower correlation than those with 2% and 5% soy protein. The recovery rate in commercial meat products showed difference individually. The correlation of some products with raw vegetable and wheat starch was relatively low.

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