• Biomolecules & Therapeutics
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• 제23권5호
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• pp.465-470
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• 2015

Chrysin is a 5,7-dihydroxyflavone and was recently shown to potently inhibit enterovirus 71 (EV71) by suppressing viral 3C protease ($3C^{pro}$ activity. In the current study, we investigated whether chrysin also shows antiviral activity against coxsackievirus B3 (CVB3), which belongs to the same genus (Enterovirus) as EV71, and assessed its ability to prevent the resulting acute pancreatitis and myocarditis. We found that chrysin showed antiviral activity against CVB3 at $10{\mu}M$, but exhibited mild cellular cytotoxicity at $50{\mu}M$, prompting us to synthesize derivatives of chrysin to increase the antiviral activity and reduce its cytotoxicity. Among four 4-substituted benzyl derivatives derived from C(5) benzyl-protected derivatives 7, 9-11 had significant antiviral activity and showed the most potent activity against CVB3 with low cytotoxicity in Vero cells. Intraperitoneal injection of CVB3 in BALB/c mice with $1{\times}10^6TCID_{50}$ (50% tissue culture infective dose) of CVB3 induced acute pancreatitis with ablation of acinar cells and increased serum CXCL1 levels, whereas the daily administration of 9 for 5 days significantly alleviated the pancreatic inflammation and reduced the elevation in serum CXCL1 levels. Collectively, we assessed the anti-CVB3 activities of chrysin and its derivatives, and found that among 4-substituted benzyl derivatives, 9 exhibited the highest activity against CVB3 in vivo, and protected mice from CVB3-induced pancreatic damage, simultaneously lowering serum CXCL1 levels.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.127-140
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• 2019

Since 1990, many nucleic acid expression platforms consisting of DNA or RNA have been developed. However, although RNA expression platforms have been relatively neglected, several such platforms capped at the 5' end of RNA by an anti-reverse cap analog have now been developed. At the same time, the capping reaction is a bottleneck in the production of such platforms, with high cost and low efficiency. Here, we investigated several viral and eukaryotic internal ribosome entry sites (IRESs) to develop an optimal RNA expression platform, because IRES-dependent translation does not require a capping step. RNA expression platforms constructed with IRESs from the 5' untranslated regions of the encephalomyocarditis virus (EMCV) and the intergenic region of the cricket paralysis virus (CrPV) showed sufficient expression efficiency compared with cap-dependent RNA expression platforms. However, eukaryotic IRESs exhibited a lower viral IRES expression efficiency. Interestingly, the addition of a poly(A) sequence to the 5' end of the coxsackievirus B3 (CVB3) IRES (pMA-CVB3) increased the expression level compared with the CVB3 IRES without poly(A) (pCVB3). Therefore, we developed two multiexpression platforms (termed pMA-CVB3-EMCV and pCrPV-EMCV) by combining the IRESs of CVB3, CrPV, and EMCV in a single-RNA backbone. The pMA-CVB3-EMCV-derived RNA platform showed the highest expression level. Moreover, it clearly exhibited expression in mouse muscles in vivo. These RNA expression platforms prepared using viral IRESs will be useful in developing potential RNA-based prophylactic or therapeutic vaccines, because they have better expression efficiency and do not need a capping step.

• 생약학회지
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• 제48권3호
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• pp.173-178
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• 2017

We investigated the efficacy of single compound of plant extract in coxsackievirus B3 (CVB3) infection. CVB3 is a main cause of Hand-foot-mouth diseases (HFMD) and viral myocarditis in children and adult. Several single compounds of plant extract were purified by HPLC and tested as antiviral drug candidate. Among them, kaempferol was selected to effective anti-enterovirus compound by HeLa cells survival assay. CVB3 infected HeLa cells were treated with kaempferol ($100{\mu}g/ml-100ng/ml$) and their antiviral effect was confirmed. After 16 hours of treatment, HeLa cells were lysed and proteins were extracted for western blot analysis. CVB3 viral capsid protein VP1 production and transcription factor eIF4G-1 cleavage was significantly decreased in $100{\mu}g/ml$ kaempferol treatment. Virus replication was observed by virus RNA amplification. Kaempferol strongly reduced virus positive and negative strand RNA amplification. Moreover, MAPK signal induced by CVB3 infection, pERK and pmTOR, kaempferol treatment significantly inhibited the activity. Plant extract single compound, kaempferol, is a strong candidate to be developed non-toxic anti-enterovirus treatment agent.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.2012-2018
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• 2016

