a small molecule screen in yeast identifies inhibitors targeting protein-protein interactions within the vaccinia virus replication complex.genetic and biochemical data have identified at least four viral proteins essential for vaccinia virus (vacv) dna synthesis: the dna polymerase e9, its processivity factor (the heterodimer a20/d4) and the primase/helicase d5. these proteins are part of the vacv replication complex in which a20 is a central subunit interacting with e9, d4 and d5. we hypothesised that molecules able to modulate protein-protein interactions within the replication complex may represent a new class of compounds with ...201222884885
prevalence of coxsackievirus a6 and enterovirus 71 in hand, foot and mouth disease in nanjing, china in 2013.although hand, foot and mouth disease (hfmd) has been strongly associated with enterovirus 71 (ev71), coxsackievirus a16 (cva16) and other enteroviruses, studies regarding coxsackievirus a6 (cva6) infection in hfmd are limited. the aim of this study was to identify the major etiological agents causing hfmd in nanjing in 2013 and explore the clinical and genetic characteristics of the prevalent enterovirus (ev) types in hfmd.201526090576
dynamic constitution of the pathogens inducing encephalitis in hand, foot and mouth disease in kunming, 2009-2011.hand, foot and mouth disease (hfmd), caused by various viral pathogens, is an emerging infectious disease in children in asia. understanding the composition of these pathogens is necessary to prevent and control this disease. in the present study, the pathogens in 436 hfmd patients (from 2009 to 2011) with concurrent clinical indications of encephalitis, meningoencephalitis, or both, were defined using the semi-nested pcr. a systematic analysis of the composition of these pathogens was performed ...201525866107
a novel enterovirus and parechovirus multiplex one-step real-time pcr-validation and clinical the number of new enteroviruses and human parechoviruses seems ever growing, the necessity for updated diagnostics is relevant. we have updated an enterovirus assay and combined it with a previously published assay for human parechovirus resulting in a multiplex one-step rt-pcr assay. the multiplex assay was validated by analysing the sensitivity and specificity of the assay compared to the respective monoplex assays, and a good concordance was found. furthermore, the enterovirus assay was ab ...201323845901
enterovirus 71 blocks selectively type i interferon production through the 3c viral protein in mice.type i interferons (ifns) represent an essential innate defense mechanism for controlling enterovirus 71 (ev 71) infection. mice inoculated with ev 71 produced a significantly lower amount of type i ifns than those inoculated with poly (i:c), adenovirus type v, or coxsackievirus b3 (cb3). ev 71 infection, however, mounted a proinflammatory response with a significant increase in the levels of serum and brain interleukin (il)-6, monocyte chemoattractant protein-1, tumor necrosis factor, and ifn-γ ...201222997081
human sentinel surveillance of influenza and other respiratory viral pathogens in border areas of western cambodia.little is known about circulation of influenza and other respiratory viruses in remote populations along the thai-cambodia border in western cambodia. we screened 586 outpatients (median age 5, range 1-77) presenting with influenza-like-illness (ili) at 4 sentinel sites in western cambodia between may 2010 and december 2012. real-time reverse transcriptase (rrt) pcr for influenza was performed on combined nasal and throat specimens followed by viral culture, antigenic analysis, antiviral suscept ...201627028323
endocytosis of integrin-binding human picornaviruses.picornaviruses that infect humans form one of the largest virus groups with almost three hundred virus types. they include significant enteroviral pathogens such as rhino-, polio-, echo-, and coxsackieviruses and human parechoviruses that cause wide range of disease symptoms. despite the economic importance of picornaviruses, there are no antivirals. more than ten cellular receptors are known to participate in picornavirus infection, but experimental evidence of their role in cellular infection ...201223227048
prolonged viral rna detection in the central nervous system of one-week-old swiss albino mice following coxsackievirus b4 and echovirus 9 infection.type b coxsackieviruses (cv-b), together with echoviruses (e), are among the most common pathogens encountered in aseptic meningitis and meningoencephalitis. they frequently infect the central nervous system (cns). the mechanisms of virus spreading in the cns are poorly understood. in the present study, we investigated cv-b4 and e-9 spreading and neurotropism within intraperitoneally inoculated one-week-old swiss albino mice.201222398876
molecular identification of human enteroviruses associated with aseptic meningitis in yunnan province, southwest china.human enteroviruses (evs) are the major causative agents of aseptic meningitis. in this study, a total of 524 children were admitted to the children kunming hospital (continental china) for aseptic meningitis manifestations in 2009 and 2010. an ev infection was diagnosed in 85/524 children (16.2 %) and the viruses detected were assigned to 16 serotypes. most serotypes belonged to the enterovirus b species. echovirus 9 was predominant (24.7 %), followed by coxsackievirus b5 (23.5 %) and then echo ...201627652088
