• Title/Summary/Keyword: Cytopathic effect(CPE)

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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.

Production and Characteriuation of Monoclonal Antibodies against Human Interferon-$\alpha$ (인터페론 알파에 대한 단세포 군항체의 제조 및 특성)

  • Park, Kyung-Hee;Lee, Ihn-Sook
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.1-7
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    • 1992
  • Seven monoclonal antibodies were produced by fusing splenocytes from Balb/C mouse immunized with partially purified human interferon-a (HUIFN-a) with NSO plasmacytoma cells. aery were identified as five IgG class (432.22: IgG2b/n, 460.52: IgG2b/a , 548.46: IgG2a/n , 573.10: IgG2b/h , 625.12: IgG2b/n ), one IgA class (460.50: IgA/n ) and one IsM class (465.27: IgA/n ), and all of them revealed highly sensitive to HUIFN- a IgG class monoclonal antibodies have pts ranged from 8.2 to 8.6. Ascites fluids produced from primed Balb/c mice and were purified through column chromatography. The cytopathic effect (CPE) inhibition assay to examine neutralization of HuIFU-a by IgG class monoclonal antibodies, gave that MAbs 460.52, 548.46, 573.10 can neutralize HUIFU- a arith varying degrees except 432.22. Therefore, it is deduced that these various monoclonal antibodies may recognize the distinct epitopes on HUIFN-a.

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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.

Effects of substitution of viral hemorrhagic septicemia virus genotype IVa glycoprotein with vesicular stomatitis virus (VSV) glycoprotein on cell line preference

  • Kim, Min Sun;Choi, Tae-Jin;Kim, Ki Hong
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.71-78
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    • 2017
  • The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

A Simple Method for the Concentration of Fish Pathogenic Virus in Sea Water (한외여과막을 이용한 해수내 어류 병원바이러스 농축법)

  • Oh, Myung-Joo;Kim, Suk-Ryul;Jung, Sung-Ju;Kim, Hyeung-Rak;Kim, Heung-Yun;Yeo, In-Kyu
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.61-66
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    • 2000
  • A method was developed for concentrating fish pathogenic virus from sea water using membrane ultrafiltration system and centricon. The method consists of passing large volumes (Ca. 20 liter) of sea water through ultrafiltration (PAN) filter followed by cross-flow filtration method and centrifugation use the centricon (Plus-20). This procedure permitted the processing of 20 liter of sea water which resulted in a 20,000-fold reduction in the volume of water and greater than 90% recovery of the seeded MABV.

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Evaluation of the virulence genes and Shiga toxin-producing abilities of Escherichia coli field isolates causing edema disease in pigs (국내 분리 돼지 부종병 대장균의 병원성 유전자 및 시가독소 생성 검증)

  • Seo, Byoung-Joo;Jeong, Chang-Gi;Kang, A-Rum;Cho, Ho-Seong;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.87-92
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    • 2016
  • Porcine edema disease (ED) is a communicable disease of pigs caused by infection with Shiga toxin (Stx)-producing Escherichia coli (STEC) which expresses F18 fimbriae and/or Stx type 2e (Stx2e). While STEC causes a severe illness including hemorrhagic colitis and hemolytic-uremic syndrome in humans, it induces damage to the vascular endothelium, which results in edema, hemorrhage, and microthrombosis, leading in high mortality in pigs. In the present study, we cultured Stx2e-producing E. coli field isolates from conventional pig farms that experienced sudden deaths previously with symptoms similar to porcine edema disease, which were further investigated with Shiga toxin profiles. A total of 43 strains were identified from the collected samples by F18 or Stx2e specific PCR. Based on the PCR, 42 isolates out of 43 isolates were proved to carry one of F18 or Stx2e genes and 14 isolates to carry both F18 and Stx2e genes. All of the 30 isolates that harbored Stx2e gene induced the cytopathic effect (CPE) in vero cells and especially, the isolate 150229 produced the highest level of Shiga toxin. Therefore, we identified the virulence genes (F18 and Stx2e) and demonstrated Shiga toxin-producing abilities from porcine edema disease causing E. coli filed isolates. These results suggested that one of the isolates could be a vaccine antigen candidate against STEC through further investigating to elicit an immune response.

