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

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Isolation, identification and epidemiological study of akabane virus on Jeiu-do (제주지역의 akabane virus에 대한 역학조사 및 원인체 분리동정)

  • 강완철;김은주;현관종;전창익;김희석;이두식
    • Korean Journal of Veterinary Service
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    • v.23 no.1
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    • pp.93-102
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    • 2000
  • In this experiment, we studied the sero-positive rate of akabane virus in cattle from Jeju-do and analyzed the seroepidemiological features. In an analysis of 1,051 samples, the positive rate for neutralizing antibody in sera collected in nine regions on Jeju-do was 56.7%. The rate varied with the region. The positive rate was 69.6% in Aewol, 63.1% in Jeju city, 54.4% in Anduck, 51.0% in Hallim, 69.8% in Jocheun, 47.6% in Pyosun, 40% in Daejeong, 30.0% in Harkyung, 71.6% in Namwon, 24.5% in Sungsan, 133.,3% in Seokypo and 44.5% in Gujwa, respectively The rate also depended on the age of the cattle. The positive rate was 67.2% in calves 0- to 12-month old, 48.3% in cattle 13- to 24-month old, 65.4% in cattle 25- to 36-month old, and 65.4% in cattle more than ,B7 months old. To isolate the virus from calves with malformations including arthrogryposis and hydranencephaly, cerebral homogenates were inoculated into Vero cells, which were determined for cytopathic effect (CPE). Vero cells with CPE were examined for Akabane virus using an electron microscope (EM) and indirect immunofluorescent antibody test (EM). Typical virus particles with a width of 90-130nm and specific immunofluorescence in the cytoplasm of infected cells were sought for identification.

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Cytyotoxicity and Anti-Malassezia Activity of Limonene (Limonene의 세포독성과 항Malassezia 활성)

  • Lee, Jeong-Hyun;Lee, Jae-Sug
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.387-389
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    • 2011
  • A previous study of ours indicated that Citrus auranifoli oil possesses antifungal activity against Malassezia furfur and Malassezia pachydermatis. In this study, we evaluated the anti-M. furfur and M. pachydermatis activities of limonene, which is a major component of C. aurantifolia oil, using the disk diffusion method. We also examined cytotoxicity against human normal epithelial (Beas-2B) cells using the cytopathic effect reduction (CPE) method. The results revealed that the minimum fungicidal concentration (MFC) value of limonene is lower than the value for itraconazole. The MFC value of limonene was seen to be 7.81 ${\mu}g$/mL against M. furfur and 3.90 ${\mu}g$/mL against M. pachydermatis. MFC values of itraconazole against M. furfur and M. pachydermatis were 62.50 ${\mu}g$/mL and 31.25 ${\mu}g$/mL, respectively. In addition, it was noted that limonene was not toxic to Beas-2B cells with normal morphology at a concentration of 100 ${\mu}g$/mL. However, itraconazole exhibited weak toxicity at the same concentration. Therefore, our results indicate that limonene could potentially be effective at controlling M. furfur and M. pachydermatis infections with no cytotoxicity.

Anti-Porcine Epidemic Diarrhea Virus (PEDV) Activity and Antimicrobial Activities of Artemisia dubia Essential Oil (참쑥(Artemisia dubia) 오일의 돼지 유행성 설사 바이러스(Porcine Epidemic Diarrhea Virus)에 대한 항바이러스 항균활성)

  • Kim, Jong-Im
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.396-402
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    • 2012
  • The chemical composition, anti-porcine epidemic diarrhea virus (PEDV) activity and antimicrobial activity of Artemisia dubia essential oil were evaluated in this study. Fifty eight compounds from A. dubia essential oil were identified through analysis by gas chromatography-mass spectrometry (GC-MS). The major constituents of the oil were camphor (17.18 %), germacrene-D (15.70%), trans (${\beta}-$) racaryophyllene (6.79%), ene thujones (6.57%), 1, 8-cineole (5.94%) and camphene (5.08%). The essential oil was evaluated for antiviral activity against PEDV in Vero cells using a cytopathic effect (CPE) reduction method. The oils actively inhibited PEDV replication with a 50% inhibitory concentration ($IC_{50}$) of 43.7 ${\mu}^3/mL$. The 50% cytotoxicity concentration ($CC_{50}$) of the oils was over 100 ${\mu}/mL$ and the derived therapeutic index was >2.3. Similar analysis of the ribavirin revealed that they have a relatively weaker efficacy when compared to the oils. The antimicrobial activity of the essential oil against 5 microorganisms was evaluated by the disc diffusion method. The essential oil exhibited antimicrobial activity against 5 tested microorganisms with a clear zone of 8-22 mm. Among the tested microorganisms, Streptococcus pyogenes was the most sensitive and Candida albicans the least. Therefore, in can be concluded that essential oils of A. dubia may have interesting applications for microbial control or the control of PEDV-derived diseases.

