• Osong Public Health and Research Perspectives
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• v.9 no.6
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• pp.348-353
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• 2018

Objectives: This study was conducted to determine whether essential oils had anti-influenza A/WS/33 virus activity and whether there were specific compounds associated with this activity. Methods: There were 63 essential oils evaluated for anti-influenza (A/WS/33 virus) activity using a cytopathic effect reduction method. The chemical composition of the anti-influenza essential oils was phytochemically analyzed by gas chromatography-mass spectrometry. Results: The antiviral assays demonstrated that 11 of the 62 essential oils ($100{\mu}g/mL$) possessed anti-influenza activity, reducing visible cytopathic effects of influenza A/WS/33 virus activity by > 30%. Furthermore, marjoram, clary sage and anise oils exhibited anti-influenza A/WS/33 virus activity of > 52.8%. However, oseltamivir (the anti-influenza A and B drug), showed cytotoxicity at the same concentration ($100{\mu}g/mL$) as the essential oils. The chemical composition detected by GC-MS analysis, differed amongst the 3 most potent anti-viral essential oils (marjoram, clary sage and anise oils) except for linalool, which was detected in all 3 essential oils. Conclusion: This study demonstrated anti-influenza activity in 11 essential oils tested, with marjoram, clary sage and anise essential oils being the most effective at reducing visible cytopathic effects of the A/WS/33 virus. All 3 oils contained linalool, suggesting that this may have anti-influenza activity. Further investigation is needed to characterize the antiviral activity of linalool against influenza A/WS/33 virus.

• Korean Journal of Pharmacognosy
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• v.30 no.3
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• pp.244-249
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• 1999

In order to find less toxic antiviral agents from Basidiomycetes, EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata(Pers.) Karst. Antiviral activity of EA against vesicular stomatitis virus [Indiana serotype, VSV(IND)] was examined in Vero cells using plaque reduction assay in vitro. And the combined antiviral effects of EA with interferon (IFN) alpha or gamma were examined on the multiplication of VSV(IND). EA caused a concentration-dependent reduction in the plaque formation of VSV(IND) with 50% effective concentration $(EC_{50})$ of $104.02\;{\mu}g/ml$. The results of combination assay were evaluated by the combination index (CI) that was analysed by the multiple drug effect analysis. All cases of the combination of EA with IFN alpha or IFN gamma showed potent synergism with CI values of $0.38{\sim}0.52$ for $50{\sim}90%$ effective levels.

• 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.

• Korean Journal of Pharmacognosy
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• v.34 no.1 s.132
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• pp.28-32
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• 2003

In order to search for the anti-HIV agents from natural products, eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. Repeated column chromatoghaphy of the methylene chloride fraction of A. altissima afforded compounds 1$({\beta}-sitosterol-3-O-{\beta}-D-glucoside)$, 2(tetramethoxycoumarin), and 3(ocotillone). Virus-cell fusion inhibitory activity of compound 3(ocotillone) was $70.76{\pm}4.09%$ at the concentration of $100\;{\mu}g/ml$.

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.32-38
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• 2001

Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.

• Journal of Veterinary Science
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• v.22 no.6
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• pp.87.1-87.12
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• 2021

Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5' untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10⁰ copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions: The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.

• Korean Journal of Veterinary Research
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• v.30 no.1
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• pp.65-71
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• 1990

Rabbits were experimentally infected with rabbit hemorrhagic disease virs and the viral pathogenicity, hemagglutinability, and the effect of physicochemical factors were studied. The experimental results were summariaed as follows: 1. Mean rectal temperature of 11 infected rabbits was $40.0{\pm}0.47^{\circ}C$ prior to the virus inoculation, and $39.9{\pm}0.75^{\circ}C$ after 12hrs., $40.2{\pm}0.65^{\circ}C$ after 24hrs., $40.1{\pm}0.77^{\circ}C$ after 36hurs, and $40.6{\pm}0.56^{\circ}C$ just before the death. 2. Mean death time of infected rabbits was $40.3{\pm}22.0$ honrs and its range was 24 to 93 hours. 3. O, B, AB and A type of human erythrocytes were shown their HA in the order, but rabbit and chicken erythrocytes were not hemagglutinated by the virus. 4. In the hemagglutination, less than 0.25 per cent of a final concentration of erythrocytes and 0.2 per cent of BSA in PBS resulted in the best hemagglutination. Phosphate concentration in a range of 0.01M to 0.10M in PBS was not influenced on the hemagglutination reaction, and its pH 7.0 resulted in a better HA. 5. The hemagglutinating titers, in log 2 scale, of organs and tissues of the virus infected rabbits were $9.3{\pm}3.8$ (liver), $9.1{\pm}3.9$ (blood), $6.2{\pm}2.6$ (spleen) and $5.0{\pm}2.5$ (kidney). 6. The physicochemical factors such as heating ($50^{\circ}C$, 10 min.), trypsin treatment (0.05 pre cent, 5 min.), acid treatment (pH 3.0, 20 min.) and ether extraction (3 times) were not affective to the stability of virus and viral HA activities.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.728-732
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• 2000