Coxsackievirus B3 (CVB3) is the main cause of acute myocarditis and dilated cardiomyopathy. Plant extracts are considered as useful materials to develop new antiviral drugs. We had previously selected candidate plant extracts, which showed anti-inflammatory effects. We examined the antiviral effects by using a HeLa cell survival assay. Among these extracts, we chose the Amomi Cardamomi (Amomi) extract, which showed strong antiviral effect and preserved cell survival in CVB3 infection. We investigated the mechanisms underlying the ability of Amomi extract to inhibit CVB3 infection and replication. HeLa cells were infected by CVB3 with or without Amomi extract. Erk and Akt activities, and their correlation with virus replication were observed. Live virus titers in cell supernatants and viral positive- and negative-strand RNA amplification were measured. Amomi extract significantly increased HeLa cell survival in different concentrations ($100-10{\mu}g/ml$). CVB3 capsid protein VP1 expression (76%) and viral protease 2A-induced eIF4G1 cleavage (70%) were significantly decreased in Amomi extract ($100{\mu}g/ml$) treated cells. The levels of positive- (20%) and negative-strand (80%) RNA were dramatically decreased compared with the control, as revealed by reverse transcription-PCR. In addition, Amomi extract improved mice survival (51% vs 26%) and dramatically reduced heart inflammation in a CVB3-induced myocarditis mouse model. These results suggested that Amomi extract significantly inhibited Enterovirus replication and myocarditis damage. Amomi may be developed as a therapeutic drug for Enterovirus.

• 대한바이러스학회지
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• 제30권1호
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• pp.71-81
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• 2000

We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.

• 생약학회지
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• 제47권2호
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• pp.137-142
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• 2016

Enterovirus is a common cause of several severe diseases such as myocarditis, hand-foot-mouth disease, and meningitis in children and adult. There are many try to develop new antiviral drug for direct treatment in virus infection. However, synthetic chemical antiviral drug is not working. To overcome this limitation, we examined plant extracts. The antiviral effect of plant extracts was screened by HeLa cell survival assay in coxsackievirus B3 (CVB3) infection. We observed a strong antiviral effect of Poria cocos extract in a dose-dependent manner (1 mg/ml~0.01 mg/ml). P. cocos extract (1 mg/ml) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and virus capsid protein VP1 production. CVB3 positive and negative strand RNA amplification were significantly reduced in P. cocos extract treatment. P. cocos extract completely blocked early time activation of ERK and AKT activity in CVB3 infection. Taken together these data indicate that the treatment of P. cocos extract strongly inhibit CVB3 replication. Poria cocos extract may possible to developed as a therapeutic agent for enterovirus.

• 한국동물위생학회지
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• 제35권4호
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• pp.255-261
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• 2012

Foot and mouth disease (FMD) is highly infectious disease of cloven-hoofed animals, particularly cattle, sheep, pigs and goats. Last outbreak reported in November, 2010 induced the enormous social and economical impacts. Culling of infected animals, movement control, and vaccination are the major control measures of FMD. The aim of this study was to detection foot-and-mouth disease virus (FMDV) in the soil and leachate from modeling burial for pig carcass as measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). FMDV and Coxsakievirus B1 (CVB1) were detected in soil by week 16 and Coxsakievirus B1 (CVB1) by weeks 12, respectively. FMDV and CVB1 also detected by weeks 8 in the leachate. Results from this study provides an evidence that FMDV could be inactivated for safe of pig carcasses infected with FMDV within 4 month in the carcass burial site.

