Publications

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cytoplasmic proteome and secretome profiles of differently stimulated human dendritic cells.dendritic cells (dcs), the most potent and specialized antigen-presenting cells, play a key role in the regulation of the adaptive immunity. immature dcs were generated by in vitro culturing of peripheral blood monocytes and functionally activated with the classical pathogen-associated molecular pattern lipopolysaccharide (lps). alternative activation resulting in th-2 polarization was induced with lipid oxidation products derived from 1-palmitoyl-2-arachidoyl-sn-glycerol-3-phosphorylcholin (oxp ...200919351150
expression and purification of the calcium binding photoprotein mitrocomin using zz-domain as a soluble partner in e. coli cells.we constructed a vector for soluble protein expression in the cytoplasm of escherichia coli cells using the cold induced system. the vector, named pcold-zz-p-x, consists of a histidine tag sequence, igg binding domain of protein a (zz domain), the cleavage site of human rhinovirus 3c protease followed by the multiple cloning sites under the controlled of the cold shock protein a (cspa) promoter and the lac operator. using this expression vector, the calcium binding photoprotein mitrocomin from l ...200919367679
synthesis and antirhinovirus activity of new 3-benzyl chromene and chroman derivatives.a series of 3-benzyl chromenes and chromans were synthesized and tested in vitro against human rhinovirus (hrv) 1b and 14, two representative serotypes for rhinovirus group b and a, respectively. all the new compounds, with the exception of 3-benzyl-2h-chromene (3a), showed a potent activity against hrv serotype 1b within micro or submicromolar range (ic(50)s from 0.11 to 6.62 microm). the low cytotoxicity of all the derivatives resulted in compounds with high therapeutic index (ti). on the cont ...200919398344
antibodies to the buried n terminus of rhinovirus vp4 exhibit cross-serotypic neutralization.development of a vaccine for the common cold has been thwarted by the fact that there are more than 100 serotypes of human rhinovirus (hrv). we previously demonstrated that the hrv14 capsid is dynamic and transiently displays the buried n termini of viral protein 1 (vp1) and vp4. here, further evidence for this "breathing" phenomenon is presented, using antibodies to several peptides representing the n terminus of vp4. the antibodies form stable complexes with intact hrv14 virions and neutralize ...200919403680
detection of human rhinovirus c in children with acute lower respiratory tract infections in south korea.recently, hrv-c was identified as a new species of hrv, but its spectrum of clinical disease is still not clear. the purpose of this study was to investigate the molecular epidemiology of hrvs in children with acute lower respiratory tract infections (lrtis). a total of 54 hrv-positive samples that were negative for other respiratory viruses were sequenced. hrv-a was detected in 33, hrv-b in 4, and hrv-c in 17 of these samples. all hrv-c-positive patients showed favorable clinical outcomes. we c ...200919415451
pneumonia and pericarditis in a child with hrv-c infection: a case report.human rhinovirus type c is a recently discovered species that has been associated with respiratory tract infections of unusual severity in some cases. however, the precise type of diseases associated with this new human rhinovirus needs to be investigated. in the present report, we used adapted real-time pcr assays to screen different clinical specimens collected from a 14-month-old boy presenting an acute lower respiratory tract disease complicated by a severe pericarditis. rt-pcr identified pi ...200919427260
human rhinoviruses: coming in from the cold.abstract : rhinovirus infections cause at least 70% of virus-related wheezing exacerbations and cold and flu-like illnesses. infections are also associated with otitis media, sinusitis and pneumonia. the annual impact of human rhinovirus (hrv) infections costs billions of healthcare dollars. to date, 100 serotyped hrv or 'classical' strains have been divided between two genetically distinct species based on subgenomic sequences, but many more, apparently novel strains remain un-characterized, ci ...200919439028
robust production of a peptide library using methodological synchronization.peptide libraries have proven to be useful in applications such as substrate profiling, drug candidate screening and identifying protein-protein interaction partners. however, issues of fidelity, peptide length, and purity have been encountered when peptide libraries are chemically synthesized. biochemically produced libraries, on the other hand, circumvent many of these issues due to the fidelity of the protein synthesis machinery. using thioredoxin as an expression partner, a stably folded pep ...200919457455
structure-activity relationships of heteroaromatic esters as human rhinovirus 3c protease inhibitors.human rhinovirus 3c protease (hrv 3c(pro)) is known to be a promising target for development of therapeutic agents against the common cold because of the importance of the protease in viral replication as well as its expression in a large number of serotypes. to explore non-peptidic inhibitors of hrv 3c(pro), a series of novel heteroaromatic esters was synthesized and evaluated for inhibitory activity against hrv 3c(pro), to determine the structure-activity relationships. the most potent inhibit ...200919464175
sensing picornaviruses using molecular imprinting techniques on a quartz crystal microbalance.molecular imprinting techniques were adapted to design a sensor for the human rhinovirus (hrv) and the foot-and-mouth disease virus (fmdv), which are two representatives of picornaviruses. stamp imprinting procedures lead to patterned polyurethane layers that depict the geometrical features of the template virus, as confirmed by afm for hrv. quartz crystal microbalance (qcm) measurements show that the resulting layers incorporate the template viruses reversibly and lead to mass effects that are ...200919469532
clinical severity and molecular typing of human rhinovirus c strains during a fall outbreak affecting hospitalized patients.the circulation rate and the clinical severity of infections caused by members of the new human rhinovirus c (hrv-c) species remain to be defined.200919473873
streptococcus pneumoniae coinfection is correlated with the severity of h1n1 pandemic influenza.initial reports in may 2009 of the novel influenza strain h1n1pdm estimated a case fatality rate (cfr) of 0.6%, similar to that of seasonal influenza. in july 2009, however, argentina reported 3056 cases with 137 deaths, representing a cfr of 4.5%. potential explanations for increased cfr included virus reassortment or genetic drift, or infection of a more vulnerable population. virus genomic sequencing of 26 argentinian samples representing both severe and mild disease indicated no evidence of ...200920046873
