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group 2 coronaviruses prevent immediate early interferon induction by protection of viral rna from host cell recognition.many viruses encode antagonists to prevent interferon (ifn) induction. infection of fibroblasts with the murine hepatitis coronavirus (mhv) and sars-coronavirus (sars-cov) did not result in nuclear translocation of interferon-regulatory factor 3 (irf3), a key transcription factor involved in ifn induction, and induction of ifn mrna transcription. furthermore, mhv and sars-cov infection could not prevent ifn induction by poly (i:c) or sendai virus, suggesting that these covs do not inactivate irf ...200717316733
severe acute respiratory syndrome--retrospect and lessons of 2004 outbreak in china.to summarize lessons learned from an outbreak of severe acute respiratory syndrome (sars) in china during the spring of 2004.200617319269
challenges faced by hospital healthcare workers in using a syndrome-based surveillance system during the 2003 outbreak of severe acute respiratory syndrome in taiwan.because the severe acute respiratory syndrome (sars) outbreak in taiwan in 2003 was worsened by hospital infections, we analyzed 229 questionnaires (84.8% of 270 sent) completed by surveyed healthcare workers who cared for patients with sars in 3 types of hospitals, to identify surveillance problems. atypical clinical presentation was the most often reported problem, regardless of hospital type, which strongly indicates that more timely syndromic surveillance was needed.200717326030
review of bats and sars.bats have been identified as a natural reservoir for an increasing number of emerging zoonotic viruses, including henipaviruses and variants of rabies viruses. recently, we and another group independently identified several horseshoe bat species (genus rhinolophus) as the reservoir host for a large number of viruses that have a close genetic relationship with the coronavirus associated with severe acute respiratory syndrome (sars). our current research focused on the identification of the reserv ...200617326933
long-term psychological and occupational effects of providing hospital healthcare during sars outbreak.healthcare workers (hcws) found the 2003 outbreak of severe acute respiratory syndrome (sars) to be stressful, but the long-term impact is not known. from 13 to 26 months after the sars outbreak, 769 hcws at 9 toronto hospitals that treated sars patients and 4 hamilton hospitals that did not treat sars patients completed a survey of several adverse outcomes. toronto hcws reported significantly higher levels of burnout (p = 0.019), psychological distress (p<0.001), and posttraumatic stress (p<0.0 ...200617326946
sars molecular epidemiology: a chinese fairy tale of controlling an emerging zoonotic disease in the genomics era.severe acute respiratory syndrome (sars) was the first natural disaster that challenged the chinese people at the beginning of the twenty-first century. it was caused by a novel animal coronavirus, never recognized or characterized before. this sars coronavirus (sars-cov) exploited opportunities provided by 'wet markets' in southern china to adapt to the palm civet and human. under the positive selection pressure of human host, certain mutated lineages of the virus became readily transmissible b ...200717327210
heterologous viral rna export elements improve expression of severe acute respiratory syndrome (sars) coronavirus spike protein and protective efficacy of dna vaccines against sars.the sars-cov spike glycoprotein (s) is the main target of the protective immune response in humans and animal models of sars. here, we demonstrated that efficient expression of s from the wild-type spike gene in cultured cells required the use of improved plasmid vectors containing donor and acceptor splice sites, as well as heterologous viral rna export elements, such as the cte of mazon-pfizer monkey virus or the pre of woodchuck hepatitis virus (wpre). the presence of both splice sites and wp ...200717331558
heterologous viral rna export elements improve expression of severe acute respiratory syndrome (sars) coronavirus spike protein and protective efficacy of dna vaccines against sars.the sars-cov spike glycoprotein (s) is the main target of the protective immune response in humans and animal models of sars. here, we demonstrated that efficient expression of s from the wild-type spike gene in cultured cells required the use of improved plasmid vectors containing donor and acceptor splice sites, as well as heterologous viral rna export elements, such as the cte of mazon-pfizer monkey virus or the pre of woodchuck hepatitis virus (wpre). the presence of both splice sites and wp ...200717331558
[further analysis and study based on a visualized method for sars rna sequences].this paper proposed a new kind of visualized method of genome. using cellular automation theory, the visual method transfers one-dimensional rna sequence into two-demension visual image. applying this method to sars rna sequence analysis, the characteristic of sars-cov differing from non-sars is discovered. this paper extracts characteristic genome fragment, visualize them, and study them with some pattern recognition method such as pca and svm. the result shows that the characteristic of sars-c ...200717333886
synthesis and biological evaluation of nucleoside analogues having 6-chloropurine as anti-sars-cov agents.nucleoside analogues that have 6-chloropurine as the nucleobase were synthesized and evaluated for anti-sars-cov activity by plaque reduction and yield reduction assays in order to develop novel anti-sars-cov agents. among these analogues, two compounds, namely, 1 and 11, exhibited promising anti-sars-cov activity that was comparable to those of mizoribine and ribavirin, which are known anti-sars-cov agents. moreover, we observed several sar trends such as the antiviral effects of the 6-chloropu ...200717336519
severe acute respiratory syndrome coronavirus accessory protein 6 is a virion-associated protein and is released from 6 protein-expressing cells.analysis of severe acute respiratory syndrome coronavirus (scov) by either sucrose gradient equilibrium centrifugation or a virus capture assay using an anti-scov s protein antibody demonstrated that the scov 6 protein, which is one of the accessory proteins of scov, was incorporated into virus particles. coexpression of the scov s, m, e, and 6 proteins was sufficient for incorporation of the 6 protein into virus-like particles. cells transfected with plasmid expressing the 6 protein released sc ...200717344286
