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prospective identification of enteroviruses involved in meningitis in 2006 through direct genotyping in cerebrospinal fluid.enterovirus infections were investigated with special emphasis on performing rapid molecular identification of enterovirus serotypes responsible for aseptic meningitis directly in cerebrospinal fluid (csf). enterovirus genotyping was carried out directly with specimens tested for the diagnostic procedure, using two seminested pcr assays designed to amplify the complete and partial gene sequences encoding the vp1 and vp4/vp2 capsid proteins, respectively. the method was used for identifying the e ...200817977989
tissue uptake of mercury is changed during the course of a common viral infection in mice.mercury (hg) has been shown to have immunotoxic effects and to influence the severity of infection. however, the impact of infection on the normal hg homeostasis in different target organs involved in the disease process has not been studied. in this study, hg was measured through inductively coupled plasma-mass spectrometry (icp-ms) in the intestine, serum, liver, and brain on days 3, 6, and 9 of coxsackievirus b3 (cvb3) infection in female balb/c mice. the severity of the infection was assesse ...200817888900
rapid and differential diagnosis of foot-and-mouth disease, swine vesicular disease, and vesicular stomatitis by a new multiplex rt-pcr assay.a highly sensitive and specific one-step multiplex rt-pcr assay has been developed and standardised for the simultaneous and differential detection of the most important vesicular viruses affecting livestock: foot-and-mouth disease virus (fmdv), swine vesicular disease virus (svdv), and vesicular stomatitis virus (vsv). the method uses three primer sets, each one specific for the corresponding virus, selected to detect of all serotypes of fmd and vs. the detection range was confirmed by examinat ...200817964668
recombinant coxsackievirus vectors for prevention and therapy of virus-induced heart disease.cardiovascular diseases are the major cause of human death and have been linked to many different risk factors. among them, coxsackievirus b3 (cvb3), as a member of the enterovirus group, is one of the most important infectious agents of virus-induced myocarditis. despite the fact that the molecular structure of this pathogen has been characterized very precisely, there is no virus-specific preventive or therapeutic procedure against cvb3-induced heart disease in clinical use today. a promising ...200817897883
a one-step reverse transcriptase loop-mediated isothermal amplification assay for simple and rapid detection of swine vesicular disease virus.this report describes the development of a one-step reverse transcriptase loop-mediated isothermal amplification (rt-lamp) assay for the detection of swine vesicular disease virus (svdv). the assay detects the virus rapidly, within 30-60 min and the result is visualised either by gel-electrophoresis or by the naked eye through the addition of sybrgreen. a collection of 28 svdv isolates were tested positive, while heterologous viruses such as foot-and-mouth disease virus and vesicular stomatitis ...200817920701
using ifn-gamma as a biomarker for detecting exposure to viral pathogens.to determine whether interferon gamma (ifn-gamma) can be used as a biomarker of exposure to viral pathogens, 12-week-old balb/c mice were injected intraperitoneally with coxsackievirus b3 (cvb3) or coxsackievirus b4 (cvb4) diluted in sterilized phosphate-buffered saline (pbs). control mice were injected with pbs only. four months after viral infection, mouse spleen cells were harvested and assayed for the release of ifn-gamma by memory t cells after in vitro stimulation with viral antigens, phyt ...200817926092
1a and 3d gene sequences of coxsackievirus b3 strain cc: variation and phylogenetic analysis.coxsackievirus b3 (cvb3) was thought to be the most common causative agent of life-threatening viral myocarditis. coxsackievirus b3 strain cc (cvb3-cc) was isolated in china; however, no sequence data are available. the 1a and 3d regions of cvb3-cc were sequenced and phylogenetic analysis was done with reference to ten other cvb3 strains and all 36 prototype strains of human enterovirus b (hev-b). sequence analysis showed that the 1a gene region of cvb3-cc consisted of 207 nucleotides, encoding ...200817852360
role of gnra motif mutations within stem-loop v of internal ribosome entry segment in coxsackievirus b3 molecular attenuation.the lengthy 5' nontranslated region of coxsackievirus b3 (cvb3) forms a highly ordered secondary structure containing an internal ribosome entry segment (ires), which plays an important role in controlling viral translation and pathogenesis. the stem-loop v (sl-v) of this ires contains a large lateral bulge loop which encompasses two conserved gnra motifs. in this study, we analyzed the effects of point mutations within the gnra motifs of the cvb3 ires. we characterized in vitro virus production ...200817693702
molecular epidemiological study of hev-b enteroviruses involved in the increase in meningitis cases occurred in spain during 2006.human enteroviruses are one of the main etiological agents of aseptic meningitis and other central nervous system infections, particularly the serotypes included in the enterovirus b species. molecular methods have proved useful to identify serotypes in clinical samples, facilitating the epidemiological study of these viruses. in the spring of 2006, there was a significant increase in meningitis cases caused by enteroviruses in spain. in the present study, 138 enteroviruses directly detected in ...200818428125
development of robust antiviral assays for profiling compounds against a panel of positive-strand rna viruses using atp/luminescence readout.the development of antiviral assays using an atp/luminescence-based readout to profile antiviral compounds against the positive-strand rna viruses: yellow fever virus (yfv), west nile virus (wnv), sindbis virus, and coxsackie b virus, representing three virus families, is described. this assay readout is based upon the bioluminescent measurement of atp in metabolically active cells. antiviral efficacy was determined by measuring the atp level in cells that were protected from the viral cytopathi ...200818433887
a raft-derived, pak1-regulated entry participates in alpha2beta1 integrin-dependent sorting to caveosomes.we have previously shown that a human picornavirus echovirus 1 (ev1) is transported to caveosomes during 2 h together with its receptor alpha2beta1 integrin. here, we show that the majority of early uptake does not occur through caveolae. alpha2beta1 integrin, clustered by antibodies or by ev1 binding, is initially internalized from lipid rafts into tubulovesicular structures. these vesicles accumulate fluid-phase markers but do not initially colocalize with caveolin-1 or internalized simian vir ...200818448666
