• Journal of fish pathology
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• v.5 no.1
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• pp.1-7
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• 1992

CHSE(Chinook Salmon Embryo)-214 fish cell lines was cultured in Eagle's minimal medium (MEM) supplemented with 10% fetal bovine serum and 2mM-glutamin. Optimum growth temperature of CHSE-214 cell line was $20^{\circ}C$. Infectious pancreatic necrosis virus(IPNV) was successfuly multiplied and showed the cytopathic effect in CHSE-214 cell line. Infection symptom of IPNV was observed with inverted phase contrast microscopy. At 6h-12hrs post-infection, the cells infected with IPNV were similer to normal cells. At 18-24hrs post-infection, the cells were somewhat round form and a little swollen form than normal cells. At 30hs post-infection, the cells were becoming more abnormal cells. At 48-68 post-infection, the infected cells were lysed and showed the severe cytopathic effect.

• Korean Journal of Veterinary Research
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• v.32 no.1
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• pp.77-82
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• 1992

A noncytopathic bovine viral diarrhea virus(NC BVDV) strain isolated and purified from persistently infected primary bovine fetal lung(BFL) cells was studied by biophysical methods. The buoyant density of particles of the NC BVDV strain was shown to be between 1,090 and $1,114g/cm^3$ and the maximum virus infectivity occured at $1,098g/cm^3$. Immunoelectron microscopic examination by using the partially purified virus revealed regular spherical particles 30~80nm in diameter. Differences in the genomic size of cytopathic and noncytopathic BVDV from infected cells were not found. A comparison of viral proteins of a noncytopathic and cytopathic strain(NADL) by immunoprecipitation using monoclonal antibody indicated that NC BVDV, compared to cytopathic NADL, was cell associated.

• Parasites, Hosts and Diseases
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• v.49 no.3
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• pp.285-290
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• 2011

Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A $^{51}Cr$ release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-${\alpha}$, IL-6, and IL-$1{\beta}$, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response.

• Parasites, Hosts and Diseases
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• v.57 no.3
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• pp.217-223
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• 2019

Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of $IL-1{\alpha}$, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of $IL-1{\alpha}$, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

• Journal of Applied Biological Chemistry
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• v.51 no.3
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• pp.123-127
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• 2008

Anti-enterovirus 71 (EV 71) activities of fifteen herb plant species extracts were examined by SRB assay, among which Origanum vulgare and Rosmarinus officinalis (Anna Rosemary) extracts exhibited the activities with $IC_{50}$ of 8.28 and $8.17\;{\mu}g/mL$, respectively. Their 50% cytotoxicity concentrations ($CC_{50}$) were 691.89 and $1104.19\;{\mu}g/mL$, and the therapeutic indices were 83.56 and 135.15, respectively. Amantadine (positive control) showed anti-EV 71 activity with 50% inhibitory concentration and $CC_{50}$ of 4.46 and $145.22\;{\mu}g/mL$, respectively. Addition of the methanol extracts of O. vulgare and R. officinalis (Anna Rosemary) in EV 71-infected Vero cells strongly inhibited the formation of visible cytopathic effects without changing the normal morphology of the cells. These results indicate that methanol extracts of O. vulgare and R. officinalis (Anna Rosemary) may contain antiviral compound inhibiting the EV 71 replication.

• The Journal of Internal Korean Medicine
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• v.21 no.2
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• pp.291-297
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• 2000

Objective : In order to find antiviral compounds against Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, a convenient virus-induced cytopathic effect(CPE) inhibition assay was introduced. Methods : Fourteen purchased herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Results : Cytotoxic concentration (CC) of the $H_2O$ extracts of Hyongbangpaedoksan against HSV-1 and HSV-2 was 181.12. This is high level cytotoxic concentration compared with the contrast. Therefore, we assumed that the high level cytotoxic concentration of herbal medicine play a major role in improvement of antiviral activity at the first infective cell. But antiviral effect was unable to figure out for selective index(Sl)=CC50/EC50. The other herbal medicines were unable to showed potent anti-HSV activity. Conclusions : The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.

