• Title/Summary/Keyword: cytopathic

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Use of Nested Polymerase Chain Reaction for Identification of Rickettsia tsutsugamushi Serotype Cultured in Human Embryonic Lung Cells (Nested PCR을 이용한 사람 유래 태아 폐세포에서 배양된 Rickettsia tsutsugamushi의 혈청형 동정)

  • An, Chang-Nam;Woo, Gyu-Jin;Kim, Tae-Yeon;Shin, Kwang-Soon;Kim, Chul-Joong;Baek, Luck-Ju
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.235-244
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    • 1996
  • We selected the adequate cell line to be used for propagation and plaquing of R. tsutsugamushi in laboratory and identified R. tsutsugamushi serotype cultured in LuMA cells by nested PCR. As in this study, we concluded that. 1. LuMA cell was suitable for the study of the biology of rickettsiae-host cell interaction. 2. The plaque-forming unit (PFU) per ml of R. tsutsugamushi Karp strain propagated in embryonated egg yolk sacs was $10^{8.8}$ and the PFU/ml of Gilliam strain was $10^{7.1}$. 3. The rate and extent of cytopathic changes depended on the PFU titer of R. tsutsugamushi. 4. PCR with nested primer pairs was useful for identification of R. tsutsugamushi serotype cultured in human embryonic lung cells.

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Isolation and Characterization of Jeju Island Bovine Rotavirus (JBR) (제주도 Bovine Rotavirus의 분리와 그 특성)

  • Yu, Jae-Hyeun;Lee, Young-Kun;Ju, Ji-Sun;Kim, You-Seong;Kim, Soon-Jae;Ebina, T.;Nakagomi, O.;Urasawa, S.;Taniguchi, K.;Greenberg, Harry B.
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.181-189
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    • 1996
  • Fecal samples of calf diarrhea were taken on farms in Jeju island, rotavirus was isolated and cytopathic effect (CPE) was determined after infection to MA104 cell. Morphological evaluation on electron microscopy proved it as rotavirus. Also, its infection to MA104 cell was reidentified using a fluorescence antibody method. Genotype of Jeju island bovine rotavirus (JBR) analyzed through PAGE was 4: 2: 3: 2 pattern, which was unique in bovine and that analyzed through general PAGE was somewhat different from NCDV, UK, KK3, A5-3A, 61A, B223 and similar to N stool-5, N culture-5 and Kawatabi (Japan). By titration after plaquing, the level was $1-3\;{\times}\;10^6\;PFU/ml$, which was lower than those of NCDV and UK. Electrophoresis analysis of RNA-RNA hybridization, ELISA, and first and second PCR products of VP7 and VP4 in 1% agarose ($TAE+1{\mu}l$ EtBr) revealed that the rotavirus was a serotype of G6P11.

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Inhibition of Herpesvirus-6B RNA Replication by Short Interference RNAs

  • Yoon, Jong-Sub;Kim, Sun-Hwa;Shin, Min-Chul;Lee, Dong-Gun;Hong, Seong-Karp;Jung, Yong-Tae;Khang, In-Gu;Shin, Wan-Shik;Kim, Chun-Choo;Paik, Soon-Young
    • BMB Reports
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    • v.37 no.3
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    • pp.383-385
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    • 2004
  • RNA interference (RNAi) is a process of sequence-specific gene silencing, which is initiated by double-stranded RNA (dsRNA). RNAi may also serve as an antiviral system in vertebrates. This study describes the inhibition of herpesvirus-6B (HHV-6B) replication by short interference RNAs (siRNAs) that are targeted to the U38 sequence that encodes DNA polymerase. When virus-infected SupT1 cells were treated by siRNA, these cells blocked the cytopathic effect (CPE) and detected the HHV-6B antibody-negative in indirect immunofluorescence assays (IFA). Our result suggests that RNAi can efficiently block Herpesvirus-6B replication.

