• Title/Summary/Keyword: Immunodiffusion

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Enzymeimmunoassay for the Plasma Vitellogenin and Early Determination of Ovarian Maturation in Red Seabream, Pagrus major (참돔(Pagrus major)의 혈장 난황단백전구체에 대한 효소면역측정법과 난소성숙의 조기판정)

  • Han Chang-Haa;Yang Mun-Ho;Paek Jae-Min;Lim Sang-Koo;Kim Kwang-Hyun
    • Journal of Aquaculture
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    • v.8 no.1
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    • pp.1-19
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    • 1995
  • In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

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A Pneumococcal Conjugate Vaccine Formula Induces Protection in Mice Against Disseminated Disease due to Streptococcus pneumoniae (페렴구균 전신감염에 대한 협막. 표면단백질 접합백신의 효과)

  • Han , Yong-Moon;Lee , Jue-Hee
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.345-351
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    • 2004
  • ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

Pathological and serological detection of bovine viral leukosis in a dairy farm in Jeonbuk province (유우농장에서 발생한 소바이러스성 백혈병의 병리학적 및 혈청학적 조사)

  • Jo Young-Suk;Jang Sae-Gun;Chu Keum-Suk;Choi Eun-Young;Chon Hee-Woong;Hong Jae-Hee;Lim Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.89-96
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    • 2006
  • Bovine viral leukosis is a viral disease of cattle characterized by the development of tumors in the lymphatic tissue. A female Holstein, 3-year-old, was submitted for diagnosis at the Diagnostic laboratory, Chonbuk National University. Clinical sign of the affected animal showed emaciation, enlargement of superficial lymph node and mild diarrhea. Remarkable lesions were enlargement of many internal lymph nodes. Histopathology revealed excessive neoplastic lymphoid cells characteristic of BVL infection. Subsequently, serums from all cattle were collected and serological examination was done where a 85% seropositive rate was detected using ELISA test. ELISA method showed a comparatively 75% higher detection rate than the agar gel immunodiffusion (AGID) test (85% vs 40%). Serologically positive cattle were variably detected in all ages from under 1 year to over 6 year of age. Hematological examination consistently showed leukocytosis and a differential lymphocytosis of seropositive cattle. Detailed comparative pathological and serological data diagnosed the presence of bovine viral leukosis.

Survey for Antibodies to Bovine Leukemia Virus in Dairy Cattle (유우(乳牛) 백혈병(白血病)바이러스에 대(對)한 혈청항체(血淸抗體) 조사(調査))

  • Park, No-chan;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.26 no.1
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    • pp.61-68
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    • 1986
  • This paper described the distribution and transmissibility of BLV(bovine leukemia virus), the relationship between antibodies against BLV and lymphocyte count in 313 dairy cattle from 36 herds, the clinical signs and hematological findings of 2 lymphosarcomatous cattle in the northern area of Kyungpook. Eighty three (26.5%) of 313 cattle from 36 herds were positive for BLV antibodies and 19 (52.8%) of 36 herds were infected with BLV by the immunodiffusion test with BLV-gp antigen. The rate of BLV infection in cattle varied from 9.5 to 87.5% in 19 positive herds, it was higher in herds pastured during summer and included lymphosarcomatous onset than the other and also higher with the age. Eight (88.9%) out of 9 cattle which showed persistent lymphocytosis by the hematological test were positive for BLV antibodies. After 5 to 14 months, 13 (31.0%) of 42 cattle being negative for BLV antibodies in the positive herds converted into positive. Two lymphosarcomatous cattle were identified to be EBL (enzootic bovine leukemia) by the clinical sign, hematological examination and serological test.

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Characterization of Mouse Hybridoma Producing Monoclonal Antibody Paragonimus westermani (폐디스토마에 대한 Mmonoclonal Antibody생산 세포주개발에 대한 연구)

  • 고광삼;이숙영;이근배
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.64-68
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    • 1985
  • In this study, hybridoma techniques were applied to produce monoclonal antobodies to Paragonimus westermani, commonly known as lung distoma. Balb/c mice were immunized intraperitoneally every week with increasing doses of purfied protein of Paragonimus westermani adult worms begining with 0.3/mg/mouse/7 days and ending with 0.5mg at 28th day. Spleen cells from these immunized mice were hybridized with myeloma cells (NS-1) and the hybridized cells were selected in HAT media. The antibody secreting cells among the hybrid cells were initially selected by ELISA. Those initially selected cells were further screened by the criteria of antibody producing activity, and seneral cell lines among them were further tested with immunodiffusion method. One hybridoma clone produced IgM and another clone produced IgG1. The supernatant of the hybridoma clone producing IgM had titer 1:64 and the hybridoma clone producing IgG1 had titer 1:256 measured by immunofluorescence technique.