remission of cvb3-induced viral myocarditis by in vivo th2 polarization via hydrodynamics-based interleukin-4 gene transfer.regulation of th polarization was critical for the prevention of coxsackievirus b3 (cvb3) induced myocarditis. in the present study, interleukin (il)-4 was over-expressed by hydrodynamics-based gene transfection (hgt) to induce the in vivo th2 bias and evaluate the influence of th polarization on the pathogenesis of cvb3-myocaditis.200818512734
a laboratory study of survival of selected microorganisms after heat treatment of biowaste used in biogas plants.the aim of the study was to assess the effect of pasteurisation, as set by the european regulation ec 1774/2002, on selected pathogens and indicator organisms. unpasteurised substrate (biowaste), including animal by-products from a full-scale biogas plant was heat treated under laboratory conditions at 70 degrees c and 55 degrees c for 30 min and 60 min. heat treatment at 55 degrees c for 60 min was not sufficient to achieve a hygienically acceptable product. heat treatment at 70 degrees c for 3 ...200818513960
proteasome inhibition attenuates coxsackievirus-induced myocardial damage in mice.coxsackievirus b3 (cvb3) is one of the most prevalent pathogens of viral myocarditis, which may persist chronically and progress to dilated cardiomyopathy. we previously demonstrated a critical role of the ubiquitin-proteasome system (ups) in the regulation of coxsackievirus replication in mouse cardiomyocytes. in the present study, we extend our interest to an in vivo animal model to examine the regulation and role of the ups in cvb3-induced murine myocarditis. male myocarditis-susceptible a/j ...200818515649
the structure and function of a cis-acting element located upstream of the ires that influences coxsackievirus b3 rna translation.we have investigated the importance of a conserved hexa-nucleotide stretch in the apical loop within stem-loop c (sl c, nt 104-180), upstream of the ribosome landing site, on cvb3 ires function. the deletion or substitution mutation at this apical loop resulted in significant decrease in ires activity. both the mutant ires rnas failed to interact with certain trans-acting factors. furthermore, expression of cvb3 2a protease significantly enhanced ires activity of the wild type, but the effect wa ...200818533219
fatal neonatal myocarditis caused by a recombinant human enterovirus-b variant.we report a case of fatal myocarditis in a newborn infant who was infected with a human enterovirus detected by throat culture and rt-pcr for viral rna in plasma. whole genome sequence analysis revealed the virus to be a genomic chimera that likely arose from recombination between coxsackievirus b3 and two recently identified enteroviruses, ev 86 and ev97.200818536618
cardiac-targeted rna interference mediated by an aav9 vector improves cardiac function in coxsackievirus b3 cardiomyopathy.rna interference (rnai) has potential to be a novel therapeutic strategy in diverse areas of medicine. in this paper, we report on targeted rnai for the treatment of a viral cardiomyopathy, which is a major cause of sudden cardiac death or terminal heart failure in children and young adults. rnai therapy employs small regulatory rnas to achieve its effect, but in vivo use of synthetic small interfering rnas is limited by instability in plasma and low transfer into target cells. we instead evalua ...200818548221
comparison of automated nucleic acid extraction methods with manual extraction.automated nucleic acid extractors can improve workflow and decrease variability in the clinical laboratory. we evaluated qiagen ez1 (valencia, ca) and biomérieux (durham, nc) easymag extractors compared with qiagen manual extraction using targets and matrices commonly available in the clinical laboratory. pooled samples were spiked with various organisms, serially diluted, and extracted in duplicate. the organisms/matrices were bordetella pertussis/bronchoalveolar lavage, herpes simplex virus ii ...200818556770
cleavage of eukaryotic initiation factor eif5b by enterovirus 3c proteases.the enteroviruses poliovirus (pv), coxsackie b virus (cvb) and rhinovirus (hrv) are members of picornaviridae that inhibit host cell translation early in infection. enterovirus translation soon predominates in infected cells, but eventually also shuts off. this complex pattern of modulation of translation suggests regulation by a multifactorial mechanism. we report here that eif5b is proteolytically cleaved during pv and cvb infection of cultured cells, beginning at 3 hours post-infection and in ...200818572216
coxsackievirus b3-induced myocarditis: infection of females during the estrus phase of the ovarian cycle leads to activation of t regulatory cells.transgenic female mice expressing the tnfalpha gene under the cardiac myosin promoter (tnf1.6) develop substantially increased myocarditis and increased numbers of cd4+th1 (interferon gamma+) cells when infected with coxsackievirus b3 (cvb3) during the diestrus and proestrus phases of the estrus cycle compared to females infected during the estrus and metestrus phases. cardiac virus titers were increased in females infected in estrus compared to females infected during the other phases. t regula ...200818586295
autophagosome supports coxsackievirus b3 replication in host cells.recent studies suggest a possible takeover of host antimicrobial autophagy machinery by positive-stranded rna viruses to facilitate their own replication. in the present study, we investigated the role of autophagy in coxsackievirus replication. coxsackievirus b3 (cvb3), a picornavirus associated with viral myocarditis, causes pronounced intracellular membrane reorganization after infection. we demonstrate that cvb3 infection induces an increased number of double-membrane vesicles, accompanied b ...200818596087
mirror syndrome due to coxackie b virus associated to maternal peripartum cardiomyopathy. 200818601628
ubiquitination is required for effective replication of coxsackievirus b3.protein ubiquitination and/or degradation by the ubiquitin/proteasome system (ups) have been recognized as critical mechanisms in the regulation of numerous essential cellular functions. the importance of the ups in viral pathogenesis has become increasingly apparent. using murine cardiomyocytes, we have previously demonstrated that the ups plays a key role in the replication of coxsackievirus b3 (cvb3), an important human pathogen associated with various diseases. to further elucidate the under ...200818612413
an enterovirus strain isolated from diabetic child belongs to a genetic subcluster of echovirus 11, but is also neutralised with monotypic antisera to coxsackievirus a9.an enterovirus strain (designated d207) isolated from a slovakian diabetic child and originally serotyped as coxsackievirus a9 (cav-9) was found to cause rapid cytolysis coinciding with severe functional damage of the surviving cells in primary cultures of human pancreatic islets. this finding prompted us to clone the isolate for full-length genome sequencing and molecular characterization as the prototype strain of cav-9 is known to cause only minimal damage to insulin-producing beta-cells. bas ...200818632967
