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duration of rt-pcr positivity in severe acute respiratory syndrome.severe acute respiratory syndrome (sars) is a highly infectious respiratory infection with a high mortality. the duration of infectivity is unknown. the rt-pcr positivity for sars-associated coronavirus (sars-cov) was followed in 45 virologically confirmed sars patients. serial rt-pcrs for sars-cov were performed in the nasopharyngeal aspirate, stool and urine of 45 sars patients who survived until discharge. all patients had at least one site that was positive for sars-cov on presentation. time ...200515640317
severe acute respiratory syndrome and critical care medicine: the toronto experience.the 2003 global outbreak of severe acute respiratory syndrome (sars) provided numerous challenges to the delivery of critical care. the toronto critical care community has learned important lessons from sars, which will help in preparation for future disease outbreaks.200515640680
structure and intracellular targeting of the sars-coronavirus orf7a accessory protein.the open reading frame (orf) 7a of the sars-associated coronavirus (sars-cov) encodes a unique type i transmembrane protein of unknown function. we have determined the 1.8 a resolution crystal structure of the n-terminal ectodomain of orf7a, revealing a compact seven-stranded beta sandwich unexpectedly similar in fold and topology to members of the ig superfamily. we also demonstrate that, in sars-cov- infected cells, the orf7a protein is expressed and retained intracellularly. confocal microsco ...200515642263
generation of predictive pharmacophore model for sars-coronavirus main proteinase.pharmacophore-based virtual screening is an effective, inexpensive and fast approach to discovering useful starting points for drug discovery. in this study, we developed a pharmacophore model for the main proteinase of severe acute respiratory syndrome coronavirus (sars-cov). then we used this pharmacophore model to search nci 3d database including 250, 251 compounds and identified 30 existing drugs containing the pharmacophore query. among them are six compounds that already exhibited anti-sar ...200515642409
prokaryotic expression, refolding, and purification of fragment 450-650 of the spike protein of sars-coronavirus.the spike (s) glycoprotein is one of the major structure proteins of sars-associated coronavirus (cov). fragment 450-650 (s450-650) of the s protein contains receptor-binding domain and neutralizing epitopes. in this study, s450-650 was expressed with a histidine tag in escherichia coli bl21. bacterial inclusion bodies containing the recombinant s450-650 were solubilized with 8 m urea and then applied onto a ni-nitrilotriacetic acid column. on-column refolding and purification was performed. red ...200515642467
evasion of antibody neutralization in emerging severe acute respiratory syndrome coronaviruses.molecular characterization of the severe acute respiratory syndrome coronavirus has revealed genetic diversity among isolates. the spike (s) glycoprotein, the major target for vaccine and immune therapy, shows up to 17 substitutions in its 1,255-aa sequence; however, the biologic significance of these changes is unknown. here, the functional effects of s mutations have been determined by analyzing their affinity for a viral receptor, human angiotensin-converting enzyme 2 (hace-2), and their sens ...200515642942
monoclonal antibody-based antigen capture enzyme-linked immunosorbent assay reveals high sensitivity of the nucleocapsid protein in acute-phase sera of severe acute respiratory syndrome patients.accurate and timely diagnosis of severe acute respiratory syndrome coronavirus (sars-cov) infection is a critical step in preventing another global outbreak. in this study, 829 serum specimens were collected from 643 patients initially reported to be infected with sars-cov. the sera were tested for the n protein of sars-cov by using an antigen capture enzyme-linked immunosorbent assay (elisa) based on monoclonal antibodies against the n protein of sars-cov and compared to 197 control serum sampl ...200515642998
susceptibility of different eukaryotic cell lines to sars-coronavirus.in order to define and characterize target cells of sars-coronavirus (sars-cov) we studied the susceptibility of 23 different permanent and primary eukaryotic cell lines to sars-coronavirus. beneath vero e6 cells sars- coronavirus infection could also be demonstrated in two pig cell lines (poek, ps) and one human cell line (huh-7) using the indirect immunofluorescence assay and a newly established quantitative real-time pcr. in all susceptible cell lines mrna of the angiotensin-converting enzyme ...200515645376
high-dose hydrocortisone reduces expression of the pro-inflammatory chemokines cxcl8 and cxcl10 in sars coronavirus-infected intestinal cells.clinical observations and our high-density oligonucleotide microarray results demonstrated increased expression of proinflammatory chemokines after sars-cov infection. here, we investigated the influence of sars-cov infection on cxcl8 (interleukin 8) and cxcl10 (interferon-gamma-inducible protein 10) in human intestinal epithelial (caco2) cells. rt-pcr and elisa showed time-dependent up-regulation of both chemokines after sars-cov infection. electric mobility shift assay revealed increased dna b ...200515647850
inhibition of sars-cov replication by sirna.serious outbreaks of severe acute respiratory syndrome (sars), caused by the newly discovered coronavirus sars-cov, occurred between late 2002 and early 2003 and there is an urgent need for effective antiviral agents. rna interference in animals and post-transcriptional gene silencing plants is mediated by small double-stranded rna molecules named small interfering rna (sirna). recently, sirna-induced rna interference(rnai) may provide a new approach to therapy for pathogenic viruses, e.g. hiv a ...200515652970
severe acute respiratory syndrome associated coronavirus is detected in intestinal tissues of fatal cases.a significant percentage of confirmed severe acute respiratory syndrome (sars) patients experienced gastrointestinal symptoms, and the viral sequence was detectable in the stool of most patients. at present, the knowledge of the pathology of the digestive system in sars patients is limited. because a resurgence of the sars epidemic is constantly possible, there is an urgent need to understand the involvement of the digestive system in this new disease.200515654797
development and characterization of a severe acute respiratory syndrome-associated coronavirus-neutralizing human monoclonal antibody that provides effective immunoprophylaxis in mice.severe acute respiratory syndrome (sars) remains a significant public health concern after the epidemic in 2003. human monoclonal antibodies (mabs) that neutralize sars-associated coronavirus (sars-cov) could provide protection for exposed individuals.200515655773