recent developments in the virology and antiviral research of severe acute respiratory syndrome coronavirus.this article summarizes the significant developments and new discoveries in both the virology and antiviral research associated with the severe acute respiratory syndrome coronavirus (sars cov) that were reported in 2005 and 2006. areas reviewed include genomic studies and the identification of bat-sars cov, spike protein and host cell entry, nucleocapsid protein, accessory proteins, non-structural proteins of the replicase complex, viral proteases and their inhibitors, and clinical treatment of ...200717346209
antiviral strategies against human coronaviruses.since the mid 60's the human coronaviruses (hcov), represented by hcov-oc43 and hcov-229e, were generally considered relatively harmless viruses. this status changed dramatically with the emergence of sars-cov in 2002/2003. the sars-cov pandemic took 774 lives around the globe and infected more than 8000 people in 29 countries. sars-cov is believed to be of zoonotic origin, transmitted from its natural reservoir in bats through several animal species (e.g., civet cats, raccoon dogs sold for huma ...200717346212
carbohydrate-binding agents: a potential future cornerstone for the chemotherapy of enveloped viruses?carbohydrate-binding agents (cbas) inhibit hiv-1 and it is proposed that therapy with such agents may have important implications for the future of anti-hiv therapy. examples of cbas include the procaryotic cyanovirin-n (cv-n), plant lectins such as hha, gna, npa, ca and uda, the monoclonal antibody 2g12 directed against a glycan-containing epitope on hiv envelope gp120, and the mannose-specific non-peptidic antibiotic pradimicin a, which inhibits the entry of hiv-1 into its target cells. cbas p ...200717354647
characterisation of cyclin d1 down-regulation in coronavirus infected cells.the positive strand rna coronavirus, infectious bronchitis virus (ibv), induces a g2/m phase arrest and reduction in the g1 and g1/s phase transition regulator cyclin d1. quantitative real-time rt-pcr and western blot analysis demonstrated that cyclin d1 was reduced post-transcriptionally within infected cells independently of the cell-cycle stage at the time of infection. confocal microscopy revealed that cyclin d1 decreased in ibv-infected cells as infection progressed and inhibition studies i ...200717359980
a dominant antigenic epitope on sars-cov spike protein identified by an avian single-chain variable fragment (scfv)-expressing phage.severe acute respiratory syndrome (sars) is a newly emergent human disease, which requires rapid diagnosis and effective therapy. among antibody sources, immunoglobulin y (igy) is the major antibody found in chicken eggs and can be used as an alternative to mammalian antibodies normally used in research and immunotherapy. in this study, phage-expressing chicken monoclonal scfv antibody was chosen and characterized with phage display antibody technology. truncated fragments of sars-cov spike prot ...200717360045
bioactive coumarins from boenninghausenia sessilicarpa.bioassay guided fractionation of boenninghausenia sessilicarpa (rutaceae) resulted in the isolation of a new dimeric coumarin glucoside 9'-methoxyl rutarensin (1) and a cytotoxic compound rutamarin (4), as well as an antivirus component leptodactylone (8), together with six known coumarins. their structures were elucidated by 1d- and 2d nmr spectroscopy and esi-ms analyses, respectively. rutamarin (4) showed significant inhibitory activities against a-549, bel-7402, hepg-2 and hct-8 tumour cell ...200717365191
positive rate of serum sars-cov immunoglobulin g antibody among healthcare workers.we evaluated the positive rate for severe acute respiratory syndrome (sars) -cov igg antibody among taiwanese healthcare workers (hcws). a retrospective cohort research was performed in 2 teaching hospitals and 2 non-teaching hospitals. sars-cov igg antibodies were checked and a structured questionnaire was prepared. a total of 2512 sets of questionnaires was distributed with a return rate of 87.5% (2197 sets). 882 of the respondents had contact with sars patients. among 2197 subjects studied, a ...200717366033
computational simulation of interactions between sars coronavirus spike mutants and host species-specific receptors.as a critical adaptive mechanism, amino acid replacements on the severe acute respiratory syndrome coronavirus (sars-cov) spike protein could alter the receptor-binding specificity of this envelope glycoprotein and in turn lead to the emergence or reemergence of this viral zoonosis. based on the x-ray structures of sars-cov spike receptor-binding domain (rbd) in complex with its functional receptor (angiotensin-converting enzyme 2, ace2), we perform computational simulations of interactions betw ...200717368104
[emerging viral infectious diseases and pregnancy].various emerging diseases have been reported in recent years as epidemiologic monitoring improves. these infections can compromise the pregnancy, through adverse maternal outcomes, as well as the outcome for the child following transmission. we report a review of the literature concerning the most quoted emergent virus (h5n1, sars coronavirus, chikungunya, west nile virus), and relate the perinatal management.200717368948
human immunopathogenesis of severe acute respiratory syndrome (sars).progressive immune-associated injury is a hallmark of severe acute respiratory syndrome (sars). viral evasion of innate immunity, hypercytokinemia and systemic immunopathology in the sars coronavirus (sars cov) infected host have been suggested as possible mechanisms for the cause of severe pathology and morbidity in sars patients. the molecular and cellular basis for how sars cov impacts the host immune system resulting in severe sars, however, has not been elucidated. the variable clinical cou ...200817374415
computational characterization and design of sars coronavirus receptor recognition and antibody neutralization.the sequential determination of crystal structures of the sars coronavirus spike receptor-binding domain (rbd) in complex with its cellular receptor or neutralizing antibody opened a door for the design and development of antiviral competitive inhibitors. based on those complex structures, we conduct computational characterization and design of rbd-mediated receptor recognition and antibody neutralization. the comparisons between computational predictions and experimental evidences validate our ...200717374510
structure of the sars coronavirus nucleocapsid protein rna-binding dimerization domain suggests a mechanism for helical packaging of viral rna.coronavirus nucleocapsid proteins are basic proteins that encapsulate viral genomic rna to form part of the virus structure. the nucleocapsid protein of sars-cov is highly antigenic and associated with several host-cell interactions. our previous studies using nuclear magnetic resonance revealed the domain organization of the sars-cov nucleocapsid protein. rna has been shown to bind to the n-terminal domain (ntd), although recently the c-terminal half of the protein has also been implicated in r ...200717379242