Cytotoxic Effects of an Oncolytic Adenoviral Vector AdLPCDIRESE1A in Hepatocellular Carcinoma Cells (암세포 용해성 AdLPCDIRESE1A 벡터의 간암 세포독성효과)

  • Chung, In-Jae
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.75-79
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    • 2011
  • The replication competent adenoviral vector (AV), AdLPCDIRESE1A was generated and reported previously to have cytotoxic effects in some cell lines. In AdLPCDIRESE1A, the expression of cytosine deaminse (CD) and E1A genes are under the control of tumor-specific L-plastin promoter. CD enzyme can deaminate the nontoxic prodrug 5-fluorocytosine (5-FC) to the toxic 5-fluorouracil (5-FU). E1A gene is essential for viral replication. Primary liver cancer, most of which is hepatocellular carcinoma (HCC), is the third common leading cancer in Korea. Thus, we have conducted in vitro preclinical study to evaluate effectiveness of AdLPCDIRESE1A on HCC. The efficacy of cytotoxicity was measured by generation of cytopathic effect (CPE) and cell counting. We infected HepG2 cells with various MOI of vector alone or concurrent with 5-FC. Exposure of cells to AdLPCDIRESE1A generated a significant cytotoxic effect as compared to the control. Almost 83% of the cell had manifested the characteristic cytotoxic effect on day 9 after infection of cells with 10 MOI of vector. We also observed the additive cytotoxic effects when AdLPCDIRESE1A vector had been coadministrated with 5-FC. The results suggest that the use of AdLPCDIRESE1A/5FC may be value in treatment of liver cancer. Further animal studies are needed for clinical trial.

The Combined Anti-apoptotic Effect from Tamiflu and Pinoresinol of Forsythia fructus Extract Against Influenza Virus Infection (연교 추출물 Pinoresinol와 Tamiflu의 병용효과로부터 Influenza Virus 감염에 의한 세포사멸 억제효과)

  • Kim, Sang-Tae;Kim, Jang-Soo;Choe, Young-Uung;Kim, Young-Kyoon
    • Korean Journal of Pharmacognosy
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    • v.42 no.1
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    • pp.9-14
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    • 2011
  • The fruit body of Forsythiae Fructus (Oleaceae), a common Korean medical herb, is widely used in the treatment of cold and inflammation. In order to elucidate the action mechanism and the active principles from the plant against anti-influenza virus, the influenza virus hemagglutinin (HA) and neuraminidase (NA) gene RT-PCR and Viral Screening & Identification (VSI) assay were conducted, and the activity against viral replication was also investigated. Consequently, one active constituent, namely pinoresinol showed the in vitro antiviral principle using a cytopathic effect (CPE) reduction method, indicating pinoresinol possessed anti-influenza viral activity. Furthermore, combination of pinoresinol and Tamiflu exhibited higher activities than Tamiflu alone against influenza virus (H3N2) infection. The results suggested that combination of pinoresinol with Tamiflu could be a better candidate for an ant-H3N2 viral agent in the treatment of the influenza.

Anticancer and Antimicrobial Activities of 13(E)-labd-13-ene-8α,15-diol from Brachyglottis monroi

  • Kim, Jong-Im;Choi, Hwa-Jung;Lee, Jae-Sook
    • Journal of Applied Biological Chemistry
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    • v.56 no.1
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    • pp.49-51
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    • 2013
  • In a previous study, we reported that 13(E)-labd-13-ene-$8{\alpha}$,15-diol (13E) possesses antiviral and anticancer activities. In this study, the anticancer and antimicrobial activities of 13(E) were evaluated against 4 cancer cell lines and 6 bacteria. 13(E) showed inhibitory effect on a variety of cancer cell lines. The $IC_{50}$ values was 8.3-21.3 ${\mu}g/mL$. 13(E) was the most effective growth inhibitor of murine leukaemia cell lines P388, producing approximately 8.3 ${\mu}g/mL$ of $IC_{50}$ in the cytopathic effect (CPE) method. 13(E) also inhibited the growth of the gram-positive bacteria (Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes) and gram-negative bacteria (Vibrio parahaemolyticus, Escherichia coli and Salmonella enteritidis) with a range of minimum inhibitory concentration (MIC) values from 0.092 to 0.598 mg/mL and gram-negative bacteria were more sensitive to the compound (MIC, 0.092 mg/mL).

