• 약학회지
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• 제43권4호
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• pp.464-468
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• 1999

In order to find less toxic antiviral agents from basidiomycetes, EA, the water soluble substance, was isolated from the carpophores of Elfvingia applanata (Pers.) Karst. Anti-encephalomyocarditis (EMC) virus activity of EA was examined in Vero cells by plaque reduction assay in vitro. And the combined antiviral effects of EA with interferon (IFN) alpha and gamma were examined on the multiplication of EMC virus. EA exhibited a concentration-dependent reduction in the plaque formation of EMC virus with 50% effective concentration ($EC_{50}$) of 2.12 mg/ml. The results of combination assay were evaluated by the combination index (CI) that was analysed by the multiple drug effect analysis. The combination of EA with IFN alpha showed potent synergism with CI values of 0.40~0.60 for 50%, 70% and 90% effective levels, but that with IFN gamma showed antagonism with CI values of 2.16~2.83.

• 한국잠사곤충학회지
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• 제16권2호
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• pp.55-75
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• 1974

가잠에 있어서의 세포질다각체 바이러스의 경난전달감락을 구명하기 위하여 육각형 세포직다각체 바이러스를 전대잠에 접종하고 이에 감염된 자아의 교배조와 비감염된 자아의 교배에서 얻어진 대잠에 있어 유발현상의 차이, 유발.간섭현상의 차이를 관찰하고 분석하여 경난전달감염적 가능성을 밝혔다. 여기에서 얻어진 실험결과는 다음과 같다. 1. 이형 바이러스인 사각형 세포질다각체 바이러스에 의한 육각형 세포질다각체 바이러스의 유발실험에서 감염된 바이러스의 종류를 육각형의 단일감염과 육각형과 사각형의 혼합감염을 모두 기준삼을 때 감염자아겨배군에서는 대조군보다 40내지 60%가 더 높은 유발정도를 보였다. 2. formalin 첨식에 의한 육각형 세포질다각체 바이러스의 우발실험에서 육각형 세포질다각체 바이러에 의한 유발만을 기준 삼을 때 실험군은 대조군에 비햐여 40%의 높은 유발정도를 보였다. 이 실험에서 첨식한 2 또는 3%의 formalin농도는 우발정도에 있어서 차이를 가져오지 못했다. 3. 냉장처리에 의한 육각형 세포질다각체 바이러스의 유발실험에서 육각형 세포질다각체 바이러스에 의한 유발만을 기준삼을 때 실험군 대조군에 비하여 약 50%의 높은 유발정도를 보였다. 이 실험에서 적용한 12 또는 24시간의 첨식처리군간에 의의있는 차이가 인정되지 않았다. 4. 유발.간섭현상에 관한 실험에서 유발과 간섭되는 바이러스를 육각형 세포질다각체 바이러스에 국한시켰을 때 이차 바이러스인 사각형 세포질다각체 바이러스의 처리농도가 높아질수록 일차 바이러스인 육각형 세포질다각체 바이러스 량이 1$\times$$10^{8}$/ml에 이르러서는 대조군과 동일한 수준에 놓이게 되었다. 한편 유발과 간섭되는 바이러스를 사각형 세포질다각체 바이러스에 국한시켰을 때는 얕은 농도의 이차 바이러스는 유발된 일차 바이러스에 의하여 간섭받았고, 이차 바이러스 량이 1$\times$$10^{7}$ /ml에서부터 이차 바이러스에 감염된 가잠이 증가되었으나 대조군보다는 약 30%가 미달되는 감염률을 보였다. 5. 유발.병원성 증진현상에 간한 실험에서 유발과 증진되는 바이러스를 사각형과 육각형 세포질다각체 바이러스의 혼합감염 된것에 국한시켜 보면 이차 바이러스인 사각형 세포질다각체 바이러스의 농도가 높아져서 1$\times$$10^{7}$ /ml에 이르면 혼합감염은 최고도에 이르러 실험군과 대조군간의 차이는 40%를 넘게 되었다. 여기에서 대조군은 이차 바이러스의 농도과 관계없이 아무런 영향을 받지 않았다. 사사 ： 이 연구를 수행함에 있어서 시종 지도해 주신 김낙정 교수님, 최병희 교수님 그리고 박광의 교수님께 심심한 사의를 드리며 본 논문이 완성될 수 있도록 노고를 아끼지 않고 지도하여주신 전윤성 교수님과 정길택 교수님께 또한 심심한 사의를 드립니다. 그리고 본 논문을 위한 실험을 수행함에 있어 전자현미경사용에 적극 협조해 주신 이순형 학형과 누에 사육시험을 도와 준 금근영, 임대준군에게 아울러 감사드립니다. 본 실험에 공시한 세식질다각체 바이러스를 분양하여 주신 동경대학 농학부 도부인 교수님께 또한 감사드립니다.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.924-929
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• 2005