quick identification of effective small interfering rnas that inhibit the replication of coxsackievirus a16.coxsackievirus a16 (ca16) is a major causative agent of hand, foot, and mouth disease (hfmd). it can cause myocarditis, pericarditis and fatal shock. there is no effective therapy against ca16. rna interference (rnai) is a powerful tool to silence gene expression. the small interfering rna (sirna) that induces rna degradation has recently been used as an anti-virus agent to inhibit virus replication. in this study, we established the complete nucleotide sequence of ca16 strain shzh05-1, and then ...200818662724
design of lna-modified sirnas against the highly structured 5' utr of coxsackievirus b3.this study describes a strategy to develop lna-modified small interfering rna (sirnas) against the highly structured 5' utr of coxsackievirus b3 (cvb-3), which is an attractive target site due to its high degree of conservation. accessible sites were identified based on structural models and rnase h assays with dna oligonucleotides. subsequently, lna gapmers, sirnas, silnas and small internally segmented interfering rna (sisilnas) were designed against sites, which were found to be accessible in ...200818691577
identification of hla-a*01- and hla-a*02-restricted cd8+ t-cell epitopes shared among group b enteroviruses.acute enteroviral infections ranging from meningitis, pancreatitis to myocarditis are common and normally well controlled by the host immune system comprising virus-specific cd8+ cytotoxic t lymphocytes (ctl). however, in some patients enteroviruses and especially coxsackieviruses of group b are capable of inducing severe chronic forms of diseases such as chronic myocarditis. currently, it is not known whether divergences in the ctl-related immune response may contribute to the different outcome ...200818753217
coxsackievirus b4 uses autophagy for replication after calpain activation in rat primary neurons.coxsackievirus is the most important cause of meningitis and encephalitis in infants; an infection is sometimes fatal or may lead to neurodevelopmental defects. here, we show that coxsackievirus b4 (cvb4) induces an autophagy pathway for replication in rat primary neurons. notably, calpain inhibitors reduce autophagosome formation. conversely, the inhibition of the autophagy pathway with 3-methyladenine inhibits calpain activation. this work reveals, for the first time, that calpain is essential ...200818799585
sicam-1 intrathecal synthesis and release during the acute phase in children suffering from coxsackie a9 and s. pneumoniae meningoencephalitis.the intercellular adhesion molecule is a transmembrane glycoprotein belonging to the immunoglobulin superfamily. serum and cerebrospinal fluid (csf) soluble intercellular adhesion molecule 1 (sicam-1) from normal control children as well as from children with guillain-barré syndrome (gbs), with coxsackie a9 virus meningoencephalitis and with streptococcus pneumoniae meningoencephalitis were studied. sicam-1 was quantified using an immunoenzimatic assay and albumin using the immunodiffusion techn ...200818813709
role of class i human leukocyte antigen molecules in early steps of echovirus infection of rhabdomyosarcoma cells.several echoviruses use decay accelerating factor (daf) as a cell surface receptor. however, most of them require additional cell surface coreceptors. we investigated the respective roles of daf and class i human leukocyte antigen (hla) molecules in the early steps of the echovirus 11 (ev11) lifecycle in rhabdomyosarcoma (rd) cells. ev11 infection was inhibited at an early stage by anti-beta2-microglobulin (beta2m) and anti-hla monoclonal antibodies and by a soluble monochain hla class i molecul ...200818823925
coxsackievirus b3 induction of nfat: requirement for myocarditis susceptibility.ultraviolet (u.v.) inactivated coxsackievirus b3 (cvb3) induces rapid calcium flux in naïve balb/c cd4+ t cells. cd4+ cells lacking decay accelerating factor (daf-/-) show little calcium flux indicating that virus cross-linking of this virus receptor protein is necessary for calcium signaling in cvb3 infection. interaction of cvb3 with cd4+ cells also activates nfat dna binding. to show that nfat activation is crucial to cvb3 induced disease, wild-type mice and transgenic mice expressing dominan ...200818829062
[rapid enterovirus genotyping in cerebrospinal fluids: a two-year prospective study in a virology laboratory setting].enterovirus (ev - 68 serotypes) infections comprise a wide spectrum of clinical presentations including infections of the central nervous system. in severe clinical presentation or epidemics, the precise identification of the involved serotype is necessary.200818835107
increased gelatinase b/matrix metalloproteinase 9 (mmp-9) activity in a murine model of acute coxsackievirus b4-induced pancreatitis.infection of mice with coxsackievirus b4 results within days in a severe acute necrotizing pancreatitis, which resolves completely within weeks. gelatinase b or matrix metalloproteinase 9 (mmp-9) has previously been shown to be involved in several models of pancreatitis, but its role in virus-induced pancreatitis has never been investigated. we here report that mmp-9 levels are markedly increased in the pancreas of mice that developed acute pancreatitis following infection with coxsackievirus b4 ...200818929380
role of coxsackievirus b4 in the pathogenesis of type 1 diabetes.environmental factors, especially viruses, are thought to play an important role in the initiation or acceleration of the pathogenesis of type 1 diabetes (t1d). data from retrospective and prospective epidemiological studies strongly suggest that enteroviruses, such as coxsackievirus b4 (cv-b4), may be associated with the development of t1d. it has also been shown that enterovirus infections are significantly more prevalent in at-risk individuals such as the siblings of diabetic patients, when t ...200818951821
visualizing the dynamics of viral replication in living cells via tat peptide delivery of nuclease-resistant molecular beacons.in this study, we describe the use of nuclease-resistant molecular beacons (mbs) for the real-time detection of coxsackievirus b6 replication in living buffalo green monkey kidney (bgmk) cells via tat peptide delivery. a nuclease-resistant mb containing 2'-o-methyl rna bases with phosphorothioate internucleotide linkages was designed to specifically target an 18-bp 5' noncoding region of the viral genome. for intracellular delivery, a cell-penetrating tat peptide was conjugated to the mb by usin ...200818988730
shedding light on virus replication. 200818997007
