• Title/Summary/Keyword: Virus concentration

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The Change of Cytosolic Free Calcium Concentration Following Herpes Simplex Virus Type-1 (HSV-1) Infection (Herpes Simplex Virus Type-1 (HSV-1) 감염에 따른 세포내 유리 $Ca^{2+}$농도의 변화)

  • 남윤정;이규철;이찬희
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.306-311
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    • 2000
  • Infection of Vero cells with herpes simplex virus type-1 (HSV-1) resulted in a series of changes in intra-cellular free calcium concentration $([Ca^{2+}]_i)$. A significant and maximal decrease $[Ca^{2+}]_i$ was observed at 4 hours postinfection (hr p.i.) in HSV-1-infected in Vero cells. Inactivation of HSV-1 with UV irradiation and heat treatment abolished HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells. And the degree of the decrease in $[Ca^{2+}]_i$ was dependent on the amount of input virus. Taxol, which stabilizes the polymerization of microtubule blocked HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i., suggesting that microtubule may mediate the transport of HSV-1 nucleocapsid to the nucleus of infected cell. Treatment of HSV-1-infected Vero cells with metabolic inhibitors such as cycloheximide, cordycepin, or acyclovir partially reversed the decrease in $[Ca^{2+}]_i$ at 4 hr p.i.. Thus, it is suggested that HSV-1 induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells may play an important role in the multiplication of HSV-1.

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Studies on the production of Japanese encephalitis virus vaccine (일본뇌염백신 제조에 관한 연구)

  • 유건희;이용재
    • Korean Journal of Microbiology
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    • v.9 no.4
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    • pp.175-178
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    • 1971
  • Because of the cases of Japanese Encephalitis(J.E.) were reported every year in Korea. We, Dong-A Pharmaceutical Co., Ltd., produced J.E. virus vaccine, with lower price, since 1970 in order to prevent ourselves from being infected by the disease. And inoculated the J.E. virus vaccine for the children with a great success. We are going to report several questions which brought about in producing the J.E. virus vaccine by alcohol precipitation, protamine sulfate treatment method. The results obtained were as folows ; 1) In process treated with 40% alcohol, we used to ethanol made in Germany, but it was too expensive to use it. As the result which we had studied about it, we were satisfied with J.E. virus vaccine which produced with alcohol made in Korea, and then, we treated with accurate specific gravity of 40% ethanol for the precipitation of the virus. And also, we knew that it was the best method to be treated it for 3hrs, $13^{\circ}C$. 2) When we treated with protamine sulfate (0.025mg/ml), we acquired the highest potent titer, and suited into purpose for the nitrogen concentration. 3) The filtration of the purified J.E. virus vaccine, in case of millipore filter paper of large pore size was not suitable for the sterility. Therefore the pore size less than 0.8.$\mu$ (AA filter paper) in millipore filter paper was very suitable. But it seemed to be important subhects that the smaller was the pore size, the lower was the potent titer.

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Study on the Anti-influenza Virus A type Activity of Citrus junos (유자의 항 Influenza 바이러스 A형 활성에 관한 연구)

  • Kim, Ho-Kyoung;Ko, Byoung-Seob;Jeon, Won-Kyung
    • Korean Journal of Pharmacognosy
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    • v.31 no.1
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    • pp.82-86
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    • 2000
  • To evaluate anti-influenza virus activity of 113 specimens of Korean traditional medicine both water and methanol extracts were examined using haemagglutination inhibition test. The water extract from Citrus junos was found to inhibit influenza virus A/Taiwan/l/86(H1N1). The survival rates of virus were determined by in situ cellular enzyme-linked immunosorbent assay. The water extract of Citrus junos was fractionated by chromatographic separating using Amberlite XAD-4, 40% MeOH and 60% MeOH layer had antiviral activity. The half inhibition concentration $(IC_{50})$ of 40% MeOH layer on survival of influenza virus was $MIC>361.5{\mu}g/ml$ and $IC_{50}$ value of fr. 40-4 fractionated from 40% MeOH layer was $677.19{\mu}g/ml$. These results suggested that the fractions of Citus junos have potent anti-influenza A virus activity.

