• Title/Summary/Keyword: Immunofluorescence Assay

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Studies on the Characters of Avian Infectious Bronchitis Virus Isolated in Korea. (국내 분리 닭 전염성 기관지염 바이러스 성상에 관한 연구)

  • 이청산;조우영;최윤식;김순재
    • Korean Journal of Veterinary Service
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    • v.14 no.1
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    • pp.27-40
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    • 1991
  • In order to investigate the biological properties, pathogenicity and immune responses in artficially infected SPF chickens with Avian infectious bronchitis virus that was isolated from chickens showing IB like signs in southern region of Chung buk. Results obtained throuth the experiments are summarized as follows. 1. From 15 IB suspected cases, two strains of IB virus were isolated, one each from the tracheas and lungs. 2. Infectious bronchitis specific embryo lesions were observed after four serial passages of the isolates in chicken embryos. 3. The field isolates and M-41 strain of IB virus interfered with the replication of Newcastle disease virus in chicken embryos. 4. When specific pathogen free chickens, two week old, were inoculated with the IB virus isolates, clinical respiratory signs as dyspnea, coughing were observed. Airsacculitis was observed by necropsy. 5. AGP antibody positive rates of inoculated SPF chickens were highest on day 14 and lowest on day 36, while HI antibody responses were detected on day 14 in all Groups, the reinoculated Group was shown highest titers. 6. By Indirect immunofluorescence antibody assay of artificially infected SPF chickens, the viral antigens were detected in tissues of larynx, trachea and lung on the 4 th to 7 th days post inoculation.

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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.133-141
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    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

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Characterization of Akabane virus (KV0505) from cattle in Korea

  • Yang, Dong-Kun;Kim, Yeon-Hee;Kim, Byoung-han;Kweon, Chang-Hee;Yoon, Soon-Seek;Song, Jae-Young;Lee, Sang-Hun
    • Korean Journal of Veterinary Research
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    • v.48 no.1
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    • pp.61-66
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    • 2008
  • Akabane disease is caused by an arthropod-borne viral pathogen and leads to congenital abnormalities of the central nervous system in infected ruminants. One isolate, KV0505, showed cytopathic effect in Vero cells. The KV0505 isolate was obtained from plasma, which was collected from a cattle raised on Jeju Island in May 2005. Jeju Island is located near the southern part of the Korean peninsula. The isolate was confirmed as Akabane virus (AKAV) by immunofluorescence assay using AKAV specific monoclonal antibodies and reverse transcription polymerase chain reaction (RTPCR). Suckling mice inoculated with the isolate showed signs of paralysis and died within 10 days postinoculation. Comparisons of the KV0505 N gene sequence with 39 other known AKAV strains revealed nucleotide homologies ranging from 83.6% (MP496 strain) to 99.7% (M171 strain). When compared with the K-9 strain, which was isolated from a cow in Korea in 1994, the nucleotide sequence homology with the N gene was 99.7%. Thus, genes of the KV0505 isolate were closely related to those of the M171 strain, which were clustered into the Ic group of AKAV.

Effects of hypoxia inducible factors-$1{\alpha}$ on autophagy and invasion of trophoblasts

  • Choi, Jong-Ho;Lee, Hyun-Jung;Yang, Tae-Hyun;Kim, Gi Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.39 no.2
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    • pp.73-80
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    • 2012
  • Objective: This study was undertaken to determine the effect of hypoxia inducible factor (HIF)-$1{\alpha}$ on the cell death, autophagy, and invasion of trophoblasts. Methods: To understand the effect of HIF-$1{\alpha}$, we inhibited HIF-$1{\alpha}$ using siRNA under normoxia and hypoxia conditions. Invasion assay and zymography were performed to determine changes in the invasion ability of HIF-$1{\alpha}$. Western blotting and immunofluorescence were performed to determine some of the signal events involved in apoptosis and autophagy. Results: There was no difference in cell death through the inhibition of HIF-$1{\alpha}$ expression by siRNA; however, the expression of LC3 and autophagosome formation increased. On the other hand, autophagy was increased, and the invasive ability of trophoblast cells decreased according to the inhibition of HIF-$1{\alpha}$ expression by siRNA. These experimental results mean that HIF-$1{\alpha}$ genes regulate the invasive ability of trophoblasts by increasing autophagy. Conclusion: This study contributes important data for understanding the mechanism of early pregnancy implantation and the invasive ability of trophoblasts by defining the relationship between the roles of HIF-$1{\alpha}$ and autophagy.

Prevalence, infection intensity and histopathology ofPerkinsus in Venus clam, Protothaca jedoensis from Yo-su, Korea

  • Ngo, Thao T.T.;Park, Kyung-Il;Lim, Na-Rae;Choi, Sang-Duk;Choi, Kwang-Sik
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2004.05a
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    • pp.512-512
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    • 2004
  • Venus clam, Protothaca jedoensis was collected in lune 2003 from Yosu, Korea. The prevalence and infection intensity of Perkinsus was determined by Ray's Fluid thioglycollate medium technique and Choi's 2 M NaOH digestive assay. The prevalence of infection was 80% in Yosu; 54.2% in Taejon and 15.5% in Sachon. Consequently, infection intensity was highest in clams from Yosu (6250${\pm}$8188 Perkinsus cells/g of tissue) whereas lower levels were found in Taejon (5072${\pm}$10862 Perkinsus cells/g of tissue) and in Sachon (2452${\pm}$15300 Perhnsus cells/g of tissue). Histological observation showed that Perkinsus presented in mantle, gills, digestive tubes and gonad of clams. The hemocytic infiltrations and tissue damages were found in heavy Perkinsus-infected clams. Results after immunofluorescence staining with antibody developed from Perkinsus atlanticus suggested that there is closed relationship between Perkinsus found in Venus clam and Manila clam.