letter by pinto and heymans regarding article, "ablation of matrix metalloproteinase-9 increases severity of viral myocarditis in mice". 200819001029
[matrine's inhibitory effect on apoptosis of cardiomyocytes induced by coxsackie virus b(3) and on calcium overload]. 200819099839
[intracellular clearance of coxsakievirus b3 infection by short interfering rna and its mechanism study].to evaluate the possibility of short interfering rna (sirna) inhibiting coxsackievirus b3 (cvb3) infection in vitro, and discover the mechanism initially.200819105337
[relationship between eb virus, cytomegalo virus, herpes simplex virus and coxsackievirus infection and relapse of multiple sclerosis].to study the relation between the recent active infection with epstein-barr virus, cytomegalovirus,herpes simplex virus-1, coxsackievirus b i-iv and the relapse of relapsing-remitting multiple sclerosis (rr ms).200819544649
inhibition of coxsackievirus b3 and related enteroviruses by antiviral short interfering rna pools produced using phi6 rna-dependent rna polymerase.coxsackievirus b3 (cbv3) is a member of the human enterovirus b species and a common human pathogen. even though much is known about the enteroviral life cycle, no specific drugs are available to treat enterovirus infections. rna interference (rnai) has evolved to be an important tool for antiviral experimental therapies and gene function studies. we describe here a novel approach for rnai against cbvs by using a short interfering (sirna) pool covering 3.5 kb of cbv3 genomic sequence. the rna-de ...200919553393
[acute myocarditis due to coxsackie virus b3 in two infants].in the spring and summer of 2008 two seriously ill male infants were admitted to a paediatric intensive care unit. initially, both had a fever, were drinking less and were pale complexioned. physical examination revealed tachycardia, slow capillary filling and liver enlargement. within a few hours, both infants developed circulatory and respiratory failure. a chest radiograph showed that the heart was enlarged and echocardiography revealed that the pump function of both ventricles was severely d ...200919785892
coxsackievirus b3 inhibits antigen presentation in vivo, exerting a profound and selective effect on the mhc class i pathway.many viruses encode proteins whose major function is to evade or disable the host t cell response. nevertheless, most viruses are readily detected by host t cells, and induce relatively strong t cell responses. herein, we employ transgenic cd4(+) and cd8(+) t cells as sensors to evaluate in vitro and in vivo antigen presentation by coxsackievirus b3 (cvb3), and we show that this virus almost completely inhibits antigen presentation via the mhc class i pathway, thereby evading cd8(+) t cell immun ...200919834548
usefulness of real-time reverse transcription-polymerase chain reaction for the diagnosis of echovirus aseptic meningitis using cerebrospinal fluid.quantitative real-time reverse transcription-polymerase chain reaction (q-rt-pcr) was used to diagnose echovirus infection and the results were compared to those obtained with the viral culture rate. cerebrospinal fluid (csf) from a total of 40 aseptic meningitis patients was used. positive csf samples, determined by viral culture (n=29), contained significantly higher echovirus genome copy numbers (mean, 329 copies/microl) than did culture-negative csf samples (n=11) (mean, 34.2 copies/microl; ...200919934538
[analysis of 62 adult patients with viral meningitis].to investigate the clinical manifestation, cerebrospinal fluid (csf) and auxiliary examination findings of adult viral meningitis.200920104783
[oxidative stress in patients with type i diabetes mellitus and persistent coxsackie virus b infection as the reason of dysfunction of the immune system].a chronic inflammatory process takes place in patients with diabetes mellitus type 1. numerous disorders of the immune status and complications testify the present of this process. the presence of chronic inflammation at diabetes mellitus enhances free radical reactions which are accompanied by oxidative stress.200920455442
[molecular characterization of coxsackievirus b3 isolated from an outbreak of aseptic meningitis in shandong province, china].to identify the pathogen caused an outbreak of aseptic meningitis in tancheng county of shandong province in 2008, and to analyze the molecular characterization of vp1 gene of the coxsackievirus b3(cvb3) isolates.200920718355
il-10 is pathogenic during the development of coxsackievirus b4-induced chronic pancreatitis.using a mouse model of coxsackievirus b4 (cvb4-v)-induced chronic pancreatitis, we investigated whether cytokines are involved in the progression of acute disease to chronic inflammatory disease. we show that il-10 contributed to the development of chronic pancreatitis since acute disease resolved when il-10 was absent or when il-10 signaling was disrupted. we explored the underlying mechanisms by which il-10 affected disease progression, using a novel approach to assess immunological events occ ...200919800092
[extraction, selenium-nanoparticle preparation and anti-virus bioactivity determination of polysaccharides from caulerpa taxifolia].to extract polysaccharides from caulerpa taxifolia and determine its anti-virus bioactivity.200920432908
amino acids of coxsackie b5 virus are critical for infection of the murine insulinoma cell line, min-6.it was shown recently that 15 successive passages of a laboratory strain of the coxsackie b virus 5 in a mouse pancreas (cbv-5-mpp) resulted in apparent changes in the virus phenotype, which led to the capacity to induce a diabetes-like syndrome in mice. for further characterization of islet cell interactions with a passaged virus strain, a murine insulinoma cell line, min-6, was selected as an experimental model. the cbv-5-mpp virus strain was not able to replicate in min-6 cells in vitro but r ...200919107967
epidemiology of enterovirus types causing neurological disease in austria 1999-2007: detection of clusters of echovirus 30 and enterovirus 71 and analysis of prevalent genotypes.between 1999 and 2007 1,388 stool specimens from patients with acute flaccid paralysis or aseptic meningitis were submitted to the austrian reference laboratory for poliomyelitis. samples (201) yielded non-poliovirus enterovirus in culture. one hundred eighty-one viruses were available for typing and 78 isolates which remained serologically untyped were further analyzed by codehop-pcr and sequencing of the vp1 gene and the 5'-untranslated region (5'-utr). typing revealed an echovirus 30 outbreak ...200919107980
in vivo and in vitro antiviral activities of calycosin-7-o-beta-d-glucopyranoside against coxsackie virus b3.calycosin-7-o-beta-d-glucopyranoside (ccgr) is the main isoflavonoid compound isolated from astragalus membranaceus var. mongholicu (bge.) hsiao, a chinese herb medicine traditionally used to treat viral myocarditis. in this study, its antiviral activities against coxsackie virus b3 (cvb3) causing myocarditis were investigated. in vitro assay showed that ccgr displayed a low cytotoxicity and effectively inhibited cvb3-mediated cytopathic effects on vero cells with an ic(50) value of 25 microg/ml ...200919122283