• 디지털융복합연구
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• 제5권2호
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• pp.91-98
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• 2007

Destination marketing organizations (DMO) have been building up the cyber city in the WWW. Website for DMO is a core channel to promote regional attractions. This research suggests the issue of criteria for evaluating DMO's performance in the Internet. The method of evaluation focuses on the structure in perspective of linkage based on small world theory and direct network. Convention & Visitors & Bureau (CVB) in tourism and travel industry playa role to promote and held the international meeting and exhibitions. CVB's websites evaluated according to web link structure and performance.

• 식물병연구
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• 제28권4호
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• pp.237-244
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• 2022

국화(Dendranthema morifolium)는 한국에서 경제적으로 중요한 화훼식물에 포함되고, 바이러스와 바이로이드 병들에 의해 경제적 피해를 받고 있다. 본 연구에서는 충청남도 예산군에서 재배된 국화 350개체를 대상으로 7종의 바이러스들과 2종의 바이로이드들(Chrysanthemum chlorotic mottle viroid, CChMVd; Chrysanthemum stunt viroid, CSVd; Cucumber mosaic virus, CMV; Chrysanthemum virus B, CVB; Chrysanthemum stem necrosis orthotospovirus, CSNV; Impatiens necrotic spot orthotospovirus, INSV; Potato virus X, PVX; Tomato aspermy virus, TAV; Tomato spotted wilt orthotospovirus, TSWV)을 검정하였다. 그 결과 2종의 바이러스들(CVB-CN-Y, TAV-CN-Y)과 2종의 바이로이드들(CChMVd-CN-Y, CSVd-CN-Y)이 검정되었다. CVBCN-Y는 6개의 국화 식물체들에서 검정되었고, TAV-CN-Y는 한개가 국화 식물체에서만 검정되었다. CChMVd-CN-Y는 97개의 국화 식물체들에서 검정되었으며, CSVd-CN-Y는 21개의 국화 식물체들에서 검정되었다. CVB-CN-Y는 CVB-GS1과 86.9% 염기서열 상동성을 보였으며, TAV-CN-Y는 3개의 분리주들(TAV-Chj, TAV-P, TAV-V)과 100% 염기서열이 일치하였다. CChMVd-CN-Y는 CChMVd-Horst와 99.5% 염기서열 상동성을 보였으며, CSVd-CN-Y는 4개의 분리주들(Au1.1, K4pop, Sagae, Tochigi)과 99.7% 염기서열 상동성을 보였다. 본 연구는 2021년 예산군에서 재배된 국화들을 대상으로 바이러스 및 바이로이드들을 검정하고 그들의 감염률에 관한 보고서이다.

• 생약학회지
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• 제45권4호
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• pp.282-287
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• 2014

The ORI2 (3-[3,4-dihydroxyphenyl]acrylic acid 1-[3,4-dihydroxyphenyl]-2-methoxycarbonylethyl ester) was purified from the extract of Isodon excisus. We confirmed the antiviral effect of ORI2 in a coxsackievirus-induced pancreatitis model. Coxsackievirus B4 (CVB4) is a common cause of pancreatitis and may be reason of the type-1 diabetes. Anti-enteroviral compounds were screened by HeLa cell survival assay. Purified natural compounds were added to HeLa cells cultured 96-well plates after $10^4PFU/ml$ CVB4 pre-incubation for 30 min. ORI2 significantly improved HeLa cell survival in a dose-dependent manner. In addition, ORI2 (1 mM) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and viral VP1 capsid protein production. HeLa cell virus titers and viral RNA replication were significantly decreased in ORI2-treatment in a dose dependent manner (1 mM~0.001 mM). These results demonstrate that ORI2 has a strong antiviral effect. It was significantly decreased virus replication. ORI2 may be developed as a potential therapeutic agent for CVB4.