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Staphylococcusaureus protein A as a means of assessing sperm penetrability in cervical mucus in vitro

  • Al-Daghistani, Hala I.
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.3
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    • pp.186-193
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    • 2020
  • Objective: The effectiveness of Staphylococcus protein A (SPA) in improving the penetration ability of sperm and reducing antisperm antibody (ASA) titers in immunologically infertile males was evaluated. Methods: Seminal fluid samples were obtained from 15 infertile men, and ASA titers were assessed with the latex agglutination test. Identification of immunoglobulin (Ig) classes and characterization of the antigens involved in the immune response were performed using indirect immunofluorescence. Local ASAs typically present as a mixture of IgG and IgA classes. The capillary tube penetration method was used to assess the capability of spermatozoa to penetrate the cervical mucus (CM). Results: ASAs associated with the neck region of sperm showed a significantly lower migration distance in the CM of infertile females than ASAs associated with the head or tail segments. ASA-positive seminal fluid exhibited significant increases in the mean migration distance (2.6 ± 1.4 cm vs. 1.54 ± 1.1 cm, respectively; p< 0.001) and sperm concentration (174 ± 121.0 × 103/mL vs. 101 ± 93.7 × 103/mL, respectively; p= 0.033) after treatment with SPA compared to pre-treated samples. A significant reduction (p< 0.01) in the recorded ASA titer was detected. Conclusion: These results indicate that SPA can be used as a sorting regimen for insemination programs. However, further studies are warranted to assess its influence on pregnancy rate.

Longevity of Antibodies to Live Orientia tsutsugamushi Inoculated in Sprague Dawley Rats

  • An, Chang-Nam;Kim, Sung-Min;Kim, Tae-Yeon;Bae, Luck-Ju;Kim, Chul-Joong;Shin, Kwang-Soon;Park, Song-Yong
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.193-201
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    • 1998
  • In Sprague Dawley (SD) rats, antibodies against strains of Orientia tsutsugamushi, Kato, Karp and Gilliam, were produced in order to investigate their longevity and cross-reactivities to their corresponding homologous and heterologous antigens. By immunofluorescence assay (IFA) of IgG and IgM, it was shown that the immunity to the homologous strains persisted at a higher level (longevity of at least 34 weeks with higher IFA titers). On the other hand, the immunity to the heterologous strains persisted at a lower level (longevity of 10 to 34 weeks with lower IFA titers). Since infection with one strain of O. tsutsugamushi does not preclude reinfection with other strains, understanding of the antigenic diversity of O. tsutsugamushi and duration of the immunity to both homologous and heterologous strain is very important in diagnosis of scrub typhus.

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Potential Interaction of Plasmodium falciparum Hsp60 and Calpain

  • Yeo, Seon-Ju;Liu, Dong-Xu;Park, Hyun
    • Parasites, Hosts and Diseases
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    • v.53 no.6
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    • pp.665-673
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    • 2015
  • After invasion of red blood cells, malaria matures within the cell by degrading hemoglobin avidly. For enormous protein breakdown in trophozoite stage, many efficient and ordered proteolysis networks have been postulated and exploited. In this study, a potential interaction of a 60-kDa Plasmodium falciparum (Pf)-heat shock protein (Hsp60) and Pf-calpain, a cysteine protease, was explored. Pf-infected RBC was isolated and the endogenous Pf-Hsp60 and Pf-calpain were determined by western blot analysis and similar antigenicity of GroEL and Pf-Hsp60 was determined with anti-Pf-Hsp60. Potential interaction of Pf-calpain and Pf-Hsp60 was determined by immunoprecipitation and immunofluorescence assay. Mizoribine, a well-known inhibitor of Hsp60, attenuated both Pf-calpain enzyme activity as well as P. falciparum growth. The presented data suggest that the Pf-Hsp60 may function on Pf-calpain in a part of networks during malaria growth.

Porcine growth hormone induces the nuclear localization of porcine growth hormone receptor in vivo

  • Lan, Hainan;Liu, Huilin;Hong, Pan;Li, Ruonan;Zheng, Xin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.499-504
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    • 2018
  • Objective: Recent studies have challenged the traditional paradigm that growth hormone receptor (GHR) displays physiological functions only in the cell membrane. It has been demonstrated that GHR localizes to the cell nucleus and still exhibits important physiological roles. The phenomenon of nuclear localization of growth hormone (GH)-induced GHR has previously been described in vitro. However, until recently, whether GH could induce nuclear localization of GHR in vivo was unclear. Methods: In the present study, we used pig as an animal model, and porcine growth hormone (pGH) or saline was injected into the inferior vena cava. We subsequently observed the localization of porcine growth hormone receptor (pGHR) using multiple techniques, including, immunoprecipitation and Western-blotting, indirect immunofluorescence assay and electronmicroscopy. Results: The results showed that pGH could induce nuclear localization of pGHR. Taken together, the results of the present study provided the first demonstration that pGHR was translocated to cell nuclei under pGH stimulation in vivo. Conclusion: Nuclear localization of pGHR induced by the in vivo pGH treatment suggests new functions and/or novel roles of nuclear pGHR, which deserve further study.

Application of PCR for diagnosis of porcine circovirus type 2 (돼지 써코바이러스 2형의 진단을 위한 PCR법 적용)

  • Park Hyo-Sun;Lee Hyo-Sang;Na Ki-Bok;Lee Kwan-Bok;Kang Su-Jung;Moon Sun-Hwa
    • Korean Journal of Veterinary Service
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    • v.29 no.1
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    • pp.1-8
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    • 2006
  • Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.