Boiled-water extracts from 101 Korean medicinal plants were tested in vitro for their inhibitory activity against influenza virus type A by means of a modified hemagglutination inhibition test. Thirteen of the 101 extracts exhibited strong anti-influenza virus type A activity at concentrations of less than $780\;{\mu}g/ml$. Out of the above 13 extracts, MW-40 (Chaenomeles speciosa), MW-88 (Citrus junos), and MW-100 (Zingiber officinale) exhibited marked antiviral activity in the concentration range of $0.195\;{\mu}g/ml$ to 100 mg/ml, $0.0487\;{\mu}g/ml$ to 100 mg/ml, and $0.0487\;{\mu}g/ml$ to 100 mg/ml, respectively. The extracts MW-88 and MW-100 were not cytotoxic to red blood cells, whereas MW-40 showed very weak cytotoxicity in the concentration range of 50 mg/ml to 100 mg/ml. Therefore, the present results demonstrate that boiled water extracts of 2 Korean medicinal plants, MW-88 and MW-100, have strong anti-influenza virus type A activity and no cytotoxic effects, and they may inhibit attachment of the virus to the cell and may be used for prophylaxis.

• Journal of Microbiology
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• 제34권3호
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• pp.253-269
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/)] in CHSE, gradual decrease in [Ca$\^$2+/] in FHM, and no significant change in RTG cells. The degree of [Ca$\^$2+/] increase or decrease was dependent on the amount of infectious virus, and these [Ca$\^$2+/] variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in [Ca$\^$2+/] was not observed. Thus, infectivity of IHNV appears to correlate with changes in [Ca$\^$2+/] in virus-infected cells. These IHNV-induced [Ca$\^$2+/] changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced Ca$\^$2+/ variations were more related with protein synthesis than RNA synthesis. Various Ca$\^$2+/ related drugs were used in search for the mechanisms of the [Ca$\^$2+/], changes following IHNV infection of CHSE cells. Decreasing extracellular Ca$\^$2+/ concentration or blocking Ca$\^$2+/ influx from extracellular media inhibited the IHNV-induced increase in [Ca$\^$2+/], in CHSE cells. Similar results were obtained with intracellular Ca$\^$2+/ sources are important in IHNV-induced [Ca$\^$2+/] increase in CHSE cells.

• 미생물학회지
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• 제14권3호
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• pp.135-145
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• 1976

Confluent AGMK cells were infected by large plaque SV40 virus. Levels of DNA polymeras $({\alpha}\;and\;{\beta})$ were measured in the cytoplasm and the cell nucleus. The activities of DNA $polymerase-{\alpha}$ which found in both the cell nucleus and the cytoplasm were increased approximately eight folds at 48 hours after infection of SV40 virus. Only insignificant but constant amounts of DNA $polymerase-{\beta}$ were found either in the nucleus of the SV40 infected cell or of the uninfected cell. The characteristics of the SV40 virus induced DNA polymerases were compared with that of the uninfected cellular DNA polymerase in regard of the effects of pH, salt concentration, NEM concentration and temperature on those enzyme activities. No differential effect was found between both enzymes. Endouclease activities wre examined in the purified DNA $polymerase-{\alpha}\;and\;{\beta}$. The low level of endonuclease activity which might cut SV40 DNA 1 at one site was observed in the DNA $polymerase-{\alpha}$ whereas high but nonspecific endonuclease activities were found in the DNA $polymerase-{\beta}$.