human rhinoviruses, allergy, and asthma: a clinical approach.the prevalence of allergic diseases is increasing in lithuania as in the world. the prevalence of allergic sensitization is often higher than 50% of the population. the "hygiene hypothesis" proposed that reduced immune-stimulation by infections may have resulted in the more widespread clinical expression of atopic disease. however, it alone does not provide an adequate explanation for the observed increase of allergic diseases. human rhinovirus infections are the major infections with a worldwid ...200920051716
antiviral activity of the zinc ionophores pyrithione and hinokitiol against picornavirus infections.we have discovered two metal ion binding compounds, pyrithione (pt) and hinokitiol (hk), that efficiently inhibit human rhinovirus, coxsackievirus, and mengovirus multiplication. early stages of virus infection are unaffected by these compounds. however, the cleavage of the cellular eukaryotic translation initiation factor eif4gi by the rhinoviral 2a protease was abolished in the presence of pt and hk. we further show that these compounds inhibit picornavirus replication by interfering with prop ...200918922875
nitric oxide inhibits human rhinovirus-induced transcriptional activation of cxcl10 in airway epithelial cells.human rhinovirus (hrv) infections trigger exacerbations of asthma and chronic obstructive pulmonary disease. nitric oxide (no) inhibits hrv replication in human airway epithelial cells and suppresses hrv-induced epithelial production of several cytokines and chemokines.200918986693
multiple receptors involved in human rhinovirus attachment to live cells.minor group human rhinoviruses (hrvs) attach to members of the low-density lipoprotein receptor family and are internalized via receptor-mediated endocytosis. the attachment of hrv2 to the cell surface, the first step in infection, was characterized at the single-molecule level by atomic force spectroscopy. sequential binding of multiple receptors was evident from recordings of characteristic quantized force spectra, which suggests that multiple receptors bound to the virus in a timely manner. u ...200818997008
amiloride derivatives inhibit coxsackievirus b3 rna replication.amiloride derivatives are known blockers of the cellular na(+)/h(+) exchanger and the epithelial na(+) channel. more recent studies demonstrate that they also inhibit ion channels formed by a number of viral proteins. we previously reported that 5-(n-ethyl-n-isopropyl)amiloride (eipa) modestly inhibits intracellular replication and, to a larger extent, release of human rhinovirus 2 (hrv2) (e. v. gazina, d. n. harrison, m. jefferies, h. tan, d. williams, d. a. anderson and s. petrou, antiviral re ...200818032495
enhanced severity of virus associated lower respiratory tract disease in asthma patients may not be associated with delayed viral clearance and increased viral load in the upper respiratory tract.viral respiratory infections, particularly human rhinovirus (hrv) infections, are the most common cause of asthma exacerbation. hrv infections usually lead to more severe and longer duration of lower respiratory tract (lrt) symptoms in asthmatics than in otherwise healthy individuals. however, the exact mechanism by which viruses contribute to exacerbation of asthma is unknown.200818096430
growth phenotypes and biosafety profiles in poliovirus-receptor transgenic mice of recombinant oncolytic polio/human rhinoviruses.the use of oncolytic recombinant polioviruses has an important therapeutic potential in the treatment of human gliomas. this study was carried out to assess parameters of the utility of the oncolytic poliovirus/human rhinovirus type 2 chimeras (pv/hrv2). the prototype pv/hrv2 chimera was constructed containing the complete genome of wild-type pv type 1 (mahoney) [pv1(m)] in which the cognate ires was replaced with that of hrv2 [called pv1(ripo)]. a derivative of pv1(ripo) is pv1(ripos) in which ...200818098139
syk associates with clathrin and mediates phosphatidylinositol 3-kinase activation during human rhinovirus internalization.human rhinovirus (hrv) causes the common cold. the most common acute infection in humans, hrv is a leading cause of exacerbations of asthma and chronic obstruction pulmonary disease because of its ability to exacerbate airway inflammation by altering epithelial cell biology upon binding to its receptor, icam-1. icam-1 regulates not only viral entry and replication but also signaling pathways that lead to inflammatory mediator production. we recently demonstrated the syk tyrosine kinase to be an ...200818178826
structure-based virtual screening for the discovery of natural inhibitors for human rhinovirus coat protein.inhibitors of the human rhinovirus (hrv) coat protein are promising candidates to treat and prevent a number of upper respiratory diseases. the aim of this study was to find antiviral compounds from nature, focusing on the hrv coat protein. through computational structure-based screening of an in-house 3d database containing 9676 individual plant metabolites from ancient herbal medicines, combined with knowledge from traditional use, we selected sesquiterpene coumarins from the gum resin asafeti ...200818247552
new anti-viral drugs for the treatment of the common cold.human rhinovirus (hrv) is the most important aetiologic agent of common cold in adults and children. hrv is a single-stranded, positive sense rna virus and, despite the high level of conservation among different serotypes, sequence alignment of viral protease 3c with mammalian protease reveals no homology. thus, protease 3c is an optimal target for the development of anti-hrv agents. in the present work we investigated the design, the synthesis and the development of new potential reversible inh ...200818248816
bronchial response pattern of antigen presenting cells and regulatory t cells in children less than 2 years of age.in early childhood, the ability to mount protective immune responses in the airways is impaired, with increased risk of allergic sensitisation to inhaled allergens. antigen presenting cells (apc) and regulatory t cells (treg) are important modifiers of t cell immunity but little is known about their distribution in bronchial mucosa at this age. here the subset distribution of apc and the appearance of foxp3(+) treg and bronchus associated lymphoid tissue (balt) were examined immunohistochemicall ...200818250182
recombinant vp4 of human rhinovirus induces permeability in model membranes.in common with all nonenveloped viruses, the mechanism of picornavirus membrane penetration during cell entry is poorly understood. the small, myristylated capsid protein vp4 has been implicated in this process. here we show that recombinant vp4 of human rhinovirus 16 has the ability to associate with and induce membrane permeability in otherwise intact liposomes. this provides further evidence that vp4 plays a key role in picornavirus cell entry.200818256154
