• Title/Summary/Keyword: cytopathic effects

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Helicobacter pylori Vacuolating Toxin Exhibits Polar Activity of $Cl^-$ Secretion and Secretory Response to Carbachol in T84 Cells

  • Jin, Nan-Ge;Jin, Yong-Ri;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.5
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    • pp.289-293
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    • 2004
  • To investigate whether VacA (vacuolating toxin) produced by Helicobacter pylori Korean stain 99 induces intestinal secretion, purified VacA was added to T84 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of low pH-pretreated VacA increased short circuit current (Isc). The effect was time- and dose-dependent and saturable. The time-to-peak Isc was concentration-dependent. Chloride channel inhibitors, niflumic acid or 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), inhibited VacA-stimulated Isc. Carbachol (CCh)-induced increase of Isc was prolonged by the addition of VacA to the mucosal side only. The effect was unaltered by the addition of niflumic acid. VacA did not show cytopathic effects. These studies indicate that VacA is a nonlethal toxin that acts in a polar manner on T84 monolayers to potentiate $Cl^-$ secretion and the response to CCh secretion without decrease in monolayer resistance. VacA may contribute to diarrhea diseases in human intestinal epithelial cells.

Ultrastructural Aspects of the Mixed Infections of Watermelon Mosaic Potyvirus and Cucumber Green Mottle Mosaic Tobamovirus Isolated from Watermelon

  • Kim, Jeong-Soo;Cho, Jeom-Deog;Park, Hong-Soo;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.211-215
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    • 2000
  • Symptoms on 4 varieties of watermelons inoculated with watermelon mosaic potyvirus II isolated from watermelon (WMV-W) were severe mosaic and leaf malformation while those inoculated with cucumber green mottle mosaic tobamovirus from watermelon (CGMMV-W) were mild mosaic and chlorotic spots. Inoculation of the mixture of WMV-W and CGMMV-W produced extremely severe mosaic along with necrotic spots and general necrosis. Doubly infected plants were also stunted. Cells infected with WMV-W or CGMMV-W alone exhibited the intrinsically ultra-structural properties of each virus infection. WMV-W induced potyvirus-characteristic cylindrical inclusions in the cytosol. Virus particles were orderly aligned along the tonoplasts. CGMMV-W induced tobamovirus-characteristic stacked crystalline arrays of virus particles in the cytosol. Cells infected doubly with WMV-W and CGMMV-W contained striking cytopathic effects that were not present in single infection of each virus. The unique ring structure, nonagon, was that a single potyvirus particle was surrounded by 9 CGMMV-W tobamovirus particles.

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Repression of Type-1 Fimbriae in Shiga Toxin-Producing Escherichia coli O91:H21 Isolated from Asymptomatic Human Carriers in Korea

  • Kim, Jung-Beom;Oh, Kyung-Hwan;Park, Mi-Sun;Cho, Seung-Hak
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.731-737
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    • 2013
  • Seventy-four Shiga toxin-producing Escherichia coli (STEC) isolates belonging to the serotype O91:H21 were isolated from 1,643 asymptomatic human carriers in a STEC outbreak at Gwangju in Korea. Although the isolates did not cause any symptoms, all of them produced Shiga toxins 1 (Stx1) and 2 (Stx2). In order to determine why these strains cause no symptoms, we explored the differences in virulence potential between the asymptomatic STEC O91:H21 isolates and symptomatic STEC O91:H21 strains (ATCC 51435 and ATCC 51434). The asymptomatic STEC O91:H21 isolates showed strongly reduced cytopathic effects compared with the symptomatic strains when intact bacterial cells were used as an inoculant. Moreover, we found a reduced adherence phenotype when testing asymptomatic strains on HeLa cells. Real-time quantitative PCR results suggest that transcriptional repression of the genes encoding type-1 fimbriae occurs in the asymptomatic isolates but not in the symptomatic strains.

