• Korean Journal of Pharmacognosy
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• v.48 no.3
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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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• v.19 no.9
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• pp.1055-1064
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• 2009

Enteroviruses were identified and characterized from patients with aseptic meningitis and other enterovirus-related diseases in Chungnam, Korea from 2005 to 2006. Enteroviruses were isolated from 79 of 519 cases (15.2%) in 2005, and 37 of 386 cases (9.6%) in 2006. Based on partial VP1 sequencing, a total of 116 enterovirus isolates were resolved into 13 types. Prevalent among the Chungnam isolates were echovirus 18 and coxsackievirus B5 in 2005, and echoviruses 5 and 25 in 2006. This is the first time echoviruses 5 and 18 have been identified in Korea since enterovirus surveillance began there in 1993. The temporal distribution of enterovirus epidemics in Chungnam showed a remarkable seasonal pattern, with cases occurring during most of the three months of the summer from June to August. The highest rate of enterovirus-positive cases occurred in patients less than 1 year of age. The ratio of male to female enterovirus-positive patients was approximately 1.8:1. Comparison of the VP1 amino acid sequences of the 15 coxsackievirus B5 isolates with reference strains revealed that all Chungnam isolates are substituted at positions 23 (V231), 19 (S19G), 75 (Y75F), and 95 (N95S). Upon comparing the nine ECV5 isolates with foreign strains, it was found that only the Chungnam isolates, with the exception of Kor06-ECV5-239cn, have P at position 153 and F at position 146. The three ECV9 isolates from 2006 show alterations at amino acids 36, 148, and 154 outside of the BC-loop and at position 84 in the BC-loop, whereas the seven isolates from 2005 and the other ECV9 strains in the database only show the alteration at position 84 (D, I, N, S). The five ECV25 isolates have an S residue at position 134, whereas most of the foreign strains have an N residue.

• Korean Journal of Veterinary Research
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• v.25 no.2
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• pp.103-112
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• 1985

A total of 1-0 colostrum-deprived pigs (1 or 2-day-old) and 6 pigs (35-day-old), which had been raised by natural maternal nursing, were used to study the pathogenicity of the porcine enteroviruses by the intracerebral and intramuscular routes of inoculation, which the enterovirus were isolated from the diseased pigs in Korea. The porcine enteroviruses produced an identical polioencephalomyelitis in colostrum-deprived pigs and 35-day-old pigs, which manifested clinical signs and histopathological changes. Clinically it was characterized by incoordination, rise in rectal temperature, ataxia, flaccid paralysis in all the experimental groups. Histopathologically, the lesions were present in both grey and white matter at all levels of central nervous system, though usually more severe in the grey matter. These changes were characterized by meningeal infiltration, degeneration of nerve cells, neuronophagia, diffuse and focal gliosis, glial nodules and perivascular lymphocytic infiltrations. Ganglionitis of the dorsal root ganglia was frequently observed. On the basis of the clinical and histopathological changes mentioned above, it was concluded that porcine enteroviruses isolated in Korea were pathogenic strains which could produce polioencephalomyelitis in pigs. The most severe Jisease was prcduced by the inoculation of both enterovirus and hog cholera vaccine in the 35-day-old pigs at a time when colostral immunity presumably was low. The porcine enterovirus infections seemed to be associated with certain stress factor such as hog cholera vaccine in or immediately following the weanling period.

• Journal of Microbiology
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• v.42 no.4
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• pp.299-304
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• 2004