gas-phase electrophoretic molecular mobility analysis of size and stoichiometry of complexes of a common cold virus with antibody and soluble receptor molecules.attachment of a nonaggregating monoclonal antibody and of a soluble recombinant receptor molecule to the icosahedral nonenveloped human rhinovirus serotype 2 was studied with a nanoelectrospray ionization gas-phase electrophoretic molecular mobility analyzer (nesi-gemma). the virus mass, as determined via nesi-gemma, was within instrument accuracy (+/-6%) close to the theoretical value (8 x 10(6) da) calculated from the sum of all constituents of one virus particle (60 copies of each of the four ...200818260676
attenuation of the type i interferon response in cells infected with human rhinovirus.the type i interferon (ifn) response requires the coordinated activation of the latent transcription factors nf-kappab, irf-3 and atf-2 which in turn activate transcription from the ifn-beta promoter. here we have examined the type i interferon response in rhinovirus type 14-infected a549 cells, with particular emphasis on the status of the transcription factor irf-3. our results indicate that although rhinovirus type 14 (rv14) infection induces the activation of nf-kappab and atf-2, only very l ...200818272195
human rhinovirus 1b exposure induces phosphatidylinositol 3-kinase-dependent airway inflammation in mice.infection with rhinovirus (rv) triggers exacerbations of asthma and chronic obstructive lung disease.200818276942
the presence of rhinovirus in lower airways of patients with bronchial asthma.the common cold virus, human rhinovirus (hrv), is the most frequent cause of asthma exacerbations. however, a possible contribution of hrv to the pathogenesis of chronic, persistent asthma has not been defined.200818276945
phasing of the triatoma virus diffraction data using a cryo-electron microscopy reconstruction.the blood-sucking reduviid bug triatoma infestans, one of the most important vector of american human trypanosomiasis (chagas disease) is infected by the triatoma virus (trv). trv has been classified as a member of the cripavirus genus (type cricket paralysis virus) in the dicistroviridae family. this work presents the three-dimensional cryo-electron microscopy (cryo-em) reconstruction of the trv capsid at about 25 a resolution and its use as a template for phasing the available crystallographic ...200818308357
a morpholino oligomer targeting highly conserved internal ribosome entry site sequence is able to inhibit multiple species of picornavirus.members of the genera enterovirus and rhinovirus (family picornaviridae) cause a wide range of human diseases. an established vaccine is available only for poliovirus, and no effective therapy is available for the treatment of infections caused by any pathogenic picornavirus. peptide-conjugated phosphorodiamidate morpholino oligomers (ppmo) are single-stranded dna-like antisense agents that readily enter cells. a panel of ppmo was tested for their antiviral activities against various picornaviru ...200818347107
human rhinovirus infection enhances airway epithelial cell production of growth factors involved in airway remodeling.childhood human rhinovirus (hrv) infections are associated with an increased risk of asthma. we reasoned that hrv infections might be important in the pathogenesis of airway remodeling, thereby providing a mechanism by which these children are at risk of asthma.200818355907
respiratory viruses in hiv-infected patients with suspected respiratory opportunistic infection.to assess the incidence and epidemiological pattern of respiratory viruses in hiv-infected patients and to evaluate their potential clinical impact.200818356599
induction of b7-h1 and b7-dc expression on airway epithelial cells by the toll-like receptor 3 agonist double-stranded rna and human rhinovirus infection: in vivo and in vitro studies.t-cell infiltration of the epithelium is a key feature of chronic rhinosinusitis and asthma. viral infections are an important cause of disease exacerbations. we have found virus-induced expression of t cell-interacting ligands, b7 homolog costimulatory molecules, on airway epithelium.200818378285
distinguishing molecular features and clinical characteristics of a putative new rhinovirus species, human rhinovirus c (hrv c).human rhinoviruses (hrvs) are the most frequently detected pathogens in acute respiratory tract infections (artis) and yet little is known about the prevalence, recurrence, structure and clinical impact of individual members. during 2007, the complete coding sequences of six previously unknown and highly divergent hrv strains were reported. to catalogue the molecular and clinical features distinguishing the divergent hrv strains, we undertook, for the first time, in silico analyses of all availa ...200818382652
serial viral infections in infants with recurrent respiratory illnesses.to better understand the viral aetiology of recurrent and prolonged illnesses, nasal secretions were prospectively collected from 285 infants at increased risk of developing asthma. of these, 27 infants had recurrent (at least five) moderate-to-severe respiratory illnesses (msis). the viral aetiology of the 150 msis and 86 scheduled visits was analysed by molecular diagnostics. the demographic and clinical data were compared with infants who had 0-4 msis. frequently ill infants had higher exposu ...200818448489
ifn-gamma regulation of icam-1 receptors in bronchial epithelial cells: soluble icam-1 release inhibits human rhinovirus infection.abstract:200818534017
the cis-acting replication elements define human enterovirus and rhinovirus species.replication of picornaviruses is dependent on vpg uridylylation, which is linked to the presence of the internal cis-acting replication element (cre). cre are located within the sequence encoding polyprotein, yet at distinct positions as demonstrated for poliovirus and coxsackievirus-b3, cardiovirus, and human rhinovirus (hrv-a and hrv-b), overlapping proteins 2c, vp2, 2a, and vp1, respectively. here we report a novel distinct cre element located in the vp2 region of the recently reported hrv-a2 ...200818541697
atomic force microscopy-derived nanoscale chip for the detection of human pathogenic viruses.native-protein nanolithography (npnl) was used to fabricate stable bioactive arrays of viral receptor spots. the arrays were specific for the cognate virus and devoid of nonspecific protein and virus adsorption under physiologic conditions. the spot size ranged from 200 nm x 200 nm to 2 microm x 2 microm and up to 3 x 3 spots were arranged per array. with proper force adjustment in the patterning experiments, his(6)-tagged bovine serum albumin (bsa) molecules were selectively removed from the un ...200818561273
