• 한국어병학회지
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• 제15권1호
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• pp.9-16
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• 2002

The objective of the study was to clarify the mechanism of persistent infection of marine birnavirus (MABV) in various nonpermissive cell lines. It was observed in CHSE-214, RTG-2 and RSBK-2 that the virus produced at high yield with typical cytopathic effect (CPE). On the contrary, the CPE was not produced in EPC, FHM and BF-2 cells. However amount of virus protein in both permissive and nonpermissive cell lines detected by ELISA was almost the same. Electron microscopy showed virions in permissive cells but not in nonpermissive cells. From the results, it is clear that virus protein and RNA were produced in nonpermissive cells as observed in permissive cells; however, assembly of the virus particles did not occur in nonpermissive cells.

• 미생물학회지
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• 제22권2호
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• pp.85-90
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• 1984

Two viruses were isolated from kidney and spleen tissues of goldfish(Carasius auratus) and the ovarian fluid of chum salmon(Oncorhynchus keta). Both viruses replicated and produced cytopathic effect in EPC, CHSE-214, and CHH-1 cell lines at $15^{\circ}C$. The isolates were resistant to pH 3 and choloroform. Antiserum to infections pancreatic necrosis virus(IPNV) serotype VR 299 neutralized the infectivity of both of the isolates. Electron microscopy showed that the particles had typical IPNV particle morphology with average diameters of 55nm, This paper describes the first isolation of viruses infecting cultured fish in Korea.

• 한국미생물·생명공학회지
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• 제40권4호
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• pp.396-402
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• 2012

이 연구에서 참쑥(Artemisia dubia) 오일의 화학적 조성, 돼지 유행성 설사 바이러스(porcine epidemic diarrhea virus, PEDV)에 대한 항바이러스 활성 및 항균 활성을 평가하였다. 참쑥 오일로부터 질량 분석기가 장착된 가스 크로마토 그래피(gas chromatography-mass spectrometry, GC-MS)에 의해 58개의 화합물이 분석되어졌다. 그 오일의 주요한 성분은 각각 camphor (17.18%), germacrene-D (15.70%), trans(${\beta}-$)caryophyllene (6.79%), ${\beta}$-thujones (6.57%), 1, 8-cineole (5.94%) and camphene (5.08%) 순으로 나타났다. 참쑥 오일의 돼지 유행성 설사 바이러스에 대한 항바이러스능을 세포독성 감소 방법 [cytopathic effect (CPE) reduction method]에 의해 베로 세포(원숭이 신장세포, Vero cells)에서 평가하였다. 그 오일은 43.7 ${\mu}g/ml$의 50% 저해농도 ($IC_{50}$)와 함께 돼지 유행성 설사 바이러스 복제를 억제하였다. 또한 그 오일은 100 ${\mu}g/ml$ 이상에서 세포독성을 나타내지 않았으며, 그 치료지수(therapeutic index, TI value)는 2.3 이상이였다. 리바비린(rivabirin)은 그 오일에 비해 상대적으로 낮은 항바이러스능을 나타내었다. 다섯개 미생물(Staphyloscoccus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Propionibacterium acnes, Candida albicans)에 대한 참쑥 오일의 항균 활성은 페이퍼 디스크 방법(paer disck diffusion method)에 의해 평가하였다. 그 결과 그 오일은 5가지 미생물에 대해 8-22 mm의 투명환(clear zone)을 나타내며 항균활성을 보였다. 테스트된 미생물들 가운데 S. pyogenes가 가장 높은 활성을 나타냈으며, C. albicans가 가장 낮은 활성을 나타냈다. 이상의 결과로부터 참쑥 오일은 항균 및 돼지 유행성 설사 바이러스 유래 질병을 조절하는데 응용 가능할 것이라 사료된다.

• 미생물학회지
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• 제38권3호
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• pp.221-229
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• 2002

일본뇌염바이러스(Japanese encephalitis virus, JEV) Nakayama strain을 초기 multiplicity of infection 5.0으로 Vero세포에 감염하여 1년 이상 안정적으로 바이러스를 방출하는 지속감염(persistently-infected)세포주를 확립하였다. 지속감염 세포에서 지속적으로 방출되는 총 11 개의 Plaque 형태 변이바이러스(morphology mutants)클론을 확보하였다. 분리된 변이바이러스의 복제효율을 분석한 결과 생물학적 표현형과 복제효율은 유의하게 상관하였다. 변이바이러스 RNA 게놈 양 말단의 non-coding region 및 envelop 단백질의 ORF에서는 유의한 염기서열 변화가 관찰되지 않아 JEV 약독화에 새로운 인자가 추가로 관여할 가능성을 제시하였다. 변이바이러스에 감염된 신선한 Vero세포는 wild-type JEV의 일반적 감염성상과 다르게 대다수의 세포가 유의할 만한 세포병변현상을 나타내지 않았다. 감염된 Vero세포에서 wild-type JEV 및 large plaque을 형성하는 변이바이러스의 경우 mRNA와 함께 Bcl-2의 발현은 모두 유의하게 감소하였으며 p53은 뚜렷하게 증가하였다. 반면 small plaque을 형성하는 변이바이러스의 감염세포에서는 Bcl-2와 p53 모두 유의한 변화를 볼 수 없었다. 이상의 결과들과 함께, 감염된 Vero세포의 internucleosomal DNA fragmentation과DNA profile의 유형분석에 따르면 궁극적 인 세포병변효과의 변화는 변이바이러스의 복제효율과 더불어 p53에 비의존적인 apoptosis 수위의 전반적인 감소에 기인하는 것으로 판단된다.