design, synthesis, and evaluation of inhibitors for severe acute respiratory syndrome 3c-like protease based on phthalhydrazide ketones or heteroaromatic esters.the 3c-like protease (3clpro), which controls the severe acute respiratory syndrome (sars) coronavirus replication, has been identified as a potential target for drug design in the treatment of sars. a series of tetrapeptide phthalhydrazide ketones, pyridinyl esters, and their analogs have been designed, synthesized, and evaluated as potential sars 3clpro inhibitors. some pyridinyl esters are identified as very potent inhibitors, with ic50 values in the nanomolar range (50-65 nm). electrospray m ...200717381079
using patient-collected clinical samples and sera to detect and quantify the severe acute respiratory syndrome coronavirus (sars-cov).severe acute respiratory syndrome (sars) caused a large outbreak of pneumonia in beijing, china, in 2003. reverse transcriptase polymerase chain reaction (rt-pcr) was used to detect and quantify sars-cov in 934 sera and self-collected throat washes and fecal samples from 271 patients with laboratory-confirmed sars managed at a single institution.200717386116
new and emerging infectious diseases.new emerging infectious diseases include "severe acute respiratory distress syndrome" (sars) and avian influenza a h5n1. first cases of sars, induced by a new strain of coronavirus, were described in china in 2002 and by may 2003 8360 cases and 764 deaths were reported by the world health organization. the disease can be transmitted from person to person and at the onset is characterized by nonspecific symptoms such as a fever of > 38 degrees c, dry cough, myalgia, and dyspnea. adults can develo ...200717390754
pathology and pathogenesis of severe acute respiratory syndrome.severe acute respiratory syndrome (sars) is an emerging infectious viral disease characterized by severe clinical manifestations of the lower respiratory tract. the pathogenesis of sars is highly complex, with multiple factors leading to severe injury in the lungs and dissemination of the virus to several other organs. the sars coronavirus targets the epithelial cells of the respiratory tract, resulting in diffuse alveolar damage. several organs/cell types may be infected in the course of the il ...200717392154
identification and characterization of dominant helper t-cell epitopes in the nucleocapsid protein of severe acute respiratory syndrome coronavirus.by using a series of overlapping synthetic peptides covering 98% of the amino acid sequence of the nucleocapsid protein (np) of severe acute respiratory syndrome coronavirus (sars-cov), four helper t-cell (th) epitopes (np11, residues 11 to 25; np51, residues 51 to 65; np61, residues 61 to 75; and np111, residues 111 to 125) in c57bl mice (h-2(b)), four (np21, residues 21 to 35; np91, residues 91 to 105; np331, residues 331 to 345; and np351, residues 351 to 365) in c3h mice (h-2(k)), and two (n ...200717392374
evaluating the 3c-like protease activity of sars-coronavirus: recommendations for standardized assays for drug discovery.although the initial outbreaks of the deadly coronavirus that causes severe acute respiratory syndrome (sars-cov) were controlled by public health measures, the development of vaccines and antiviral agents for sars-cov is essential for improving control and treatment of future outbreaks. one potential target for sars-cov antiviral drug development is the 3c-like protease (3clpro). this enzyme is an attractive target since it is essential for viral replication, and since there are now a number of ...200817397958
sars coronavirus replicase proteins in pathogenesis.much progress has been made in understanding the role of structural and accessory proteins in the pathogenesis of severe acute respiratory syndrome coronavirus (sars-cov) infections. the sars epidemic also brought new attention to the proteins translated from orf1a and orf1b of the input genome rna, also known as the replicase/transcriptase gene. evidence for change within the orf1ab coding sequence during the sars epidemic, as well as evidence from studies with other coronaviruses, indicates th ...200817397959
[study on the dynamic prevalence of serum antibody against severe acute respiratory syndrome coronavirus in employees from wild animal market in guangzhou].to investigate the dynamic trend of specific antibody against severe acute respiratory syndrome (sars)-cov in serum collected at various periods among employees in guangzhou xinyuan animal market.200617402195
[study on the dynamic prevalence of serum antibody against severe acute respiratory syndrome coronavirus in employees from wild animal market in guangzhou].to investigate the dynamic trend of specific antibody against severe acute respiratory syndrome (sars)-cov in serum collected at various periods among employees in guangzhou xinyuan animal market.200617402195
knowledge of and attitudes toward severe acute respiratory syndrome among a cohort of dental patients in hong kong following a major local outbreak.to assess the knowledge of and attitudes toward severe acute respiratory syndrome (sars) among patients attending a teaching dental hospital and private dental practices in hong kong during a major local sars outbreak.200717405470
proteomic analysis of up-regulated proteins in human promonocyte cells expressing severe acute respiratory syndrome coronavirus 3c-like protease.the pathogenesis of severe acute respiratory syndrome coronavirus (sars cov) is an important issue for treatment and prevention of sars. previously, sars cov 3c-like protease (3clpro) has been demonstrated to induce apoptosis via the activation of caspase-3 and caspase-9 (lin, c. w., lin, k. h., hsieh, t. h., shiu, s. y. et al., fems immunol. med. microbiol. 2006, 46, 375-380). in this study, proteome analysis of the human promonocyte hl-cz cells expressing sars cov 3clpro was performed using 2- ...200717407183
crystal structure of a monomeric form of severe acute respiratory syndrome coronavirus endonuclease nsp15 suggests a role for hexamerization as an allosteric switch.mature nonstructural protein-15 (nsp15) from the severe acute respiratory syndrome coronavirus (sars-cov) contains a novel uridylate-specific mn2+-dependent endoribonuclease (nendou). structure studies of the full-length form of the obligate hexameric enzyme from two covs, sars-cov and murine hepatitis virus, and its monomeric homologue, xendou from xenopus laevis, combined with mutagenesis studies have implicated several residues in enzymatic activity and the n-terminal domain as the major dete ...200717409150