toll-like receptor 3 signaling on macrophages is required for survival following coxsackievirus b4 infection.toll-like receptor 3 (tlr3) has been proposed to play a central role in the early recognition of viruses by sensing double stranded rna, a common intermediate of viral replication. however, several reports have demonstrated that tlr3 signaling is either dispensable or even harmful following infection with certain viruses. here, we asked whether tlr3 plays a role in the response to coxsackievirus b4 (cb4), a prevalent human pathogen that has been associated with pancreatitis, myocarditis and diab ...200919122812
gonadectomy of male balb/c mice increases tim-3(+) alternatively activated m2 macrophages, tim-3(+) t cells, th2 cells and treg in the heart during acute coxsackievirus-induced myocarditis.the incidence of cardiovascular disease, including inflammatory heart diseases like myocarditis, is increased in men. similarly, male balb/c mice infected with coxsackievirus b3 (cvb3) develop more severe acute inflammation in the heart compared to females. to better understand the effect of male sex hormones on cardiac inflammation, we gonadectomized (gdx) male balb/c mice and examined acute cvb3-induced myocarditis compared to sham controls. viral replication in the heart was not significantly ...200919126426
direct identification of human enterovirus serotypes in cerebrospinal fluid by amplification and sequencing of the vp1 region.human enteroviruses (hev) are a major cause of meningitis and other neurological disease. identification of hev serotypes in clinical cases is important for monitoring emergence of more pathogenic variants, epidemiological surveillance and investigating sources of infection. serotype identification is currently problematic following the widespread adoption of polymerase chain reaction (pcr)-based methods for hev detection in place of virus culture.200919135410
regional administration of oncolytic echovirus 1 as a novel therapy for the peritoneal dissemination of gastric cancer.the dissemination of malignant gastric cells to the peritoneum occurs frequently, usually as an early event in disease, and results in poor patient prognosis. surgery and chemotherapy offer limited therapeutic success. the low-pathogenic human enterovirus, echovirus 1 (ev1), is an oncolytic virus that selectively targets and destroys malignant prostate and ovarian cancer xenografts in vivo. lytic ev1 infection requires the cell surface expression of alpha(2)beta(1), an integrin involved in the d ...200919139835
inflammatory rather than infectious insults play a role in exocrine tissue damage in a mouse model for coxsackievirus b4-induced pancreatitis.infection with coxsackievirus b4 (cvb4) may result in an acute severe necrotizing pancreatitis that mostly remains restricted to the acini of the exocrine parenchyma. the mechanisms responsible for this tissue damage, however, remain poorly understood. we here report that coam, a polyanionic carboxylic acid, provides marked protection against cvb4-induced pancreatitis in a mouse model. despite the fact that coam largely reduced disease severity, as detected by serum amylase and lipase levels as ...200919142976
[effect of a c-type cpg odn on cvb3-infected mice].to investigate the potential effect of a self-designed c-type cpg odn (d-sl01) on coxsackie virus b3 (cvb3) infection.200919174007
antiviral activity of highly potent sirnas against echovirus 30 and its receptor.rna interference (rnai) has been shown to be suitable to inhibit viruses in experimental setups and is considered a promising antiviral strategy that is currently being tested in various clinical trials. the present study provides an approach to design sirnas with high potency against a virus-specific target gene. in recent years, several outbreaks of aseptic meningitis caused by an echovirus 30 (ev-30) infection have been described. based on an initial set of 30 in silico designed sirnas, six s ...200919187792
impact of myocardial inflammation on cytosolic and mitochondrial creatine kinase activity and expression.the disturbance of myocardial energy metabolism has been discussed as contributing to the progression of heart failure. little however is known about the cardiac mitochondrial/cytosolic energy transfer in murine and human inflammatory heart disease. we examined the myocardial creatine kinase (ck) system, which connects mitochondrial atp-producing and cytosolic atp-consuming processes and is thus of central importance to the cellular energy homeostasis. the time course of expression and enzymatic ...200919190956
beta-androstenes and resistance to viral and bacterial infections.this report illustrates that the beta-androstenes are indeed able to upregulate the host immune response to a level that enables the host to resist lethal infection by viruses or bacteria. these agents consist of a subgroup of steroids, which also mediates a rapid recovery of hematopoietic precursor cells after whole-body lethal radiation injury. in vivo, the androstenes increase the levels of the th1 cytokines such as il-2, il-3, and ifn. similar to hydrocortisone, they suppress inflammation, b ...200919212128
bosentan enhances viral load via endothelin-1 receptor type-a-mediated p38 mitogen-activated protein kinase activation while improving cardiac function during coxsackievirus-induced myocarditis.reduced cardiac output is one of the consequences of myocarditis. bosentan, an endothelin-1 receptor (et1r) antagonist, could be useful to reduce cardiac afterload, preserving cardiac output. in this study, we investigated the potential therapeutic use of bosentan in an animal model of viral myocarditis. using a mouse model of coxsackievirus b3 (cvb3)-induced myocarditis, we demonstrated preserved ejection fraction (ef) and fractional shortening (fs) by treatment with bosentan (68+/-5.8% ef and ...200919213955
the evidence of coxsackievirus b3 induced myocarditis as the cause of death in a sichuan snub-nosed monkey (rhinopithecus roxellana).a 5-year-old female sichuan snub-nosed monkey died at the zoological garden from infection with coxsackievirus b3.200919220685
internalization of swine vesicular disease virus into cultured cells: a comparative study with foot-and-mouth disease virus.we performed a comparative analysis of the internalization mechanisms used by three viruses causing important vesicular diseases in animals. swine vesicular disease virus (svdv) internalization was inhibited by treatments that affected clathrin-mediated endocytosis and required traffic through an endosomal compartment. svdv particles were found in clathrin-coated pits by electron microscopy and colocalized with markers of early endosomes by confocal microscopy. svdv infectivity was significantly ...200919225001