To investigate the presence of infectious agents in human atherosclerotic arterial tissues. Atherosclerotic plaques were removed from 128 patients undergoing carotid endarterectomy or other bypass proce­dures for occlusive disease, and from twenty normal arterial wall samples, obtained from transplant donors with no history of diabetes, hypertension, smoking, or hyperlipidemia. Using the polymerase chain reaction (PCR) or reverse transcription-PCR, these samples were analyzed for the presence of Chlamydia pneumoniae, cytomegalovirus, enterovirus, adenovirus, herpes simplex viruses types 1 and 2, and Epstein-Barr virus. The amplicons were then sequenced, and phylogenetic analyses were per­formed. Enteroviral RNA was found in 22 of 128 atherosclerotic vascular lesions $(17.2\%),$ and C. pneu­moniae and cytomegalovirus were each found in 2 samples $(1.6\%).$ In contrast, adenovirus, herpes simplex viruses, and Epstein-Barr virus were not identified in any of the atherosclerotic samples. Enterovirus was detected in 6/24 $(25.0\%)$ aortas, 7/33 $(21.2\%)$ carotid arteries, 6/40 $(15.0\%)$ femoral arteries, and 3/31 $(9.7\%)$ radial arteries of patients with chronic renal failure. There were no infectious agents detected in any of the control specimens. Using phylogenetic analysis, the enterovirus isolates were clustered into 3 groups, arranged as echovirus 9 and coxsackieviruses Bl and B3. Enteroviral RNA was detected in $17.2\%$ of atherosclerotic plaques, but was not observed in any of the control spec­imens. This suggests a connection between enteroviral infection and atherosclerosis. These findings dif­fer from those of other studies, which found more frequent incidence of C. pneumoniae and cytomegalovirus infection in atherosclerotic plaques.

• Pediatric Infection and Vaccine
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• v.19 no.1
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• pp.28-36
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• 2012

Purpose : This study was conducted to evaluate epidemiological data of the viral pathogens obtained from stool exams and provide information on the regional prevalence of infectious diarrheal disease west in Gyeonggi Province, Korea. Methods : We enrolled a cohort of children <10 years of age admitted for treatment of acute diarrhea at Bucheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea. In total, 310 fecal specimens, documented to be free of common bacterial pathogens, were collected from pediatric patients during a 12-month period from January to December 2009 and were tested for the presence of rotavirus, parechovirus, adenovirus, astrovirus, enterovirus, and norovirus using polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) assay. Results : The most common virus was parechovirus (16%), followed by adenovirus (15%), astrovirus (14%), rotavirus (13%), and enterovirus (5%). Interestingly, only one of the specimens was positive for norovirus. Single infection cases were detected in 173 (55.8%) of the 310 children, whereas mixed viral infections were detected in 10 (3.2%) of the same children. Viral gastroenteritis generally showed a double peak of incidence. Parechovirus, rotavirus, and adenovirus shared a similar pattern of peak incidence with overall viruses; however, astrovirus infections occurred more frequently in the spring. Eighty-five percent of the confirmed viral gastroenteritis cases developed in under 24 months. Conclusion : The results support the importance of parechovirus, adenovirus, astrovirus, and enterovirus as causative agents of diarrhea in children, which may be underestimated by current routine diagnostic testing.

• Journal of Life Science
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• v.26 no.10
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• pp.1182-1188
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• 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.

• Pediatric Infection and Vaccine
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• v.30 no.2
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• pp.91-96
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• 2023

Infection with enterovirus (EV) 71 is usually associated with hand-foot-and-mouth disease and herpangina. The most frequent neurologic complication is brainstem encephalitis. A 30-month-old boy visited the pediatric emergency department with fever, lethargy, and abnormal eye contact. His mental status was slightly drowsy. On hospitalization day 2, the patient experienced respiratory arrest with apnea. Brain magnetic resonance imaging revealed bilateral symmetric T2-high signal lesions without enhancement in the posterior aspect of the brainstem and left medial temporal lobe. Electroencephalography was indicative of diffuse cerebral dysfunction with diffuse high amplitude and irregular delta activities. He underwent a gene study and was diagnosed with myoclonic epilepsy with ragged red fibers syndrome. We report a case of EV 71 brainstem encephalitis by polymerase chain reaction for nasopharyngeal aspirates and feces with rapid progression within one day of fever without the manifestation of throat and skin lesions because of his underlying mitochondrial disease.