simultaneous detection of different respiratory virus by a multiplex reverse transcription polymerase chain reaction combined with flow-through reverse dot blotting assay.cell culture and immunofluorescence (if) assays have been traditionally used for the laboratory diagnosis of respiratory viral infections, but these assays have a low sensitivity and are time consuming. we developed a multiplex reverse transcription polymerase chain reaction combined with flow-through reverse dot blotting (mrt-pcr-ft-rdb) assay for the simultaneous detection of influenza virus type a including h5 subtype and h9 subtype, influenza virus type b, parainfluenza virus types 1 and 3, ...200818639996
the linker domain of poly(rc) binding protein 2 is a major determinant in poliovirus cap-independent translation.poliovirus, a member of the enterovirus genus in the family picornaviridae, is the causative agent of poliomyelitis. translation of the viral genome is mediated through an internal ribosomal entry site (ires) encoded within the 5' noncoding region (5' ncr). ires elements are highly structured rna sequences that facilitate the recruitment of ribosomes for translation. previous studies have shown that binding of a cellular protein, poly(rc) binding protein 2 (pcbp2), to a major stem-loop structure ...200818656221
gene expression profiles during in vivo human rhinovirus infection: insights into the host response.human rhinovirus infections cause colds and trigger exacerbations of lower airway diseases.200818658112
assay for 5' noncoding region analysis of all human rhinovirus prototype strains.increasing recognition of the association of rhinovirus with severe lower respiratory tract illnesses has clarified the need to understand the relationship between specific serotypes of rhinovirus and their clinical consequences. to accomplish this, a specific and sensitive assay to detect and serotype rhinovirus directly from clinical specimens is needed. traditional methods of serotyping using culture and serum neutralization are time-consuming, limited to certain reference laboratories, and c ...200818753359
human rhinovirus group c infection in children with lower respiratory tract infection. 200818826844
does isoflurane enhance rhinovirus replication and virus-induced cytokine secretion in airway epithelial cells?there have been no previous studies regarding the effect of volatile anesthetics on human rhinovirus (rv) infection in airway epithelial cells of patients with an upper respiratory infection (uri). we have therefore evaluated in vitro the effect of isoflurane on rv infection in airway epithelial cells.200818840102
in vitro evaluation of the antiviral effects of the homeopathic preparation gripp-heel on selected respiratory viruses.gripp-heel is a homeopathic preparation frequently used in the treatment of respiratory viral infections such as various types of influenza and the common cold. the antiviral activity of gripp-heel was studied in vitro on human pathogenic enveloped and nonenveloped rna and dna viruses. before the antiviral assays, in vitro cytotoxicity of gripp-heel was determined with cells used for the infection experiments (hela, hep-2, mdck, bgm) as well as with mitogen-stimulated peripheral blood mononuclea ...200718066110
[study on the status of human rhinovirus infections in infants and young children with acute respiratory infections in beijing, from 2002 to 2006].to understand the relationship between human rhinovirus (hrv) and acute respiratory infections in infants and young children in beijing.200718069559
acute otitis media and respiratory viruses.the present study was performed to elucidate the clinical outcome, and etiology of acute otitis media (aom) in children based on virologic and bacteriologic tests. the study group consisted of 120 children aged 6 to 144 months with aom. middle ear fluid (mef) was tested for viral pathogens by reverse transcriptase polymerase chain reaction (rt-pcr) and for bacteria by gram-staining and culture. clinical response was assessed on day 2 to 4, 11 to 13, 26 to 28. respiratory viruses were isolated in ...200716967296
protein kinase r, ikappab kinase-beta and nf-kappab are required for human rhinovirus induced pro-inflammatory cytokine production in bronchial epithelial cells.rhinovirus infections cause the majority of acute exacerbations of airway diseases such as asthma and chronic obstructive pulmonary disease, with increased pro-inflammatory cytokine production by infected bronchial epithelial cells contributing to disease pathogenesis. theses diseases are a huge cause of morbidity worldwide, and contribute a major economic burden to healthcare costs. current steroid based treatments are only partially efficient at controlling virus induced inflammation, which re ...200716989899
understanding human rhinovirus infections in terms of qsar.the human rhinoviruses (hrvs) are the single most important cause of common colds. the widespread nature of this affliction, the economic consequences, and the well-known impracticality of vaccine development due to the large number of hrv serotypes (>100) have justified the search for chemotherapeutic agents. the interest in the application of quantitative structure-activity relationships has steadily increased in recent decades and we hope it may be useful in the search for anti-hrv agents. in ...200717045322
attenuation of herpes simplex virus neurovirulence with picornavirus cis-acting genetic elements.viral pathogenesis depends on a suitable milieu in target host cells permitting viral gene expression, propagation, and spread. in many instances, viral genomes can be manipulated to select for propagation in certain tissues or cell types. this has been achieved for the neurotropic poliovirus (pv) by exchange of the internal ribosomal entry site (ires), which is responsible for translation of the uncapped plus-strand rna genome. the ires of human rhinovirus type 2 (hrv2) confers neuron-specific ...200717079296
vp4 protein from human rhinovirus 14 is released by pressure and locked in the capsid by the antiviral compound win.rhinoviruses are the major causative agents of the common cold in humans. here, we studied the stability of human rhinovirus type 14 (hrv14) under conditions of high hydrostatic pressure, low temperature, and urea in the absence and presence of an antiviral drug. capsid dissociation and changes in the protein conformation were monitored by fluorescence spectroscopy, light scattering, circular dichroism, gel filtration chromatography, mass spectrometry and infectivity assays. the data show that h ...200717161425
rapid dna mapping by fluorescent single molecule detection.dna mapping is an important analytical tool in genomic sequencing, medical diagnostics and pathogen identification. here we report an optical dna mapping strategy based on direct imaging of individual dna molecules and localization of multiple sequence motifs on the molecules. individual genomic dna molecules were labeled with fluorescent dyes at specific sequence motifs by the action of nicking endonuclease followed by the incorporation of dye terminators with dna polymerase. the labeled dna mo ...200717175538