characterization of cytokine/chemokine profiles of severe acute respiratory syndrome.there is currently no optimal treatment or effective drug for severe acute respiratory syndrome (sars), because the immunopathologic mechanism is poorly understood. objectives: to explore the immune mechanism underlying the pathogenesis of sars, we studied the expression profile of cytokines/chemokines in the blood and the immunopathology of the lung and lymphoid tissues.200515657466
enzymatic activity characterization of sars coronavirus 3c-like protease by fluorescence resonance energy transfer technique.to characterize enzymatic activity of severe acute respiratory syndrome (sars) coronavirus (cov) 3c-like protease (3cl(pro)) and its four site-directed mutants.200515659121
a molecular beacon, bead-based assay for the detection of nucleic acids by flow cytometry.molecular beacons are dual-labelled probes that are typically used in real-time pcr assays, but have also been conjugated with solid matrices for use in microarrays or biosensors. we have developed a fluid array system using microsphere-conjugated molecular beacons and the flow cytometer for the specific, multiplexed detection of unlabelled nucleic acids in solution. for this array system, molecular beacons were conjugated with microspheres using a biotin-streptavidin linkage. a bridged conjugat ...200515659574
expression of sars-coronavirus nucleocapsid protein in escherichia coli and lactococcus lactis for serodiagnosis and mucosal vaccination.the nucleocapsid (n) protein of the severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) is an important antigen for the early diagnosis of sars and the development of vaccines. it was expressed in escherichia coli as a fusion with human glutathione s-transferase (hgst) and was confirmed by western blotting analysis. this recombinant n protein (hgst-n) was purified and used to measure the sars-cov n-specific antibody in the sera of eight sars patients by enzyme-linked immun ...200515660214
molecular mechanisms of severe acute respiratory syndrome (sars).severe acute respiratory syndrome (sars) is a new infectious disease caused by a novel coronavirus that leads to deleterious pulmonary pathological features. due to its high morbidity and mortality and widespread occurrence, sars has evolved as an important respiratory disease which may be encountered everywhere in the world. the virus was identified as the causative agent of sars due to the efforts of a who-led laboratory network. the potential mutability of the sars-cov genome may lead to new ...200515661082
molecular and biological characterization of human monoclonal antibodies binding to the spike and nucleocapsid proteins of severe acute respiratory syndrome coronavirus.human monoclonal antibodies (mabs) were selected from semisynthetic antibody phage display libraries by using whole irradiated severe acute respiratory syndrome (sars) coronavirus (cov) virions as target. we identified eight human mabs binding to virus and infected cells, six of which could be mapped to two sars-cov structural proteins: the nucleocapsid (n) and spike (s) proteins. two mabs reacted with n protein. one of the n protein mabs recognized a linear epitope conserved between all publish ...200515650189
identification of the membrane-active regions of the severe acute respiratory syndrome coronavirus spike membrane glycoprotein using a 16/18-mer peptide scan: implications for the viral fusion mechanism.we have identified the membrane-active regions of the severe acute respiratory syndrome coronavirus (sars cov) spike glycoprotein by determining the effect on model membrane integrity of a 16/18-mer sars cov spike glycoprotein peptide library. by monitoring the effect of this peptide library on membrane leakage in model membranes, we have identified three regions on the sars cov spike glycoprotein with membrane-interacting capabilities: region 1, located immediately upstream of heptad repeat 1 ( ...200515650199
identification of two neutralizing regions on the severe acute respiratory syndrome coronavirus spike glycoprotein produced from the mammalian expression system.the spike (s) protein of the severe acute respiratory syndrome-associated coronavirus (sars-cov) plays important roles in viral pathogenesis and potentially in the development of an effective vaccine against this virulent infectious disease. in this study, the codon-optimized s gene of sars-cov was synthesized to construct dna vaccine plasmids expressing either the full-length or segments of the s protein. high titer s-specific immunoglobulin g antibody responses were elicited in rabbits immuniz ...200515650214
nitric oxide inhibits the replication cycle of severe acute respiratory syndrome coronavirus.nitric oxide (no) is an important signaling molecule between cells which has been shown to have an inhibitory effect on some virus infections. the purpose of this study was to examine whether no inhibits the replication cycle of the severe acute respiratory syndrome coronavirus (sars cov) in vitro. we found that an organic no donor, s-nitroso-n-acetylpenicillamine, significantly inhibited the replication cycle of sars cov in a concentration-dependent manner. we also show here that no inhibits vi ...200515650225
virtual screening of novel noncovalent inhibitors for sars-cov 3c-like proteinase.the sars coronavirus 3c-like proteinase is considered as a potential drug design target for the treatment of severe acute respiratory syndrome (sars). owing to the lack of available drugs for the treatment of sars, the discovery of inhibitors for sars coronavirus 3c-like proteinase that can potentially be optimized as drugs appears to be highly desirable. we have built a "flexible" three-dimensional model for sars 3c-like proteinase by homology modeling and multicanonical molecular dynamics meth ...200515667124
programmed ribosomal frameshifting in decoding the sars-cov genome.programmed ribosomal frameshifting is an essential mechanism used for the expression of orf1b in coronaviruses. comparative analysis of the frameshift region reveals a universal shift site u_uua_aac, followed by a predicted downstream rna structure in the form of either a pseudoknot or kissing stem loops. frameshifting in sars-cov has been characterized in cultured mammalian cells using a dual luciferase reporter system and mass spectrometry. mutagenic analysis of the sars-cov shift site and mas ...200515680415
