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characterization of the n-terminal part of the neutralizing antigenic site i of coxsackievirus b4 by mutation analysis of antigen chimeras.coxsackievirus b3 (cvb3) as a potential rna virus vector for the presentation of foreign antigenic epitopes was further characterized. insertion mutagenesis of infectious cvb3 cdna yielded viable antigen chimeras containing variant bc loops of vp1 of coxsackievirus b4 (cvb4). analysis of three antigen chimeras allowed the mapping of the n-terminal part of the neutralizing antigenic site 1 (n-ag1) of cvb4 which is located in the bc loop of the structural protein vp1. a significant neutralization ...19947531922
molecular mimicry, anti-coxsackievirus b3 neutralizing monoclonal antibodies, and myocarditis.molecular mimicry has been suggested as one mechanism to explain chronic myocarditis in some murine strains in the postinfectious period following induction of acute myocarditis by coxsackievirus b3 (cvb3). to test this hypothesis, neutralizing mabs were generated against a highly myocarditic cvb3 virus (cvb3m). these mabs neutralized several myocarditic and amyocarditic cvb3 variants by cytopathic effects inhibition assays. data from several experiments suggest that these mabs recognize discont ...19957533190
echovirus 1 interaction with the isolated vla-2 i domain.the isolated i domain of the integrin vla-2, produced as a bacterial fusion protein, specifically bound echovirus 1 and prevented virus attachment to cells. these results demonstrate that the receptor structures critical for virus attachment are contained solely within the vla-2 i domain and that soluble receptor fragments are capable of preventing infection.19957535868
transient elevation of granulocyte colony-stimulating factor levels in cerebrospinal fluid at the initial stage of aseptic meningitis in children.at the early stage of aseptic meningitis, there is a transient increase in neutrophil counts in the cerebrospinal fluid. some factors in the cerebrospinal fluid might induce migration of neutrophils into the cerebrospinal fluid. granulocyte colony-stimulating factor (g-csf) plays an important role, not only as a hemopoietic factor but also as a regulating factor for a biologic defense system by neutrophils in the foci of infection. to analyze the role of g-csf on accumulating neutrophils in the ...19957537365
coxsackieviruses b1, b3, and b5 use decay accelerating factor as a receptor for cell attachment.receptor binding and subsequent cell-mediated internalization or disassembly are the initial steps in virus replication. cell surface molecules that participate in this process are the primary determinants of virus tissue tropism. monoclonal antibody blockade, immunoprecipitation, and dna transfection were used to identify decay accelerating factor as a major cell attachment receptor for coxsackieviruses b1, b3, and b5. however, expression of human decay acceleration factor on the surface of non ...19957538177
detection of picornavirus genomic and template rna strands by a novel semi-nested polymerase chain reaction-technique and agarose-gel electrophoresis.attempts to evaluate the relative levels of enteroviral genomic and template rna strands in small biopsy tissue samples from patients have yielded ambiguous data, largely due to the limited amount of rna available. a novel semi-nested polymerase chain reaction (pcr) technique was developed to enable rna levels to be examined more accurately. pcr products were visualised by horizontal agarose gel electrophoresis. this technique was demonstrated on linear single-stranded plasmid dna; viral rna iso ...19957541748
characterization of the echovirus 7 receptor: domains of cd55 critical for virus binding.cd55, or decay-accelerating factor (daf), is a cell surface glycoprotein which regulates complement activity by accelerating the decay of c3/c5 convertases. recently, we and others have established that this molecule acts as a cellular receptor for echovirus 7 and related viruses. daf consists of five domains: four short consensus repeats (scrs) and a serine/threonine-rich region, attached to the cell surface by a glycosylphosphatidyl inositol anchor. chinese hamster ovary cells stably transfect ...19957543583
coxsackie virus and diabetes revisited. 19957585050
deficient diet evokes nasty heart virus. 19957585080
rapid genomic evolution of a non-virulent coxsackievirus b3 in selenium-deficient mice results in selection of identical virulent isolates.previous work from our laboratory demonstrated that selenium deficiency in the mouse allows a normally benign (amyocarditic) cloned and sequenced coxackievirus to cause significant heart damage. furthermore, coxsackievirus recovered from the hearts of selenium-deficient mice inoculated into selenium-adequate mice still induced significant heart damage, suggesting that the amyocarditic coxsackievirus had mutated to a virulent phenotype. here we report that sequence analysis revealed six nucleotid ...19957585090
henoch-schonlein purpura associated with coxsackie-virus b1 infection. 19957586994
comparison of coxsackie b neutralisation and enteroviral pcr in chronic fatigue patients.coxsackie b enteroviruses have been implicated repeatedly as agents associated with chronic fatigue syndrome (cfs). the objective of this study was to compare the serological evidence for the presence of coxsackie b virus neutralising antibody, with the polymerase chain reaction (pcr) detecting a portion of the 5' nontranslated region (ntr) of the enterovirus genome. serum samples from 100 chronic fatigue patients and from 100 healthy comparison patients were used in this study. in the cfs study ...19957595406
[the molecular pathogenesis of enterovirus myocarditis. virus persistence and chronic inflammation]. 19957601616
inhibition of herpes simplex virus type 1, respiratory syncytial virus and echovirus type 11 by peroxidase-generated hypothiocyanite.the human mouth is an important route of viral transmission and evidence exists that human saliva can neutralize some viruses, e.g. herpes simplex type 1 (hsv-1) and human immunodeficiency virus (hiv) in vitro. however, little is known of the actual antiviral agents in saliva. we have analyzed how hypothiocyanite (hoscn/-oscn) ions, present in human saliva and generated by salivary peroxidase systems, affect the viability of three different types of viruses; hsv-1 (capable of inducing oral lesio ...19957605114
a single amino acid substitution in the capsid protein vp1 of coxsackievirus b3 (cvb3) alters plaque phenotype in vero cells but not cardiovirulence in a mouse model.we previously described a large plaque attenuant (p14v-1) derived from a cardiovirulent coxsackievirus b3 (cvb3) and showed that there were no major determinants of either attenuation or plaque phenotype in the 5' nontranslated region (5'ntr). part of the region encoding the last 124 amino acids of vp3 and the first 106 amino acids of vp1 of the attenuant was then sequenced and compared to the wild-type. three nucleotide changes were found in the vp1 coding region: a silent single base change at ...19957605207