nmr structure of stem-loop d from human rhinovirus-14.the 5'-cloverleaf of the picornavirus rna genome is essential for the assembly of a ribonucleoprotein replication complex. stem-loop d (sld) of the cloverleaf is the recognition site for the multifunctional viral protein 3cpro. this protein is the principal viral protease, and its interaction with sld also helps to position the viral rna-dependent rna polymerase (3dpol) for replication. human rhinovirus-14 (hrv-14) is distinct from the majority of picornaviruses in that its sld forms a cuaug tri ...200717194719
identification of viruses in patients with postviral olfactory dysfunction.causative viruses of postviral olfactory dysfunction (pvod) have not yet been identified. the aim of this study was to investigate causative viruses in patients with pvod.200717277621
dynamic force microscopy imaging of plasmid dna and viral rna.plasmid dna and viral rna were imaged in a liquid environment by dynamic force microscopy (dfm) and fine structures of dna with heights of 1.82+/-0.66 nm were obtained in topographical images. in simultaneously acquired phase images, dna could be imaged with better contrast at lower imaging forces. by splitting the cantilever oscillation signal into lower and upper parts, the contribution of the adhesion between tip and sample to the topographical images was eliminated, resulting in better signa ...200717291581
mimicking early events of virus infection: capillary electrophoretic analysis of virus attachment to receptor-decorated liposomes.the attachment of human rhinovirus serotype 2 to an artificial cell membrane was followed by capillary electrophoresis. the cell membrane was mimicked by liposomes (average diameter of about 190 nm) containing a lipid with a nitrilotriacetic acid (nta) group. this group, in the presence of ni(2+) ions, served as anchor for the his(6)-tags of recombinant derivatives of the very-low-density lipoprotein (vldl) receptor comprising either modules 1, 2, and 3 (v123) or five tandem copies of module 3 ( ...200717297964
human rhinovirus type 54 infection via heparan sulfate is less efficient and strictly dependent on low endosomal ph.k-type major-group human rhinoviruses (hrvs) (including hrv54) share a prominent lysine residue in the hi surface loop of vp1 with all minor-group hrvs. despite the presence of this residue, they cannot use members of the low-density lipoprotein receptor family for productive infection. reexamining all k-type viruses for receptor usage, we noticed that hrv54 is able to replicate in rd cells that lack the major-group receptor intercellular adhesion molecule 1 (icam-1). by using receptor blocking ...200717301156
human rhinovirus 3c protease as a potential target for the development of antiviral agents.as the major cause of the common cold in children and adults, human rhinoviruses (hrvs) are a group of small single-stranded positive-sense rna viruses. hrvs translate their genetic information into a polyprotein precursor that is mainly processed by a virally encoded 3c protease (3cpro) to generate functional viral proteins and enzymes. it has been shown that the enzymatic activity of hrv 3cpro is essential to viral replication. the 3cpro is distinguished from most other proteases by the fact t ...200717305557
pdtc inhibits picornavirus polyprotein processing and rna replication by transporting zinc ions into cells.previously, it was shown that pyrrolidine dithiocarbamate (pdtc) inhibits proteolytic polyprotein processing and replication of human rhinovirus by transporting metal ions into cells. here, it is shown that pdtc also inhibits replication of two other picornaviruses: coxsackievirus b3 (cvb3), a closely related virus that belongs to the genus enterovirus, and mengovirus, an encephalomyocarditis virus strain that belongs to the genus cardiovirus, and that this inhibition is due to the dithiocarbama ...200717374764
the leader region of laminin b1 mrna confers cap-independent translation.translation initiation of eukaryotic mrnas generally occurs by cap-dependent ribosome scanning. however, certain mrnas contain internal ribosome entry sites (ires) allowing cap-independent translation. several of these ires-competent transcripts and their corresponding proteins are involved in tumourigenesis. this study focused on ires-driven translation control during the epithelial to mesenchymal transition (emt) of hepatocytes that reflects crucial aspects of carcinoma progression. expression ...200717395640
pocket factors are unlikely to play a major role in the life cycle of human rhinovirus.human rhinovirus 14 (hrv14) is a member of the rhinovirus genus, which belongs to the picornavirus family, which includes clinically and economically important members, such as poliovirus, foot-and-mouth disease virus, and endomyocarditis virus. capsid stability plays an important role in the viral infection process, in that it needs to be stable enough to move from cell to cell and yet be able to release its genetic material upon the appropriate environmental cues from the host cell. it has bee ...200717428846
models of infection and exacerbations in copd.despite optimal use of currently available therapy for stable copd, acute exacerbations are common events that constitute a major health burden. the development of highly sensitive diagnostic tools highlighted the role of viral infections in inducing copd exacerbations, with rhinoviruses being the most frequently identified virus type. so far, little is known about the mechanisms of virus-induced exacerbations. the recent development of the first human rhinovirus-induced copd exacerbation model ...200717475555
genome-wide diversity and selective pressure in the human rhinovirus.the human rhinoviruses (hrv) are one of the most common and diverse respiratory pathogens of humans. over 100 distinct hrv serotypes are known, yet only 6 genomes are available. due to the paucity of hrv genome sequence, little is known about the genetic diversity within hrv or the forces driving this diversity. previous comparative genome sequence analyses indicate that recombination drives diversification in multiple genera of the picornavirus family, yet it remains unclear if this holds for h ...200717477878
characterisation of a newly identified human rhinovirus, hrv-qpm, discovered in infants with bronchiolitis.human rhinoviruses (hrvs) are some of the earliest identified and most commonly detected viruses associated with acute respiratory tract infections (artis) and yet the molecular epidemiology and genomic variation of individual serotypes remains undefined.200717482871
development of potent inhibitors of the coxsackievirus 3c protease.coxsackievirus b3 (cvb3) 3c protease (3cp) plays essential roles in the viral replication cycle, and therefore, provides an attractive therapeutic target for treatment of human diseases caused by cvb3 infection. cvb3 3cp and human rhinovirus (hrv) 3cp have a high degree of amino acid sequence similarity. comparative modeling of these two 3cps revealed one prominent distinction; an asn residue delineating the s2' pocket in hrv 3cp is replaced by a tyr residue in cvb3 3cp. ag7088, a potent inhibit ...200717485072