viral rna kinetics is associated with changes in trace elements in target organs of coxsackie virus b3 infection.trace elements are pivotal for the host defense, as well as potentially important for viral replication and virulence. studies of sequential changes in viral replication in target organs of infection are sparse and a possible association with changes in specific trace elements is unknown. in this study balb/c mice were infected with coxsackie virus b3 (cvb3). results indicated that sequential changes in viral replication (rt-pcr) were related to changes in trace element (arsenic, copper, iron, s ...200919233309
enhanced resistance to coxsackievirus b3-induced myocarditis by intranasal co-immunization of lymphotactin gene encapsulated in chitosan particle.coxsackievirus b3 (cvb3) is a gastrointestinal virus causing myocarditis in human and mice. an ideal vaccine for cvb3-myocarditis requires both humoral and cellular immunity at systemic and mucosal compartments. we described here an enhancing strategy for chitosan-pvp1 vaccine by co-immunizing with lymphotactin (ltn) gene, a t cell-attractive-chemokine, encapsulated in chitosan particle to provide more protection against cvb3. mice were intranasally co-immunized with 4 doses of chitosan-dna vacc ...200919233446
mutations in the nonstructural protein 3a confer resistance to the novel enterovirus replication inhibitor ttp-8307.a novel compound, ttp-8307, was identified as a potent inhibitor of the replication of several rhino- and enteroviruses. ttp-8307 inhibits viral rna synthesis in a dose-dependent manner, without affecting polyprotein synthesis and/or processing. drug-resistant variants of coxsackievirus b3 were all shown to carry at least one amino acid mutation in the nonstructural protein 3a. in particular, three mutations located in a nonstructured region preceding the hydrophobic domain (v45a, i54f, and h57y ...200919237651
dideoxy fluoro-ketopyranosyl nucleosides as potent antiviral agents: synthesis and biological evaluation of 2,3- and 3,4-dideoxy-3-fluoro-4- and -2-keto-beta-d-glucopyranosyl derivatives of n(4)-benzoyl cytosine.the synthesis of the dideoxy fluoro ketopyranonucleoside analogues, 1-(2,3-dideoxy-3-fluoro-6-o-trityl-beta-d-glycero-hexopyranosyl-4-ulose)-n(4)-benzoyl cytosine (7a), 1-(3,4-dideoxy-3-fluoro-6-o-trityl-beta-d-glycero-hexopyranosyl-2-ulose)-n(4)-benzoyl cytosine (13a) and their detritylated analogues 8a and 14a, respectively, is described. condensation of peracetylated 3-deoxy-3-fluoro-d-glucopyranose (1) with silylated n(4)-benzoyl cytosine, followed by selective deprotection and isopropyliden ...200919246130
osteopontin: a fibrosis-related marker molecule in cardiac remodeling of enterovirus myocarditis in the susceptible host.the characteristics of dilated cardiomyopathy (dcm) resulting from chronic viral myocarditis are remodeling processes of the extracellular matrix. based on our findings of enhanced osteopontin (opn) expression in inflamed human hearts, we further investigated in the murine model of acute and chronic coxsackievirus (cv)b3-myocarditis the role of opn regarding its involvement in resolution of cardiac virus infection and fibrosis. in hearts of a.by/snj mice susceptible to chronic cvb3-myocarditis, ...200919246678
[neuropathogenecity of coxsackie b virus strains isolated from children with clinical manifestations of aseptic meningitis in different years].the neuropathogenicity of coxsackie b3, b1, and b5 virus strains was studied in experimentally infected suckling balb/c mice and in contact animals from the same nest and litter infected due to the natural virus spread. the similar neurological disorders were found in both groups of animals. the data on pathological and morphological changes in the medulla oblongata as dystrophic and necrotic changes in the neurons with their lysis and deletion, brain tissue edema, and hemodynamic changes confir ...200919253729
[mapping of point mutations leading to loss of virus ech011 affinity for receptor daf (cd55)].comparative sequence of the structural part of genomes pf the original daf+ clone and two derived daf- mutant clones (101 and 103) was analyzed to map mutations responsible for the loss of affinity for the receptor glycoprotein daf (cd55). the obtained nucleotide sequences (eu167520, eu167521, eu167522) were deposited in the genbank. there were single point mutations causing amino acid changes in vp2 protein: s145 --> n in clone 101 and g162 --> e in clone 103.200919253731
erk map kinase-activated arf6 trafficking directs coxsackievirus type b3 into an unproductive compartment during virus host-cell entry.clathrin- and caveolae-mediated endocytosis have been implicated in the productive entry of many viruses into host cells. adp-ribosylation factor 6 (arf6)-dependent endocytosis is another endocytosis pathway that traffics from the cell surface and it is the only arf that traffics at the plasma membrane. however, little is known about arf6-dependent trafficking during virus entry. this study showed that coxsackievirus type b3 (cvb3) associated with decay-accelerating factor in non-polarized hela ...200919264606
indolylarylsulfones bearing natural and unnatural amino acids. discovery of potent inhibitors of hiv-1 non-nucleoside wild type and resistant mutant strains reverse transcriptase and coxsackie b4 virus.new potent indolylarylsulfone (ias) hiv-1 nnrtis were obtained by coupling natural and unnatural amino acids to the 2-carboxamide and introducing different electron-withdrawing substituents at position 4 and 5 of the indole nucleus. the new iass inhibited the hiv-1 replication in human t-lymphocyte (cem) cells at low/subnanomolar concentration and were weakly cytostatic. against the mutant l100i, k103n, and y181c rt hiv-1 strains in cem cells, sulfones 3, 4, 19, 27, and 31 were comparable to efv ...200919281225
antiviral activity of arbidol against coxsackie virus b5 in vitro and in vivo.we investigated the antiviral activity of arbidol, an antiviral chemical compound, against coxsackie virus b5 (cvb(5)) in vitro and in vivo. arbidol not only prevented the cytopathic effect (cpe) of cvb(5), as demonstrated in an mtt colorimetric assay, when added during or after viral infection, with a 50% inhibitory concentration (ic(50)) from 2.66 to 6.62 microg/ml, but it also decreased the cvb(5)-rna level in infected host cells, as shown in semi-quantitative rt-pcr. balb/c mice were used as ...200919291363
persistent infection of a carcinoma thyroid cell line with coxsackievirus b.viral infections are described as environmental factors that are implicated in various thyroid diseases. the role of enteroviruses (ev) in the pathogenesis of thyroid diseases has been suspected. recently, we found that ev rna could be detected in postoperative thyroid specimens. we decided to investigate the infection of a human thyroid cell line with coxsackievirus b4 (cvb4).200919292692