Propagation of Anagrapha falcifera nuclear polyhedrosis virus(AfNPV) was investigated using well-plates and split-flow air-lift bioreactors. In well-plate experiments, the effects of pH, cell density at a point of infection, serum concentration, DEAE-dextran, and lipid on virus propagation were all closely examined. The AfNPV titer in well-plates was optimal at pH 6.8 and $3{\times}10^6$ cells/$cm^2$. The virus titer was not dramatically affected when the fetal bovine serum concentration was reduced from 10% to 5%. The addition of cholesterol at AfNPV infection of Sf21 cells enhanced the virus titer, whereas the addition of DEAE-dextran did not improve the titer. The AfNPV titer ($3.8{\times}10^7$ $TCID_{50}/ml$) at optimized conditions for well-plate experiments was 2.5-fold higher than for the control. In bioreactor experiments, the AfNPV titer showed its maximum level at air flow rates of 20-40 ml/min. In a split-flow air-lift bioreactor, AfNPV titer ($2.3{\times}10^7\;TCID_{50}/ml$) was 1.5-fold higher than the control when the culture was at pH 6.8 and supplemented with 0.34 mM cholesterol.

• BMB Reports
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• v.53 no.3
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• pp.166-171
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• 2020

A chemical library comprising 2,354 drug-like compounds was screened using a transcription and replication-competent viruslike particle (trVLP) system implementing the whole Ebola virus (EBOV) life cycle. Dose-dependent inhibition of Ebola trVLP replication was induced by 15 hit compounds, which primarily target different types of G protein-coupled receptors (GPCRs). Based on the chemical structure, the compounds were divided into three groups, diphenylmethane derivatives, promazine derivatives and chemicals with no conserved skeletons. The third group included sertindole, raloxifene, and ibutamoren showing prominent antiviral effects in cells. They downregulated the expression of viral proteins, including the VP40 matrix protein and the envelope glycoprotein. They also reduced the amount of EBOV-derived tetracistronic minigenome RNA incorporated into progeny trVLPs in the culture supernatant. Particularly, ibutamoren, which is a known agonist of growth hormone secretagogue receptor (GHSR), showed the most promising antiviral activity with a 50% effective concentration of 0.2 μM, a 50% cytotoxic concentration of 42.4 μM, and a selectivity index of 222.8. Here, we suggest a strategy for development of anti-EBOV therapeutics by adopting GHSR agonists as hit compounds.

• Archives of Pharmacal Research
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• v.24 no.1
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• pp.74-78
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• 2001

A preparation of water soluble components (EA) was made from carpophores of Elfvingia applanata (Pers.) Karst and its in vitro antiviral activity on vesicular stomatitis virus [(Indiana serotype, VSV(IND)] was investigated by plaque reduction assay. EA exhibited potent antiviral activity on VSV(IND) growth and negligible cytotoxicity on Vero cells, 50% effective concentration ($EC_{50}C$/) of 104$ug\textrm\/ml$ and 50% cytotoxic concentration ($CC_{50}C$) of 3,793$ug\textrm\/ml$, respectively. Selectivity index (Sl $CC_{50}C$/$EC_{50}C$) of EA on Vero cell and VSV(IND) was about 36.5. EA did not display either a direct virucidal effect on V5V(IND) or induction of antiviral substance by Vero cells upon its treatment. Thus, the mode of antiviral activity of EA was studied at steps of viral adsorption onto cell. When both EA and virus were added to cell monolayers, titer of cell-free virus in culture supernatant increased in ca. 30-40% compared with that of control group and titer of cell-associated virus was 60-100% higher than that of control group. These results suggested that antiviral activity of EA on VSV(IND) might be due to the hindrance of viral entry to cells at eITher endocytosis or loss of envelope.