• Title/Summary/Keyword: peroxidase-antiperoxidase(PAP)

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Immunohistochemical diagnosis of hog cholera with peroxidase-antiperoxidase(PAP) complex method (Peroxidase-antiperoxidase(PAP) 복합체법을 이용한 돼지콜레라의 면역조직화학적 진단)

  • Moon, Oun-gyeong;Cho, Hee-tack;Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.215-221
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    • 1990
  • The present study was intended to use the peroxidase-antiperoxidase method for the identification of hog cholera virus(HCV) in the lymphatic organs of HCV-infected pigs. Sections were incubated with primary antibody (rabbit anti-HCV polyclonal or mouse anti-HCV monoclonal), followed by incubation with linkserum (goat anti-rabbit IgG) in excess and rabbit or mouse PAP complex. The viral antigen was localized mainly in the cytoplasms of lymphoid cells and macrophages. Positive reaction cells were frequently detected in the marginal areas of the germinal centers of the spleens, and also found in the tensils and lymph nodes. The method approved to be highly specific for the identification of the virus and allowed a precise localization of the viral antigen in infected cells.

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Demonstration of Newcastle Disease Virus Antigens in Paraffin Embedded Tissues of Experimentally Infected Chickens Using Peroxidase-antiperoxidase(PAP) Technique (Peroxidase-antiperoxidaes법을 이용한 실험감염 계의 조직내 뉴캣슬병 바이러스 항원동정)

  • 노환국;신종백;임기재;김병지
    • Korean Journal of Veterinary Service
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    • v.15 no.2
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    • pp.184-194
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    • 1992
  • This study was done to identify Newcastle disease virus(NDV) antigens in paraffin sections of various organs from experimentally NDV-infected chicken using peroxidase-antiperoxidase(PAP) technique. Sections were Incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit IgG conjugate and peroxidase anti-peroxidase ( PAP ). Positive reactions were often detected in the epithelim of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. The viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the indetification of NDV and allowed a precise localization of the viral antigens in infected cells.

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Purification of the Yolk Protein, and Identification of the Synthetic Site of Its Precursor in Eriocheir japonicus (Decapoda, Brachiura) (동남참게(Eriocheir japonicus)의 난황단백질 정제와 그 전구체의 합성부위 구명)

  • HAN Chang-Hee;BAE Hyun-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.5
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    • pp.432-442
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    • 1992
  • To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

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Immunohistochemistry of Gastrointestinal Endocrine Cells in the Meckel′s Diverticulum of the Bean Goose, Anser fabalis Latham

  • Ku, Sae-Kwang;Lee, Hyeung-Sik;Park, Ki-Dae;Lee, Jae-Hyun
    • Animal cells and systems
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    • v.4 no.4
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    • pp.375-379
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    • 2000
  • The appearance of some gastrointestinal endocrine cells in the Meckel's diverticulum (MD) of the bean goose, Anser fabalis Latham was observed using specific antisera against serotonin, gastrin, cholecystokinin (CCK)-8, glucagon, secretin, somatostatin and human pancreatic polypeptide (HPP) with the peroxidase antiperoxidase (PAP) method. Among these specific antisera, serotonin-, gastrin-, CCK-8-, somatostatin- and HPP-immunoreactive cells were demonstrated in this study. Serotonin-, gastrin- and somatostatin-immunoreactive cells were detected at moderate frequency and CCK-8- and HPP-immunoreactive cells was rare and low frequencies, respectively. These immunoreactive cells were located in the superficial epithelium, intestinal crvpt and intestinal glands with spherical or spindle shaped cells having long cytoplasmic processes (open typed-cell). Mucosal layer of MD was composed of simple columnar epithelium and numerous intestinal glands. In addition, numerous lymphatic tissues were also demonstrated. In conclusion, histological profiles of MD were similar to any parts of the large intestine, especially the cecum, but the appearance, distribution and relative frequency of gastrointestinal endocrine cells were similar to those of upper parts of the small intestine. Although the exact digestive functions were unknown, the finding that the appearance, distribution and relative frequency of gastrointestinal endocrine cells in MD is similar to small intestine may be considered as distinct evidence that this organ may have some digestive functions.

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Immunohistochemical Studv on the Gastrin, Somatostatin and Serotonin Cells in the Gastric and Small Intestinal Mucosa of Rat during Development (발생기 흰쥐 위와 소장점막의 gastrin, somatostatin 및 serotonin세포에 대한 면역조직화학적연구)