the sars coronavirus spike glycoprotein is selectively recognized by lung surfactant protein d and activates macrophages.the severe acute respiratory syndrome coronavirus (sars-cov) infects host cells with its surface glycosylated spike-protein (s-protein). here we expressed the sars-cov s-protein to investigate its interactions with innate immune mechanisms in the lung. the purified s-protein was detected as a 210 kda glycosylated protein. it was not secreted in the presence of tunicamycin and was detected as a 130 kda protein in the cell lysate. the purified s-protein bound to vero but not 293t cells and was its ...200717412287
g1 phase cell cycle arrest induced by sars-cov 3a protein via the cyclin d3/prb pathway.sars-cov 3a is a structural protein, mainly localizing to golgi apparatus and co-localizing with sars-cov m in co-transfected cells. here we observed that transient expression of 3a inhibited cell growth and prevented 5-bromodeoxyuridine incorporation, suggesting that 3a deregulated cell cycle progression. cell cycle analysis demonstrated that 3a expression was associated with blockage of cell cycle progression at g1 phase in hek 293, cos-7, and vero cells 24-60 h after transfection. mutation an ...200717413032
community acquired pneumonia in the tropics.due to the exponential growth of international exchange, millions of travelers are exposed to respiratory pathogens in the tropics and may return ill. community-acquired pneumonia is one of the more prevalent infections.200717414123
vaccines to prevent severe acute respiratory syndrome coronavirus-induced disease.an important effort has been performed after the emergence of severe acute respiratory syndrome (sars) epidemic in 2003 to diagnose and prevent virus spreading. several types of vaccines have been developed including inactivated viruses, subunit vaccines, virus-like particles (vlps), dna vaccines, heterologous expression systems, and vaccines derived from sars-cov genome by reverse genetics. this review describes several aspects essential to develop sars-cov vaccines, such as the correlates of p ...200817416434
plant lectins are potent inhibitors of coronaviruses by interfering with two targets in the viral replication cycle.we describe the antiviral activity of plant lectins with specificity for different glycan structures against the severe acute respiratory syndrome coronavirus (sars-cov) and the feline infectious peritonitis virus (fipv) in vitro. the sars-cov emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and fipv infection causes a chronic and often fatal peritonitis in cats. a unique collection of 33 plant lectins with different specificities were evaluated. the p ...200717428553
new structure model for the packaging signal in the genome of group iia coronaviruses.a 190-nucleotide (nt) packaging signal (ps) located in the 3' end of open reading frame 1b in the mouse hepatitis virus, a group iia coronavirus, was previously postulated to direct genome rna packaging. based on phylogenetic data and structure probing, we have identified a 95-nt hairpin within the 190-nt ps domain which is conserved in all group iia coronaviruses but not in the severe acute respiratory syndrome coronavirus (group iib), group i coronaviruses, or group iii coronaviruses. the hair ...200717428856
induction of apoptosis by the severe acute respiratory syndrome coronavirus 7a protein is dependent on its interaction with the bcl-xl protein.the severe acute respiratory syndrome coronavirus (sars-cov) 7a protein, which is not expressed by other known coronaviruses, can induce apoptosis in various cell lines. in this study, we show that the overexpression of bcl-xl, a prosurvival member of the bcl-2 family, blocks 7a-induced apoptosis, suggesting that the mechanism for apoptosis induction by 7a is at the level of or upstream from the bcl-2 family. coimmunoprecipitation experiments showed that 7a interacts with bcl-xl and other prosur ...200717428862
[cell entry mechanism of coronaviruses: implication in their pathogenesis].coronaviruses infect many species of animals, including humans. among them, murine coronavirus, mouse hepatitis virus (mhv) has been well studied as a model of human diseases, such as hepatitis and demyelinating disease. an agent causing severe acute respiratory disease (sars), sars coronavirus (sars-cov), is a newcomer in this genus, however, it is now one of the most studied coronaviruses due to its medical impact. the receptors of those two viruses have been identified and their cell entry me ...200617446665
intracellular localization of the sars coronavirus protein 9b: evidence of active export from the nucleus.open reading frame 9b (orf 9b) encodes a 98 amino acid group-specific protein of severe acute respiratory syndrome (sars) coronavirus (cov). it has no homology with known proteins and its function in sars cov replication has not been determined. the n-terminal part of the 9b protein was used to raise polyclonal antibodies in rabbits, and these antibodies could detect 9b protein in infected cells. we analyzed the sub-cellular localization of recombinant 9b protein using fluorescence microscopy of ...200717448558
peptide reagent design based on physical and chemical properties of amino acid residues.it has tremendous values for both drug discovery and basic research to develop a solid bioinformatical tool for guiding peptide reagent design. based on the physical and chemical properties of amino acids, a new strategy for peptide reagent design, the so-called aabpd (amino acid based-peptide design), is proposed. the peptide samples in a training dataset are described by a series of hmlp (heuristic molecular lipophilicity potential) parameters and other physicochemical properties of amino acid ...200717450553
sars coronavirus and innate immunity.the emergence of the highly pathogenic sars coronavirus (sars-cov) has reignited interest in coronavirus biology and pathogenesis. an emerging theme in coronavirus pathogenesis is that the interaction between specific viral genes and the host immune system, specifically the innate immune system, functions as a key determinant in regulating virulence and disease outcomes. using sars-cov as a model, we will review the current knowledge of the interplay between coronavirus infection and the host in ...200817451827
sars-cov replication and pathogenesis in an in vitro model of the human conducting airway epithelium.sars 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 severe and sudden onset of symptoms, resulting in an atypical pneumonia with dry cough and persistent high fever in cases of severe acute respiratory virus brought to light the importance of coronaviruses as potentially lethal human pathogens and the identificat ...200817451829