진흙버섯의 항인플루엔자 활성 및 활성성분 규명

  • Hwang, Byung Soon;Yun, Bong-Sik
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.41-41
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    • 2016
  • Influenza viruses are RNA viruses that belong to the Orthomyxoviridae family, and those can be divided into three types; A, B, and C, which based on the differences of the inner nucleoproteins and genomic structures. All three genera differ in their genomic structure and nucleoprotein content, they are further classified into various serotypes based on the two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). These glycoproteins play crucial roles in viral infection and replication. Hemagglutinin mediates binding of virions to sialic acid receptors on the surfaces of target cells at the initial stage of infection. Neuraminidase cleaves the glycosidic bonds of sialic acids from the viral and cell surfaces to release the mature virions from infected cells, after viral replication. Because NA plays an important role in the viral life cycle, it is considered an attractive therapeutic target for the treatment of influenza. The methanolic extracts of Phellinus baumii and Phellinus igniarius exhibited significant activity in the neuraminidase inhibition assay. Polyphenolic compounds were isolated from the methanolic extracts. The structures of these compounds were determined to be hispidin, hypholomine B, inoscavin A, davallialactone, phelligridin D, phelligridin E, and phelligridin G by spectroscopic methods. Compounds inhibited the H1N1 neuraminidase activity in a dose-dependent manner with $IC_{50}$ values of 50.9, 22.9, 20.0, 14.2, 8.8, 8.1 and $8.0{\mu}M$, respectively. Moreover, these compounds showed anti-influenza activity in the viral cytopathic effect (CPE) reduction assay using MDCK cells. These results suggests that the polyphenols from P. baumii and P. igniarius are promising candidates for prevention and therapeutic strategies against viral infection.

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Distribution of Waterborne Enteric Viruses in Raw Water and Tap Water in Busan Metropolitan City (부산시 상수원수와 수돗물에서의 수인성 장관계 바이러스 분포조사)

  • 박홍기;정은영;이유정;정종문;최동훈;손희종;권기원;홍용기
    • Journal of Life Science
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    • v.13 no.2
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    • pp.197-205
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    • 2003
  • We detested waterborne enteric viruses from the raw water and tap water in Busan metropolitan city by the total culturable virus assay of EPA standard method. According to the results of survey from July 2001 to November 2002, thirteen out of twenty one in raw water samples were positive (61.9%) for enteric viruses and all of the treated water and tap water samples were negative. The enteric viruses in raw water were mainly distributed through the summer to the earl y winter, suggesting the seasonal characteristics of virus distribution in water The titer of enteric viruses per 100 liters of the raw water was ranged from 1.92 to 9.70 MPN by TCVA-MPN program. The isolated viruses were identified as either human poliovirus type 1 or enteroviruses by the immunofluorescent assay.

Prophylactic Effects of Bifidobacterium longum HY8001 against Escherichia coli O157:H7 and Salmonella typhimurium DT104 Enteric Infection and Evaluation of Vero Cytotxin Neutralizing Effects (Bifidobacterium longum HY8001 균주의 Escherichia coli O157:H7과 Salmonella typhimurium DT104 장곤 내 감염 예방효과 및 Vero cytotoxin 중화효과)

  • 양수진;윤장원;서근석;구혜정;김소현;배형석;백영진;박용호
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.419-425
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    • 1999
  • Prophylactic effects of Bifidobacterium longum HY8001, Korean isolate, against Escherichia coli O157:H7 and Salmonella typhimurium DT104 enteric infection were examined at four groups of specific pathogen free(SPF)-ICR mouse for each pathogen. B. longum HY8001+B. typhimurium DT104+B. longum HY8001(BL+ST+BL) group and B. longum HY8001+E. coli O157:H7+B. longum HY8001(BL+E+BL) group were fed with B. longum HY8001 before and after E. coli O157:H7 or s. typhimurium DT104 challenge, while B. longum HY8001+S. typhimurium DT104(BL+ST) and B. longum HY8001+e. coli O157:H7(BL+E) groups were fed with B. longum HY8001 only before E. coli O157:H7 or S. typhimurium DT104 challenge. E. coli O157:H7(E) and S. typhimurium DT104(ST) groups were challenged with each pathogen without B. longum HY8001 administration and control groups were administered with phosphate buffered solution(PBS). After the oral administration with B. longum HY8001(109cfu), th emice were challenged with E. coli O157:H7(2$\times$1010cfu) or S. typhimurium DT104(108cfu) and the mortality rate and the fecal shedding of challenged pathogen were also examined define the reactivity of the B. longum HY8001. Production of toxin neutralizing substance(s) of B. longum HY8001 was determined by cell cytotoxicity assay using Vero cells. Fecal shedding of th eS. typhimurium DT104 was significantly decreased in BL+ST+BL group fed with B. longum HY8--1 before and after challenge(p<0.05), while the fecal shedding s of S. typhimurium DT104 in BL+ST and St groups remained more than 106cfu. the protective effect of the B. longum HY8001 against E. coli O157:H7 was significantly high only in BL+E+BL group fed with b. longum Hy8001 before and after E. coli O157:H7 challenge from the result of fecal E. coli O157:H7 isolation rate, mortality rate, and intestinal contents culture to detect E. coli O157:H7. the mortality rate of the BL+e and E groups. The cytopathic effect (CPE) of the Vero cytotoxin (Shiga like toxin I & II) in Vero cell was neutralized in B. longum HY8001 culture supernatant added wells which indicate the presence of soluble Vero cytotxin neutralizing substance(s) in B. longum HY8001 culture suprnatant.

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