• Microbiology and Biotechnology Letters
• /
• 제32권2호
• /
• pp.179-183
• /
• 2004

Enterovirus isolation was attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2002. Enteroviruses were found in 83 of 703 cases. Echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 29, 30 were isolated in 72 cases while coxsackievirus serotypes B3 and B4 were isolated in 11 cases. The occurrence was found to be distributed from April thru November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Age distribution of patients, 61 patients in Enteroviruses and 11 patients in coxsackievirus, respectively, occurred in 0 to 10 year-olds. The sex distribution of the patients is as follows, 52 males (62.7％), 31 females (37.3％). Occurrence rate was found to be higher in male patients. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 437 bp single band in all the strains.

• Pediatric Infection and Vaccine
• /
• 제5권1호
• /
• pp.115-120
• /
• 1998

Purpose : In the summer 1997, there was an outbreak of aseptic meningitis in Pusan area. We report the clinical features and viral studies of aseptic meningitis in Pusan area in 1997. Methods : 265 children with aseptic meningitis who had been admitted to Department of Pediatrics, St. Benedict Hospital between April to October 1997 were included. Results : 1) Male-to-female ratio was 1.7:1. 2) Mean age was $6.2{\pm}3.29$ years. 3) It occured mostly April to October. 4) Clinical manifestations were fever 99.6%, vomiting 99.2%, headache 99.2%, rash 6.0%. 5) The duration of fever was $3.34{\pm}2.21$ days. 6) The duration of admission was $5.3{\pm}3.21$ days. 7) WBC count in peripheral blood were $11,200{\pm}4,163/mm^3$. 8) WBC count in CSF were $156.1{\pm}394.7/mm^3$. 9) Causative agents were coxsackievirus B5, echovius 6, 30, type nonspecific enterovirus. Conclusion : Aseptic meningitis in 1997 compared with that in 1996 had clinical feature of increase in age, decrease in duration of fever and incidende of rash. It occurred mostly April to October in 1997 and May to October in 1996. Causative agents were coxackievirus B5, echovirus 6, 30, type nonespecific enterovirus in 1997, and echovirus 9, coxsackievirus A24, type nonespecific enterovirus in 1996.

• Korean Journal of Pharmacognosy
• /
• 제48권3호
• /
• pp.173-178
• /
• 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.

• Korean Journal of Pharmacognosy
• /
• 제47권2호
• /
• pp.137-142
• /
• 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.

• Korean Journal of Microbiology
• /
• 제39권4호
• /
• pp.248-252
• /
• 2003

Enteroviruses isolation were attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2000-2002. Enteroviruses were found in 2 of 292 cases in 2000, 4 of 371 cases in 2001, 83 of 703 cases in 2002. In 2000, the isolated viruses were found to be echovirus serotype 11 and coxsackievirus serotype B2. Coxsackievirus serotype B5 was isolated in 2001 and in 2002, echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 30 were isolated in 70 cases while coxsackievirus serotypes B3 and B4 were isolated in 10 cases. Various specimens tended to emerge over the years. The occurrence in 2000 tended to be mostly focus during the cold months, December through January, while in 2001, it occurred in May. In 2002, occurrence was found to be distributed from April to November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 436 bp single band in all the strains. The serotype was sent to the Department of Virology at the Korean National Institute of Health for identification.

• Journal of Life Science
• /
• 제26권10호
• /
• pp.1182-1188
• /
• 2016

Membrane-associated guanylate kinase inverted-3 (MAGI-3) is a member of the family of membrane-associated guanylate kinases (MAGUKs). MAGI-3 modulates the kinase activity of protein kinase B (PKB)/AKT through interactions with phosphatase and tensin homolog (PTEN)/MMAC. Coxsackievirus B3 (CVB3) is a common causative agent of acute myocarditis and chronic dilated cardiomyopathy. Activation of AKT and extracellular signal-regulated kinases 1/2 (ERK1/2) is essential for CVB3 replication, but the relation between MAGI-3 signaling and CVB3 replication is not well understood. This study investigated the role of MAGI-3 in CVB3 infection and replication. MAGI-3 was overexpressed in HeLa cells by polyethylenimine (PEI) transfection. To optimize the transfection conditions, different ratios of plasmid DNA to PEI concentrations were used. MAGI-3 and empty plasmid DNA were transfected into the HeLa cells. MAGI-3 overexpression alone was not sufficient to efficiently activate AKT. However, expression of the CVB3 capsid protein VP1 dramatically increased in the HeLa cells overexpressing MAGI-3 24 h after CVB3 infection. In addition, the activities of AKT and ERK were significantly induced in the CVB3-infected MAGI-3 cells overexpressing HeLa. These results demonstrate that MAGI-3 expression upregulates CVB3 replication through AKT and ERK signaling activation. MAGI-3 may be an important target to control CVB3 replication.