sars-cov replication and pathogenesis in an in vitro model of the human conducting airway epithelium.sars 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 severe and sudden onset of symptoms, resulting in an atypical pneumonia with dry cough and persistent high fever in cases of severe acute respiratory virus brought to light the importance of coronaviruses as potentially lethal human pathogens and the identificat ...200817451829
a review of studies on animal reservoirs of the sars coronavirus.in this review, we summarize the researches on animal reservoirs of the sars coronavirus (sars-cov). masked palm civets were suspected as the origin of the sars outbreak in 2003 and was confirmed as the direct origin of sars cases with mild symptom in 2004. sequence analysis of the sars-cov-like virus in masked palm civets indicated that they were highly homologous to human sars-cov with nt identity over 99.6%, indicating the virus has not been circulating in the population of masked palm civets ...200817451830
a review of studies on animal reservoirs of the sars coronavirus.in this review, we summarize the researches on animal reservoirs of the sars coronavirus (sars-cov). masked palm civets were suspected as the origin of the sars outbreak in 2003 and was confirmed as the direct origin of sars cases with mild symptom in 2004. sequence analysis of the sars-cov-like virus in masked palm civets indicated that they were highly homologous to human sars-cov with nt identity over 99.6%, indicating the virus has not been circulating in the population of masked palm civets ...200817451830
sars-cov nucleocapsid protein induced apoptosis of cos-1 mediated by the mitochondrial pathway.to investigate the apoptosis effect of sars coronavirus nucleocapsid protein on cultured cell lines and to explore the possible pathway of apoptosis. pcdna3.1(-)/his-myc vector containing the sars coronavirus nucleocapsid gene (n), matric gene (m), spike gene (s) were transfected into cos-1, huh-7 and hepg2 cells. apoptosis induced by sars coronavirus n protein under starvation of serum of cos-1 cells was monitored by annexin v and electron microscopy assays. intracellular reactive oxygen specie ...200717453707
severe acute respiratory syndrome (sars) coronavirus.the global severe acute respiratory syndrome (sars) outbreak was the first pandemic of the 21st century. although the outbreak was successfully controlled, evidence that sars emerged from an animal reservoir has raised concerns that another pandemic could occur. this review discusses the likelihood of another sars pandemic and reviews the epidemiological and clinical features of the disease with an emphasis on the clinical presentation, diagnosis, and management of sars.200717458774
in silico prediction of sars protease inhibitors by virtual high throughput screening.a structure-based in silico virtual drug discovery procedure was assessed with severe acute respiratory syndrome coronavirus main protease serving as a case study. first, potential compounds were extracted from protein-ligand complexes selected from protein data bank database based on structural similarity to the target protein. later, the set of compounds was ranked by docking scores using a electronic high-throughput screening flexible docking procedure to select the most promising molecules. ...200717461975
the emerging role of ace2 in physiology and disease.the renin-angiotensin-aldosterone system (raas) is a key regulator of systemic blood pressure and renal function and a key player in renal and cardiovascular disease. however, its (patho)physiological roles and its architecture are more complex than initially anticipated. novel raas components that may add to our understanding have been discovered in recent years. in particular, the human homologue of ace (ace2) has added a higher level of complexity to the raas. in a short period of time, ace2 ...200717464936
sars-cov: lessons for global health. 200817467837
proceedings of a meeting entitled emerging viral infectious diseases, april 4-5, 2005, hanoi, vietnam. 200717470907
the molecular biology of sars coronavirus.severe acute respiratory syndrome (sars) is the first emerging infectious disease of the 21st century that has been highly transmissible and fatal and was caused by a previously unknown coronavirus (sars-cov). the sars epidemic in 2003 resulted in more than 8400 sars cases and approximately 800 deaths. existing in non-identified animal reservoirs, sars-cov continues to represent a threat to humans although more than four years have passed since a large outbreak of sars, and no new cases have bee ...200717470909
signal transduction in sars-cov-infected cells.severe acute respiratory syndrome (sars) is a newly found infectious disease that is caused by a novel human coronavirus, sars coronavirus (sars-cov). because the mortality rate of sars patients is very high, understanding the pathological mechanisms of sars not only in vivo but in vitro is important for the prevention of sars. activation of signaling pathways caused by sars-cov infection leads to the phosphorylation and activation of downstream molecules. two conflicting cellular programs, apop ...200717470913
a fluorogenic peptide containing the processing site of human sars corona virus s-protein: kinetic evaluation and nmr structure elucidation.human severe acute respiratory syndrome coronavirus (hsars-cov) is the causative agent for sars infection. its surface glycoprotein (spike protein) is considered to be one of the prime targets for sars therapeutics and intervention because its proteolytic maturation by a host protease is crucial for host-virus fusion. using intramolecularly quenched fluorogenic (iqf) peptides based on hsars-cov spike protein (abz-(755)glu-gln-asp-arg-asn-thr-arg-glu-val-phe-ala-gln(766)-tyx-nh(2)) and in vitro s ...200717471479
comparison of immunoglobulin g responses to the spike and nucleocapsid proteins of severe acute respiratory syndrome (sars) coronavirus in patients with sars.both the nucleocapsid (n) and the spike (s) proteins of severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) are able to induce strong humoral responses in humans following an infection. to compare the immunoglobulin g (igg) responses to the s and n proteins of sars-cov in sars patients during the manifestation/convalescent period with those during the postinfection period, serum samples were collected from hospitalized sars patients within 6 weeks after the onset of illnes ...200717475765
the 3a accessory protein of sars coronavirus specifically interacts with the 5'utr of its genomic rna, using a unique 75 amino acid interaction domain.more than four years have passed since the outbreak of the severe acute respiratory syndrome (sars) epidemic, and still very little is known about the molecular biology and pathogenesis of this deadly virus. among the accessory proteins of the sars coronavirus (sars-cov), the 3a protein has been shown to interact with the spike, envelope, and membrane glycoprotein and has recently been established to be a structural component of capsid. recent studies suggest that the 3a protein may function as ...200717488094