analysis of protein synthesis by two-dimensional gel electrophoresis in t cells persistently infected with coxsackie b virus.coxsackie b viruses (cbv) have been implicated in various human diseases that present as either limited acute infections or prolonged chronic infections. a number of investigations have suggested that a virus-induced immune dysfunction might play a role in in vivo pathogenesis. in the current study, we describe cbv infection of two human t cell-derived cell lines (jurkat and molt-4 cells) as potential models for cbv infection of lymphocytes. short term (up to 144 h post-infection) cbv infection ...19957607163
shigella flexneri adherence to and multiplication in coxsackie b1 virus-infected hep-2 cells.coxsackie b1 virus infection enhances the susceptibility of in vitro cultured hep-2 cells to invasiveness by shigella flexneri. we have studied the effect of viral infection on two phases of the invasiveness. only a minor part was mediated by enhanced bacterial adherence to the cells, and the intracellular multiplication was unaffected by the virus. enhanced adherence was not dependent on the presence of the gene product of the 140 md virulence associated plasmid. our data indicate that enhanced ...19957612255
coxsackie b virus infection and onset of childhood diabetes.infection with coxsackie b viruses has been linked to insulin-dependent diabetes mellitus. nine of 14 serum samples (64%) taken from children at the onset of diabetes were positive for enterovirus rna by pcr. all of the children were under age six, and five were under age three. by contrast, enterovirus sequences were detected in only two of 45 serum samples from appropriate comparison children (4%). sequences from six of the positive patients showed strong homology with coxsackie b3 and b4 viru ...19957616803
comparison of pcr and cell culture for detection of enteroviruses in sludge-amended field soils and determination of their transport.pcr and cell culture assays for enteroviruses were conducted on soil samples collected from an experimental farm that had received mesophilic anaerobically digested sludge for the past 7 years. of 24 samples assayed, 21 samples were positive by pcr, implying that at least some viral nucleic acid sequences remained intact. however, these viral particles were unable to infect the buffalo green monkey cell line used in subsequent cell culture assays. it is significant that positive pcr detection of ...19957646051
therapy with immunoglobulin suppresses myocarditis in a murine coxsackievirus b3 model. antiviral and anti-inflammatory effects.the treatment of some inflammatory diseases (eg, kawasaki disease) with immunoglobulin has been demonstrated to be effective. accordingly, to elucidate the mechanism underlying such actions of immunoglobulin, we examined its effects on murine coxsackievirus b3 (cb3) myocarditis.19957664447
gamma/delta t lymphocytes in viral infections.t lymphocyte progenitors differentiate into two distinct t cell lineages. although the alpha beta and gamma delta t cell lineages resemble each other phenotypically and functionally, there are some striking differences. some gamma delta t cells recognize, similarly to alpha beta t cells, peptides presented by major histocompatibility complex (mhc) proteins or mhc-like molecules. however, there are gamma delta t cells that recognize mhc molecules in a fundamentally different manner in comparison ...19957665983
requirement of mip-1 alpha for an inflammatory response to viral infection.macrophage inflammatory protein-1 alpha (mip-1 alpha) is a chemokine that has pro-inflammatory and stem cell inhibitory activities in vitro. its biologic role in vivo was examined in mice in which the gene encoding mip-1 alpha had been disrupted. homozygous mip-1 alpha mutant (-/-) mice were resistant to coxsackievirus-induced myocarditis seen in infected wild-type (+/+) mice. influenza virus-infected -/- mice had reduced pneumonitis and delayed clearance of the virus compared with infected +/+ ...19957667639
differential diagnosis and genetic analysis of the antigenically related swine vesicular disease virus and coxsackie viruses.monoclonal antibodies directed against an isolate of swine vesicular disease virus (svdv), characterized by virus neutralization tests and competition assays, were used to compare svdv isolates and isolates of the antigenically related coxsackie viruses by elisa. svdv-specific reaction patterns and one specific for coxsackie viruses were observed. this provided a method for distinguishing between these enteroviruses. in addition, rt-pcrs were undertaken with coxsackie virus and svdv genomes. dif ...19957673387
the role of cd8+ t lymphocytes in coxsackievirus b3-induced myocarditis.coxsackievirus infections have previously been shown to cause acute or chronic myocarditis in humans, and several mouse models have been established to study the pathology of this disease. myocardial injury may result from direct viral effects and/or may be immune mediated. to determine the relative roles of these processes in pathogenesis, we have compared coxsackievirus b3 (cvb3) infections of normal and immuno-compromised transgenic knockout (ko) mice. cvb3 was able to infect all strains used ...19957474082
characterization of a 100-kilodalton binding protein for the six serotypes of coxsackie b viruses.viral infection of host cells primarily depends on binding of the virus to a specific cell surface protein. in order to characterize the binding protein for group b coxsackieviruses (cvb), detergent-solubilized membrane proteins of different cell lines were tested in virus overlay protein-binding assays. a prominent virus-binding protein with a molecular mass of 100 kda was detected in various cvb-permissive human and monkey cell lines but was not detected in nonpermissive cell lines. the specif ...19957474086
colony-stimulating factors and coxackievirus b3 myocarditis in mice: macrophage colony-stimulating factor suppresses acute myocarditis with increasing interferon-alpha.the effects of macrophage colony-stimulating factor (m-csf) and granulocyte colony-stimulating factor (g-csf) on murine coxsackievirus b3 myocarditis were investigated. a total of 4 x 10(6) u/kg/day m-csf and 20 micrograms/kg/day g-csf were injected subcutaneously every day on day 0 to day 14 starting simultaneously with virus inoculation. serum interferon-alpha was measured periodically. the survival rate of the m-csf group was higher than that of the untreated control group (p < 0.05). on days ...19957484779
analysis of picornavirus 2a(pro) proteins: separation of proteinase from translation and replication functions.the poliovirus (pv) genome was manipulated by replacing its 2a-encoding sequence with the corresponding sequence of coxsackie b4 virus (cbv4) or human rhinovirus type 2 (hrv2). in vitro translation of the resulting chimeric pv genomes revealed a normal cis-cleavage activity for both heterologous 2a(pro) proteinases in the chimeric pv polyproteins. however, only the genome containing the 2a-encoding sequence of cbv4 (pv/cbv4-2a) yielded viable virus in transfected cells, producing a mixture of la ...19957494250