inhibition of beta interferon induction by severe acute respiratory syndrome coronavirus suggests a two-step model for activation of interferon regulatory factor 3.severe acute respiratory syndrome (sars) is caused by a novel coronavirus termed sars-cov. we and others have previously shown that the replication of sars-cov can be suppressed by exogenously added interferon (ifn), a cytokine which is normally synthesized by cells as a reaction to virus infection. here, we demonstrate that sars-cov escapes ifn-mediated growth inhibition by preventing the induction of ifn-beta. in sars-cov-infected cells, no endogenous ifn-beta transcripts and no ifn-beta promo ...200515681410
civets are equally susceptible to experimental infection by two different severe acute respiratory syndrome coronavirus isolates.severe acute respiratory syndrome (sars) was caused by a novel virus now known as sars coronavirus (sars-cov). the discovery of sars-cov-like viruses in masked palm civets (paguma larvata) raises the possibility that civets play a role in sars-cov transmission. to test the susceptibility of civets to experimental infection by different sars-cov isolates, 10 civets were inoculated with two human isolates of sars-cov, bj01 (with a 29-nucleotide deletion) and gz01 (without the 29-nucleotide deletio ...200515681462
civets are equally susceptible to experimental infection by two different severe acute respiratory syndrome coronavirus isolates.severe acute respiratory syndrome (sars) was caused by a novel virus now known as sars coronavirus (sars-cov). the discovery of sars-cov-like viruses in masked palm civets (paguma larvata) raises the possibility that civets play a role in sars-cov transmission. to test the susceptibility of civets to experimental infection by different sars-cov isolates, 10 civets were inoculated with two human isolates of sars-cov, bj01 (with a 29-nucleotide deletion) and gz01 (without the 29-nucleotide deletio ...200515681462
folding of the sars coronavirus spike glycoprotein immunological fragment (sars_s1b): thermodynamic and kinetic investigation correlating with three-dimensional structural modeling.spike glycoprotein of sars coronavirus (s protein) plays a pivotal role in sars coronavirus (sars_cov) infection. the immunological fragment of the s protein (ala251-his641, sars_s1b) is believed to be essential for sars_cov entering the host cell through s protein-ace-2 interaction. we have quantitatively characterized the thermally induced and guhcl-induced unfolding features of sars_s1b using circular dichroism (cd), tryptophan fluorescence, and stopped-flow spectral techniques. for the therm ...200515683230
[prokaryotic expression of sars coronavirus spike protein and construction of its dna vaccine].to study the immunological characteristics of the spike (s) protein of sars coronavirus (sars-cov) and analyze the feasibility of using this protein as the component for sars vaccine development.200515683993
function of hab18g/cd147 in invasion of host cells by severe acute respiratory syndrome coronavirus.to identify the function of hab18g/cd147 in invasion of host cells by severe acute respiratory syndrome (sars) coronavirus (cov), we analyzed the protein-protein interaction among hab18g/cd147, cyclophilin a (cypa), and sars-cov structural proteins by coimmunoprecipitation and surface plasmon resonance analysis. although none of the sars-cov proteins was found to be directly bound to hab18g/cd147, the nucleocapsid (n) protein of sars-cov was bound to cypa, which interacted with hab18g/cd147. fur ...200515688292
molecular modeling and chemical modification for finding peptide inhibitor against severe acute respiratory syndrome coronavirus main proteinase.severe acute respiratory syndrome (sars) is a respiratory disease caused by a newly found virus, called sars coronavirus. in this study, the cleavage mechanism of the sars coronavirus main proteinase (mpro or 3clpro) on the octapeptide nh2-avlq downward arrowsgfr-cooh was investigated using molecular mechanics and quantum mechanics simulations based on the experimental structure of the proteinase. it has been observed that the catalytic dyad (his-41/cys-145) site between domains i and ii attract ...200515691506
1h, 13c, and 15n resonance assignments of the n-terminal domain of the sars cov nucleocapsid protein. 200515692748
sars-cov protease inhibitors design using virtual screening method from natural products libraries.two natural products databases, the marine natural products database (mnpd) and the traditional chinese medicines database (tcmd), were used to find novel structures of potent sars-cov protease inhibitors through virtual screening. before the procedure, the databases were filtered by lipinski's rof and xu's extension rules. the results were analyzed by statistic methods to eliminate the bias in target-based database screening toward higher molecular weight compounds for enhancing the hit rate. e ...200515693056
cross-host evolution of severe acute respiratory syndrome coronavirus in palm civet and human.the genomic sequences of severe acute respiratory syndrome coronaviruses from human and palm civet of the 2003/2004 outbreak in the city of guangzhou, china, were nearly identical. phylogenetic analysis suggested an independent viral invasion from animal to human in this new episode. combining all existing data but excluding singletons, we identified 202 single-nucleotide variations. among them, 17 are polymorphic in palm civets only. the ratio of nonsynonymous/synonymous nucleotide substitution ...200515695582
detection of severe acute respiratory syndrome coronavirus rna in plasma during the course of infection.we examined severe acute respiratory syndrome-associated coronavirus (sars-cov) rna in plasma of 32 patients (probable sars cases) by a quantitative real-time reverse transcription-pcr assay and reported that the highest detection rate, 75%, was found between day 5 and day 7 of illness, followed by rates of 64, 50, and 38% found between day 8 and day 11, day 2 and day 4, and day 12 and day 16, respectively. analysis of sequential sars-cov load in plasma from six cases revealed different patterns ...200515695719
novel immunofluorescence assay using recombinant nucleocapsid-spike fusion protein as antigen to detect antibodies against severe acute respiratory syndrome coronavirus.severe acute respiratory syndrome (sars) is caused by a novel and highly infectious virus named sars coronavirus (sars-cov). among the serological tests currently available for the detection of sars-cov, a whole-virus-based immunofluorescence assay (ifa) was considered one of the most sensitive assays and served as a "gold standard" during the sars epidemic in singapore in 2003. however, the need to manipulate live sars-cov in the traditional ifa limits its wide application due to the requiremen ...200515699428