nucleocapsid protein of sars-cov activates interleukin-6 expression through cellular transcription factor nf-kappab.high levels of interleukin-6 (il-6) in the acute stage associated with lung lesions were found in sars patients. to evaluate the mechanisms behind this event, we investigated the roles of sars-cov proteins in the regulation of il-6. results showed that the viral nucleocapsid (n) protein activated il-6 expression in a concentration-dependent manner. promoter analyses suggested that nf-kappab binding element was required for il-6 expression regulated by n protein. further studies demonstrated that ...200717490702
induction of specific immune responses by severe acute respiratory syndrome coronavirus spike dna vaccine with or without interleukin-2 immunization using different vaccination routes in mice.dna vaccines induce humoral and cellular immune responses in animal models and humans. to analyze the immunogenicity of the severe acute respiratory syndrome (sars) coronavirus (cov), sars-cov, spike dna vaccine and the immunoregulatory activity of interleukin-2 (il-2), dna vaccine plasmids pcdna-s and pcdna-il-2 were constructed and inoculated into balb/c mice with or without pcdna-il-2 by using three different immunization routes (the intramuscular route, electroporation, or the oral route wit ...200717494640
antigenic characterization of severe acute respiratory syndrome-coronavirus nucleocapsid protein expressed in insect cells: the effect of phosphorylation on immunoreactivity and specificity.the nucleocapsid (n) protein of severe acute respiratory syndrome-coronavirus (sars-cov) is involved in the pathological reaction to sars and is a key antigen for the development of a sensitive diagnostic assay. however, the antigenic properties of this n protein are largely unknown. to facilitate the studies on the function and antigenicity of the sars-cov n protein, 6x histidine-tagged recombinant sars-cov n (rsars-n) with a molecular mass of 46 and 48kda was successfully produced using the re ...200717499376
animal models and vaccines for sars-cov infection.we summarize findings of sars-cov infections in several animal models each of which support viral replication in lungs accompanied by histopathological changes and/or clinical signs of illness to varying degrees. new findings are reported on sars-cov replication and associated pathology in two additional strains (c57bl/6 and 129s6) of aged mice. we also provide new comparative data on viral replication and associated pathology following infection of golden syrian hamsters with various sars-cov s ...200817499378
epidemic modeling in complex realities.in our global world, the increasing complexity of social relations and transport infrastructures are key factors in the spread of epidemics. in recent years, the increasing availability of computer power has enabled both to obtain reliable data allowing one to quantify the complexity of the networks on which epidemics may propagate and to envision computational tools able to tackle the analysis of such propagation phenomena. these advances have put in evidence the limits of homogeneous assumptio ...200717502293
interaction between respiratory syncytial virus and glycosaminoglycans, including heparan sulfate.glycosaminoglycans (gags), including heparan sulfate (hs), are expressed on the surface of nearly all cells, linked to transmembrane proteins. these gags are sulfated to varying extents, lending a negative charge, and are used by a large number of viruses to initiate infection of immortalized cell lines. here we describe the rationale and methods for analyzing gag usage by one such virus, respiratory syncytial virus (rsv). the protocols presented allow the determination of which gag(s) is employ ...200717502668
expression of the severe acute respiratory syndrome coronavirus 3a protein and the assembly of coronavirus-like particles in the baculovirus expression system.the bac-to-bac baculovirus expression system was used to generate a recombinant baculovirus capable of expressing the severe acute respiratory syndrome (sars)-coronavirus (cov) 3a protein. using the same expression system, two structural proteins, membrane (m) and envelope (e), were co-expressed to form sars-cov virus-like particles (vlps) within an insect cell. expression of viral proteins was confirmed by western blot analysis and the formation of vlps was studied by transmission electron micr ...200717502669
expression, glycosylation, and modification of the spike (s) glycoprotein of sars cov.the spike (s) glycoprotein of coronaviruses is known to be essential in the binding of the virus to the host cell at the advent of the infection process. to study the maturation pathway of the s glycoprotein of the severe acute respiratory syndrome (sars)-coronavirus (cov) within the host cell, a t7/vaccinia virus-based expression system coupled to immunoprecipitation with anti-s antibodies was used to test and analyze different forms of the s glycoprotein. the state of maturity of the s glycopr ...200717502675
the two faces of the tissue renin-angiotensin systems: implication in cardiovascular diseases.the implication of the renin-angiotensin system (ras) in the regulation of the cardiovascular system has been well known for many years. accordingly, many pharmaceutical inhibitors have been developed to treat several pathologies, like hypertension and heart failure, and angiotensin converting enzyme (ace) became one of the major target in the treatment of these cardiovascular diseases. in the last decade however, it has become apparent that the classical view of the ras was not quite accurate. ...200717504232
gxxxg motif of severe acute respiratory syndrome coronavirus spike glycoprotein transmembrane domain is not involved in trimerization and is not important for entry.recently, a paper was published in which it was proposed that the gxxxg motif of the severe acute respiratory syndrome (sars) coronavirus spike (s) protein transmembrane domain plays a vital role in oligomerization of the protein (e. arbely, z. granot, i. kass, j. orly, and i. t. arkin, biochemistry 45:11349-11356, 2006). here, we show that the gxxxg motif is not involved in sars s oligomerization by trimerization analysis of s gxxxg mutant proteins. in addition, the capability of s to mediate e ...200717507478
synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice.the severe acute respiratory syndrome (sars) epidemic was characterized by high mortality rates in the elderly. the molecular mechanisms that govern enhanced susceptibility of elderly populations are not known, and robust animal models are needed that recapitulate the increased pathogenic phenotype noted with increasing age. using synthetic biology and reverse genetics, we describe the construction of a panel of isogenic sars coronavirus (sars-cov) strains bearing variant spike glycoproteins tha ...200717507479
synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice.the severe acute respiratory syndrome (sars) epidemic was characterized by high mortality rates in the elderly. the molecular mechanisms that govern enhanced susceptibility of elderly populations are not known, and robust animal models are needed that recapitulate the increased pathogenic phenotype noted with increasing age. using synthetic biology and reverse genetics, we describe the construction of a panel of isogenic sars coronavirus (sars-cov) strains bearing variant spike glycoproteins tha ...200717507479
simultaneous detection and high-throughput identification of a panel of rna viruses causing respiratory tract infections.clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, high-throughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. we evaluated two multiplex reverse transcription-pcr (rt-pcr) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens. the ngen respiratory virus analyte-specific assay (nanogen, san diego, ca) detects influenza a virus (flu-a) ...200717507510
[loop-mediated isothermal amplification].we have developed the loop-mediated isothermal amplification (lamp) method, an innovative gene amplification technique, which has the potential to become important in genetic testing. this method allows highly efficient amplification by a single enzyme at a constant temperature. it also has features such as high specificity, rapidity, and simplicity of detection. therefore, application to various life sciences, such as clinical medicine, is expected.200717511240
formalin-treated uv-inactivated sars coronavirus vaccine retains its immunogenicity and promotes th2-type immune responses.the demand for rapid and simple development of a vaccine against a newly emerging infectious disease is increasing worldwide. we previously revealed that uv-inactivated severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) virions (uv-v) elicited high levels of humoral immunity and a weak th0 response in mice immunized subcutaneously. to ensure the safety of such a whole inactivated sars-cov vaccine, we additionally treated the uv-v vaccine with formalin, resulting in the uv ...200717515642
a retrospective serological study of severe acute respiratory syndrome cases in guangdong province, china. 200717517191
human immunodeficiency viral vector pseudotyped with the spike envelope of severe acute respiratory syndrome coronavirus transduces human airway epithelial cells and dendritic cells.the human severe acute respiratory syndrome coronavirus (sars-cov) is a highly infectious virus that causes severe respiratory infections in humans. the spike envelope glycoprotein of sars-cov, the main determinant of sars-cov tropism, was isolated and used to pseudotype a human immunodeficiency virus (hiv)-based vector. spike-pseudotyped hiv vector was generated and evaluated in vitro on well-differentiated human airway epithelial cells and bronchial explants and in vivo in murine airways. the ...200717518614
analysis of intraviral protein-protein interactions of the sars coronavirus orfeome.the severe acute respiratory syndrome coronavirus (sars-cov) genome is predicted to encode 14 functional open reading frames, leading to the expression of up to 30 structural and non-structural protein products. the functions of a large number of viral orfs are poorly understood or unknown. in order to gain more insight into functions and modes of action and interaction of the different proteins, we cloned the viral orfeome and performed a genome-wide analysis for intraviral protein interactions ...200717520018
clathrin-dependent entry of severe acute respiratory syndrome coronavirus into target cells expressing ace2 with the cytoplasmic tail deleted.the penetration of various viruses into host cells is accomplished by hijacking the host endocytosis machinery. in the case of severe acute respiratory syndrome coronavirus (sars-cov) infection, viral entry is reported to require a low ph in intracytoplasmic vesicles; however, little is known about how sars-cov invades such compartments. here we demonstrate that sars-cov mainly utilizes the clathrin-mediated endocytosis pathway for its entry to target cells by using infectious sars-cov, as well ...200717522231
trafficking motifs in the sars-coronavirus nucleocapsid protein.the severe acute respiratory syndrome-coronavirus nucleocapsid (n) protein is involved in virus replication and modulation of cell processes. in this latter respect control may in part be achieved through the sub-cellular localisation of the protein. n protein predominately localises in the cytoplasm (the site of virus replication and assembly) but also in the nucleus/nucleolus. using a combination of live-cell and confocal microscopy coupled to mutagenesis we identified a cryptic nucleolar loca ...200717524366
[severe acute respiratory syndrome]. 200717526201
the nonstructural protein 8 (nsp8) of the sars coronavirus interacts with its orf6 accessory protein.severe acute respiratory syndrome (sars) coronavirus (sars-cov) caused a severe outbreak in several regions of the world in 2003. the sars-cov genome is predicted to contain 14 functional open reading frames (orfs). the first orf (1a and 1b) encodes a large polyprotein that is cleaved into nonstructural proteins (nsp). the other orfs encode for four structural proteins (spike, membrane, nucleocapsid and envelope) as well as eight sars-cov-specific accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b an ...200717532020
up-regulation of il-6 and tnf-alpha induced by sars-coronavirus spike protein in murine macrophages via nf-kappab pathway.the clinical picture of severe acute respiratory syndrome (sars) is characterized by an over-exuberant immune response with lung lymphomononuclear cells infilteration and proliferation that may account for tissue damage more than the direct effect of viral replication. to understand how cells response in the early stage of virus-host cell interaction, in this study, a purified recombinant s protein was studied for stimulating murine macrophages (raw264.7) to produce proinflammatory cytokines (il ...200717532082
cleavage of spike protein of sars coronavirus by protease factor xa is associated with viral infectivity.the spike (s) protein of sars coronavirus (sars-cov) has been known to recognize and bind to host receptors, whose conformational changes then facilitate fusion between the viral envelope and host cell membrane, leading to viral entry into target cells. however, other functions of sars-cov s protein such as proteolytic cleavage and its implications to viral infection are incompletely understood. in this study, we demonstrated that the infection of sars-cov and a pseudovirus bearing the s protein ...200717533109