human rhinovirus attenuates the type i interferon response by disrupting activation of interferon regulatory factor 3. 200717496231
antiviral activity of arbidol against influenza a virus, respiratory syncytial virus, rhinovirus, coxsackie virus and adenovirus in vitro and in vivo.arbidol, ethyl-6-bromo-4-[(dimethylamino)-methyl]-5-hydroxy-1-methyl-2-[(phenylthio)methyl]-in dole-3-carboxylate hydrochloride monohydrate, is an antiviral chemical agent. in this report, we studied the antiviral activity of arbidol against a panel of human respiratory viruses, namely influenza a virus (flu-a, a/pr/8/34 h1n1), respiratory syncytial virus (rsv), human rhinovirus type 14 (hrv 14), coxsackie virus b3 (cvb3) and adenovirus type 7 (adv-7) in vitro in cell culture. arbidol was found ...200717497238
a new, simple method for linking of antibodies to atomic force microscopy tips.functionalization of atomic force microscope (afm) tips with bioligands converts them into monomolecular biosensors which can detect complementary receptor molecules on the sample surface. flexible peg tethers are preferred because the bioligand can freely reorient and locally palpate the sample surface while the afm tip is moved along. in a well-established coupling scheme [hinterdorfer et al. (1996) proc. natl. acad. sci. u.s.a. 93, 3477-3481], a heterobifunctional peg linker is used to tether ...200717516625
high-throughput, sensitive, and accurate multiplex pcr-microsphere flow cytometry system for large-scale comprehensive detection of respiratory viruses.human respiratory viruses are a diverse group of pathogens composed of hundreds of virus strains, and this presents a major challenge for diagnostic laboratories. to efficiently detect numerous viruses in a large epidemiologic study, we developed a fast, multitarget, sensitive, and specific assay named the respiratory multicode-plx assay (rma). the rma utilizes improved multiplex pcr chemistry (eragen multicode-plx technology) coupled with high-throughput microsphere flow cytometry (luminex). ei ...200717537928
interleukin-17a modulates human airway epithelial responses to human rhinovirus infection.human rhinovirus (hrv) infections are associated with exacerbations of asthma and chronic obstructive pulmonary disease that are characterized by a selective neutrophil infiltration. il-17a, a cytokine derived primarily from activated t cells, has been linked to neutrophilic inflammation of the airways. we hypothesized that il-17a alters the response of hrv-infected epithelial cells to modulate airway inflammatory cell populations. il-17a synergistically enhanced hrv-16-induced epithelial produc ...200717545490
upper and lower respiratory tract infections by human enterovirus and rhinovirus in adult patients with hematological malignancies.the impact of human enterovirus (hev) and human rhinovirus (hrv) respiratory tract infections in adult patients with hematological malignancies has been infrequently reported. we retrospectively studied 31 patients with an upper or lower respiratory tract infection (urti/lrti) by hev (n = 18) or hrv (n = 15). at onset, a lrti was present in 6 (33%) and 2 (13%) episodes of hev and hrv infections, respectively, with or without an urti. progression to lrti (pneumonia) from prior urti was seen in 1 ...200717563077
infections with human rhinovirus induce the formation of distinct functional membrane domains.the plasma membrane contains distinct domains that are characterized by a high concentration of sphingolipids and cholesterol. these membrane microdomains also referred to as rafts, seem to be intimately involved in transmembranous signaling and often initiate interactions of pathogens and the host cell membranes. here, we investigated the further reorganization of membrane rafts in cultured epithelial cells and ex vivo isolated nasal cells after infection with rhinoviruses. we demonstrate the f ...200717595532
multicode-plx system for multiplexed detection of seventeen respiratory viruses.the multicode-plx system (eragen biosciences, inc., madison, wi) for the detection of respiratory viruses uses an expanded genetic alphabet, multiplex pcr chemistry, and microsphere flow cytometry to rapidly detect and specifically identify 17 different respiratory viruses directly in clinical specimens. the multicode-plx system was tested in parallel with direct fluorescent-antibody (dfa) staining and rapid shell vial culture (r-mix cells; diagnostic hybrids, inc. athens, oh) with 354 respirato ...200717596361
capillary electrophoresis of viruses, subviral particles and virus complexes.cze and cief were so far applied to the analysis of tobacco mosaic virus, semliki forest virus, human rhinovirus, adenovirus, norovirus and the bacteriophages t5 and ms2. the concentration of viral or subviral particles, of capsid proteins and viral genomes were determined, their electrophoretic mobilities and pi values were measured and bioaffinity reactions between viruses and antibodies, antibody fragments and receptor fragments were assessed. the role of detergents added to the bge to obtain ...200717623450
quantitative structure-activity relationship studies of [(biphenyloxy)propyl]isoxazole derivatives. inhibitors of human rhinovirus 2 replication.the 50% cytotoxic concentration (cc50) in hela cells, the 50% inhibitory concentration (ic50) against human rhinovirus 2 (hrv-2), and the selectivity index (si = cc50/ic50) of [(biphenyloxy)propyl]isoxazole derivatives were used to develop quantitative structure-activity relationships (qsar) based on simplex representation of molecular structure. statistic characteristics for partial least-squares models are quite satisfactory (r2 = 0.838 - 0.918; q2 = 0.695 - 0.87) for prediction of cc50, ic50, ...200717665898
induction of autophagy does not affect human rhinovirus type 2 production.induction of autophagy has been shown to be beneficial for the replication of poliovirus, a phenomenon that might also apply for other picornaviruses. we demonstrate that de novo synthesis of human rhinovirus type 2 (hrv2), an hrv of the minor receptor group, is unaffected by tamoxifen, rapamycin, and 3-methyladenine (3-ma), drugs either stimulating (tamoxifen and rapamycin) or inhibiting (3-ma) autophagic processes. furthermore, lc3-positive vesicles (i.e., autophagosomes) are not induced upon ...200717670838
human rhinovirus models in asthma.in both children and adults, rhinovirus (rv) infections remain a major cause of exacerbations in asthma. with the use of both in vitro models of rv infection and experimental models of asthma exacerbation in humans, insight into the precise role of rv in this process has been obtained. rv infects the lower airways, and the virus itself, together with the immune response to the virus, leads to increased airway obstruction in some patients with asthma. defects in the immune response to rv in these ...200717684329