  • 최병태;조운복
    • The Korean Journal of Zoology
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    • v.37 no.4
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    • pp.478-487
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    • 1994
  • The developmental changes of three enteroendocrine cells, i.e. gastrln, somatostatin and serotonin, of gastric and small intestinal mucosa in pre- and postnatal rat were examined by peroxidase-antiperoxidase (PAP) method. In the course of development, gastrin cells were obsenred in the pyloric gland region and the whole part of small intestine, while somstostatin and serotonin cells in the whole gastric gland region and small intestine. More entroendocrine cells were detected in the pyloric gland region and duodenum than in the other portion. In the stomach, gastrin, somatostatin and serotonin ceils were first obsenred in the pyloric Bland region on 17, 19 and 19 days of gestation respectively. The small intestinal gastrin and serotonin cells were first appeared in the duodenum and iriunum on 17 and 15 days of gestation respectively, and somatostBtin cells in duodenum on 17 days of gestation. The number of cells examined from the stomach were increased from fetal to weanling period and showed a decrease during adult period: the notable increase was shown at the end of suckling period or at early weanling period. The cells of the small intestine increased from fetal to suckling period, especially, these cells markedly increased at the end of fetal period or at early suckling period, and decreased from weanling period. The shape of these cells was oval or fusiform during fetal period. In the stomach, most of gastrin cells turned out to be oval and open-type from suckling period, while the remaining two tripes of cells were oval and open- or closed-type. In the small intestine, 311%Ves of cells examined were changed to fusiform and open-type from the end of fetal period. Three types of cell were distributed over the stratified epithelium on 15 and 17 days of gestation. In the stomach, these cells were distributed lower gastric pit and gland from the following fetal period, and were detected mainly on the upper part of gland from suckling period, and then obsenred on the whole part of gland. In the small intestine, most of cells distributed over only between epithelium of villi on 19 days of gestation, increased in number on the crypt from following fetal period, and also observed abundantly in the crypt at adult period.

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Tissue origin of soluble component proteins in salin extract of adult Paragonimus westermani (폐흡충 성충 수용성 단백질의 성분별 생성위치)

  • Yoon Kong;Chi- Yong Park;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.91-100
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    • 1992
  • Tissue origin of individual component proteins in crude extract of adult Paragonimus westermani was investigated. Major soluble component proteins were separated by disc-PAGE in 8% slab gel. By predefined Rf values, strips of gel containing each band protein was cut out. Each band protein was eluted by electrophoresis. Monospecific antibodies were prepared by immunizing rabbits with each band protein. When peroxidase-antiperoxidase (PAP) staining was done, antiserum to Band 1 reacted to content of eggs both in the worm and in the infected lung tissue. Antiserum to Band 2 reacted to parenchymal tissue of the worm. Antiserum to Band 4 showed the positive reaction at intestinal content while that to Band 5 reacted to the intestinal epithelial border. Antiserum to combined proteins of Bands 617 and that to Band 8 reacted to parenchymal tissue of the worm respectively. From the results, the origin of individual proteins in crude extract of adult P. westermani could be differentiated.

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Immunohistochemical Study of the Endocrine Cells in the Pancreas of the Korean Aucha Perch, Serranidae (Coreoperca herzi) (Serranidae (Coreoperca herzi) 췌장 내분비세포에 대한 면역조직화학적 연구)

  • Lee, Jae-hyun;Ku, Sae-kwang;Lee, Hyeung-sik;Ham, Tae-su
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.339-347
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    • 2003
  • The regional distribution and relative frequency of some endocrine cells in the pancreas of the Korean aucha perch, Coreoperca herzi Herzenstein belonging to the family Serranidae in order Perciformis, were observed using specific mammalian antisera against serotonin, insulin, glucagon, somatostatin and human pancreatic polypeptide (hPP) by peroxidase antiperoxidase (PAP) method. The pancreas was divided into four portions (principal and secondary islets, exocrine and pancreatic duct regions). In addition, the pancreatic islet regions were further subdivided into three regions (central, mantle and peripheral regions). Spherical to spindle or occasionally round to oval immunoreactive (IR) cells were demonstrated in the pancreatic islets and exoccrine portions, but no cells were detected in the pancreatic duct portions. In the principal islets, serotonin-IR cells were not detected but most of insulin-IR cells were located in the central regions and they were also demonstrated in the mantle and peripheral regions in moderate and rare frequencies, respectively. Glucagon- and hPP-IR cells were mainly situated in the mantle regions but the cells were also demonstrated in the peripheral regions in relatively lower frequency. Somatostatin-IR cells were evenly distributed in the central and mantle regions in a few frequency and cells were also demonstrated in the peripheral regions in rare frequency. Cell clusters were consisted of hPP-IR cells that were situated in the peripheral to mantle regions. In the secondary islet portions, serotonin-IR cells were randomly distributed throughout the whole pancreatic islet regions but lower frequency was detected in the peripheral regions compared to that in central and mantle regions where cells were detected in a few frequency, respectively. Insulin-IR cells were restricted to the central regions in numerous frequency and glucagon-IR cells were evenly distributed in the mantle and peripheral regions in moderate frequencies, respectively. Somatostatin-IR cells were observed in the central and mantle regions in moderate and a few frequencies, respectively. In addition, hPP-IR cells showed similar distributional patterns to those of glucagon-IR cells except cells were also located in the central regions in rare frequency. In the exocrine portions, only glucagon- and hPP-IR cells were demonstrated in rare and a few frequencies, respectively. In conclusion, the regional distribution and relative frequency of pancreatic endocrine cells of the Korean aucha perch showed general patterns, which were observed in other teleost. However, some species-dependent different distributional patterns and/or relative frequencies were also demonstrated especially to serotonin-IR cells. In pancreas of the Korean aucha perch, insulin-IR cells were the most predominant cell type followed by glucagon-, somatostatin-, hPP- and serotonin-IR cells.