systemic analysis of a novel coxsackievirus gene delivery system in a mouse model.in order to systemically investigate the possibility of using coxsackievirus b3 (cvb3) to deliver foreign genes in vivo, a recombinant strain of cvb3 encoding the renilla gene (cvb3- renilla) was constructed. the recombinant cvb3 resulted in extensive and transient expression of the renilla protein within mouse organs, especially the pancreas. the level of expression was generally dependent upon the viral titer present. moreover, the cvb3-renilla strain was completely attenuated. interestingly, ...200919349757
detection of human viruses in rivers of a densly-populated area in germany using a virus adsorption elution method optimized for pcr analyses.transmission of viruses via surface water is a major public health concern. to determine the viral concentration in rivers of a densely-populated area in germany, the virus adsorption elution (viradel) method was optimized for downstream pcr applications. using a high-salt alkaline phosphate buffer for elution, the median recovery efficiency from spiked 1l water samples ranged from 21.3% to 100% for jc polyomavirus, human adenovirus type 5, echovirus 11, and norovirus genogroup i. analyses of 41 ...200919361832
a case of fulminant type 1 diabetes with coxsackie b4 virus infection diagnosed by elevated serum levels of neutralizing antibody.a 39-year-old woman with hyperglycemia and ketonuria but with normal hba1c level was diagnosed as having fulminant type 1 diabetes. the patient had 8-fold increase in the plasma titer of coxsackie b4 virus neutralizing antibody. infection with coxsackie b4 virus was associated with fulminant type 1 diabetes.200919362384
anti-coxsackie virus b3 norsesquiterpenoids from the roots of phyllanthus emblica.three new norsesquiterpenoid glycosides, 4'-hydroxyphyllaemblicin b (1) and phyllaemblicins e (2) and f (3), were isolated from the roots of phyllanthus emblica, together with three known compounds, phyllaemblic acid (4), phyllaemblicin b (5), and phyllaemblicin c (6). of these, 3 is a new norsesquiterpenoid dimer. the structures of 1-3 were established by spectroscopic data information and by acidic hydrolysis. the isolated compounds, together with two other known analogues, phyllaemblic acid m ...200919374435
packaging limits and stability of hiv-1 sequences in a coxsackievirus b vector.enteroviruses elicit protective mucosal immune responses that could be harnessed as part of a strategy to prevent sexual transmission of the human immunodeficiency virus-1 (hiv-1). we report the construction of replication-competent recombinant vectors of coxsackievirus b3 (cvb3) that express one or more portions of the hiv-1 gag protein. vectors containing the capsid domain of gag were initially genetically unstable with protein expression lost after brief passage in tissue culture. codon modif ...200919389440
pathogenesis of coxsackievirus-b5 acquired from intra-renal porcine islet cell xenografts in diabetic mice.we previously demonstrated the ability of a human isolate of coxsackievirus-b5 (cvb5) to infect productively adult porcine islet cells (pics) in vitro. pics infected with cvb5 remain viable, and upon transplantation reversed diabetes in c56bl/6 mice for up to 5 days.200919392724
circulating adhesion molecules in patients with keshan disease and their relationship with coxsackie b virus infection.this study determined the levels of serum soluble intercellular adhesion molecule-1 (sicam-1) and soluble vascular cell adhesion molecular-1 (svcam-1) in patients with different types of keshan disease (kd), examined the relationship between coxsackie b virus-specific igm antibody (cbv-igm) and sicam-1 or svcam-1 in kd patients, and investigated the role of these adhesion molecules in the pathogenesis of kd and their clinical implications. the levels of serum sicam-1, svcam-1 and cbv-igm were me ...200919399399
echocardiographic detection of early myocardial calcification in acute neonatal myocarditis due to coxsackie virus type b. 200919412563
a novel program to design sirnas simultaneously effective to highly variable virus genomes.a major concern of antiviral therapy using small interfering rnas (sirnas) targeting rna viral genome is high sequence diversity and mutation rate due to genetic instability. to overcome this problem, it is indispensable to design sirnas targeting highly conserved regions. we thus designed capsid (convenient application program for sirna design), a novel bioinformatics program to identify sirnas targeting highly conserved regions within rna viral genomes. from a set of input rnas of diverse sequ ...200919422797
sequential changes in serum cytokines reflect viral rna kinetics in target organs of a coxsackievirus b infection in mice.the pattern of cytokine responses related to viral replication during the course of the common human coxsackievirus b3 (cvb3) infection is not known.200919430896
advances in viral disease diagnostic and molecular epidemiological technologies.the early and rapid detection and characterization of specific nucleic acids of medico-veterinary pathogens have proven invaluable for diagnostic purposes. the integration of amplification and signal detection systems, including online real-time devices, have increased speed and sensitivity and greatly facilitated the quantification of target nucleic acids. they have also allowed for sequence characterization using melting or hybridization curves. the newer-generation molecular diagnostic techno ...200919435457
coxsackie b meningoencephalitis in a patient with acquired immunodeficiency syndrome and a multiple sclerosis-like illness.both coxsackie infection and multiple sclerosis (ms) are rare in human immunodeficiency virus (hiv) infection. we report a 35-year-old woman with known hiv infection of 12 years' duration and a clinical illness of 4 years' duration consistent with ms. the latter was characterized by optic neuritis, bilateral abducens palsies, recurrent bell's palsy, hemiparesis, and ataxia coupled with white matter abnormalities on magnetic resonance imaging (mri). autopsy revealed coxsackie b meningoencephaliti ...200919444695
altered interactions between stem-loop iv within the 5' noncoding region of coxsackievirus rna and poly(rc) binding protein 2: effects on ires-mediated translation and viral infectivity.coxsackievirus b3 (cvb3) is a causative agent of viral myocarditis, meningitis, pancreatitis, and encephalitis. much of what is known about the coxsackievirus intracellular replication cycle is based on the information already known from a well-studied and closely related virus, poliovirus. like that of poliovirus, the 5' noncoding region (5' ncr) of cvb3 genomic rna contains secondary structures that function in both viral rna replication and cap-independent translation initiation. for poliovir ...200919446305