Propagation of lymphocystis disease virus (LCDV) in the FIN cell line originated from olive flounder Paralichthys olivaceus fin

  • Hossain, Mosharrof;Kim, Wi-Sik;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.24 no.2
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    • pp.47-51
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    • 2011
  • The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

Inhibitory Effects of Norwogonin, Oroxylin A, and Mosloflavone on Enterovirus 71

  • Choi, Hwa Jung;Song, Hyuk-Hwan;Lee, Jae-Sug;Ko, Hyun-Jeong;Song, Jae-Hyoung
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.552-558
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    • 2016
  • Severe complications associated with EV71 infections are a common cause of neonatal death. Lack of effective therapeutic agents for these infections underlines the importance of research for the development of new antiviral compounds. In the present study, the anti-EV71 activity of norwogonin, oroxylin A, and mosloflavone from Scutellaria baicalensis Georgi was evaluated using a cytopathic effect (CPE) reduction method, which demonstrated that all three compounds possessed strong anti-EV71 activity and decreased the formation of visible CPEs. Norwogonin, oroxylin A, and mosloflavone also inhibited virus replication during the initial stage of virus infection, and they inhibited viral VP2 protein expression, thereby inhibiting viral capsid protein synthesis. However, ribavirin has a relatively weaker efficacy compared to the other drugs. Therefore, these findings provide important information that will aid in the utilization of norwogonin, oroxylin A, and mosloflavone for EV71 treatment.

Isolation of chicken anemia agent (virus) from naturally infected chickens (자연감염된 닭으로부터 chicken anemia agent (virus)의 분리)

  • Seong, Hwan-woo;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.471-477
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    • 1991
  • Attempts to isolate chicken anemia agent (CAA) were made by inoculating tissue homogenates into MDCC-MSBl or LSCC-1104B1 cell lines and passaging the cells serially. CAA was isolated from the liver and thymus of 11 weeks old layer chickens and from the liver of 10 weeks old broiler breeder chickens. The layer flock experienced approximately 45% mortality during 9 to 14 week of age from gangrenous dermatitis and lymphoid organs of affected chickens were severely atrophied. The broiler breeder flock experienced approximately 7% mortality during 7 to 9 weeks of age and affected birds showed lesions of colibacillosis, staphylococcal arthritis, and coccidiosis together with atrophied lymphoid organs. The isolated viruses were identified as CAA by the indirect fluorescent antibody test and virus neutralization test using CAA immune sera including one to Gifu-1 strain of CAA. The CAA isolate 89-69, when inoculated into susceptible 1 day old SPF chicks, induced anemia 14 to 16 days after inoculation. It did not induce any cytopathic effects in chicken embryo liver and chicken embryo fibroblast cell cultures. Infectivity of the isolate was not affected by the treatment of chloroform or heat ($70^{\circ}C$ for 15 minutes).

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Isolation and characterization of fowl adenovirus serotype 4 from chickens with hydropericardium syndrome in Korea

  • Park, Hong-Su;Lim, Il-Soo;Kim, Sang-Kyu;Kim, Toh-Kyung;Yeo, Sang-Geon
    • Korean Journal of Veterinary Research
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    • v.51 no.3
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    • pp.209-216
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    • 2011
  • Four strains of fowl adenovirus (FAdV) were isolated from 4 flocks of broiler or layer chickens affected by hydropericardium syndrome in Korea. These FAdVs were classified as serotype 4 by restriction fragment length polymorphism patterns of hexon genes and whole genomes. The virus exhibited cytopathic effects consisting of rounding, ballooning and clustering in primary chicken embryo liver cell cultures. In transmission electron microscopy, virus particles in hexagonal shape aggregated exclusively in the nuclei of hepatocytes of the chickens as the typical appearances in adenovirus infections. Buoyant density of the virus in cesium chloride (CsCl) was 1.34 g/mL. The virus was stable to chloroform, ether, 50~70% ethanol, acidic condition at pH 3, 0.25% trypsin (1 : 250), heat at $50^{\circ}C$ for 30 min, but labile to 100% ethanol, heat at $52{\sim}60^{\circ}C$ for 30 min, 1 M $MgCl_2$ at $50^{\circ}C$ for 1 h, 1 : 2,000 formalin (37%). All of the physicochemical properties pertained to the characteristics of adenoviruses. Eight viral polypeptides were determined in CsCl-purified virus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Isolation and identification of infectious pancreatic necrosis virus form rainbow trout cultured in Korea (Isolation and Identification of Infectious Pancreatic Necrosis Virus from Rainbow Trout Cultured in Korea)