using cellular automata to generate image representation for biological sequences.a novel approach to visualize biological sequences is developed based on cellular automata (wolfram, s. nature 1984, 311, 419-424), a set of discrete dynamical systems in which space and time are discrete. by transforming the symbolic sequence codes into the digital codes, and using some optimal space-time evolvement rules of cellular automata, a biological sequence can be represented by a unique image, the so-called cellular automata image. many important features, which are originally hidden i ...200515700108
the 3a protein of sars-coronavirus induces apoptosis in vero e6 cells.an outbreak of severe acute respiratory syndrome (sars) occurred in china and the first case emerged in mid november 2002. the etiologic agent of this disease was found to be a previously unknown coronavirus, sars-cov. the detailed pathology of sars-cov infection and the host response to the viral infection are still not known. the 3a gene encodes a non-structural viral protein which is predicted to be a transmembrane protein. in this study, we showed that the 3a protein was localized to the end ...200517282011
investigation of interactions between two monoclonal antibodies and sars virus with a label-free protein array.the investigation of interactions between two kinds of monoclonal antibodies and sars virus with a label-free protein array technique were presented in this paper. the performance consists of three parts: a surface modification for ligand immobilization/surface, a protein array fabrication with an integrated microfluidic system for patterning, packaging and liquid handling, and a protein array reader of imaging ellipsometer. this revealed the technique could be used as an immunoassay for qualita ...200517282435
[receptor-binding ability of fragments 260-600 and 397-796 of sars-associated coronavirus spike protein].to investigate the interaction between the host cell and the truncated s fragments to identify the receptor-binding domain of the spike (s) protein of sars-associated coronavirus (sars-cov).200516415994
[detection of sars coronavirus rna]. 200516416820
virus safety of intravenous immunoglobulin: future challenges.patients with immunodeficiencies or some types of autoimmune diseases are dependent on safe therapy with intravenous immunoglobulins. state-of-the-art manufacturing processes provide a high safety standard by incorporating virus elimination procedures into the manufacturing process. based on their mechanism, these procedures are grouped into three classes: partitioning, inactivation, and removal based on size. because of current socioeconomic and ecological changes, emerging pathogens continue t ...200516391410
anti-hiv, anti-poxvirus, and anti-sars activity of a nontoxic, acidic plant extract from the trifollium species secomet-v/anti-vac suggests that it contains a novel broad-spectrum antiviral.enveloped animal viruses such as human immunodeficiency virus (hiv), hepatitis b virus, hepatitis c virus, human papillomavirus, marburg, and influenza are major public health concerns around the world. the prohibitive cost of antiretroviral (arv) drugs for most hiv-infected patients in sub-saharan africa and the serious side effects in those who have access to arv drugs make a compelling case for the study of complementary and alternative therapies. such therapies should have scientifically pro ...200516387696
history and recent advances in coronavirus discovery.human coronaviruses, first characterized in the 1960s, are responsible for a substantial proportion of upper respiratory tract infections in children. since 2003, at least 5 new human coronaviruses have been identified, including the severe acute respiratory syndrome coronavirus, which caused significant morbidity and mortality. nl63, representing a group of newly identified group i coronaviruses that includes nl and the new haven coronavirus, has been identified worldwide. these viruses are ass ...200516378050
[development of infection pathology in china]. 200516383292
clinical manifestations and inflammatory cytokine responses in patients with severe acute respiratory syndrome.severe acute respiratory syndrome (sars) is a highly transmissible disease with significant morbidity and mortality. death from sars is most often due to rapidly progressive respiratory compromise (acute respiratory distress syndrome, ards) and subsequent multi-organ dysfunction. however, the mechanisms evoking respiratory distress and a fulminant systemic response remain unclear. in order to elucidate the pathogenic mechanisms of sars, we analyzed clinical manifestations and levels of serum cyt ...200516385373
[sars coronavirus].sars is a new type of infectious pneumonia that emerged in china in november 2002. it spread around the world through an outbreak in hong kong in march 2003. patients were reported in 29 countries, and around 800 people died, although a majority of these cases were in asian countries. within a month after the virus that causes sars was isolated, its entire genome sequence became available. the result is the acknowledgement of a novel coronavirus, now called sars coronavirus (scov), that is diffe ...200516363682
a practical protocol for carbohydrate microarrays.we have established a high-throughput biochip platform for constructing carbohydrate microarrays. using this technology, carbohydrate-containing macromolecules of diverse structures, including polysaccharides, natural glycoconjugates, and mono- and oligosaccharides coupled to carrier molecules, can be stably immobilized on a glass chip without chemical modification. here, we describe a practical protocol for this technology. we hope that anyone who has access to a standard cdna microarray facili ...200516350957
coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus.coronaviruses are a family of enveloped, single-stranded, positive-strand rna viruses classified within the nidovirales order. this coronavirus family consists of pathogens of many animal species and of humans, including the recently isolated severe acute respiratory syndrome coronavirus (sars-cov). this review is divided into two main parts; the first concerns the animal coronaviruses and their pathogenesis, with an emphasis on the functions of individual viral genes, and the second discusses t ...200516339739
coronavirus pathogenesis and the emerging pathogen severe acute respiratory syndrome coronavirus.coronaviruses are a family of enveloped, single-stranded, positive-strand rna viruses classified within the nidovirales order. this coronavirus family consists of pathogens of many animal species and of humans, including the recently isolated severe acute respiratory syndrome coronavirus (sars-cov). this review is divided into two main parts; the first concerns the animal coronaviruses and their pathogenesis, with an emphasis on the functions of individual viral genes, and the second discusses t ...200516339739