• BMB Reports
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• v.38 no.1
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• pp.34-40
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• 2005

GLPG (Ganoderma lucidum proteoglycan) was a bioactive fraction obtained by the liquid fermentation of the mycelia of Ganoderma lucidum, EtOH precipitation, and DEAE-cellulose column chromatography. GLPG was a proteoglycan with a carbohydrate: protein ratio of 10.4: 1. Its antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were investigated using a cytopathic inhibition assay. GLPG inhibited cell death in a dose-dependent manner in HSV-infected cells. In addition, it had no cytotoxic effect even at 2 mg/ml. In order to study the mode of action of the antiviral activity of GLPG, cells were treated with GLPG before, during, and after infection, and viral titer in the supernatant of cell culture 48 h post-infection was determined using a $TCID_{50}$ assay. The antiviral effects of GLPG were more remarkable before viral treatment than after treatment. Although the precise mechanism has yet to be defined, our work suggests that GLPG inhibits viral replication by interfering with the early events of viral adsorption and entry into target cells. Thus, this proteoglycan appears to be a candidate anti-HSV agent.

• Korean Journal of Veterinary Service
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• v.20 no.4
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• pp.331-338
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• 1997

The purpose of this study was to establish early diagnosis for bovine virus diarrhea-mucosal disease(BVD-MD) Two Holstein among 22 breeding cows were shown ulcer in the mouth and watery diarrhea. Diarrheal feces and ulcerous lesion of the mouth from 2 cows were sampled for detection of viral antigen. BVD virus was isolated by inoculation of the samples to MDBK cells, and the cytopathic effects were observed in cultured MDBK cells which inoculated with virus isolates from the feces. Viral antigens were detected in the feces and ulceruous lesion by immunogold staining. The serum neutralization titers were shown 1 : 64 or greater in 8 blood samples by using BVD virus (NADL strain). By the RT-PCR, using reverse primer 5'-ACTCCATGTGCCATGTACAG-3', forward primer 5'-ACTCCATGTGCCATGTACAG-3', 285 base pair band specific to BVD virus was detected. In conclusions, the results of above tests which executed using the diarrheal feces and ulcerous lesion of the mouth and the isolates were conformed as BVD virus.

• Archives of Pharmacal Research
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• v.28 no.11
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• pp.1293-1301
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• 2005

Flavonoids, a group of low molecular weight phenylbenzopyrones, have various pharmacological properties including antioxidant, anticancer, bactericidal, and anti-inflammatory. We carried out anti-herpetic assays on 18 flavonoids in five classes and a virus-induced cytopathic effect (CPE) inhibitory assay, plaque reduction assay, and yield reduction assay were performed. When flavonoids were applied at various concentrations to Vero cells infected by HSV-1 and 2, most of the f1avonoids showed inhibitory effects on virus-induced CPE. Among the flavonoids, EC, ECG (flavanols), genistein (isoflavone), naringenin (flavanone), and quercetin (flavonol) showed a high level of CPE inhibitory activity. The antiviral activity of flavonoids were also examined by a plaque reduction assay. EC, ECG, galangin, and kaempferol showed a strong antiviral activity, and catechin, EGC, EGCG, naringenin, chrysin, baicalin, fisetin, myricetin, quercetin, and genistein showed moderate inhibitory effects against HSV-1. In these experiments, flavanols and flavonols appeared to be more active than flavones. Furthermore, treatment of Vero cells with ECG and galangin (which previously showed strong antiviral activities) before virus adsorption led to a slight enhancement of inhibition as determined by a yield reduction assay, indicating that an intracellular effect may also be involved.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.207-210
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• 2014

Viral infections are capable of inducing reactive oxygen species (ROS) production in the infected cells and antioxidants have been reported to have antiviral activities against many viruses. In this study, an antiviral assay using the cytopathic effect (CPE) reduction method revealed that gallic acid possesses good anti-coxsackievirus B3 (CB3) and coxsackievirus B4 (CB4) activities, reducing the formation of visible CPE. However, ribavirin did exhibit weak anti-CB3 and CB4 activities and was unable to prevent CPE. Therefore, we conclude that the inhibition of CB3 or CB4 production by gallic acid may be due to its general action as an antioxidant.