Chrysin is a 5,7-dihydroxyflavone and was recently shown to potently inhibit enterovirus 71 (EV71) by suppressing viral 3C protease (3Cpro) 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 muM, but exhibited mild cellular cytotoxicity at 50 muM, 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 1x106 TCID50 (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.
Acinar Cells ; Animals ; Enterovirus ; Inflammation ; Injections, Intraperitoneal ; Mice ; Myocarditis ; Pancreatitis ; Vero Cells

Sturgeon aqua-cultured in Korea is mainly Acipenser ruthenus and its culture began in the early 2000's. In this study, Carnobacterium sp. was isolated from unprocessed caviar of aqua-cultured Acipenser ruthenus. The 16s rRNA nucleotide sequence obtained from Carnobacterium sp. isolate (accession no. KM236206) was deposited with GenBank and homologous with Carnobacterium divergens DSM 20623 and NBRC15683 strain. In conclusion, this is first report of isolation of Carnobacterium sp. from caviar of Acipenser ruthenus aqua-cultured in Korea. In the future, it must be ascertained whether Carnobacterium sp. degenerate of caviar or cause diseases in sturgeon.
Base Sequence ; Carnobacterium* ; Databases, Nucleic Acid ; Korea

Objectives: Coxsackievirus A group 16 strain (CVA16) is one of the predominant causative agents of hand, foot, and mouth disease (HFMD). Methods: Using a specimen from a male patient with HFMD, we isolated and performed sequencing of the Korean CVA16 strain and compared it with a G10 reference strain. Also, we were investigated the effects of medicinal plant extract on the cytopathic effects (CPE) by CPE reduction assay against Korean CVA16. Results: Phylogenetic analysis showed that the Korean CVA16 isolate belonged to cluster B-1 and was closely related to the strain PM-15765-00 isolated in Malaysia in 2000. The Korean CVA16 isolate showed 73.2% nucleotide identity to the G10 prototype strain and 98.7% nucleotide identity to PM-15765-00. Next, we assessed whether the Korean CVA16 isolate could be used for in vitro screening of antiviral agents to treat HFMD infection. Vero cells infected with the Korean CVA16 isolate showed a cytopathic effect 2 days after the infection, and the treatment of cells with Cornus officinalis, Acer triflorum, Pulsatilla koreana, and Clematis heracleifolia var. davidiana Hemsl extracts exhibited strong antiviral activity against CVA16. Conclusion Collectively, our work provides potential candidates for the development of vaccine and novel drugs to treat the CVA16 strain isolated from a Korean patient.

Noroviruses (NoV) are the major viral pathogen that causes epidemic acute gastroenteritis and outbreaks of food-borne illness. The major genotypes responsible for the epidemics of NoV are GII.4 and GII.3. However, most studies of NoV have been associated with GII.4 genotype and only few studies have been done with GII.3 genotype. Here, we selected 18 GII.3 strains, which recently circulated in Korea, and determined the partial sequences of the capsid gene. Phylogenetic analysis comparing these sequences with 29 reference strains from the GenBank database was performed using the MEGA program. All NoV GII.3 strains formed 2 distinct genetic lineages and variant groups. Lineage A showed 94.1~97.6%, 90.2~94.6% nucleotide identities from lineage B and variant group, respectively. Lineage B showed 90.2~94.6% nucleotide identities from variant group. These different genetic lineages based on the phylogenetic analysis of capsid sequences imply that the circulating Korean NoV GII.3 strains have potential antigenic variation.
Antigenic Variation ; Capsid ; Databases, Nucleic Acid ; Disease Outbreaks ; Gastroenteritis ; Genotype ; Korea ; Norovirus

Echovirus 6 (ECV6) is the prevalent serotype detected in aseptic meningitis cases in Korea. To analyze the genetic variation of ECV6 isolates recently circulating in Korea, we determined the partial sequence of the VP1 capsid gene from 22 Korean ECV6 isolates and performed pairwise analysis against 42 reference strains from the GenBank database using MegAlign. The 22 Korean ECV6 isolates formed 3 distinct genetic clusters: Kor-lineage I, II, and III. The Korean ECV6 strains showed significant genetic diversity with 14.8~22.8% nucleotide divergence among the 3 different lineages. These ECV6 Kor-lineages were demonstrated to belong to different genetic clusters using VP1 sequence-based phylogenetic analysis, implying that the recently circulating Korean ECV6 strains have potential antigenic variation.
Antigenic Variation ; Capsid ; Databases, Nucleic Acid ; Echovirus 6, Human ; Enterovirus B, Human ; Genetic Variation ; Korea ; Meningitis, Aseptic