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Effect of Metals on Tobacco Mosaic Virus Infection (담배모자이크 바이러스 감염성에 대한 금속의 영향)

  • Choi, C.W
    • The Journal of Natural Sciences
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    • v.10 no.1
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    • pp.23-26
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    • 1998
  • The efficacy of various concentration of divalent copper and zinc ions was evaluated separately for the infectivity of tobacco mosaic virus. Infectivity of TMV was more enhanced by addition of zinc, while it was decreased by addition of copper. The number of local lesions were more produced on tobacco leaves inoculated with inoculum sap containing zinc than those inoculated with sap only. The effect of copper inhibited the infectivity of TMV is dependent on copper concentration. TMV particles treated with various concentration of zinc and copper, respectively, analyzed by electrophoresis, and appeared to be altered in electrophoretic behavior. When TMV was exposed to zinc concentration at more than 200mM, the viral particles were completely degraded, and at 40-20 mM they were barely detectable, but at 2 mM they were quite stable. When TMV was exposed at less than concentration of 20 mM of copper were degraded.

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Detection of antibodies in swine serum to Aujeszky's disease virus using agar-gel immunodiffusion test (Agar-gel immunodiffusion test를 이용한 돼지 혈청중 Aujeszky's disease virus 항체 검출에 관한 연구)

  • Cho, Hyo-gueon;Jun, Moo-hyung
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.297-307
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    • 1990
  • To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

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Antiviral activity of Herba Patrinea (a Chinese medicinal herb) against respiratory syncytial virus in vitro

  • Li, Hong-Yuan;Li, Shan-Shan;Liu, Dian-Li;Dong, Yan-Mei;Tian, Wen-Jing
    • Advances in Traditional Medicine
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    • v.3 no.2
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    • pp.106-110
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    • 2003
  • Respiratory syncytial virus (RSV) has long been considered an important cause of severe lower respiratory tract infection in infants and young children throughout the world. Unfortunately, no effective treatment of RSV exists. Therefore, New agents are needed to reduce the impact of RSV. We have studied the anti-viral effect of traditional Chinese midicinal herbs for over ten years and find Herba Patrinea (a Chinese medicinal herb) has the anti-RSV effect in vitro. In this study, the Herba Patrinea was extracted with hot water, condensed and sterilized. The cytotoxicity of the aqueous extract was tested by adding the diluted extract directly to HeLa cells and its effect on anti-RSV was estimated by the CPEI assay. As a result, the median cytotoxic concentration $(CC_{50})$ of Herba Patrinea was 32 mg/ ml by morphological observation, the median effective concentration (50% effective concentration, $EC_{50}$) of the Herba Patrinea against replication of the Long strain of RSV in HeLa cells were 1.25 mg/ml. The selectivity index $(SI=CC_{50}/EC{50})$ is 25.6. Moreover, Herba Patrinea gave a dose-dependent response in inhibiting RSV. In time of addition experiment, Herba Patrinea inhibited replication of RSV in HeLa cells when it was added at 0h, 2h, and 4h after virus infection. In summary, the results of this study suggest Herba Patrinea may be a novel anti-RSV drug and it is worthy of further studying.

Effects of fetal bovine serum concentrations on viral infectivity titers of infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus (Fetal bovine serum의 농도에 따른 infectious hematopoietic necrosis virus와 infectious pancreatic necrosis virus의 감염가 변화)

  • Kim, Hyoung Jun;Park, Jeong Su;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.81-85
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    • 2018
  • Fetal bovine serum (FBS) is an essential element of cell growth and can also affect the viral replication. In this study, we tried to find out whether FBS concentration affects the viral infectivity titer of IHNV and IPNV. EPC cells were suspended with MEM supplemented with various concentrations of FBS (MEM0, MEM2, MEM5 and MEM10) and cultured in 96-well plate. Each virus was 10-fold diluted virus and inoculated in 96-well plate. The highest infectivity titer of IHNV was $10^{7.88}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{7.30}\;TCID_{50}/mL$ in 96-well plate using MEM10. The highest infectivity titer of IPNV was $10^{7.47}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{6.97}\;TCID_{50}/mL$ in 96-well plate using MEM10. This study showed that not only 0% FBS but 10% FBS leads low infectivity titer of IHNV and IPNV. Therefore, it is considered that the desirable concentration of FBS is 2% or 5% for measurement of infectivity titer of IHNV and IPNV.