a human rhinovirus model of chronic obstructive pulmonary disease exacerbations.chronic obstructive pulmonary disease (copd) exacerbations are common events that punctuate the natural history of copd contributing to disease severity progression and being the major cause of copd-related morbidity and mortality. currently available pharmacological strategies are only partially effective at reducing or preventing copd exacerbations. viral infections are the most frequent cause of copd exacerbations. the recent development of a human experimental model of rhinovirus-induced cop ...200717684335
pan-viral screening of respiratory tract infections in adults with and without asthma reveals unexpected human coronavirus and human rhinovirus diversity.between 50% and 80% of asthma exacerbations are associated with viral respiratory tract infections (rtis), yet the influence of viral pathogen diversity on asthma outcomes is poorly understood because of the limited scope and throughput of conventional viral detection methods.200717703411
anti-rhinovirus-specific activity of the alpha-sympathomimetic oxymetazoline.oxymetazoline (cas 2315-02-8, omz, nasivin) known as the active ingredient in nose drops and sprays demonstrates excellent efficacy in the treatment of rhinitis symptoms that are mainly caused by rhinovirus infections. to elucidate possible modes of action, the antiviral activity of omz was studied in vitro on human pathogenic viruses. no in vitro effects were detected against enveloped rna viruses, parainfluenza virus and respiratory syncytial virus and against adenovirus, a non-enveloped dna-v ...200717803062
clinical features and complete genome characterization of a distinct human rhinovirus (hrv) genetic cluster, probably representing a previously undetected hrv species, hrv-c, associated with acute respiratory illness in children.although human rhinoviruses (hrvs) are common causes of respiratory illness, their molecular epidemiology has been poorly investigated. despite the recent findings of new hrv genotypes, their clinical disease spectrum and phylogenetic positions were not fully understood. in this study, 203 prospectively collected nasopharyngeal aspirates (npas), negative for common respiratory viruses (83 were human bocavirus [hbov] positive and 120 hbov negative), from hospitalized children during a 1-year peri ...200717804649
human rhinovirus type 14 gain-of-function mutants for orii utilization define residues of 3c(d) and 3dpol that contribute to assembly and stability of the picornavirus vpg uridylylation complex.vpg linkage to the 5' ends of picornavirus rnas requires production of vpg-pupu. vpg-pupu is templated by an rna stem-loop (the cre or orii) found at different locations in picornavirus genomes. at least one adaptive mutation is required for human rhinovirus type 14 (hrv-14) to use poliovirus type 3 (pv-3) or pv-1 orii efficiently. one mutation changes leu-94 of 3c to pro; the other changes asp-406 of 3dpol to asn. by using an in vitro vpg uridylylation system for hrv-14 that recapitulates biolo ...200717855535
effect of detergent on electromigration of proteins: ce of very low density lipoprotein receptor modules and viral proteins.the different electrophoretic behavior of the members of two groups of proteins with respect to the absence or presence of detergent additives in the bge was explored. recombinant soluble concatemers of repeat 3 of the very low density lipoprotein (vldl)-receptor fused at their n-terminus to maltose-binding protein (mbp) exhibited different electrophoretic mobilities in borate buffer (ph 8.3) in the absence and in the presence of dodecyl-peg ether (d-peg). this enabled the separation of the rece ...200717893950
cytoplasmic relocalization of heterogeneous nuclear ribonucleoprotein a1 controls translation initiation of specific mrnas.heterogeneous nuclear ribonucleoprotein (hnrnp) a1 is a nucleocytoplasmic shuttling protein that regulates gene expression through its action on mrna metabolism and translation. the cytoplasmic redistribution of hnrnp a1 is a regulated process during viral infection and cellular stress. here, we show that hnrnp a1 is an internal ribosome entry site (ires) trans-acting factor that binds specifically to the 5' untranslated region of both the human rhinovirus-2 and the human apoptotic peptidase act ...200717898077
in vitro resistance study of rupintrivir, a novel inhibitor of human rhinovirus 3c protease.rupintrivir (formerly ag7088) is an irreversible inhibitor of the human rhinovirus (hrv) 3c protease that has been demonstrated to have in vitro activity against all hrvs tested, consistent with its interaction with a strictly conserved subset of amino acids in the 3c protease. the potential for resistance was studied following in vitro serial passage of hrv serotypes 14, 2, 39, and hanks in the presence of increasing rupintrivir concentrations. hrv variants with reduced susceptibilities to rupi ...200717908951
nmr solution structures of the apo and peptide-inhibited human rhinovirus 3c protease (serotype 14): structural and dynamic comparison.the human rhinovirus (hrv) is a positive sense rna virus responsible for about 30% of "common colds". it relies on a 182 residue cysteine protease (3c) to proteolytically process its single gene product. inhibition of this enzyme in vitro and in vivo has consistently demonstrated cessation of viral replication. this suggests that 3c protease inhibitors could serve as good drug candidates. however, significant proteolytic substrate diversity exists within the 110+ known rhinovirus serotypes. to i ...200717944485
internal initiation of translation from the human rhinovirus-2 internal ribosome entry site requires the binding of unr to two distinct sites on the 5' untranslated region.internal initiation of translation from the human rhinovirus-2 (hrv-2) internal ribosome entry site (ires) is dependent upon host cell trans-acting factors. the multiple cold shock domain protein unr and the polypyrimidine tract-binding protein have been identified as synergistic activators of hrv-2 ires-driven translation. in order to investigate the mechanism by which unr acts in this process, we have mapped the binding sites of unr to two distinct secondary structure domains of the hrv-2 ires ...200717947529
virus analysis by electrophoresis on a microfluidic chip.exploiting the advantages of miniaturization of analytical devices we worked out conditions for the analysis of viruses, subviral particles, and virus-receptor complexes on microfluidic chips. to allow for detection via laser-induced fluorescence, the viral capsids were labelled with the fluorescent dye cy5. we analyzed human rhinovirus serotype 2 and subviral particles, followed the complexation of the virus with a synthetic fragment of the vldl-receptor, and tracked the heat-induced conversion ...200718006393