• Biomedical Science Letters
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• v.29 no.2
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• pp.81-87
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• 2023

Pan-Enterovirus (Pan-EV) infects millions of children and infants worldwide every year. As severe infections have recently been reported, the need for monitoring has consequently intensified. Pan-EV is a categorical name for waterborne enteroviruses belonging to the Picornaviridae family, and includes a wide range of pathogens including Coxsackievirus (CoxV), Echovirus (EcoV) and Enterovirus (EV). In this study, we proposed an optimal RT-nested PCR method for diagnosis of various types of Pan-EV in an aquatic environment and developed a positive control. Considering detection sensitivity, specific reaction, and final identification, one condition capable of amplifying 478 bp among the four candidates in the 1^st round PCR (RT-PCR) and one condition in the 2^nd round PCR (nested PCR) were selected. Through the detection of nucleic acids extracted from 123 groundwater samples and the detection sensitivity test based on artificial spiking in the sample, the methods are optimal for non-disinfected water samples such as groundwater. We developed a positive control for Pan-EV detection that can be amplified to different sizes under the two conditions. Accuracy could be further improved by testing for contamination from the control group. The method proposed in this study and the positive control developed are expected to be used in monitoring Pan-EV in aquatic environments including groundwater through future research using more samples.

• Clinical and Experimental Pediatrics
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• v.59 no.10
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• pp.395-401
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• 2016

Since the outbreak of the enterovirus 71 (EV71) infection in Malaysia in 1997, large epidemics of EV71 have occurred in the Asia-Pacific region. Many children and infants have died from serious neurological complications during these epidemics, and EV71 infection has become a serious public health problem in these areas. EV71 infection causes hand, foot and mouth disease (HFMD) in children, and usually resolves spontaneously. However, EV71 occasionally involves the central nervous system (CNS), and induces diverse neurological complications such as brainstem encephalitis, aseptic meningitis, and acute flaccid paralysis. Among those complications, brainstem encephalitis is the most critical neurological manifestation because it can cause neurogenic pulmonary hemorrhage/edema leading to death. The characteristic clinical symptoms such as myoclonus and ataxia, cerebrospinal fluid (CSF) pleocytosis, and brainstem lesions on magnetic resonance imaging, in conjunction with the skin rash of HFMD and the isolation of EV71 from a stool, throat-swab, or CSF sample are typical findings indicating CNS involvement of EV71 infection. Treatment with intravenous immunoglobulin and milrinone are recommended in cases with severe neurological complications from EV71 infection, such as brainstem encephalitis. Despite the recent discovery of receptors for EV71 in human cells, such as the scavenger receptor B2 and P-selection glycoprotein ligand 1, it is not known why EV71 infection predominantly involves the brainstem. Recently, 3 companies in China have completed phase III clinical trials of EV71 vaccines. However, the promotion and approval of these vaccines in various countries are problems yet to be resolved.

• Clinical and Experimental Pediatrics
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• v.52 no.11
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• pp.1234-1240
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• 2009

Purpose:Enteroviruses (EVs) are commonly known to cause infection, especially in infants and children. This report presents an overview of enterovirus epidemiology in central Korea. Methods:From the spring of 2005 to the autumn of 2006, we collected the cerebrospinal fluid (CSF) and stool samples from the pediatric patients with a febrile illness or suspected meningitis who were admitted to hospitals in central Korea. In order to test for EVs, cell lines were derived from pretreated susceptible specimen, and the cytopathic effects were observed. Seminested real time-polymerase chain reaction (RT-PCR) and direct sequencing were performed for genotypic and phylogenetic analyses. Results:Of the 305 patients examined, 51 (16.7%) tested positive for EV. Of these 51 patients, 44 showed the following serotypes: Echovirus (ECV) 18 (18 cases, 35.2%), Coxsackievirus B (CVB) 5 (13 cases, 25.4%), ECV25 (5 cases, 9.8%), ECV9 (4 cases, 7.8%), ECV5 (3 cases, 5.8%), and EV74 (1 case, 1.9%). In 2005, between June and August, ECV18 and CVB5 were mostly responsible for the enteroviral infections among the patients in central Korea. In 2006, between July and August, ECV25 was mostly the cause of enteroviral infection. Conclusions:There is a need for continuous surveillance of enteroviral infection and its clinical manifestations, particularly for EV74, which was first identified in Korea.