genetic analysis of a hydrophobic domain of coxsackie b3 virus protein 2b: a moderate degree of hydrophobicity is required for a cis-acting function in viral rna synthesis.coxsackie b virus protein 2b contains near its c terminus a hydrophobic domain with an amino acid composition that is characteristic for transmembrane regions. a molecular genetic approach was followed to define the role of this domain in virus reproduction and to study the structural and hydrophobic requirements of the domain. nine substitution mutations were introduced in an infectious cdna clone of coxsackie b3 virus. the effects of the mutations were studied in vivo by transfection of buffal ...19957494289
coxsackievirus b3 adapted to growth in rd cells binds to decay-accelerating factor (cd55).a coxsackievirus b3 (cb3) isolate adapted to growth in rd cells shows an alteration in cell tropism as a result of its capacity to bind a 70-kda cell surface molecule expressed on these cells. we now show that this molecule is the complement regulatory protein, decay-accelerating factor (daf) (cd55). anti-daf antibodies prevented cb3 attachment to the cell surface. radiolabeled cb3 adapted to growth in rd cells bound to cho cells transfected with human daf, whereas cb3 (strain nancy), the parent ...19957531780
replication and persistence of coxsackieviruses b3 in human fibroblasts.although in some cases a carrier state of coxsackieviruses in human fibroblasts was described, the persistence mechanism has remained unknown. our results demonstrate a replication of coxsackievirus b3 (cvb3) in diploid human fibroblasts that is dependent on the virus strain as well as on the cell line involved. two cvb3 nancy strains could be multiplied over more than 10 passages in cell line h, whereas cvb3 strains sh"c" and sh"w" did not longer form infectious virus following several passages ...19957734835
pathogenesis of virus-induced diabetes in mice.male cd-1 mice were challenged with a diabetogenic strain (e2) of coxsackievirus b4 (cvb4). at 7 weeks, 6 months, and 1 year after inoculation, mean histopathologic scores, postprandial blood glucose levels, and serum levels of antibody to islet cells were significantly elevated and mean fasting serum insulin levels significantly reduced in infected mice versus uninfected controls (p < .001 for each). at 7 weeks after infection, viral rna, but not protein or infectious virus, was demonstrated in ...19957751687
the genome of echovirus 11.echoviruses are the largest enterovirus subgroup consisting of 32 serotypes. they are common human pathogens causing, for example, meningitis, encephalitis and exanthema, but in spite of their clinical importance, relatively little is known about their biology. to illuminate the molecular characteristics of echoviruses, we have completed the genomic sequence of serotype 11. the rna genome is 7438 nucleotides in length and it codes for a 2195 amino acid long polyprotein. when compared to other se ...19957762294
intrauterine coxsackie virus, group b type 1, infection: viral cultivation from amniotic fluid in the third trimester.we report a case that presents clear evidence of an intrauterine infection of twin fetuses with an enterovirus. the mother had signs and symptoms of chorioamniotitis at 34 weeks' gestation. coxsackie virus, group b type 1 (cvb1), was cultured from amniotic fluid obtained by transabdominal amniocentesis when the membranes of both fetuses were still intact. delivery occurred vaginally approximately 24 hours later, following spontaneous rupture of membranes. at birth, both twins showed signs of sep ...19957779200
receptor proteins on newborn balb/c mouse brain cells for coxsackievirus b3 are immunologically distinct from those on hela cells.newborn balb/c mice are highly susceptible to infection by the six coxsackievirus serotypes of group b (cvb) and it is known that receptor for these viruses are in highest concentration in the brain as compared to other tissues. therefore, proteins from the brain tissues of these animals were solubilized (brain-ext) and characterized for the identification of mouse brain receptor (mbr) proteins. virus-blot analyses of brain-ext suggested that each of three virus variants of cvb3-(n, w and rd) re ...19957785319
left ventricular hemodynamic parameters in the course of acute experimental coxsackievirus b 3 myocarditis.in susceptible dba/2 mice coxsackievirus b 3-induced myocarditis leads to inflammatory and necrotic lesions in the myocardium 7-10 days after virus inoculation. the purpose of this study was to determine whether hemodynamic changes occur in murine coxsackievirus b 3 myocarditis and whether they are correlated to histological cardiac lesions throughout the infection. left ventricular function was determined by open chest puncture of the left ventricle in the course of acute coxsackievirus b 3 inf ...19958523420
comparison of two immunoassay procedures for detecting enterovirus igm.enteroviruses cause significant illness in man but viral diagnosis is problematic. enterovirus-specific igm tests have been developed but due to the difficulties of obtaining reliable control sera the interpretation of assay data remains mainly arbitrary and empirical. the present study was undertaken to assess the reliability of such assays by comparing two tests performed independently in two different laboratories: a mu-capture radioimmunoassay (macria) which utilizes 35s-labelled coxsackie v ...19958551255
[cuban epidemic neuropathy. ii. the antigenic characteristics of the viral isolates].with the aim of characterizing antigenically isolations producing mild cytopathogenic effect obtained from the cerebrospinal fluid of patients presenting with epidemic neuropathy, neutralization tests and western blot analysis were performed using hyperimmune sera of patients and hyperimmune sera of rabbits. it was confirmed that isolations with mild cutopathogenic effect studied have the same antigenic characteristics and that they are related to coxsackie a9 and b4 viruses. structural proteins ...19959805061
genetic mapping of the determinants of plaque morphology of coxsackievirus b4.the genetic determinants of plaque size of two variants of coxsackie-virus b4, cb4-p and cb4-v, were identified using a panel of recombinant, chimeric viruses. when grown in monkey kidney cells, cb4-v yielded large plaques with an average size of 1.0 cm while cb4-p yielded small plaques with an average size of 0.4 cm. two genetic domains, the 5' untranslated region and the vp4 gene sequence, independently influenced plaque size. recombinant viruses containing the cb4-p genetic background with po ...19958572942
cultured human myocardial fibroblasts of pediatric origin: natural human interferon-alpha is more effective than recombinant interferon-alpha 2a in carrier-state coxsackievirus b3 replication.cultured human myocardial fibroblasts of pediatric origin seem to be a useful species-specific model for studying various heart diseases which involve the myocardial interstitium, for example enterovirus heart disease. cells were propagated from small samples of human ventricular tissues (0.2 g) obtained from standard surgical procedure for the correction of fallot-tetralogy. cultured cells exhibited typical fibroblastoid morphology over a period of 4 months and were uniformly immunoreactive wit ...19958576936