Expression and characterization of RNA-dependent RNA polymerase of Ectropis obliqua virus

  • Lin, Meijuan;Ye, Shan;Xiong, Yi;Cai, Dawei;Zhang, Jiamin;Hu, Yuanyang
    • BMB Reports
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    • v.43 no.4
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    • pp.284-290
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    • 2010
  • Replication of positive-strand RNA virus is mediated by a virus-encoded RNA-dependent RNA polymerase (RdRp). To study the replication of Ectropis obliqua virus (EoV), a newly identified insect virus belonging to the family Iflaviradae, we expressed the RNA polymerase domain in Escherichia coli and purified it on a Ni-chelating HisTrap affinity column. It is demonstrated that EoV RdRp initiated RNA synthesis in a primer and poly (A)-dependent manner in vitro. Furthermore, the effect of primer concentration, temperature, metal ions ($Mg^{2+}$, $Mn^{2+}$, and $K^+$) on enzymatic activity were determined. Our study represented a first step towards understanding the mechanism of EoV replication.

Antiviral Activity of Water Soluble Substance from Elfvingia applanata (잔나비걸상버섯 수용성물질의 항바이러스효과)

  • Rym, Kyo-Hwan;Eo, Seong-Kug;Kim, Young-So;Lee, Chong-Kil;Han, Seong-Sun
    • Korean Journal of Pharmacognosy
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    • v.30 no.1
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    • pp.25-33
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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. EA was examined for antiviral activity against five strains of pathogenic viruses such as encephalomyocarditis virus (EMCV), vesicular stomatitis virus (VSV) Indiana and New Jersey strains, influenza A virus (Flu A), and varicella zoster virus (VZV) in vitro. Antiviral activity was evaluated by plaque reduction assay. Among five strains of viruses tested, EA exhibited the most potent antiviral activity against VSV Indiana strain with 50% effective concentration $(EC_{50})$ of 0.104 mg/ml in Vero cells, and its selectivity index (SI) was 36.5. EA was also examined for the virucidal activity, antiviral activity in preincubation on VSV Indiana strain in order to examine possible mode of antiviral activity. Preincubation of Vero cells with EA did not confer protection against VSV, however, prolonged exposure of cells to EA inhibited the replication of virus dose-dependently. In virucidal activity, the titer of infectious virus did not decrease significantly.

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Detection of Influenza A Virus by Interdigitated Nanogap Devices

  • Park, Jimin;Park, Dae Keun;Lee, Cho Yeon;Kang, Aeyeon;Yun, Wan Soo
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.419-419
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    • 2014
  • Interdigitated nanogap device (IND) is an attractive tool for biomolecular detection due to its huge on-off signal ratio, great tolerance to the variation in biochemical environment, and relatively simple implementation processes. Here, we report on the IND-based detection of Influneza A virus by sandwich immunoassay. The INEs were fabricated by photo lithography followed by the in-house chemical lithographic technique for the narrowing the initial gap distance. The surface of the silicon oxide between the two gold electrodes was chemically modified to immobilize primary antibodies for the immuno-specific interaction with the influenza A virus antigen. After immersing the functionalized-IND into the sample solution containing the influenza A virus, the device was exposed to the secondary antibody conjugated Au nanoparticles (Au NPs). The INDs showed a huge jump in the electric conductance when the sample solution contained the influenza A virus of the concentration as low as 10 ng/mL. We hope that this IND-based sensing can be applied to the development of simple and reliable diagnostic means of influenza viruses.

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