longitudinal analysis of severe acute respiratory syndrome (sars) coronavirus-specific antibody in sars patients.the serum antibodies to severe acute respiratory syndrome (sars) coronavirus of 18 sars patients were checked at 1 month and every 3 months after disease onset. all of them except one, who missed blood sampling at 1 month, tested positive for the immunoglobulin g (igg) antibody at 1 month. fifteen out of 17 tested positive for the igm antibody at 1 month. the serum igm antibody of most patients became undetectable within 6 months after the onset of sars. the igg antibody of all 17 patients, whos ...200516339072
subcellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid protein.the coronavirus nucleocapsid (n) protein is a viral rna-binding protein with multiple functions in terms of virus replication and modulating cell signalling pathways. n protein is composed of three distinct regions containing rna-binding motif(s), and appropriate signals for modulating cell signalling. the subcellular localization of severe acute respiratory syndrome coronavirus (sars-cov) n protein was studied. in infected cells, sars-cov n protein localized exclusively to the cytoplasm. in con ...200516298975
handwashing practice and the use of personal protective equipment among medical students after the sars epidemic in hong kong.hand hygiene is an important element of infection control. we conducted 2 surveys on hand hygiene practices and use of personal protective equipment among medical students during and after the outbreak of severe acute respiratory syndrome (sars) to study its impact on their personal hygiene practice when they contacted patients.200516330306
study on substrate specificity at subsites for severe acute respiratory syndrome coronavirus 3cl protease.autocleavage assay and peptide-based cleavage assay were used to study the substrate specificity of 3cl protease from the severe acute respiratory syndrome coronavirus. it was found that the recognition between the enzyme and its substrates involved many positions in the substrate, at least including residues from p4 to p2'. the deletion of either p4 or p2' residue in the substrate would decrease its cleavage efficiency dramatically. in contrast to the previous suggestion that only small residue ...200516331324
concentration and detection of sars coronavirus in sewage from xiao tang shan hospital and the 309th hospital of the chinese people's liberation army.a worldwide outbreak of severe acute respiratory syndrome (sars) had been reported. over 8439 sars cases and 812 sars-related deaths were reported to the world health organization from 32 countries around the world up to 5 july 2003. the mechanism of transmission of sars-cov has been limited only to close contacts with patients. attention was focused on possible transmission by the sewage system because laboratory studies showed that patients excreted coronavirus rna in their stools in amoy gard ...200516312970
neutralizing antibody response and sars severity.using the taiwan nationwide laboratory-confirmed severe acute respiratory syndrome (sars) database, we analyzed neutralizing antibody in relation to clinical outcomes. with a linear mixed model, neutralizing antibody titer was shown to peak between week 5 and week 8 after onset and to decline thereafter, with a half-life of 6.4 weeks. patients with a longer illness showed a lower neutralizing antibody response than patients with a shorter illness duration (p = 0.008). when early responders were ...200516318725
respiratory infections during sars outbreak, hong kong, 2003.the effect of community hygienic measures during the outbreak of severe acute respiratory syndrome in hong kong was studied by comparing the proportion of positive specimens of various respiratory viruses in 2003 with those from 1998 to 2002. community hygienic measures significantly reduced the incidence of various respiratory viral infections.200516318726
social factors associated with aids and sars.we conducted a survey of 928 new york city area residents to assess knowledge and worry about aids and sars. specific sociodemographic groups of persons were more likely to be less informed and more worried about contracting the diseases.200516318735
immunopathogenesis of coronavirus infections: implications for sars.at the end of 2002, the first cases of severe acute respiratory syndrome (sars) were reported, and in the following year, sars resulted in considerable mortality and morbidity worldwide. sars is caused by a novel species of coronavirus (sars-cov) and is the most severe coronavirus-mediated human disease that has been described so far. on the basis of similarities with other coronavirus infections, sars might, in part, be immune mediated. as discussed in this review, studies of animals that are i ...200516322745
update on community-acquired pneumonia. new pathogens and new concepts in treatment.recent years have witnessed the emergence of new pathogens linked to community-acquired pneumonia (cap) along with new antibiotics designed to combat them. in this article, dr mandell presents an expert's view on these developments in the context of treatment guidelines for cap. he also explores monotherapy versus combination therapy, the efficacy of rapid initiation of treatment, and short-course therapy as a hedge against antimicrobial resistance.200516296262
preparation and development of equine hyperimmune globulin f(ab')2 against severe acute respiratory syndrome coronavirus.the resurgence of severe acute respiratory syndrome (sars) is still a threat because the causative agent remaining in animal reservoirs is not fully understood, and sporadic cases continue to be reported. developing high titers of anti-sars hyperimmune globulin to provide an alternative pathway for emergent future prevention and treatment of sars.200516297347
sars: understanding the virus and development of rational therapy.in late 2002 a new disease, severe atypical respiratory syndrome (sars), emerged in china. a hitherto unknown animal coronavirus (cov) that had crossed the species barrier through close contact of humans with infected animals was identified as the etiological agent. it rapidly adapted to the new host and not only became readily transmissible between humans but also more pathogenic. air travel spread it rapidly around the world and ultimately the virus infected 8096 people and caused 774 deaths i ...200516305493