genotypic variation in coxsackievirus b5 isolates from three different outbreaks in the united states.genomic sequences in vp1/2a and 5'-non-coding region of 10 isolates of coxsackievirus b5 from three outbreaks were compared with published sequences of another coxsackievirus b5, swine vesicular disease virus, coxsackievirus b1, coxsackievirus b3, and coxsackievirus b4. isolates of coxsackievirus b5 from the same outbreak showed close relations, not exceeding 7.2% in nucleotide differences. differences were greater between isolates from different outbreaks, varying between 8.4 and 16%. we have a ...19958578854
the inhibitory effect of astragalus membranaceus on coxsackie b-3 virus rna replication.using mice infected with coxsackie b-3 virus (cvb3) as a viral myocarditis model, we observed the inhibitory effect of astragalus membranaceus (am) on cvb3-rna replication in myocardial tissue of mice by rna-rna in situ hybridization with negative-strand rna probes labelled with 35s and quantitative imaging analysis of positive signals. the mechanism of its effect on cvb3-rna replication has been investigated by detection of beta-interferon (beta-ifn) as well. results showed that the copy number ...19958580483
[effect of astragalus membranaceus on ca2+ influx and coxsackie virus b3 rna replication in cultured neonatal rat heart cells].the effect of astragalus membranaceus (am) on ca2+ influx across the myocardial plasma membrane and coxsackie virus b3(cvb3)-rna replication in cultured neonatal rat heart cells infected with cvb3 was investigated. it was found that the ca2+ influx could be inhibited significantly (p < 0.01) by am after infection of heart cells for 48 h. in addition, when the cultured heart cells infected with cvb3 and treated with am for 48 h, the ca2+ influx of infected heart cells also could be inhibited by a ...19958580697
indications that maternal coxsackie b virus infection during pregnancy is a risk factor for childhood-onset iddm.in a population-based setting, we traced serum samples collected at time of birth from 55 mothers whose children later developed insulin-dependent diabetes (iddm) and matched them pairwise to control subjects who gave birth at the same hospital during the same month. the sera were analysed for igm antibodies to coxsackie b virus serotypes 2, 3 and 4 (cbv-2, 3 and 4) using a type-specific mu-antibody-capture radioimmunoassay. despite a decreased power due to the close matching by time of birth we ...19958582549
an unseasonable winter outbreak of echovirus type 30 meningitis.during the winter of late october, 1990 to march, 1991, echovirus type 30 meningitis affected 10 children aged 3-9 years of age in mikuni, japan. five patients presented with constipation, four had diarrhoea, while five complained of severe abdominal pain, in addition to typical symptoms and signs compatible with aseptic meningitis. echovirus type 30 was isolated from the csf of nine, from the faeces of 10, and from the throats of eight children. all showed a significant antibody response to ech ...19958586842
the structure of coxsackievirus b3 at 3.5 a resolution.group b coxsackieviruses (cvbs) are etiologic agents of a number of human diseases that range in severity from asymptomatic to lethal infections. they are small, single-stranded rna icosahedral viruses that belong to the enterovirus genus of the picornavirus family. structural studies were initiated in light of the information available on the cellular receptors for this virus and to assist in the design of antiviral capsid-binding compounds for the cvbs.19958591043
effects of granulocyte colony-stimulating factor upon coxsackievirus b3 myocarditis in mice.granulocyte colony-stimulating factor is a potent activator of mature granulocytes, and subsequently enhances superoxide release. the purpose of this study was to investigate the effects of granulocyte colony-stimulating factor upon murine coxsackievirus b3 myocarditis in relation to free radical-mediated cardiac damage. two-week-old, male, c3h/he mice were inoculated intraperitoneally with coxsackievirus b3. granulocyte colony-stimulating factor 20 micrograms.kg-1.day-1, polyethylene glycol-con ...19958682024
genetics of coxsackievirus b3 cardiovirulence.the human enteroviruses, especially the coxsackie b viruses, have been established as aetiologic agents of human inflammatory heart disease, a condition which may lead to dilated cardiomyopathy and heart failure. it is clear from murine models of coxsackievirus b3-induced inflammatory heart disease that not all strains of the virus are cardiovirulent (able to cause disease). here, we present preliminary data mapping the site in a coxsackievirus b3 genome which determines a cardiovirulent phenoty ...19958682085
new aspects of murine coxsackie b3 myocarditis--focus on heavy metals.the magnitude of inflammatory lesions in the hearts of coxsackie b3 (cb3)-virus infected mice can be affected by the potentially toxic heavy metals cadmium (cd), nickel (ni) and methyl mercury (mehg). the infection is associated with a changed distribution, such as cd accumulation in the spleen and kidneys. new target organs for ni during the infection were the heart, pancreas and lungs in which inflammatory lesions were present. this increased uptake was correlated with the disturbed function o ...19958682094
the role of transgenic knockout models in defining the pathogenesis of viral heart disease.the pathogenesis of viral myocarditis involves contributions from the virus, the immune system and myocytes. in defining the molecular contributions in the disease process, modulations of the components of the immune system through transgenic knockout models provide useful insights. advantages of the transgenic knockout models are that they allow biological evaluation of the importance of a particular molecule in the physiological context of an intact organism. furthermore, the techniques of tra ...19958682095
left ventricular haemodynamics in murine viral myocarditis.in susceptible dba/2 mice infection with coxsackievirus b3 leads to severe inflammatory and necrotic lesions in the heart. there is a temporal discrepancy of peak concentrations of replicative virus in the heart and maximal cardiac inflammation. aims of this study were, first, to determine whether haemodynamic changes occur in coxsackievirus b3-induced murine myocarditis and, second, the time frame in which those alterations may be apparent. by puncture of the left ventricle, pressures and the f ...19958682096
depressed unloaded sarcomere shortening velocity in acute murine coxsackievirus myocarditis: myocardial remodelling in the absence of necrosis or hypertrophy.we used right ventricular papillary muscles to study cellular dysfunction in acute murine coxsackievirus myocarditis. we measured unloaded sarcomere shortening velocity (v0) with laser diffraction (hene, lambda = 623.8 nm) 7 days after coxsackievirus infection (m) (n = 7) and after infection + monoclonal antibodies to eliminate t cells (t) (n = 4) and in normals (n) (n = 8). a servomotor rapidly shortened a muscle until slack early in contraction and v0 was measured at the onset of zero force. v ...19958682097