Antiviral Activity of Hederasaponin B from Hedera helix against Enterovirus 71 Subgenotypes C3 and C4a

  • Song, JaeHyoung;Yeo, Sang-Gu;Hong, Eun-Hye;Lee, Bo-Ra;Kim, Jin-Won;Kim, JeongHoon;Jeong, HyeonGun;Kwon, YongSoo;Kim, HyunPyo;Lee, SangWon;Park, Jae-Hak;Ko, Hyun-Jeong
    • Biomolecules & Therapeutics
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    • v.22 no.1
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    • pp.41-46
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    • 2014
  • Enterovirus 71 (EV71) is the predominant cause of hand, foot and mouth disease (HFMD). The antiviral activity of hederasaponin B from Hedera helix against EV71 subgenotypes C3 and C4a was evaluated in vero cells. In the current study, the antiviral activity of hederasaponin B against EV71 C3 and C4a was determined by cytopathic effect (CPE) reduction method and western blot assay. Our results demonstrated that hederasaponin B and 30% ethanol extract of Hedera helix containing hederasaponin B showed significant antiviral activity against EV71 subgenotypes C3 and C4a by reducing the formation of a visible CPE. Hederasaponin B also inhibited the viral VP2 protein expression, suggesting the inhibition of viral capsid protein synthesis.These results suggest that hederasaponin B and Hedera helix extract containing hederasaponin B can be novel drug candidates with broad-spectrum antiviral activity against various subgenotypes of EV71.

Isolation and Characterization of White Spot Syndrome Baculovirus in Cultured Penaeid Shrimp (Penaeus chinensis) (양식새우(Penaeus chinensis)에서의 White Spot Baculovirus의 분리 및 특성)

  • Heo, M.S.;Sohn, S.G.;Sim, D.S.;Kim, J.W.;Park, M.A.;Lee, J.S.;Choi, D.L.;Jung, S.H.;Kim, Y.J.;Oh, M.J.
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.7-13
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    • 2000
  • Beginning in the summer of 1993, a serious mortality among cultured penaeid shrimp occurred in the western sea of Korea. The typical sign of this disease was white spots inside the surface of the carapace. Cytopathic effect (CPE) were not observed by virus in CHSE-214, RTG-2, but not by pH 11. A nonoccluded rod-shaped form virus was observed by electron microscopy in the lymphoid organ. The virion was bacilliform virus and sourrounded by a virion envelope. Its virion protein was found to be similar to hypodermal and hematopoietic necrosis virus (HHNBV) by analysis of virion proteins in SDS-PAGE. The genome of virus is double stranded DNA molecule whose full length was about 114kb. It was similar to penaeus acute viremia (PAV) of Japan.

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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.

Anti-herpetic Activity of Various Medicinal Plant Extracts

  • Woo, Eun-Rhan;Kim, Hyoung-Ja;Jong, Hwan-Kwak;Lim, Young-Kwern;Park, Seong-Kyu;Kim, Hae-Soo;Lee, Chong-Kyo;Park, Hokoon
    • Archives of Pharmacal Research
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    • v.20 no.1
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    • pp.58-67
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    • 1997
  • In order to find antiviral compounds against Herpes simplex virus type I (HSV-1) and II (HSV2) from natural products, a convenient virus-induced cytopathic effect (CPE) inhibition assay was introduced. More than 300 fractions were prepared by solvent fractionation from sixty collected plants or purchased herbal medicines, and their anti-herpetic activities were evaluated. Among them, several medicinal plants showed potent anti-herpetic activity. Selective indexes (SI) of the EtOAc extract of Caraganae Radix (Caragana sinica) against HSV-1 and HSV-2 were more than 8.06 and 24.79, SI of the MeOH extract of Acer okamotoanum leaves were 3.92 and 3.51, SI of the $CH_{2}Cl_{2}$ extract of Veratri Rhizoma et Radix (Veratrum patulum) were 5.49 and 1.31 and SI of the MeOH extract of aerial part of Osmundae Rhizoma (Osmunda japonica) were more than 3.45 and 1.25, respectively.

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