BACKGROUND: Enteroviruses are known as major pathogen for aseptic meningitis. Although rapid diagnosis for enteroviruses is very essential to exclude bacterial infections in patients with meningitis, classical diagnostic method based on virus isolation is not practicable for timely treatment of patients due to its laborious and time-consuming procedure. Recently molecular methodologies as alternatives are routinely used for rapid and sensitive diagnosis for enteroviruses infections. METHODS: Reverse transcription (RT)-PCR ELISA kit for targeting 5'non-coding region (NCR) with highly conserved genetic identity among all genotypes of enteroviruses was introduced in this investigation. RT-PCR ELISA was evaluated about sensitivity and specificity through virus isolation using clinical specimens from patients suspected of enteroviral infections and enteroviral isolates comparing with conventional RT-PCR identifying them. RESULTS: The detection limit of the RT-PCR ELISA was up to 10-100 folds higher than virus isolation using cell culture and conventional RT-PCR. On comparison between above two methods, the detection rate of RT-PCR ELISA for clinical specimens from patients with aseptic meningitis was 7% higher than that of conventional RT-PCR targeting 5'NCR (P=0.016). CONCLUSIONS: Our results suggest that RT-PCR ELISA developed in this study could be an alternative diagnostic method for the detection of enteroviral genome with high sensitivity and specificity.
5' Untranslated Regions ; Adolescent ; Child ; Child, Preschool ; Enterovirus/genetics/*isolation & purification ; Enterovirus Infections/*diagnosis ; *Enzyme-Linked Immunosorbent Assay ; Humans ; Infant ; Meningitis, Aseptic/diagnosis ; RNA, Viral/analysis ; *Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus/genetics ; Rotavirus Infections/diagnosis ; Sensitivity and Specificity

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.
Cell Line ; Child ; Enterovirus ; Enterovirus B, Human ; Humans ; Infant ; Korea ; Meningitis

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.
Cell Line ; Child ; Enterovirus ; Enterovirus B, Human ; Humans ; Infant ; Korea ; Meningitis

PURPOSE: Although the nasopharyngeal aspirate (NPA) is more commonly used because of relatively higher accuracy, the nasal swab (NS) is a less painful and easier method than NPA. A few recent reports have shown that NS is more effective than NPA for the detection of respiratory virus using immunofluorescence or viral culture. The objective of the present study was to compare the results of NPA and NS sampling specimens in children for respiratory viruses detection using multiplex RT-PCR. METHODS: From December 2008 to June 2009 Paired NPA and NS specimens were collected from 250 children admitted with symptoms of acute respiratory infections at the Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. The sensitivity and agreement of virus detection between NPA and NS using multiplex RT-PCR were compared and analyzed. RESULTS: The median age of the subjects was 1.3 years (range, 20 days to 16.5 years), and 228 patients (91.2%) were under the age of 5 years. The agreement of virus detection between NPA and NS was excellent (Cohen's kappa >0.8) for parainfluenza virus type 3 or substantial (0.6 to 0.8) for rhinovirus A, RSV A and RSV B, moderate (0.4 to 0.6) for adenovirus and metapneumovirus and poor (<0.4) for influenza A. The overall sensitivity of detection of respiratory viruses was significantly higher in NPA (0.96) than in NS (0.59, P<0.05). CONCLUSION: We recommend NPA may be more accurate specimen for detection of respiratory viruses in hospitalized children. NS might be used in limited cases at a office setting or for larger epidemiological studies. However, results obtained from NS for influenza virus type A, metapneumovirus and adenovirus, should be interpreted carefully.
Adenoviridae ; Child ; Child, Hospitalized ; Epidemiologic Studies ; Fluorescent Antibody Technique ; Humans ; Influenza, Human ; Korea ; Metapneumovirus ; Orthomyxoviridae ; Parainfluenza Virus 3, Human ; Pediatrics ; Respiratory Tract Infections ; Rhinovirus ; Viruses

PURPOSE: Although the nasopharyngeal aspirate (NPA) is more commonly used because of relatively higher accuracy, the nasal swab (NS) is a less painful and easier method than NPA. A few recent reports have shown that NS is more effective than NPA for the detection of respiratory virus using immunofluorescence or viral culture. The objective of the present study was to compare the results of NPA and NS sampling specimens in children for respiratory viruses detection using multiplex RT-PCR. METHODS: From December 2008 to June 2009 Paired NPA and NS specimens were collected from 250 children admitted with symptoms of acute respiratory infections at the Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. The sensitivity and agreement of virus detection between NPA and NS using multiplex RT-PCR were compared and analyzed. RESULTS: The median age of the subjects was 1.3 years (range, 20 days to 16.5 years), and 228 patients (91.2%) were under the age of 5 years. The agreement of virus detection between NPA and NS was excellent (Cohen's kappa >0.8) for parainfluenza virus type 3 or substantial (0.6 to 0.8) for rhinovirus A, RSV A and RSV B, moderate (0.4 to 0.6) for adenovirus and metapneumovirus and poor (<0.4) for influenza A. The overall sensitivity of detection of respiratory viruses was significantly higher in NPA (0.96) than in NS (0.59, P<0.05). CONCLUSION: We recommend NPA may be more accurate specimen for detection of respiratory viruses in hospitalized children. NS might be used in limited cases at a office setting or for larger epidemiological studies. However, results obtained from NS for influenza virus type A, metapneumovirus and adenovirus, should be interpreted carefully.
Adenoviridae ; Child ; Child, Hospitalized ; Epidemiologic Studies ; Fluorescent Antibody Technique ; Humans ; Influenza, Human ; Korea ; Metapneumovirus ; Orthomyxoviridae ; Parainfluenza Virus 3, Human ; Pediatrics ; Respiratory Tract Infections ; Rhinovirus ; Viruses