electrophoresis on a microfluidic chip for analysis of fluorescence-labeled human rhinovirus.we report the analysis of human rhinovirus serotype 2 (hrv2) on a commercially available lab-on-a-chip instrument. due to lack of sufficient native fluorescence, the proteinaceous capsid of hrv2 was labeled with cy5 for detection by the red laser (lambda ex 630 nm) implemented in the instrument. on the microdevice, electrophoresis of the labeled virus was possible in a bge without stabilizing detergents, which is in contrast to conventional ce; moreover, analysis times were drastically shortened ...200718008310
syk is downstream of intercellular adhesion molecule-1 and mediates human rhinovirus activation of p38 mapk in airway epithelial cells.the airway epithelium is the primary target of inhaled pathogens such as human rhinovirus (hrv). airway epithelial cells express icam-1, the major receptor for hrv. hrv binding to icam-1 mediates not only viral entry and replication but also a signaling cascade that leads to enhanced inflammatory mediator production. the specific signaling molecules and pathways activated by hrv-icam-1 interactions are not well characterized, although studies in human airway epithelia implicate a role for the p3 ...200617082600
transforming growth factor-beta expression induced by rhinovirus infection in respiratory epithelial cells.rhinovirus infection of the lower airways is now a recognized disease, associated with bronchiolitis and asthma. the bronchial epithelial cells are the host cells when rhinovirus infection occurs in the airway. it was hypothesized that a pro-fibrotic growth factor response may occur in these infected cells, leading to production of a key transforming growth factor, tgf-beta-1. bronchial epithelial cells were inoculated with human rhinovirus and compared at day 1, 3 and 5 to control non-infected ...200617151785
effects of picornavirus 3a proteins on protein transport and gbf1-dependent cop-i recruitment.the 3a protein of the coxsackievirus b3 (cvb3), an enterovirus that belongs to the family of the picornaviruses, inhibits endoplasmic reticulum-to-golgi transport. recently, we elucidated the underlying mechanism by showing that cvb3 3a interferes with adp-ribosylation factor 1 (arf1)-dependent cop-i recruitment to membranes by binding and inhibiting the function of gbf1, a guanine nucleotide exchange factor that is required for the activation of arf1 (e. wessels et al., dev. cell 11:191-201, 20 ...200617005635
nonneutralizing human rhinovirus serotype 2-specific monoclonal antibody 2g2 attaches to the region that undergoes the most dramatic changes upon release of the viral rna.the monoclonal antibody 2g2 has been used extensively for detection and quantification of structural changes of human rhinovirus serotype 2 during infection. it recognizes exclusively a and b subviral particles, not native virus. we have elucidated the basis of this selectivity by determining the footprint of 2g2. since viral escape mutants obviously cannot be obtained, the structures of complexes between fab fragments of 2g2 and 80s subviral b particles were determined by cryoelectron microscop ...200617005641
defining residues involved in human rhinovirus 2a proteinase substrate recognition.the 2a proteinase (2a(pro)) of human rhinoviruses (hrvs) initiates proteolytic processing by cleaving between the c-terminus of vp1 and its own n-terminus. it subsequently cleaves the host protein eif4gi. hrv2 and hrv14 2a(pro) cleave at iitta *gpsd and diksy *glgp on their respective polyproteins. the hrv2 2a(pro) cleavage site on eif4gi is tlstr *gppr. we show that hrv2 2a(pro) can self-process at the eif4gi cleavage sequence whereas hrv14 2a(pro) cannot, due to the presence of the arginine re ...200617007846
human rhinovirus induces robust ip-10 release by monocytic cells, which is independent of viral replication but linked to type i interferon receptor ligation and stat1 activation.human rhinovirus (hrv)-induced respiratory infections are associated with elevated levels of ifn-gamma-inducible protein 10 (ip-10), which is an enhancer of t lymphocyte chemotaxis and correlates with symptom severity and t lymphocyte number. increased ip-10 expression is exhibited by airway epithelial cells following ex vivo hrv challenge and requires intracellular viral replication; however, there are conflicting reports regarding the necessity of type i ifn receptor ligation for ip-10 express ...200617020930
new treatment regimes for virus-induced exacerbations of asthma.this review will focus on the role of viruses as causes of asthma exacerbations. the article will briefly review the current literature supporting this view, with a special focus on human rhinovirus (rv), the main virus associated with exacerbations of asthma. the review will then refer to possible strategies for treatment, and will include discussion on treatment with specific anti-viral therapy and type i interferon as a treatment for rv. the review will also include a discussion on current th ...200616289761
a multi-centre pilot proficiency programme to assess the quality of molecular detection of respiratory viruses.to assess the quality of molecular detection of respiratory viruses in clinical diagnostic laboratories.200616019258
alignment of capsid protein vp1 sequences of all human rhinovirus prototype strains: conserved motifs and functional domains.an alignment was made of the deduced amino acid sequences of the entire capsid protein vp1 of all human rhinovirus (hrv) prototype strains to examine conserved motifs in the primary structure. a set of previously proposed crucially important amino acids in the footprints of the two known receptor molecules was not conserved in a receptor group-specific way. in contrast, vp1 and vp3 amino acids in the minor receptor-group strains corresponding to most of the predicted icam-1 footprint definitely ...200616361425
structure and dynamics of coxsackievirus b4 2a proteinase, an enyzme involved in the etiology of heart disease.the 2a proteinases (2a(pro)) from the picornavirus family are multifunctional cysteine proteinases that perform essential roles during viral replication, involving viral polyprotein self-processing and shutting down host cell protein synthesis through cleavage of the eukaryotic initiation factor 4g (eif4g) proteins. coxsackievirus b4 (cvb4) 2a(pro) also cleaves heart muscle dystrophin, leading to cytoskeletal dysfunction and the symptoms of human acquired dilated cardiomyopathy. we have determin ...200616415022
age-dependent poliovirus replication in the mouse central nervous system is determined by internal ribosome entry site-mediated translation.mouse cells are not permissive for the replication of human rhinovirus type 2 (hrv2). to determine the role of the hrv2 internal ribosome entry site (ires) in determining species specificity, a recombinant poliovirus (p1/hrv2) was constructed by substituting the poliovirus ires with the ires from hrv2. this recombinant virus replicated in all human and murine cell lines examined, demonstrating that the hrv2 ires does not limit viral replication in transformed murine cells. p1/hrv2 replicated in ...200616501069
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