viral myocarditis: from the perspective of the virus. 200919451363
application of real-time reverse transcription polymerase chain reaction for the detection of svdv.application of real-time rt-pcr (rrt-pcr) for detection of swine vesicular disease virus (svdv) in samples of archival svdv isolates and clinical samples collected from svdv infected pigs was described. a primer set that targets the ires region of the svdv genome and taqman probe specific for a highly conserved region in svdv rna ires region were used. the assay detected viral rna in all tested archival strains of svdv isolated in europe during years 1972-73 and 1992 as well as in clinical sampl ...200919459449
magnetic purification of biotinylated cdna removes false priming and ensures strand-specificity of rt-pcr for enteroviral rnas.the detection of replicative intermediate rnas as markers of active replication of rna viruses is an essential tool to investigate pathogenesis in acute viral infections, as well as in their long-term sequelae. in this regard, strand-specific pcr has been used widely to distinguish (-) and (+) enteroviral rnas in pathogenesis studies of diseases such as dilated cardiomyopathy. it has been generally assumed that oligonucleotide-primed reverse transcription of a given rna generates only the corres ...200919524621
two clusters of mutations map distinct receptor-binding sites of echovirus 11 for the decay-accelerating factor (cd55) and for canyon-binding receptors.in this study we present the comparative sequence analysis of the parental haemagglutinating (daf+) and mutant non-haemagglutinating (daf-) clones of echovirus 11 (ev11) isolated from the prototype strain gregory. the sequence comparison revealed only a single amino acid substitution in the capsid protein vp2 of each mutant clone. these substitutions were located in the area of viral receptor-binding site for daf. since daf- mutants of ev11 did not interact with daf, they used an alternative rec ...200919540285
long-term inactivation study of three enteroviruses in artificial surface and groundwaters, using pcr and cell culture.since the transmission of pathogenic viruses via water is indistinguishable from the transmission via other routes and since the levels in drinking water, although significant for health, may be too low for detection, quantitative viral risk assessment is a useful tool for assessing disease risk due to consumption of drinking water. quantitative viral risk assessment requires information concerning the ability of viruses detected in drinking water to infect their host. to obtain insight into the ...200919074604
integrin alphavbeta6 is a high-affinity receptor for coxsackievirus a9.coxsackievirus a9 (cav9), a member of the genus enterovirus in the family picornaviridae, possesses an integrin-binding arginine-glycine-aspartic acid (rgd) motif in the c terminus of vp1 capsid protein. cav9 has been shown to utilize integrins alphavbeta3 and alphavbeta6 as primary receptors for cell attachment. while cav9 rgd-mutants (rge and rgddel) are capable of infecting rhabdomyosarcoma (rd) cell line, they grow very poorly in an epithelial lung carcinoma cell line (a549). in this study, ...200919088289
transmission networks and population turnover of echovirus 30.globally, echovirus 30 (e30) is one of the most frequently identified enteroviruses and a major cause of meningitis. despite its wide distribution, little is known about its transmission networks or the dynamics of its recombination and geographical spread. to address this, we have conducted an extensive molecular epidemiology and evolutionary study of e30 isolates collected over 8 years from a geographically wide sample base (11 european countries, asia, and australia). 3dpol sequences fell int ...200919091869
phylogeography of circulating populations of human echovirus 30 over 50 years: nucleotide polymorphism and signature of purifying selection in the vp1 capsid protein gene.a comprehensive set of 443 1d gene sequences (encoding the vp1 capsid protein) was analyzed to investigate the phylogenetic relationships and evolutionary patterns among strains of human echovirus 30 (e30; genus enterovirus, family picornaviridae) characterized over 50 years. maximum-likelihood (ml) phylogenetic trees of complete and nonredundant 1d gene sequences (total length=876 nucleotides) showed evidence of distinct lineages related to the isolation period of virus strains. virus transport ...200918595781
effect of interleukin-15 on the course of myocarditis in coxsackievirus b3-infected balb/c mice.cytokines have an important role in both the initiation and perpetuation of viral myocarditis. because a causative therapy of myocarditis is not yet well established and immunomodulation is a promising approach, the influence of interleukin (il)-15, a proinflammatory cytokine, on the course of experimental myocarditis in coxsackievirus b3 (cvb3)-infected mice was examined.200919584981
lack of il-6 during coxsackievirus infection heightens the early immune response resulting in increased severity of chronic autoimmune myocarditis.chronic myocarditis is often initiated by viral infection, the most common of which is coxsackievirus infection. the precise mechanism by which viral infection leads to chronic autoimmune pathology is poorly understood, however it is clear that the early immune response plays a critical role. previous results have shown that the inflammatory cytokine interleukin (il)-6 is integral to the development of experimental-induced autoimmune myocarditis. however, the function of il-6 during viral-mediat ...200919587788
combination of soluble coxsackievirus-adenovirus receptor and anti-coxsackievirus sirnas exerts synergistic antiviral activity against coxsackievirus b3.coxsackievirus b3 (cvb-3) is a major causative agent of chronic heart muscle infections. the present study describes a cell culture system with an ongoing virus infection to evaluate two novel inhibitory strategies, either individually or combined: (1) rna interference (rnai) to degrade cytoplasmatic cvb-3 rna and (2) a vector-delivered soluble variant of the coxsackievirus-adenovirus receptor fused to a human immunoglobulin (scar-fc), which inhibits cellular uptake of cvb-3. both approaches wer ...200919591879
depletion of gammadelta+ t cells increases cd4+ foxp3 (t regulatory) cell response in coxsackievirus b3-induced myocarditis.coxsackievirus b3 (cvb3) causes severe myocarditis in balb/c mice which depends upon cd4(+) t helper type 1 [th1; i.e. interferon-gamma(+) (ifn-gamma(+))] and gammadelta(+) cells. depleting gammadelta(+) cells using anti-gammadelta antibody suppresses myocarditis and cd4(+) ifn-gamma(+) cell numbers in the spleen and heart of infected mice while increasing cd4(+) foxp3(+) cells. mice deficient in gammadelta(+) cells have increased numbers of naïve (cd44(lo) cd62l(hi)) and fewer effector (cd44(hi ...200919604307
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