carbon nanotubes supported tyrosinase in the synthesis of lipophilic hydroxytyrosol and dihydrocaffeoyl catechols with antiviral activity against dna and rna viruses.hydroxytyrosol and dihydrocaffeoyl catechols with lipophilic properties have been synthesized in high yield using tyrosinase immobilized on multi-walled carbon nanotubes by the layer-by-layer technique. all synthesized catechols were evaluated against a large panel of dna and rna viruses, including poliovirus type 1, echovirus type 9, herpes simplex virus type 1 (hsv-1), herpes simplex virus type 2 (hsv-2), coxsackievirus type b3 (cox b3), adenovirus type 2 and type 5 and cytomegalovirus (cmv). ...201526260341
[etiology and molecular-epidemiological characteristics of viral meningoencephalitis of zhejiang province in 2013].to investigate pathogens and molecular-epidemiology characteristics of viral meningoencephalitis in the monitoring sites of zhejiang province, 2013.201525975409
echovirus serotypes circulating in malaysia from 2002 to identify the circulating serotypes of human echovirus in malaysia from 2002 to 2013.201626972396
identification and characterization of unrecognized viruses in stool samples of non-polio acute flaccid paralysis children by simplified vidisca.the use of sequence independent methods combined with next generation sequencing for identification purposes in clinical samples appears promising and exciting results have been achieved to understand unexplained infections. one sequence independent method, virus discovery based on cdna amplified fragment length polymorphism (vidisca) is capable of identifying viruses that would have remained unidentified in standard diagnostics or cell cultures.201425112200
molecular characterization of enteroviruses associated with neurological infections in spain, order to investigate the etiology of viral neurological infections in spain, a national study was performed in 2008. the results obtained have been published. enteroviruses were the most frequent cause of the aseptic meningitis and infant febrile syndromes. the present report supplements the previous study with the genotyping of the detected enteroviruses. typing was by amplification of partial vp1 region and sequencing in 70 (53%) of the 132 available cerebrospinal fluid samples positive for ...201323893817
[clinical, epidemiological, and etiological studies of aseptic meningitis in adults].from summer to autumn, we noted the occurrence of a small epidemic of aseptic meningitis in adults. over the last 10 years, we have encountered 203 male (mean age, 34.6 ± 15.0 years) and 157 female (mean age, 35.6 ± 16.3 years) patients with aseptic meningitis. we could identify the causative virus in 17 (81%) of 21 cases during the abovementioned months in 2012. identification rates of the virus in the stool, cerebrospinal fluid, throat swab, and serum samples were 71%, 67%, 42%, and 5%, respec ...201425342012
the vaccinia virus dna polymerase and its processivity factor.vaccinia virus is the prototypic poxvirus. the 192 kilobase double-stranded dna viral genome encodes most if not all of the viral replication machinery. the vaccinia virus dna polymerase is encoded by the e9l gene. sequence analysis indicates that e9 is a member of the b family of replicative polymerases. the enzyme has both polymerase and 3'-5' exonuclease activities, both of which are essential to support viral replication. genetic analysis of e9 has identified residues and motifs whose altera ...201728159613
logiq e9 shear wave elastrography for detection of liver fibrosis in patients with chronic hepatitis c virus.shear wave elastography (swe) is an emerging technology that assesses hepatic fibrosis. the aim of our study was to determine the diagnostic accuracy of general electric (ge) healthcare's logiq e9 swe in patients with chronic hepatitis c virus using liver biopsy as the reference standard.201627812720
poxvirus uracil-dna glycosylase-an unusual member of the family i uracil-dna glycosylases.uracil-dna glycosylases are ubiquitous enzymes, which play a key role repairing damages in dna and in maintaining genomic integrity by catalyzing the first step in the base excision repair pathway. within the superfamily of uracil-dna glycosylases family i enzymes or ungs are specific for recognizing and removing uracil from dna. these enzymes feature conserved structural folds, active site residues and use common motifs for dna binding, uracil recognition and catalysis. within this family the e ...201627684934
structural analysis of point mutations at the vaccinia virus a20/d4 interface.the vaccinia virus polymerase holoenzyme is composed of three subunits: e9, the catalytic dna polymerase subunit; d4, a uracil-dna glycosylase; and a20, a protein with no known enzymatic activity. the d4/a20 heterodimer is the dna polymerase cofactor, the function of which is essential for processive dna synthesis. the recent crystal structure of d4 bound to the first 50 amino acids of a20 (d4/a201-50) revealed the importance of three residues, forming a cation-π interaction at the dimerization ...201627599859
[clinical, epidemiological and etiological studies of adult aseptic meningitis: report of 13 cases with mumps meningitis].we experienced 13 cases (29.8 ± 7.0 years) of mumps meningitis and 365 cases of adult aseptic meningitis during 11 years from 2004 to 2014. a small epidemic of mumps occurred for 3-4 years, and the incidence rate of adult mumps meningitis coincided with the epidemic without seasonal fluctuation. parotitis was observed in 8 of the 13 mumps meningitis patients (61.5%) and orchitis in 2 of 7 male patients (28.6%). there were no differences in clinical manifestations, laboratory findings, and outcom ...201526156258