the papain-like protease of severe acute respiratory syndrome coronavirus has deubiquitinating activity.replication of the genomic rna of severe acute respiratory syndrome coronavirus (sars-cov) is mediated by replicase polyproteins that are processed by two viral proteases, papain-like protease (plpro) and 3c-like protease (3clpro). previously, we showed that sars-cov plpro processes the replicase polyprotein at three conserved cleavage sites. here, we report the identification and characterization of a 316-amino-acid catalytic core domain of plpro that can efficiently cleave replicase substrates ...200516306590
the papain-like protease from the severe acute respiratory syndrome coronavirus is a deubiquitinating enzyme.the severe acute respiratory syndrome coronavirus papain-like protease (sars-cov plpro) is involved in the processing of the viral polyprotein and, thereby, contributes to the biogenesis of the virus replication complex. structural bioinformatics has revealed a relationship for the sars-cov plpro to herpesvirus-associated ubiquitin-specific protease (hausp), a ubiquitin-specific protease, indicating potential deubiquitinating activity in addition to its function in polyprotein processing (t. sul ...200516306591
severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs.severe acute respiratory syndrome coronavirus (sars-cov) emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and in vitro models of the lung are needed to elucidate cellular targets and the consequences of viral infection. the sars-cov receptor, human angiotensin 1-converting enzyme 2 (hace2), was detected in ciliated airway epithelial cells of human airway tissues derived from nasal or tracheobronchial regions, suggesting that sars-cov may infect the pro ...200516306622
severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs.severe acute respiratory syndrome coronavirus (sars-cov) emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and in vitro models of the lung are needed to elucidate cellular targets and the consequences of viral infection. the sars-cov receptor, human angiotensin 1-converting enzyme 2 (hace2), was detected in ciliated airway epithelial cells of human airway tissues derived from nasal or tracheobronchial regions, suggesting that sars-cov may infect the pro ...200516306622
surface ultrastructure of sars coronavirus revealed by atomic force microscopy.atomic force microscopy has been used to probe the surface nanostructures of severe acute respiratory syndrome coronavirus (sars-cov). single crown-like virion was directly visualized and quantitative measurements of the dimensions for the structural proteins were provided. a corona of large, distinctive spikes in the envelope was measured after treatment with hydroxyoctanoic acid. high-resolution images revealed that the surface of each single sars-cov was surrounded with at least 15 spherical ...200516309462
adenovirus-mediated expression of the c-terminal domain of sars-cov spike protein is sufficient to induce apoptosis in vero e6 cells.the pro-apoptotic properties of severe acute respiratory syndrome coronavirus (sars-cov) structural proteins were studied in vitro. by monitoring apoptosis indicators including chromatin condensation, cellular dna fragmentation and cell membrane asymmetry, we demonstrated that the adenovirus-mediated over-expression of sars-cov spike (s) protein and its c-terminal domain (s2) induce apoptosis in vero e6 cells in a time- and dosage-dependent manner, whereas the expression of its n-terminal domain ...200516310778
the putative protein 6 of the severe acute respiratory syndrome-associated coronavirus: expression and functional characterization.the sars-cov open reading frame 6 (orf6) is transcribed into mrna6 and encodes a putative 7.5 kda accessory protein, sars 6, with unknown function. in this study, we have confirmed the sars 6 protein expression in lung and intestine tissues of the sars patients and in sars-cov infected vero e6 cells by immunohistochemistry. further studies by immunoblot and confocal microscopy analyses revealed the expression and the endoplasmic reticulum (er) localization of the recombinant sars 6 protein in ma ...200516310783
cathepsin enzyme provides clue to sars infection. 200516243257
severe acute respiratory syndrome (sars) in neonates and children.severe acute respiratory syndrome (sars) runs a more benign course in children during the acute phase. infants born to mothers with the disease did not acquire the infection through vertical transmission. the treatment strategy for children with sars has not been standardised and is based on adult experience. thus far, no deaths have been reported in the paediatric age group. exercise impairment and residual radiological abnormalities were present six months after diagnosis. it is important to a ...200516244207
policy conflicts: gene patents and health care in canada.several recent gene patent controversies have energized and refocused the human gene patent debate in canada. these include the use of the myriad test for breast cancer by the provinces, patenting of the severe acute respiratory syndrome virus and a recent supreme court decision rejecting the patenting of 'higher life forms'. these cases place the emerging policy conflicts between the innovation and commercialization agenda of the government and the desire to provide equitable access to health c ...200516244476
scalable transcriptional analysis routine--multiplexed quantitative real-time polymerase chain reaction platform for gene expression analysis and molecular diagnostics.we report the development of a new technology for simultaneous quantitative detection of multiple targets in a single sample. scalable transcriptional analysis routine (star) represents a novel integration of reverse transcriptase-polymerase chain reaction and capillary electrophoresis that allows detection of dozens of gene transcripts in a multiplexed format using amplicon size as an identifier for each target. star demonstrated similar or better sensitivity and precision compared to two commo ...200516237214
genomic classification using an information-based similarity index: application to the sars coronavirus.measures of genetic distance based on alignment methods are confined to studying sequences that are conserved and identifiable in all organisms under study. a number of alignment-free techniques based on either statistical linguistics or information theory have been developed to overcome the limitations of alignment methods. we present a novel alignment-free approach to measuring the similarity among genetic sequences that incorporates elements from both word rank order-frequency statistics and ...200516241900