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

• Pediatric Infection and Vaccine
• /
• 제5권1호
• /
• pp.104-114
• /
• 1998

Purpose : Enteroviruses are the most common cause of aseptic meningitis. The epidemics of aseptic meningitis in 1993 and 1996 were mostly caused by echovirus type 9. Identification of the causative virus of aseptic meningitis in epidemics, is very important not only for diagnosis but also for epidemiologic purpose. The purpose of this study was to identify the causative virus and investigate the relationship between aseptic meningitis, prevailed in Masan and surrounding areas in Kyoungsangnamdo in 1997, and its clinical manifestations. Methods : One hundred twenty eight cerebrospinal fluid(CSF) and 239 stool specimens were obtained from 239 patients(213 children and 26 adult patients) with aseptic meningitis were admitted to Masan Fatima Hospitals from March to October 1997. Viral isolation and serotype identification was performed by cell culture and immunofluorescent test. Enteroviruses not typed by immunofluorescent test was confirmed by reverse transcription-polymerase chain reaction(RT-PCR). Results : 1) The peak incidence was noted in June. 2) The age of 239 patients(pediatrics-213 cases, internal medicine-26 cases) that were diagnosed ranged from neonate to 35 years, the age of the patients of pediatrics ranged from neonate to 15years(mean 4.9 years), the age of the patients of internal medicine (above 16 years) ranged from 16 years to 35 years(mean 24.2 years). 3) Fifty-three(41.4%) of 128 CSF specimens were positive for enteroviruses, and 163(68.2%) of 239 stool specimens were positive for enteroviruses respectively. 4) Serotypes of 53 enteroviruses isolated from CSF were 16(30.2%) of echovirus type 30, 6(11.3%) of echovirus type 6, 1 of echovirus type 4, 4 of untyped echovirus, 1 of coxsackievirus type B5, and 24 isolates of untyped enteroviruses. Of 163 enterovirus isolated from stool were 72(44.2%) of echovirus type 30, 21(12.9%) of echovirus type 6, 1 of echovirus type 4, 17(10.4%) of undetermined subtyped echovirus, 1 of coxsackievirus type B5, 2 of A24, 3 of undetermined subtyped coxsackievirus type B, and 46 isolates of untyped enterovirus. Conclusion : There were epidemics of aseptic meningitis in the central areas of Kyoungsangnamdo from March to October 1997. The main causative organism was thought to be the echovirus type 30, and echovirus type 4, 6, coxsackievirus B5 and A24 were also thought to contribute to the epidemics.

• Journal of Environmental Science International
• /
• 제8권2호
• /
• pp.165-169
• /
• 1999

The incidence of aseptic meningitis infection is ensuing and threatening the health of children. Enteroviruses are the major agents of aseptic meningitis and identification of virus has been a clue to diagnosis and epidemiology. The outbreak of aseptic meningitis occurred in Pusan, 1998. Patients were concentrated from April through November. Children were more susceptible than adults. Among 306 cases of specimens from stool, throat swab tested, only 7.2% were positive on virus isolation, 12 cases from stool and 10 from throat, respectively. All isolated 7 serotypes of viruses represented cytopathic effect on cultured cells. Three types of echovirus 6.25, 30 and coxsackievirus B2, B3, B4, B6 were identified by neutralizing antibody test. Isolated coxsackievirus and echovirus were observed by an electron microscope with negative staining.

• Clinical and Experimental Pediatrics
• /
• 제49권7호
• /
• pp.745-750
• /
• 2006

Purpose : Enteroviruses are the most common cause of aseptic meningitis in patients of all ages. A definite diagnosis of enteroviral meningitis can be established by detection of virus directly in CSF specimens. But this is time-consuming and lacks sensitivity, so polymerase chain reaction(PCR) detecting of viral RNA in patient specimens such as CSF, stool has been demonstrated. But little is known about the influence of sampling time on the results of CSF PCR and stool PCR. We investigated diagnostic utility of PCR of CSF and stool according to sampling time after the onset of symptoms. Methods : PCR results were analyzed according to sampling time for 42 patients diagnosed aseptic meningits in our hospital from $11^{th}$ January to $30^{th}$ August, 2005. Results : The diagnostic yield of the test was higher of CSF specimens obtained ${\leq_-}2$ days after clinical onset(positive PCR results 9/18, 50 percent), compared with CSF collected >2 days after onset(positive PCR results 1/24, 4.2 percent)(P=0.001). Instead, positive PCR results of fecal specimens maintained highly(average 90.5 percent), 10 cases had also positive PCR results even 5-6 days after onset. 10 cases of CSF specimens had positive enterovirus PCR results containing coxsackievirus B5 (n=6), coxsackievirus B3(n=3). 38 cases of stool specimens had positive enterovirus PCR results containing echovirus 18(n=7), echovirus 9(n=3), coxsackievirus B5(n=8), coxsackievirus B3(n=3). 6 cases(coxackie B5) had positive CSF PCR and stool PCR, both. Conclusion : Stool PCR was clinically sensitive for detecting enterovirus during enteroviral meningits and could give a presumptive diagnosis throughout the disease course. A definite diagnosis was obtained by CSF PCR, but its utility was clearly lower for samples obtained >2 days after clinical onset. Therefore, it is recommended that, in addition to performance of CSF PCR, fecal samples obtained from patients with suspected enteroviral meningitis should be tested by PCR, especially when the duration of symptoms is >2 days.