• Title/Summary/Keyword: cytopathic

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The first virus isolation and partial characterization of equine herpesvirus-4 in a horse, South Korea

  • Choi, Eun-Jin;Lee, Hyun-Kyoung;Lee, Kyoung-Hyun;So, Byoung-Jae;Song, Jae-Young;Do, Jae-Chul;Yang, Seon-Joo;Lee, Hyun-Chul;Yang, Young-Jin
    • Korean Journal of Veterinary Service
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    • v.38 no.2
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    • pp.141-144
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    • 2015
  • An equine herpesvirus-4 (EHV-4) was isolated in nasal swabs collected in a horse showing respiratory clinical signs. Equine dermis cells inoculated with the sample were observed with characteristic viral cytopathic effects after 3 days of postinoculation and the infected cells exhibited bright intracelluar fluorescence by indirect immunofluorescence assay. At the nucleotide level, the partial glycoprotein B gene of the Korean EHV-4 isolate (K001) had 99.9% identity to 1942 strain (GenBank No. M26171). To author's knowledge, the report describes the first isolation and partial characterization of EHV-4 in Korea. The virus can be used for further study of EHV-4.

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.

Antiviral Effects of Sulfated Exopolysaccharide from the Marine Microalge Gyrodinium impudicum strain KG03

  • Im, Jeong-Han;Kim, Seong-Jin;Park, Gyu-Jin;An, Se-Hun;Lee, Hyeon-Sang;Lee, Hong-Geum
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.615-620
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    • 2003
  • The sulfated exopolysaccharide p-KG03, which is produced by the marine microalga Gyrodinium impudicum strain KG03, exhibited impressive antiviral activity in vitro ($EC_{50}$ = 26.9 g/ml) against the encephalomyocarditis virus (EMCV). Depending on the p-KG03 concentration, the development of cytopathic effects in EMCV-infected HeLa cells was either inhibited completely or slowed. Moreover, p-KG03 did not show any cytotoxic effects on HeLa cells, even at concentrations up to 1,000 g/ml. The polysaccharide was purified by repeated precipitation in ethanol, followed by gel filtration. The p-KG03 polysaccharide had a molecular weight of $1.87\;{\times}\;10^6$, and was characterized as a homopolysaccharide of galactose with uronic acid (2.96%, w/w) and sulfate groups (10.32% w/w). The biological activities of p-KG03 suggest that sulfated metabolites from marine organisms are a rich source of antiviral agents. This is the first reported marine source of antiviral sulfated polysaccharides against EMCV. The p-KG03 polysaccharide may be useful for the development of marine bioactive exopolysaccharides for use in biotechnological and pharmaceutical products.

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Synergistic Inhibition of Membrane ATPase and Cell Growth of Helicobacter pylori by ATPase Inhibitors

  • Ki, Mi-Ran;Yun, Soon-Kyu;Lim, Wang-Jin;Hong, Bum-Shik;Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.414-421
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    • 1999
  • Helicobacter pylori were found to be resistant to azide but sensitive to vanadate, suggesting that defect in the P-type ATPase activity rather than F-type ATPase would be lethal to cell survival or growth. To elucidate the relationship between this enzyme inhibition and H. pylori death, we determined the effect of omeprazole (OMP) plus vanadate on enzyme activity and cell growth. The minimum inhibitory concentration (MIC; ca. 0.8$\mu$mol/disk) of vanadate for H. pylori growth was lowered over l0-fold with the aid of OMP, whereby its inhibitory potential toward the P-type ATPase activity was diametrically increased. Alternatively, we found that this enzyme activity was essential for active transport in H. pylori. From these observations, we strongly suggest that the immediate cause of the growth inhibition of H. pylori cells with OMP and/or vanadate might be defective in the cell's active transport due to the lack of P-type ATPase activity. From the spectral data with circular dichroism (CD) spectroscopy, we found that activated OMP (OAS) at concentration below MIC did not disrupt helical structures of membrane proteins. Separately, we determined the cytopathic effect of OAS by SDS-PAGE, indicating the change in the production of cytoplasmic protein but not cell membrane.

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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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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.

Conditional Replication of a Recombinant Adenovirus Studied Using Neomycin as a Selective Marker

  • Xue, Feng;Qi, Yi-Peng;Joshua, Mallam Nock;Lan, Ping;Dong, Chang-Yuan
    • BMB Reports
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    • v.36 no.3
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    • pp.275-281
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    • 2003
  • An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.

Identification of a novel type of small molecule inhibitor against HIV-1

  • Kim, Byung Soo;Park, Jung Ae;Kim, Min-Jung;Kim, Seon Hee;Yu, Kyung Lee;You, Ji Chang
    • BMB Reports
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    • v.48 no.2
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    • pp.121-126
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    • 2015
  • Here we report a new chemical inhibitor against HIV-1 with a novel structure and mode of action. The inhibitor, designated as A1836, inhibited HIV-1 replication and virus production with a 50% inhibitory concentration ($IC_{50}$) of $2.0{\mu}M$ in an MT-4 cell-based and cytopathic protection antiviral assay, while its 50% cytotoxic concentration ($CC_{50}$) was much higher than $50{\mu}M$. Examination of the effect of A1836 on in vitro HIV-1 reverse transcriptase (RT) and integrase showed that neither were molecular targets of A1836. The characterization and re-infection assay of the HIV-1 virions generated in the presence of A1836 showed that the synthesis of early RT products in the cells infected with the virions was inhibited dose-dependently, due in part to abnormal protein formation within the virions, thus resulting in an impaired infectivity. These results suggest that A1836 might be a novel candidate for the development of a new type of HIV-1 inhibitor.

Study of Interferon Assay by HEp-2 : VSV system (HEp-2 : VSV system을 이용한 인터페론 역가측정 연구)

  • Jung, In Whoan;Chang, Uk;Kim, Hyun Su;Bai, Jong Chan;Lee, Won Yung
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.27-31
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    • 1985
  • A rapid assay for interferon based on reduction of cytopathic effect was developed with HEp-2 and Vesicular Stomatitis Virus. The number of manipulations and the lengths of the various incubation steps were reduced to minimum. The assay is simple to perform and can be completed with 22-24 hr. Moreover, it was precise method than CPE-reading method.

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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.