ph-dependent conformational flexibility of the sars-cov main proteinase (m(pro)) dimer: molecular dynamics simulations and multiple x-ray structure analyses.the sars coronavirus main proteinase (m(pro)) is a key enzyme in the processing of the viral polyproteins and thus an attractive target for the discovery of drugs directed against sars. the enzyme has been shown by x-ray crystallography to undergo significant ph-dependent conformational changes. here, we assess the conformational flexibility of the m(pro) by analysis of multiple crystal structures (including two new crystal forms) and by molecular dynamics (md) calculations. the md simulations t ...200516242152
[detection of sars-coronavirus in both human and animals by rt-pcr].to find the most suitable rt-pcr detection method for sars-coronavirus (sars-cov) detection in both human and animals, and to study the source of sars virus by investigating the condition of virus carried by wild, domestic animals and animals sold in market.200516229261
[detection of sars-coronavirus in both human and animals by rt-pcr].to find the most suitable rt-pcr detection method for sars-coronavirus (sars-cov) detection in both human and animals, and to study the source of sars virus by investigating the condition of virus carried by wild, domestic animals and animals sold in market.200516229261
[modified molecular beacon-based dual real-time pcr for detection of sars virus and its application].to develop the modified molecular beacon-based dual fluorescent pcr assays for detection of sars virus. the assay was applied to the early clinic diagnosis & animal tracking.200516229262
a simple and rapid approach for screening of sars-coronavirus genotypes: an evaluation study.the severe acute respiratory syndrome (sars) was a newly emerged infectious disease which caused a global epidemic in 2002-2003. sequence analysis of sars-coronavirus isolates revealed that specific genotypes predominated at different periods of the epidemic. this information can be used as a footprint for tracing the epidemiology of infections and monitor viral evolution. however, direct sequencing analysis of a large number of clinical samples is cumbersome and time consuming. we present here ...200516229749
molecular epidemiology of sars-associated coronavirus, beijing.single nucleotide variations (snvs) at 5 loci (17564, 21721, 22222, 23823, and 27827) were used to define the molecular epidemiologic characteristics of severe acute respiratory syndrome-associated coronavirus (sars-cov) from beijing patients. five fragments targeted at the snv loci were amplified directly from clinical samples by using reverse transcription-polymerase chain reaction (rt-pcr), before sequencing the amplified products. analyses of 45 sequences obtained from 29 patients showed tha ...200516229772
binding interaction of sars coronavirus 3cl(pro) protease with vacuolar-h+ atpase g1 subunit.the pathogenesis of severe acute respiratory syndrome coronavirus (sars-cov) is an important issue for treatment and prevention of sars. recently, sars-cov 3cl(pro) protease has been implied to be possible relevance to sars-cov pathogenesis. in this study, we intended to identify potential 3cl(pro)-interacting cellular protein(s) using the phage-displayed human lung cdna library. the vacuolar-h+ atpase (v-atpase) g1 subunit that contained a 3cl(pro) cleavage site-like motif was identified as a 3 ...200516226257
the nsp2 replicase proteins of murine hepatitis virus and severe acute respiratory syndrome coronavirus are dispensable for viral replication.the positive-stranded rna genome of the coronaviruses is translated from orf1 to yield polyproteins that are proteolytically processed into intermediate and mature nonstructural proteins (nsps). murine hepatitis virus (mhv) and severe acute respiratory syndrome coronavirus (sars-cov) polyproteins incorporate 16 protein domains (nsps), with nsp1 and nsp2 being the most variable among the coronaviruses and having no experimentally confirmed or predicted functions in replication. to determine if ns ...200516227261
severe acute respiratory syndrome coronavirus fails to activate cytokine-mediated innate immune responses in cultured human monocyte-derived dendritic cells.activation of host innate immune responses was studied in severe acute respiratory syndrome coronavirus (scv)-infected human a549 lung epithelial cells, macrophages, and dendritic cells (dcs). in all cell types, scv-specific subgenomic mrnas were seen, whereas no expression of scv proteins was found. no induction of cytokine genes (alpha interferon [ifn-alpha], ifn-beta, interleukin-28a/b [il-28a/b], il-29, tumor necrosis factor alpha, ccl5, or cxcl10) or ifn-alpha/beta-induced mxa gene was seen ...200516227300
insights into sars-cov transcription and replication from the structure of the nsp7-nsp8 hexadecamer.coronavirus replication and transcription machinery involves multiple virus-encoded nonstructural proteins (nsp). we report the crystal structure of the hexadecameric nsp7-nsp8 supercomplex from the severe acute respiratory syndrome coronavirus at 2.4-angstroms resolution. nsp8 has a novel 'golf-club' fold with two conformations. the supercomplex is a unique hollow, cylinder-like structure assembled from eight copies of nsp8 and held tightly together by eight copies of nsp7. with an internal dia ...200516228002
[theoretical prediction of the antigenic epitopes of severe acute respiratory syndrome (sars-cov) proteins and evaluation of their diagnostic value].analysis of the sequence of a number of proteins of the virus causing severe acute respiratory syndrome (sars-cov) was used to theoretically predict antigenic epitopes. nine sequences, encoding for the predicted epitopes in spike, nucleocapside, and membranous sars cov proteins, were synthesized by polymyrase chain reaction and cloned into the pgex 4-t2 vector. the resultant plasmids were expressed in the e. coil cells and purified by glutathione-sepharose affinity chromatography.200516250594
design and synthesis of peptidomimetic severe acute respiratory syndrome chymotrypsin-like protease inhibitors.design, synthesis, and biological evaluation of peptidomimetic severe acute respiratory syndrome chymotrypsin-like protease (sars-3clpro) inhibitors for severe acute respiratory syndrome coronavirus (sars-cov) are described. these inhibitors exhibited antiviral activity against sars-cov in infected cells in the micromolar range. an x-ray crystal structure of our lead inhibitor (4) bound to sars-3clpro provided important drug-design templates for the design of small-molecule inhibitors.200516250632