  • Lee, Jin-Hee;Heo, Gang-Joon;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.124-130
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    • 1996
  • A survey was conducted to determine the prevalance of infectious pancreatic necrosis virus (IPNV) on fish farms in Korea and the epidemiology of IPNV infection in the farmed rainbow trout. In total, 43 pools of rainbow trout with apparent signs of viral infection from five provinces were obtained and analyzed. Evident cytopathic effects, including karyopycnosis and cell destruction, were observed in CHSE (chinook samlmon embyro)-214 cells infected with the virus isolates. Of these, ten viral isolates were assumed to be IPNV based on biophysical properties. RNA analysis revealed that the isolates contained two-segmented RNA genomes, further indicating that the viral isolates are IPNV. Antigenic comparison of the IPNV isolates identified three distinct serological groups separable by the cross-neutralization test. Of the ten IPNV isolates, six could be classified as strain DRT, two as strain Ab, and two as strain VR299. We were not able to isolate new strain of IPNV or any isolate serologically similar to the standard strain Sp.poraceae and families of the Agaricales, they are genetically more related to the Polyporaceae. These results are consistent with morphological characters observed in those mushrooms. However, it is premature to conclude taxonomic status Ganoderma species in the present study employing small sample size.

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A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line

  • Demir, Ismail;Demirbag, Zihni
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1485-1490
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    • 2006
  • In this study, comparative replicational properties of Hyphantria cunea nucleopolyhedrovirus (HycuNPV) in Lymantria dispar (IPLB-LdElta) and Spodoptera frugiperda (IPLB-Sf21) cell lines were investigated. Our microscopic observations showed that cytopathic effects (CPEs) in LdElta cells appeared 12 h later than those in Sf21 cells. Whereas polyhedral inclusion bodies (PIBs) formed at 48 h postinfection (p.i.) in LdElta cells, it formed at 36 h p.i. in Sf21 cells. Extracellular virus production determined according to the 50% tissue culture infective dose ($TCID_{50}$) method in LdElta cells started about 12 h later when compared with Sf21 cells. Titers of extracellular virus in LdElta and Sf21 cells were calculated as $1.77{\times}10^9$ plaque forming units (PFU)/ml and $5.6{\times}10^9PFU/ml$, respectively, at 72 h p.i. We also showed that viral DNA replication began at 12 h p.i. in both cell lines. Viral protein synthesis was determined by SDS-polyacrylamide gel electrophoresis (PAGE) and polyhedrin synthesis was observed at 12 h p.i. in both cell lines. The results indicate that while the synthesis of macromolecules is 12 h later and production of extracellular virus is almost 3-fold lower in LdElta cells compared with those in Sf21 cells, the LdElta cell line is still a productive cell line for infection of HycuNPV.

Characterization of rock bream (Oplegnathus fasciatus) fin cells and its susceptibility to different genotypes of megalocytiviruses

  • Jeong, Ye Jin;Kim, Young Chul;Min, Joon Gyu;Jeong, Min A;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.149-159
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    • 2021
  • Genus Megalocytivirus cause red sea bream iridoviral disease (RSIVD) and scale drop disease (SDD). Based on the phylogeny of the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes, megalocytiviruses except for SDD virus (SDDV) could be three different genotypes, red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis (ISKNV), and turbot reddish body iridovirus (TRBIV). In this study, primary cells derived from the caudal fin of rock bream (Oplegnathus fasciatus) grew at 25℃ in Leibovitz's medium supplemented with 10% (v/v) fetal bovine serum and primocin (100 ㎍/mL). Rock bream fin (RBF) cells exhibited susceptibility to infections by different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV) with the appearance of cytopathic effects with an increase in the viral genome copy number. Furthermore, compared to grunt fin (GF) cells, even though 10 times lower number of RSIV genome copies were inoculated in RBF cells, viral genome copy number produced on RBF cells were 44 times higher than that of GF cells at 7 d post-inoculation. As the isolated RBF cells are sensitive to different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV), they can be used for future studies regarding in vitro viral infection and subsequent diagnosis.