The Extracts of Kalopanax pictus Nakai. for Inhibitory Effects on HIV-1 and Its Essential Enzymes (Human Immunodeficiency Virus Type Ⅰ에 대한 음나무 추출물의 억제활성)

  • Yu Young Beob;Shim Bum Sang;Ahn Kyoo Seok;Choi Seung Hoon;Park Jong Cheol;Miyashiro H.;Hattori M.
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1129-1133
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    • 2004
  • For the purpose of developing new anti-HIV agents from natural sources, the extracts of Kalopanax pictus were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT). protease and α-glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts of stem and leafstalk inhibited the HIV-1-induced cytopathic effects with Ie (inhibitory concentration) of 25 and 50㎍/㎖, respectively. Moreover water extracts (100㎍/㎖) of stem and leafstalk showed strong activity of 80% and 90% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. In the HIV-1 protease inhibition assay, aqueous stem extract inhibited the activity of the enzyme to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease with 58%, but no glucosidase inhibitory activities. We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of RT and protease. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are RT, PR and α-glucosidase inhibitors.

Isolation of novel bovine parainfluenza virus type 5 (bPIV5) and its incidence in Korean cattle

  • Yang, Dong-Kun;Nah, Jin-Ju;Kim, Ha-Hyun;Choi, Sung-Suk;Bae, You-Chan;Park, Jung-Won;Song, Jae-Young
    • Korean Journal of Veterinary Research
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    • v.54 no.2
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    • pp.107-112
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    • 2014
  • Four viruses showing cytopathic effects in MDBK cells were isolated from brains of cattle showing downer cattle syndrome in 2012. The isolates were confirmed to belong to the genus Rubulavirus of the subfamily Paramyxovirinae. Isolate QIA-B1201 had the ability to hemagglutinate red blood cells from several species of animals and was capable of adsorbing guinea pig erythrocytes on the surface of infected Vero cells. Nucleotide sequence analysis showed that two isolates (QIA-B1201 and QIA-B1204) had high similarity with other human and animal PIV5 isolates ranging from 98.1 to 99.8%. The highest sequence similarity of the two isolates corresponded to strain KNU-11 (99.8% at the nucleotide and amino acid level) isolated from suckling piglets in Korea in 2012. To evaluate the virulence of strain QIA-B1201, we inoculated bPIV5 into 5 week-old mice via both the intraperitoneal and intracranial route. Body weight was not significantly altered in mice inoculated with QIA-B1201. In this study, we isolated and characterized novel bPIV5s from brain samples showing downer cattle syndrome, but were not able to elucidate the pathogenicity of the bPIV5s in mice.

Overexpression of Fish DRG2 Induces Cell Rounding

  • Park, Jeong-Jae;Cha, Seung-Ju;Ko, Myung-Seok;Cho, Wha-Ja;Yoon, Won-Joon;Moon, Chang-Hoon;Do, Jeong-Wan;Kim, Sung-Bum;Hebok Song
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.295-300
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    • 2002
  • Previously, we reported induced expression of developmentally regulated CTP-binding protein 2 (DRG2) in fish cells at the late stage of rhabdovirus infection. To investigate the biological role of fish DRG2 (fDRG2), we transfected CHSE-214 cells with an expression vector containing complete fDRG2 fused to the N-terminal end of an enhanced green fluorescent protein (EGFP). Low level expression of fDRG2-EGFP did not induce morphological change or cell death. However, a high level expression of fDRG2-EGFP induced cell rounding and caused depletion of the cell population in FACS analysis. Several truncated fragments were fused to EGFP. FACS analysis was conducted to determine the presence of cells expressing high levels of the resulting chimera. While cells expressing a high level of N-terminus were detected, those expressing high levels of the C-terminal fragment 243-290 containing the G4 motif were absent in FACS analysis. Based on these observations, we propose that overexpression of fDRG2 may induce cell rounding, a representative cytopathic effect of virus-infected cells in the late stage of infection and the C-terminus of the fDRG2 is essential for this function.