crystal structure of the vaccinia virus uracil-dna glycosylase in complex with dna.vaccinia virus polymerase holoenzyme is composed of the dna polymerase catalytic subunit e9 associated with its heterodimeric co-factor a20·d4 required for processive genome synthesis. although a20 has no known enzymatic activity, d4 is an active uracil-dna glycosylase (ung). the presence of a repair enzyme as a component of the viral replication machinery suggests that, for poxviruses, dna synthesis and base excision repair is coupled. we present the 2.7 å crystal structure of the complex forme ...201526045555
localized exanthem due to echovirus 9. 201525919440
sepsis and meningitis caused by pasteurella multocida and echovirus 9 in a neonate. 201425386969
cell labeling and injection in developing embryonic mouse hearts.testing the fate of embryonic or pluripotent stem cell-derivatives in in vitro protocols has led to controversial outcomes that do not necessarily reflect their in vivo potential. preferably, these cells should be placed in a proper embryonic environment in order to acquire their definite phenotype. furthermore, cell lineage tracing studies in the mouse after labeling cells with dyes or retroviral vectors has remained mostly limited to early stage mouse embryos with still poorly developed organs ...201424797676
crystal structure of the vaccinia virus dna polymerase holoenzyme subunit d4 in complex with the a20 n-terminal domain.vaccinia virus polymerase holoenzyme is composed of the dna polymerase e9, the uracil-dna glycosylase d4 and a20, a protein with no known enzymatic activity. the d4/a20 heterodimer is the dna polymerase co-factor whose function is essential for processive dna synthesis. genetic and biochemical data have established that residues located in the n-terminus of a20 are critical for binding to d4. however, no information regarding the residues of d4 involved in a20 binding is yet available. we expres ...201424603707
rubella virus genotype 1g and echovirus 9 as etiologic agents of exanthematous diseases in brazil: insights from phylogenetic analysis.the aim of the present study was to identify the rubella virus (rv) and enterovirus (ev) genotypes detected during the epidemiological surveillance on exanthematic febrile diseases (vigifex) study and to perform phylogenetic analysis. ten rv- and four ev-positive oropharyngeal samples isolated from cell culture were subjected to rt-pcr and sequencing. genotype 1g and echovirus 9 (e-9) was identified in rv- and ev-positive samples, respectively. the rv 1g genotype has been persisting in brazil si ...201424327091
[complete genome sequence characteristics of human echovirus 9 strain isolated in yunnan, china].to analyze the genomic sequence characteristics of a human echovirus 9(echo-9) strain isolated from a child with hand-foot-mouth disease (hfmd) in kunming, yunnan province, in 2010. the complete genome sequence of a human echovirus 9 strain, msh-km812-2010 was determined. as other human enterovirus, its genome was 7,424 nucleotides (nts) in length and encoded for 2,203 amino acids (aas). in comparison to other human enteroviruses, msh-km812-2010 strain had the highest homology with other strains ...201323757842
a single amino acid substitution in viral vp1 protein alters the lytic potential of clone-derived variants of echovirus 9 dm strain in human pancreatic vitro studies with primary human pancreatic islets suggest that several enterovirus serotypes are able to infect and replicate in beta cells. some enterovirus strains are highly cytolytic in vitro whereas others show virus replication with no apparent islet destruction. the capability to induce islet destruction is determined only partially by the virus serotype, since strain specific differences have been detected within some serotypes including echovirus 9 (e-9). in this study, the viral ge ...201323595636
low-resolution structure of vaccinia virus dna replication machinery.smallpox caused by the poxvirus variola virus is a highly lethal disease that marked human history and was eradicated in 1979 thanks to a worldwide mass vaccination campaign. this virus remains a significant threat for public health due to its potential use as a bioterrorism agent and requires further development of antiviral drugs. the viral genome replication machinery appears to be an ideal target, although very little is known about its structure. vaccinia virus is the prototypic virus of th ...201323175373
raxl regulates chick ganglion cell development.raxl is a paired-like homeobox gene involved in vertebrate eye morphogenesis. we examined raxl protein expression patterns during chick retinal development in combination with ganglion cell markers including the ra4 antigen, cbrn-3, islet-1 and neuronal type iii beta-tubulin. double-immunostaining demonstrated that downregulation of raxl protein correlates with upregulation of ganglion cell markers in the ganglion cell layer (gcl). to explore this correlation in vivo, we performed gain- and loss ...200312963109
expression of transgenes in primary neurons from chick peripheral and central nervous systems by retroviral infection of early embryos.many cell biological studies require the expression of transgenes in cells in culture, but it is difficult to obtain uniform, stable, and efficient expression of transgenes in primary neurons. we have approached this problem by adapting from developmental biologists the avian retroviral vector, rcas. this vector allows the introduction of a transgene by infection early in chick embryonic development. transgenes that are less than 2.6 kb in size can be cloned through an adapter vector, slax 12 nc ...200312884700
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