enterovirus-infected immune cells of spleen and lymph nodes in the murine model of chronic myocarditis: a role in pathogenesis?molecular hybridization studies have demonstrated that human enteroviruses, including group b coxsackieviruses (cvb), are detectable not only in endomyocardial biopsies of patients with acute enterovirus myocarditis but also in those with chronic disease. such infections are observed in some patients with end-stage dilated cardiomyopathy, indicating the possibility of persistent heart muscle infection. enterovirus persistence in the human heart is supported by the recent discovery in various mur ...19958682099
coxsackievirus-induced chronic myocarditis in murine models.challenge of several murine strains with two highly myocarditic variants of coxsackievirus b3 (cvb3) induced acute and chronic myocarditis, detectable at 21 and 45 days post-inoculation (p.i.). in-situ hybridization of coronal heart sections showing chronic inflammation with a radiolabelled cvb3 probe detected viral genomic rna at day 7 p.i. but rarely at 21 or 45 days p.i., suggesting few murine heart cells actively replicate virus during chronic myocardial inflammation. data will be presented ...19958682103
the role of the adp/atp carrier in the pathogenesis of viral heart disease.the adp/atp carrier is an autoantigen in myocarditis and dilated cardiomyopathy, both of which are diseases related to virus infections. sera of these patients bear carrier-specific autoantibodies inhibiting transmembrane nucleotide transport on isolated mitochondria. to further assess the role of the adp/atp carrier in viral heart disease, guinea pigs were immunized with the isolated adp/atp carrier protein and a-strain mice were infected with coxsackie b3 virus. both species generated specific ...19958682105
[isolation of coxsackieviruses b5 in a fatal case of meningoencephalitis].a nine months old boy was admitted to the hospital with the diagnosis of meningoencephalitis 15 days after having a clinically diagnosed chickenpox. lumbar puncture showed clear csf with 0.23 g/l of proteins, 57 mg/dl of glucose, 30 red cells/mm3 and 5 leukocytes/mm3. blood count showed a packed red cell volume of 22%, a hemoglobin of 7 g/dl, 14800 leukocytes with 1% eosinophils, 5% band and 39% segmented neutrophils, 50% lymphocytes and 5% monocytes and a decreased platelet count. on the fourth ...19958733269
coxsackieviral proteins functionally recognize the polioviral cloverleaf structure of the 5'-ntr of a chimeric enterovirus rna: influence of species-specific host cell factors on virus growth.the 5'-non-translated region (ntr) of enteroviruses contains secondary structures which do not only serve in the initiation of translation but also in the initiation of plus-strand rna synthesis by binding of viral and cellular proteins. to investigate a very early step of enteroviral replication by cis- and trans-complementation, 220 nucleotides of the 5'-region of coxsackievirus b3 (cvb3) were exchanged with the corresponding region of poliovirus type 1 (pv1) to yield the chimeric virus cvb3[p ...19958837877
coxsackie b virus in the etiology of heart diseases in delhi.coxsackie b group of viruses have been long considered as the precipitating factor in heart diseases. their association with various heart diseases especially pericarditis, myocarditis and myopericarditis is known, but now their association is also known with cardiomyopathy and ischaemic heart diseases. a study was carried out on 87 patients suffering from various heart diseases and the role of coxackie b virus infection was studied. the study included 25 control subjects. thirty patients (34.5% ...19958866986
neurogenic diabetes insipidus in a child with fatal coxsackie virus b1 encephalitis.a 5 year-old boy presented with fever, sore throat, diarrhea, and general soreness which evolved into encephalitis. his cerebrospinal fluid showed a cell count of 3 mononuclear cells/microliters, protein 2800 mg/l, and growth of coxsackie virus b1. cardiorespiratory arrest was noted after a convulsion and infusion of diazepam. although he was immediately resuscitated, he remained unconscious with a modified glasgow coma score of 4 or 3. he developed neurogenic diabetes insipidus 169 hours after ...19958821910
permissiveness of human embryonal fibroblasts for coxsackieviruses b3. investigations on virus genetic markers in vitro and localization of virus receptor distribution by immunogold replica technique.in our previous paper (29) we could show, that the replication of four coxsackie b3 virus strains in human fibroblast lines from different origin was dependent on both the virus strain and the cell line used. although generally no cytopathic effect could be observed out of one virus strain more pathogenic virus variants could be selected able to destroy virus-sensitive as well as insensitive fibroblasts. the present study was designed to characterize these virus strains by using several in vitro ...19968837403
human enterovirus infection in stray dogs. some aspects of interest to public health.to investigate the possible role of domestic animals as reservoirs of human enteroviruses, we studied 212 stray dogs captured in different areas of the municipality of são paulo. the captured animals were divided into 19 groups of 10 to 20 dogs each; faeces of 126 of the 212 dogs were processed for enterovirus isolation. the following viruses were isolated from 12 dogs; poliovirus type 1 (2 dogs), poliovirus type 3 (1 dog), echovirus type 7 (8 dogs) and echovirus type 15 (1 dog). of the 12 infec ...19969071037
expression and distribution of the receptors for coxsackievirus b3 during fetal development of the balb/c mouse and of their brain cells in culture.this study was designed mainly to determine the relationships between the expression and distribution of the cellular receptor proteins for coxsackievirus b3 (cvb3) and susceptibility of mouse brain cells during fetal development of balb/c mice. immunoblot analysis of fetal extracts demonstrated that the cvb3 receptor proteins were first expressed at day 14 of the fetal stage, and that maximal expression of the cellular receptor occurred at near term or newborn stage. results also suggested that ...19969029788
modulation of cytokine expression by cd4+ t cells during coxsackievirus b3 infections of balb/c mice initiated by cells expressing the gamma delta + t-cell receptor.two variants of coxsackievirus b3 have been used to investigate the pathogenesis of myocarditis in balb/c mice. h3 virus induces moderate myocarditis and h310a1 virus induces minimal myocarditis, although both viruses infect and replicate in the heart. cells expressing the gamma delta+ t-cell receptor composed 5 to 13% of the lymphocytes infiltrating the hearts of h3 virus-infected mice and belonged to either the cd4- cd8+ gamma delta+- or cd4- cd8- gamma delta+-cell population. giving 5,000 gam ...19968627781
coxsackie b3 virus protein 2b contains cationic amphipathic helix that is required for viral rna replication.enterovirus protein 2b has been shown to increase plasma membrane permeability. we have identified a conserved putative amphipathic alpha-helix with a narrow hydrophilic face and an arrangement of cationic residues that is typical for the so-called lytic polypeptides. to examine the functional and structural roles of this putative amphipathic alpha-helix, we have constructed nine coxsackie b3 virus mutants by site-directed mutagenesis of an infectious cdna clone. six mutants contained substituti ...19968648724