a new lead for nonpeptidic active-site-directed inhibitors of the severe acute respiratory syndrome coronavirus main protease discovered by a combination of screening and docking methods.the coronavirus main protease, m(pro), is considered to be a major target for drugs suitable for combating coronavirus infections including severe acute respiratory syndrome (sars). an hplc-based screening of electrophilic compounds that was performed to identify potential m(pro) inhibitors revealed etacrynic acid tert-butylamide (6a) as an effective nonpeptidic inhibitor. docking studies suggested a binding mode in which the phenyl ring acts as a spacer bridging the inhibitor's activated double ...200516250642
neurological manifestations in severe acute respiratory syndrome.during the worldwide outbreak of severe acute respiratory syndrome (sars) in 2002-2003, there were 664 probable sars patients reported in taiwan. sars patients usually present with symptoms related to the respiratory system while neurological manifestations have rarely been described. there were three patients who developed axonopathic polyneuropathy 3-4 weeks after onset of sars; their clinical condition and electrophysiological studies revealed obvious improvement at follow-up. two sars patien ...200516252612
virology. what links bats to emerging infectious diseases? 200516254175
assembly of severe acute respiratory syndrome coronavirus rna packaging signal into virus-like particles is nucleocapsid dependent.the severe acute respiratory syndrome coronavirus (sars-cov) was recently identified as the etiology of sars. the virus particle consists of four structural proteins: spike (s), small envelope (e), membrane (m), and nucleocapsid (n). recognition of a specific sequence, termed the packaging signal (ps), by a virus n protein is often the first step in the assembly of viral rna, but the molecular mechanisms involved in the assembly of sars-cov rna are not clear. in this study, vero e6 cells were co ...200516254320
modulation of the immune response to the severe acute respiratory syndrome spike glycoprotein by gene-based and inactivated virus immunization.although the initial isolates of the severe acute respiratory syndrome (sars) coronavirus (cov) are sensitive to neutralization by antibodies through their spike (s) glycoprotein, variants of s have since been identified that are resistant to such inhibition. optimal vaccine strategies would therefore make use of additional determinants of immune recognition, either through cellular or expanded, cross-reactive humoral immunity. here, the cellular and humoral immune responses elicited by differen ...200516254327
real-time nasba detection of sars-associated coronavirus and comparison with real-time reverse transcription-pcr.severe acute respiratory syndrome (sars) exhibits a high mortality rate and the potential for rapid epidemic spread. additionally, it has a poorly defined clinical presentation, and no known treatment or prevention methods. collectively, these factors underscore the need for early diagnosis. molecular tests have been developed to detect sars coronavirus (sars-cov) rna using real time reverse transcription polymerase chain reaction (rt-pcr) with varying levels of sensitivity. however, rna amplifi ...200516254971
[expression of sars coronavirus nucleocapsid protein and construction of its dna vaccine].to express the nucleocapsid (n) protein of sars coronavirus (sars-cov) in e. coli and construct its dna vaccine.200516256037
molecular diagnosis of severe acute respiratory syndrome: the state of the art.severe acute respiratory syndrome (sars) first appeared in guangdong province, china, in november 2002. although virus isolation and serology were useful early in the sars outbreak for diagnosing new cases, these tests are not generally useful because virus culture requires a bsl-3 laboratory and seroconversion is often delayed until 2 to 3 weeks after infection. the first qualitative reverse transcriptase-polymerase chain reaction tests for sars-coronavirus (cov) were sensitive and capable of d ...200516258152
[preventive and therapeutic effects of recombinant ifn-alpha2b nasal spray on sars-cov infection in macaca mulata].to study the preventive and therapeutic effects of recombinant ifn-alpha2b for nasal spray on sars-cov infection in macaca mulata (rhesus monkey).200516261198
[a field trial for evaluating the safety of recombinant human interferon alpha-2b for nasal spray].to evaluate the safety of recombinant human interferon alpha-2b for nasal spray for the prevention of sars and other upper respiratory viral infections.200516261199
[a field trial of recombinant human interferon alpha-2b for nasal spray to prevent sars and other respiratory viral infections].to study the preventive effect of recombinant human interferon alpha-2b for nasal spray against sars and other common respiratory viral infections by serum-epidemiological method.200516261200
[cloning of ace-2 gene encoding the functional receptor for the sars coronavirus and its expression in eukaryotic cells].angiotensin-converting enzyme 2 (ace-2) has been identified as a functional receptor of severe acute respiratory syndrome coronavirus (sars-cov), so its gene was cloned and eukaryotic expressed for further insight into mechanisms in sars-cov entry and pathogenesis, as well as development of a safe and reliable neutralization assay for sars-cov.200516261211
[cloning, purification, and antigenic characterization of three recombinant fragments derived from sars-cov s1 domain].the present study aimed to clone and express three fragments of genomic rna derived from sars associated coronavirus (sars-cov) s1 domain and to study its immunogenicity.200516261215
cross-reactivity of antibody against sars-coronavirus nucleocapsid protein with il-11.infection of sars-associated coronavirus (sars-cov) induced a strong anti-nucleocapsid (anti-n) antibody response. however, the pathophysiological significance of the anti-n antibodies in sars pathogenesis is largely unknown. to profile the anti-n antibodies, a phage-displayed scfv library was prepared from mice immunized with heat-inactivated sars-cov-infected vero e6 cell lysate. specific anti-n scfvs were isolated by panning against a recombinant nucleocapsid protein and reactivity was confir ...200516263078
structural insights into sars coronavirus proteins.the sars coronavirus was identified as the pathogen of a global outbreak of sars (severe acute respiratory syndrome) in 2003. its large rna genome encodes four structural proteins, sixteen non-structural proteins and eight accessory proteins. the availability of structures of sars coronavirus macromolecules has enabled the elucidation of their important functions, such as mediating the fusion of viral and host cellular membranes, and in replication and transcription. in particular, the spike pro ...200516263266
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