Screening of Some Plant Extracts for Inhibitory Effects on HIV-1 and Its Essential Enzymes (Human Immunodeficiency Virus Type I에 대한 수종 식물 추출물의 억제활성 검색)

  • Park, Jong-Cheol;Lee, Jong-Ho;Kim, Gyeong-Eup;Jo, Sung-Kee;Byun, Myung-Woo;Hirotsuku, Miyashiro;Masao, Hattori;Yu, Yeong-Beob
    • Korean Journal of Pharmacognosy
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    • v.29 no.4
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    • pp.338-346
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    • 1998
  • In order to elucidate the relationship between anti-HIV-1 enzyme activity and inhibition of HIV-1 replication by natural sources, extracts from some plants using the foods and oriental medicines were tested for inhibitory effects on the viral replication, reverse transcriptase (RT), protease and ${\alpha}-glucosidase$. In the anti-RT test, water extracts of Ficus carica (leaf), Houttuynia cordata (aerial part) and Ixeris tamagawaensis (aerial part) showed more than 79% inhitibion at a concentration of $100\;{\mu}g/ml$. The protease and ${\alpha}-glucosidase-inhibiting$ samples in the screening were water extract of Syringa dilatata (leaf) and methanol extract of Hibiscus syriacus (leaf and stem), which showed more than 40% inhibition at a concentration of $100\;{\mu}g/ml$. In the primary anti-HIV-1 test, water extracts of Equisetum arvense (aerial part), Hibiscus syriacus (leaf), Ixeris tamagawaensis (aerial part) and Pueraira thunbergiana (leaf) showed the potent inhibition against HIV-1 induced cytopathic effects.

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Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case

  • Yu, Hak-Sun;Jeong, Hae-Jin;Hong, Yeon-Chul;Seol, Seong-Yong;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.11-18
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    • 2007
  • Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multi-plied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

A Role of Natural Killer Cell in Mouse Infected Herpes Simplex Virus (Herpes Simplex Virus에 감염된 Mouse의 NK세포역할)

  • Lee, Yun-Tai;Lee, Chong-Hoon
    • The Journal of the Korean Society for Microbiology
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    • v.17 no.1
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    • pp.7-14
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    • 1982
  • A model of induction of neoplasia by viruses has develpoed from experimental studies in animals and in cultured cells and oncogenic transformation of cells is the result of integration of viral genetic information into the cellular DNA. The evidence for these associations was derived primarily from seroepidemiologic investigation. However, data indicating that the relation between HSV-2 and cervical cancer fits the model derived from experimental animal studies are not yet sufficient to draw conclusion with regard to the etiologic role the virus in the development of the neoplasms. In other hand, the K562 tumor cell is highly susceptible target for natural killer cell lysis by the lymphocytes of human and murine periperal blood. The characteristics of this effector cell type has been investigated. A study on natural killer cell mediated cytotoxicity(NKMC) against $^{51}Cr$-K562 as target cell was studed in HSV-2 infected ICR mouse. We have studied for susceptibility of HSV-2 against mouse embryo fibroblast(MEF) cells and NKMC from HSV-2 infected mouse. The results obtained that the mouse embryo fibroblast cells culture, the number and size of the cells were markedly increased and formed a monolayers relatively rapid, and become complete monolayer sheet around 72 hrs. Duration of cytopathic effect on MEF cells was rapid by serial passing of HSV-2. The morphology of the HSV-2 infected cells appear to be mainly round, ovium, spindle form and some of them was forming large giant cells. The NKMC was decrease in mouse with HSV-2 and comparison between effector/target cells ratio as 25:1 and 50:1 respectively, the NKMC was found to be more significantly decreased than normal control we have concluded that the natural killer cell activity of the viral infected mouse was shown as a suppressed during the HSV-2 infection, day 7th and 14th.

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Biological and Physicochemical Properties of Porcine Epidemic Diarrhea Virus Chinju99 Strain Isolated in Korea (국내 분리 돼지 유행성설사 바이러스 Chinju99주의 생물학적 및 물리화학적 성상)

  • Lee, Hee-Kyung;Yeo, Sang-Geon
    • Journal of Veterinary Clinics
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    • v.20 no.2
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    • pp.150-154
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    • 2003
  • A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the intestines after two blind-passages in Vero cells supplemented with trypsin (10 ug/ml). and the biological and physicochemical properties of the isolate were characterized. The virion was roughly spherical in shape and had spike peplomers on its outer surface. The virus exhibited cytopathic effects such as rounding degeneration at initiation of infection and syncytia formation later in Vero cells. The virus was labile to 20% ether and 5% chloroform but stable in acid with pH 4-7 at $4^{\circ}C$. The infectivity of the virus was maintained at $50^{\circ}C$ for 180 min, and the buoyant density of the virus in sucrose was 1.180 g/ml. All biological and physicochemical properties of the virus were typical features of coronaviruses.