direct interactions of coxsackievirus b3 with immune cells in the splenic compartment of mice susceptible or resistant to myocarditis.in vitro replication of coxsackievirus b3 (cvb3) in cells of the immune system derived from uninfected adolescent a/j and c57bl/6j mice and replication of cvb3 in and association with immune cells from spleens of infected animals in vivo were assessed. nonstimulated or mitogen-stimulated spleen cells were minimally permissive for viral replication during an 8-h period. three days postinfection (p.i.), cvb3 rna was localized in vivo to b cells and follicular dendritic cells of germinal centers in ...19968676490
intratypic genome variability of echovirus type 30 in part of the vp4/vp2 coding region.the genetic relationship of 33 echovirus type 30 (e30) isolates associated with three different outbreaks of meningitis in norway and one outbreak in usa was assessed using direct sequencing of amplicons derived from a region covering part of the capsid proteins vp4 and vp2. the e30 sequences were compared to each other, and to other enteroviruses. less sequence variation was observed between the isolates from a single outbreak (2-3%) than between groups of isolates from different outbreaks (4-9 ...19968678835
coxsackievirus b3-induced chronic myocarditis in mouse: use of whole blood culture to study the activation of tnf alpha-producing cells.the pathogenesis of cvb3-induced chronic myocarditis remains unknown. activated monocytes and macrophages may maintain ongoing inflammation during a persistent cvb3 infection and possibly represent the major mechanism leading to chronic myocarditis. we decided to study the activation status of cells by studying tnf alpha secretion in vitro using whole blood culture in cvb3-induced murine chronic myocarditis. seven dba/2 +/+ mice and 18 nmri nu/nu mice were inoculated intraperitoneally with 5x10( ...19968985939
recombinant interferons beta and gamma have a higher antiviral activity than interferon-alpha in coxsackievirus b3-infected carrier state cultures of human myocardial fibroblasts.we compared the antiviral activities of three recombinant human interferons (ifn-alph2a, ifn-beta, and ifn-gamma) in cultured human myocardial fibroblasts to select a candidate for trial in heart disease induced by cardiotropic enterovirus, e.g., coxsackievirus b3 (cvb3). cells were exposed to cvb3, and after 7 days, when a persistent infection had developed, ifn was added. virus yields were measured on alternate days for the next 7 or 16 days, and ifn activity was assessed as the percentage red ...19969162521
antibody cross-reactivity induced by the homologous regions in glutamic acid decarboxylase (gad65) and 2c protein of coxsackievirus b4. childhood diabetes in finland study group.gad65 contains an amino acid sequence which is highly homologous with a sequence in the 2c protein of coxsackievirus b4 (cbv4-2c). in the present study the possibility that this region could contain an epitope capable of inducing immunological cross-reactivity between cbv4-2c and gad65 was evaluated. six out of seven rabbit sera, which were raised against seven different synthetic peptides carrying various modifications of the homology sequence, showed cross-reactivity between 2c. gad65 and gadd ...19969099922
effect of heat on the survival of infectious coxsackievirus b3 and its genome in water.the aim of this study was to estimate the resistance of the viral genome of coxsackie b3 to heat (from 25 degrees c to 95 degrees c) with respect to the infectious virus. the results show that viral genomes were much more resistant to heat than infectious virus. the infectious coxsackie b3 was undetectable following incubation at 55 degrees c for 15 min., while the viral genome was still detected after incubation at 95 degrees c for 15 min. the viral genome became undetectable by rt semi-nested ...19969353546
effect of verapamil on ca2+ influx and cvb3-rna replication in cultured neonatal rat heart cells infected with cvb3.the effect of verapamil on ca2+ influx across the myocardial plasma membrane and coxsackie virus b3 (cvb3) -rna replication in cultured neonatal rat heart cells infected with cvb3 was investigated. it was found that the ca2+ influx could be inhibited significantly (p < 0.01) by verapamil (1 mumol/l) after infection of heart cells for 48 h. however, when the cultured heart cells infected with cvb3 and treated with verapamil (1 mumol/l and 10 nmol/l) at the same time for 48 h, the amounts of cvb3- ...19969387415
serological and molecular evidence of enterovirus infection in patients with end-stage dilated cardiomyopathy.to study the relative diagnostic value of enterovirus-specific molecular biological and serological assays in patients with end-stage dilated cardiomyopathy, and to investigate the possible role of other cardiotropic viruses in dilated cardiomyopathy.19968868984
viral serologies in patients with chronic fatigue and chronic fatigue syndrome.chronic fatigue syndrome (cfs) is an illness characterized by disabling fatigue associated with complaints of fevers, sore throat, myalgia, lymphadenopathy, sleep disturbances, neurocognitive difficulties, and depression. a striking feature of cfs is its sudden onset following an acute, presumably viral, illness and the subsequent recurrent "flu-like" symptoms. it has been speculated that both cfs and debilitating chronic fatigue (cf) that does not meet strict criteria for cfs may be the direct ...19968890037
mutagenesis of the coxsackie b3 virus 2b/2c cleavage site: determinants of processing efficiency and effects on viral replication.the enterovirus 2b/2c cleavage site differs from the common cleavage site motif axxq/g by the occurrence of either polar residues at the p1' position or large aliphatic residues at the p4 position. to study (i) the putative contribution of these aberrant residues to the stability of precursor protein 2bc, (ii) the determinants of cleavage site specificity and efficiency of 3cpro, and (iii) the importance of efficient cleavage at this site for viral replication, a mutational analysis of the coxsa ...19968892883
a mutation in the puff region of vp2 attenuates the myocarditic phenotype of an infectious cdna of the woodruff variant of coxsackievirus b3.coxsackievirus b3 (cvb3) infections induce myocarditis in humans and mice. little is known about the molecular characteristics of cvb3 that activate the cellular immunity responsible for cardiac inflammation. previous experiments have identified an antibody escape mutant (h310a1) of a myocarditic variant of cvb3 (h3) that attenuates the myocarditic potential of the virus in mice in spite of ongoing viral replication in the heart. we have cloned full-length infectious cdna copies of the viral gen ...19968892902
antiviral treatment with win 54 954 reduces mortality in murine coxsackievirus b3 myocarditis.coxsackieviruses b (cbvs) are dominant causative agents in myocarditis and are associated with pathogenesis is some cases of dilated cardiomyopathy, a clinical entity with a poor survival without heart transplantation.19968901680