• 미생물학회지
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• 제9권2호
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• pp.61-68
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• 1971

Borrelina virus was inoculated into Hyphantrea cunea DRURY in the labolatory and in the field. The pathogenecity of Borrelina virus upon Bompyx mori L. and Dendrolinus spectabilis BUTLER, too, was examined with following results. 1) $10^8$/ml, $10^7$/ ml, $10^6$/ml concentration of nuclear-polyhedrosis virus was inoculated into the larvae of H.cunea at various ages. The corrected mortality of the larvae were 97.4%, 95.2%, 94.7% in the 3rd instar, and 88.6%, 73.6%, 62.5% in the 6th instar, respectively, with three different concentration of NPV. 2) The symptom of disease of the larvae appeared on 4days after inoculation and most of the larvae were dead within 18 days. 3) The youngest larvae treated with the highest concentration of NPV showed the highest mortality. With older larvae and lower concentriton treated, it appeared that the time needed for death grew longer, marking slower death curve. 4) When we sprayed NPV of $10^6$/ml concentration to H. cunea in the field, the mortality was 94.8% in the first year, 84.6% in the second year and 78.3% in third year. By this, we could admit the continuous effects of the pathogens for several years. 5) About the larvae of B. mori of 3rd and 5th instar and D.spectabilis of 3rd instar inoculated with $10^8$/ml concentration of inoclum, we could not see any pathogenic effects.

• KSBB Journal
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• 제11권4호
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• pp.438-444
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• 1996

모델 재조합 단백질인 $\beta$galactosidase를 발현하 는 Vaccinia virus를 생산하기 위하여 숙주 동물세 포의 배양조건을 관찰한 결과 감염비 5일 경우 H HeLa는 감염후 60시간 후, HeLa 83는 감염후 40 시간 후에 회수할 때 최 대 의 ${\beta}$-galactosidase 수율 을 얻을 수 있으며, 세포가 대수증식기 일 때 감염하 고 배양온도는 $37^{\circ}C$ 로 하는 것이 최적 배양조건으로 나타났다. 감염후 혈청의 농도는 단백질 수율에 크 게 영향을 미치지는 않으나 3~5% 에서 가장 높은 단백질 수율을 보였으며, 낮은 이온 농도의 용액으 로 세포층을 세척하는 것과 virus 감염시 온도를 20~∼$30^{\circ}C$ 로 낮추는 것은 Vaccinia-HeLa system에서 감염능 증대 효과를 나타내 였다. Dexamethasone 전처리는 HeLa 83에서 ViruS 복제 증대를 HeLa에 서는 virus 복제 감소를 가져왔다.

• The Plant Pathology Journal
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• 제19권3호
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• Journal of Microbiology
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• 제42권1호
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• pp.25-31
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• 2004

Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

• 대한바이러스학회지
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• 제29권4호
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• pp.211-219
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• 1999

Most of the currently licensed viral and bacterial vaccines produced in the world are in state of antigen suspension and the immunogenicity of vaccines could be maintained for one or two years only by keeping in the refrigerator, but without refrigeration vaccines would easily lose their immunogenicites. In this study, as a step to develope the method of increasing the stability of vaccines and maintaining the immunogenicity of vaccines for a long time at room temperature or higher temperature, trehalose, glucose and lactose at different concentration were added into the Hantaan virus vaccines and then kept at $37^{\circ}C$ for 12, 24, 48 hours and at room temperature for seven days respectively. Treated vaccines were then inoculated respectively into ICR mice and the titers of antibody against the antigen of Hantaan virus from the mice sera were evaluated. Vaccine without sugar lost immunogenicity completely in 24 hour at $37^{\circ}C$, but the vaccines containing trehalose could maintain some of the immunogenicity even after exposure at $37^{\circ}C$ for 48 hours and the best concentration of trehalose for maintaining the immunogenicities of vaccines was $7.5{\sim}10$ percent. The results suggest that addition of trehalose could increase the stability of Hantaan virus vaccine.