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Biochemical Characteristics of Spodoptera exigua Nuclear Polyhedrosis Virus (파밤나방 핵다각체병 바이러스의 생화학적 특성)

  • 진병래;박범석;재연호;강석권
    • Korean journal of applied entomology
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    • v.30 no.2
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    • pp.144-149
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    • 1991
  • Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

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Purification of Vitellogenin and Egg Yolk Protein, and Changes of Vitellogenin Concentration during the Ovulation Period in Elkhorn Sculpin, Alcichthys alcicornis

  • CHOI Cheol Young;CHANG Young Jin;TAKEMURA Akihiro
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.753-760
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    • 1995
  • This study was conducted to determine the serum vitellogenin (VTG) concentration changes during the ovulation period in elkhorn sculpin, Alcichthys alcicornis. The results of sepacryl S-300 showed that the molecular weight of VTG could be 380,000. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis may indicate that the purified VTG consists of three subunits with molecular weights of 180,000, 118,000 and 85,000, respectively. Yolk protein purified from the egg extracts was eluted on an equilibrated sephacryl S-300 column, and its molecular weight was estimated 250,000. The precipitation lines of the female serum against the antiserum of the egg extracts were fused completely by immunoelectrophoresis and immunodiffusion analysis. VTG was detected in the serum, and hepatocytes from males injected with $17\beta-estradiol\;(E_2).$ Furthermore, VTG was immunochemically similar to yolk proteins. The concentration of VTG was high before ovulation $(9.80\pm0.81-11.02\pm0.09 mg/ml),$ and then decreased rapidly after ovulation $(less\;than\;6.19\pm0.59 mg/ml).$ This study suggested that VTG was synthesized in the liver by the action of $E_2$ and released to blood, and then incorporated into oocytes.

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A Study on pH, Viscosity, Microorganisms and Immunoglobulin A of the Saliva (타액의 수소이온 농도, 점도, 세균 및 면역글로블린 A에 관한 연구)

  • Jin-Woo Sohn;Sung-Woo Lee
    • Journal of Oral Medicine and Pain
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    • v.17 no.2
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    • pp.87-97
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    • 1992
  • The purpose of this study is to investigate the age-and sex-related changes in the pH of resting saliva, viscosity, microorganisms and immunoglobulin A of stimulated whole saliva, and to investigate their correlations. The 120 healthy subjects were included in this study and the author used cone-and plate digital viscometer for viscosity, MSB agar for Streptococcus mutans, SL Rogosa agar for lactobacilli, and single radial immunodiffusion technique for immunoglobulinA. The obtained results were as follows : 1. There was no significant difference in pH, viscosity, Streptococcus mutans lactobacilli and immunoglobulin A of the saliva between males and females. 2. The viscosity values of stimulated whole saliva showed the increasing pattern with aging. 3. DMFS (or dmfs) rate was not correlated with pH, viscosity, Streptococcus mutans, lactobacilli and immunoglobulin A of the saliva. 4. There was a significant difference in the concentration of immunoglobulin A between the group under 10 and groups above 10. 5. The viscosity values of stimulated whole saliva showed the increasing pattern with decreasing of the number of Streptococcus mutans.

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Serotaxonomical Analyses of Some Streptomyces Strains Using Antibodies against Cell Envelope (균사 외피 항체를 이용한 Streptomyces 속 균주들의 혈청학적 유사성 분석)

  • Jo, Sung-Kee;Kim, Jae-Heon
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.137-141
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    • 2007
  • The similarities among five strains of Streptomyces were measured by the serological methods. Antigens were prepared by dissolving cell envelope fractions in Tween 20 buffer and antisera were produced from the immunized rabbits. Immunodiffusion studies and ELISA results showed that the degree of antigen-antibody reaction was not exactly matched to the taxonomic distance; i. e. the strains of the cluster group A exhibited low level of cross-reactions each other, while relatively strong cross-reactions were observed between Streptomyces lavendulae of cluster group F and Streptomyces viridochromogenes of cluster group A.

The changes of the plasma protein and the complements ($C_{3}$, $C_{4}$) after open heart surgery (개심수술후 혈장 단백 및 보체 ($C_{3}$, $C_{4}$)의 변화상 추적)

  • 남충희
    • Journal of Chest Surgery
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    • v.19 no.4
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    • pp.558-562
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    • 1986
  • The extracorporeal circulation has been much improved recently, but has yet much complex problems such as the protein denaturation and the activation of the complement system by the exposure of the blood to the foreign surface, which may result in such as the postperfusion syndrome. We studied the changes of the plasma protein fractions by the electrophoresis and the complement consumption [C3, C4] by the immunodiffusion method in the patients undergoing cardiac operation from Mar. 1, 1986 to Aug. 31, 1986. The results were summarized as follows: 1. y-globulin fraction was decreased [p<0.02 by paired t-test, N=25], but a,-globulin was increased [p<0.001 by paired t test, N=25] after operation. 2. C3,C4 were significantly reduced [p<0.001 by paired t-test, N=14] postoperatively and normalized from 24 hours after operation. 3. The consumption of C3,C4 had significant linear correlation [correlation coefficient r=0.97] and C, was more markedly reduced comparing with C3, which probably means the complement activation by classical pathway in our bubble oxygenator group.

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