detection of human viruses using primary cells immortalised by oncogene transfection, in comparison with primary cells and established cell lines.no single established cell line was found capable of substituting for primary baboon kidney (pbk) or primary rhesus macacque kidney (prk) cells for detection of human viruses. although a panel of cell lines could detect influenza, parainfluenza, and enteroviruses, which are among the most important viruses encountered in routine diagnostic laboratories, the sensitivity of this panel was not as high as that of pbk or prk cells. however, in a promising complementary approach, pbk and prk cells hav ...19968915884
cardiology casebook. coysackie virus b myocarditis presenting as a myocardial infarction in a newborn infant. 19968915943
nitric oxide and murine coxsackievirus b3 myocarditis: aggravation of myocarditis by inhibition of nitric oxide synthase.this study sought to investigate the effects of nitric oxide inhibition in a murine model of coxsackievirus b3 myocarditis.19968917278
proliferative lymphocyte responses to virus antigens homologous to gad65 in iddm.virus infection has been proposed as an initiating factor in the aetiology of insulin-dependent diabetes mellitus (iddm). we have examined lymphocyte proliferation to virus proteins which demonstrate sequence similarity to the beta-cell autoantigen glutamic acid decarboxylase (gad)65. the magnitude and frequency of response to coxsackie b viruses and adenovirus in a t-cell proliferation assay was significantly higher in a group of recently diagnosed iddm subjects than in non-diabetic control sub ...19968932998
effects of methyl mercury on cytokines, inflammation and virus clearance in a common infection (coxsackie b3 myocarditis).a myocarditic coxsackievirus b3 (cb3) infection in balb/c mice was used to investigate the effects of 12 weeks of methyl mercury (mehg) exposure (3.69 mg/g diet) on inflammatory heart lesions, virus in the heart, the cytokine response, i.e. cachectin/tnf-alpha and gamma-interferon (ifn-gamma) levels in plasma, and on disease complications and mortality. this dose of mehg did not influence mortality in this infection model. the inflammatory and necrotic lesions in the ventricular myocardium 7 day ...19968952707
pathogenesis of murine enterovirus myocarditis: virus dissemination and immune cell targets.in order to identify organ and cellular targets of persistent enterovirus infection in vivo, immunocompetent mice (swr/j, h-2q) were inoculated intraperitoneally with coxsackievirus b3 (cvb3). by use of in situ hybridization for the detection of enteroviral rna, we show that cvb3 is capable of inducing a multiorgan disease. during acute infection, viral rna was visualized at high levels in the heart muscle, pancreas, spleen, and lymph nodes and at comparably low levels in the central nervous sys ...19968971018
susceptibility of human bone marrow cells and hematopoietic cell lines to coxsackievirus b3 infection.viremia is commonly observed in association with enterovirus infections, and during this phase viruses can be transmitted to secondary target organs in the body. it is not known, however, whether blood cells play a role in the pathogenesis of enterovirus infection supporting virus replication. our earlier work (t. vuorinen, r. vainionpää, h. kettinen, and t. hyypiä, blood 84:823-829, 1994) demonstrated that coxsackievirus b3 is able to replicate in representatives of b- and t-cell lines but not ...19968971035
serial mri findings of benign poliomyelitis.a 14-year-old boy presented with incomplete transverse myelopathic symptoms associated with mild encephalitis. magnetic resonance imaging (mri) revealed involvement of the central portion of spinal cord extending from the c1 to t5 level on t2-weighted images with expansion of the cervical spinal cord. the boy made a full clinical recovery in a few weeks. cultures of the pharyngeal exudate were positive for echo virus type 11. the lesion on mri disappeared completely in a few months.19968733909
non isotopic automatable molecular procedures for the detection of enteroviruses.five microwell non isotopic hybridization assays, based on colorimetric immunoenzymatic reading, were developed and evaluated for the rapid and automatable detection of enteroviruses. virus nucleic acids and/or capture probes were covalently bound to microtiter wells, and digoxigenin-11-dutp was used as label for the detection of hybridized material. among these procedures, a reverse transcriptase polymerase chain reaction (rt-pcr) hybridization assay was the most sensitive, enabling the detecti ...19968737391
synthesis and antiviral activity evaluation of some new aminoadamantane derivatives. 2.the synthesis of some new aminoadamantane derivatives is described. the new compounds were evaluated against a wide range of viruses [influenza a h1n1, influenza a h2n2, influenza a h3n2, influenza b, parainfluenza 3, herpes simplex virus type 1 (hsv-1) and type 2 (hsv-2), thymidine kinase-deficient (tk-) hsv-1, vaccinia, vesicular stomatitis, polio 1, coxsackie b4, sindbis, semliki forest, reo 1, varicella-zoster virus (vzv), tk- vzv, human cytomegalovirus (hcmv), and human immunodeficiency vir ...19968765514
restricted usage of t-cell receptor v alpha genes in infiltrating cells in murine hearts with acute myocarditis caused by coxsackie virus b3.in murine myocarditis, it has been shown that natural killer cells first infiltrate the heart, followed by activated t-cells, which play an important role in the pathogenesis of the myocardial damage. in the same model of acute myocarditis, the repertoire of t-cell receptor (tcr) v beta genes in infiltrating cells in the heart has also been shown to be restricted. to study the nature of t-cell infiltration in more detail, the expression of tcr v alpha genes in infiltrating cells in the heart has ...19968778340
genetic determinants of susceptibility to coxsackievirus b1-induced chronic inflammatory myopathy: effects of host background and major histocompatibility complex genes.infection of outbred cd-1 mice with the tucson strain of coxsackievirus b1 (cvb1t) leads to the development of chronic hind limb weakness and associated inflammatory muscle disease. host factors that influence susceptibility have not been studied in this mouse model of chronic inflammatory myopathy (im). therefore, the pathogenesis was examined by using different inbred strains of mice. initially, seven strains of mice with either the h-2d or h-2b major histocompatibility complex (mhc) haplotype ...19968783635
echovirus type 23 observed as a nosocomial infection in infants.echovirus type 23 was isolated from 5 children, aged 1-13 months, during a 7-month period, covering a study period of several decades. the children were hospitalized for long periods because of chronic conditions. in 4 cases the virus was isolated from faecal samples, collected because of diarrhoea and/or vomiting. in one case echovirus type 23 was collected from nasopharyngeal secretions from a child with respiratory symptoms. the features of echovirus type 23 infection were similar to those of ...19968792494