• The Korean Journal of Physiology and Pharmacology
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• 제8권5호
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• pp.289-293
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• 2004

To investigate whether VacA (vacuolating toxin) produced by Helicobacter pylori Korean stain 99 induces intestinal secretion, purified VacA was added to T84 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of low pH-pretreated VacA increased short circuit current (Isc). The effect was time- and dose-dependent and saturable. The time-to-peak Isc was concentration-dependent. Chloride channel inhibitors, niflumic acid or 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), inhibited VacA-stimulated Isc. Carbachol (CCh)-induced increase of Isc was prolonged by the addition of VacA to the mucosal side only. The effect was unaltered by the addition of niflumic acid. VacA did not show cytopathic effects. These studies indicate that VacA is a nonlethal toxin that acts in a polar manner on T84 monolayers to potentiate $Cl^-$ secretion and the response to CCh secretion without decrease in monolayer resistance. VacA may contribute to diarrhea diseases in human intestinal epithelial cells.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.414-421
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• 1999

Helicobacter pylori were found to be resistant to azide but sensitive to vanadate, suggesting that defect in the P-type ATPase activity rather than F-type ATPase would be lethal to cell survival or growth. To elucidate the relationship between this enzyme inhibition and H. pylori death, we determined the effect of omeprazole (OMP) plus vanadate on enzyme activity and cell growth. The minimum inhibitory concentration (MIC; ca. 0.8$\mu$mol/disk) of vanadate for H. pylori growth was lowered over l0-fold with the aid of OMP, whereby its inhibitory potential toward the P-type ATPase activity was diametrically increased. Alternatively, we found that this enzyme activity was essential for active transport in H. pylori. From these observations, we strongly suggest that the immediate cause of the growth inhibition of H. pylori cells with OMP and/or vanadate might be defective in the cell's active transport due to the lack of P-type ATPase activity. From the spectral data with circular dichroism (CD) spectroscopy, we found that activated OMP (OAS) at concentration below MIC did not disrupt helical structures of membrane proteins. Separately, we determined the cytopathic effect of OAS by SDS-PAGE, indicating the change in the production of cytoplasmic protein but not cell membrane.

• Mycobiology
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• 제42권2호
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• pp.189-192
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• 2014

During a search for neuraminidase inhibitors derived from medicinal fungi, we found that the fermentation broth of Phellinus linteus exhibited potent neuraminidase inhibitory activity. Through bioassay-guided fractionation, two active compounds were purified from the ethyl acetate-soluble portion of the fermentation broth of P. linteus. These structures were identified as inotilone (1) and 4-(3,4-dihydroxyphenyl)-3-buten-2-one (2) by spectroscopic methods. Compounds 1 and 2 inhibited H1N1 neuraminidase activity with $IC_{50}$ values of 29.1 and 125.6 $125.6{\mu}M$, respectively, in a dose-dependent manner. They also exhibited an antiviral effect in a viral cytopathic effect reduction assay using MDCK cells. These results suggest that compounds 1 and 2 from the culture broth of P. linteus would be good candidates for the prevention and therapeutic strategies towards viral infections.

• 생약학회지
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• 제42권1호
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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.

• Journal of Applied Biological Chemistry
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• 제56권1호
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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).

• Journal of Microbiology
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• 제45권2호
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• pp.133-138
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• 2007

This study addresses the susceptibility of Spodoptera frugiperda (Sf9 and Sf21), Trichoplusia ni (Hi5), and S. exigua (Se301) cells to the Bombyx mori nucleopolyhedrovirus (BmNPV). Although these cells have classically been considered nonpermissive to BmNPV, the cytopathic effect, an increase in viral yield, and viral DNA synthesis by BmNPV were observed in Sf9, Sf21, and Hi5 cells, but not in Se301 cells. Very late gene expression by BmNPV in these cell lines was also detected via ${\beta}-galactosidase$ expression under the control of the polyhedrin promoter. Sf9 cells were most susceptible to BmNPV in all respects, followed by Sf21 and Hi5 cells in decreasing order, while the Se301 cells evidenced no distinct viral replication. This particular difference in viral susceptibility in each of the cell lines can be utilized for our understanding of the mechanisms underlying the host specificity of NPVs.