the organisms reported to cause infective myocarditis and pericarditis in england and wales.it is difficult to acquire an overall perspective of the range of organisms responsible for infective myocarditis or pericarditis, and their relative importance, as most studies have involved only case reports or case series of a single organism. this study analyses reports to the communicable disease surveillance centre, of the public health laboratory service. reports where myocarditis or pericarditis was included as the main clinical features between 1990 and 1993 were studied. between 1990 a ...19968793712
the role of sensitized t-cells in myocarditis and dilated cardiomyopathy.enteroviruses like coxsackie are known to cause myocarditis both in animals and humans and enteroviral genom was found in endomyocardial biopsies of patients with myocarditis and dilated cardiomyopathy. however, subsequent to the initial viral infection, immune mechanisms seem to play an important role in the pathogenesis of both diseases. using synthetic peptides, it was possible to identify t-cell epitopes of coxsackie b3 virus and to test their significance in the pathogenesis of myocarditis ...19968803675
the pathogenesis of postinfectious myocarditis.myocarditis is an important cause of heart failure among adolescents and young adults. a remarkable observation is the discrepancy between the limited overt evidence of myocyte injury and the global impairment of left ventricular function. this discrepancy has stimulated suggestions that immunological mechanisms contribute to cardiac damage. we have developed two murine models of myocarditis, one elicited by cardiotropic coxsackie b3 (cb3) virus infection and the other by cardiac myosin immuniza ...19968811068
expression of nitric oxide synthase in a murine model of viral myocarditis induced by coxsackievirus b3.nitric oxide is known to participate in the immune and inflammatory processes. in this study, we investigated the production of nitric oxide (no) in murine viral myocarditis induced by coxsackievirus b3. the expression of inducible no synthase (inos) mrna in the heart first appeared on day 4 after virus inoculation and it was detectable for one month by reverse transcriptase-polymerase chain reaction (rt-pcr) methods. inos activity which was determined by the conversion of l-[3h]arginine to l-[3 ...19968607880
streptococcal m protein peptide with similarity to myosin induces cd4+ t cell-dependent myocarditis in mrl/++ mice and induces partial tolerance against coxsakieviral myocarditis.immunologic similarities have been demonstrated between coxsackievirus b3 (cvb3), group a streptococcal m protein, and cardiac myosin. previous studies have also shown that t lymphocytes obtained from cvb3-infected mice expressing the h-2k mhc haplotype gave an immunodominant proliferative response to the nt4 peptide (glkteneglkteneglkte) of the streptococcal m5 protein. we now show that the nt4 peptide can induce inflammatory heart disease in mrl/++ (h-2k) mice and that induction of anergy to t ...19968617982
nitric oxide inhibits viral replication in murine myocarditis.nitric oxide (no) is a radical molecule that not only serves as a vasodilator and neurotransmitter but also acts as a cytotoxic effector molecule of the immune system. the inducible enzyme making no, inducible no synthase (inos), is transcriptionally activated by ifn-gamma and tnf-alpha, cytokines which are produced during viral infection. we show that inos is induced in mice infected with the coxsackie b3 virus. macrophages expressing inos are identified in the hearts and spleens of infected an ...19968621766
natural isolates of echo virus type 25 with extensive variations in ires sequences and different translational efficiencies.the complete nucleotide sequence of the 5'-untranslated region (5'-utr) of the genome of three echo virus type 25 strains (the jv4 reference strain and two wild isolates) was determined. the 5'-utr of the two isolates shared 85 and 82% nucleotide identity with the reference strain. the overall folding of the predicted secondary structure of the echo virus 5'-utrs showed significant conservation with that of the poliovirus. most significant differences were observed in specific domains of the pre ...19968553589
tyrosine phosphorylation events during coxsackievirus b3 replication.in order to study cellular and viral determinants of pathogenicity, interactions between coxsackievirus b3 (cvb3) replication and cellular protein tyrosine phosphorylation were investigated. during cvb3 infection of hela cells, distinct proteins become phosphorylated on tyrosine residues, as detected by the use of antiphosphotyrosine western blotting. two proteins of 48 and 200 kda showed enhanced tyrosine phosphorylation 4 to 5 h postinfection (p.i.), although virus-induced inhibition of cellul ...19978985388
kissing of the two predominant hairpin loops in the coxsackie b virus 3' untranslated region is the essential structural feature of the origin of replication required for negative-strand rna synthesis.higher-order rna structures in the 3' untranslated region (3'utr) of enteroviruses are thought to play a pivotal role in viral negative-strand rna synthesis. the structure of the 3'utr was predicted by thermodynamic calculations using the star (structural analysis of rna) computer program and experimentally verified using chemical and enzymatic probing of in vitro-synthesized rna. a possible pseudoknot interaction between the 3d polymerase coding sequence and domain y and a "kissing" interaction ...19978985400
specific detection of coxsackie viruses a by the polymerase chain reaction.most of the coxsackie virus a strains are difficult to identify using traditional diagnostic methods such as virus isolation followed by neutralization with type-specific antisera. for the laboratory diagnoses of infections with coxsackie viruses a, inoculation into newborn mice has traditional been the method of choice. however, such investigations are complicated and time-consuming.19979406647
influence of viral infection on expression of cell surface antigens in human retinal pigment epithelial cells.subacute viral infection is known to change the phenotype of infected cells, thereby causing immune-mediated tissue damage. the aim of this study was to investigate the expression of different cell surface molecules on human retinal pigment epithelial cells (rpec) following viral infection, with special emphasis on those having immune-regulatory functions.19979407229
[epidemic of echo type 30 virus meningitis in children from the lodz region in the years 1995 and 1996].two epidemics of echo virus type 30 meningitis were compared with the special emphasis on the course of the disease and results of the laboratory investigations. the first epidemic occurred in 1995 and the second one in 1996. between 20 august and 20 october 1995, thirty six children, and between 11 july and 15 october 1996, ninety one children were admitted to the department of infectious diseases, medical university of lódź. all children were 3 to 15 years old, the median age was 9.08 years in ...19979411497
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