lack of support for an association between clec4m homozygosity and protection against sars coronavirus infection. 200717534354
lack of support for an association between clec4m homozygosity and protection against sars coronavirus infection. 200717534355
newcastle disease virus, a host range-restricted virus, as a vaccine vector for intranasal immunization against emerging pathogens.the international outbreak of the severe acute respiratory syndrome-associated coronavirus (sars-cov) in 2002-2003 highlighted the need to develop pretested human vaccine vectors that can be used in a rapid response against newly emerging pathogens. we evaluated newcastle disease virus (ndv), an avian paramyxovirus that is highly attenuated in primates, as a topical respiratory vaccine vector with sars-cov as a test pathogen. complete recombinant ndv was engineered to express the sars-cov spike ...200717535926
the association of rantes polymorphism with severe acute respiratory syndrome in hong kong and beijing chinese.chemokines play important roles in inflammation and antiviral action. we examined whether polymorphisms of rantes, ip-10 and mig affect the susceptibility to and outcome of severe acute respiratory syndrome (sars).200717540042
proteinase dysbalance in pathology: the neprilysin (nep) and angiotensin-converting enzyme (ace) families.proteolytic enzymes constitute some 2% of the human genome and provide important therapeutic targets in many diseases. the identification of many novel proteases from genome sequencing programmes provides both qualitative and quantitative challenges to target identification and validation. some approaches to dealing with these questions and addressing functional correlates of proteolytic activity at the level of molecular cell biology are provided in this review focusing on two spheres of intere ...200617543197
isolation and identification of an scfv antibody against nucleocapsid protein of sars-cov.to develop reagents for early diagnosis and therapeutic drugs against sars-associated coronavirus (sars-cov), a large (3 x 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six sars convalescent patients. a single chain variable fragment antibody (n18) with high affinity against n protein of sars-cov was isolated. sequence analysis revealed that the vl gene was composed of vl3h (v lambda subgroup) and jl2 regions and the vh gene was composed of ...200717548223
the use of hepatitis c virus ns3/4a and secreted alkaline phosphatase to quantitate cell-cell membrane fusion mediated by severe acute respiratory syndrome coronavirus s protein and the receptor angiotensin-converting enzyme 2.the membrane fusion process mediated by severe acute respiratory syndrome coronavirus (sars-cov) s protein and its cellular receptor angiotensin-converting enzyme 2 (ace2) had been reconstituted using two chinese hamster ovary (cho) cell lines that constitutively express these recombinant proteins separately. this system was applied to develop a quantitative measurement of cell-cell fusion using hepatitis c virus (hcv) ns3/4a protease and a secretion-blocked egfp-4a/4b-seap (egfp: enhanced green ...200717553448
the use of hepatitis c virus ns3/4a and secreted alkaline phosphatase to quantitate cell-cell membrane fusion mediated by severe acute respiratory syndrome coronavirus s protein and the receptor angiotensin-converting enzyme 2.the membrane fusion process mediated by severe acute respiratory syndrome coronavirus (sars-cov) s protein and its cellular receptor angiotensin-converting enzyme 2 (ace2) had been reconstituted using two chinese hamster ovary (cho) cell lines that constitutively express these recombinant proteins separately. this system was applied to develop a quantitative measurement of cell-cell fusion using hepatitis c virus (hcv) ns3/4a protease and a secretion-blocked egfp-4a/4b-seap (egfp: enhanced green ...200717553448
angiotensin-converting enzyme 2 in acute respiratory distress syndrome.angiotensin-converting enzyme (ace) and ace2 are highly homologous metalloproteases that provide essential catalytic functions in the renin-angiotensin system (ras). angiotensin ii is one key effector peptide of the ras, inducing vasoconstriction and exerting multiple biological functions. ace cleaves angiotensin i to generate angiotensin ii, whereas ace2 reduces angiotensin ii levels. accumulating evidence has demonstrated a physiological and pathological role of ace2 in the cardiovascular syst ...200717558469
adenovirus-based vaccine prevents pneumonia in ferrets challenged with the sars coronavirus and stimulates robust immune responses in macaques.a ferret model of severe acute respiratory syndrome (sars)-cov infection was used to evaluate the efficacy of an adenovirus vaccine. animals were subjected to heterologous prime-boost using vectors from human serotype 5 and chimpanzee derived adenoviruses (human adhu5 and chimpanzee adc7) expressing spike protein followed by intranasal challenge with sars-cov. vaccination led to a substantial reduction in viral load and prevented the severe pneumonia seen in unvaccinated animals. the same prime- ...200717559989
adenovirus-based vaccine prevents pneumonia in ferrets challenged with the sars coronavirus and stimulates robust immune responses in macaques.a ferret model of severe acute respiratory syndrome (sars)-cov infection was used to evaluate the efficacy of an adenovirus vaccine. animals were subjected to heterologous prime-boost using vectors from human serotype 5 and chimpanzee derived adenoviruses (human adhu5 and chimpanzee adc7) expressing spike protein followed by intranasal challenge with sars-cov. vaccination led to a substantial reduction in viral load and prevented the severe pneumonia seen in unvaccinated animals. the same prime- ...200717559989
[expression of severe acute respiratory syndrome coronavirus receptors, ace2 and cd209l in different organ derived microvascular endothelial cells].to investigate the expression of the 2 severe acute respiratory syndrome coronavirus (sars-cov) receptors, angiotensin-converting enzyme 2 (ace2) and cd209l in different human organ/tissue derived microvascular endothelial cells.200717565868
a saccharomyces cerevisiae autoselection system for optimised recombinant protein expression.yeasts are attractive organisms for recombinant protein production. they combine highly developed genetic systems and ease of use with reductions in time and costs. we describe an autoselection system for recombinant protein expression in saccharomyces cerevisiae which increases yields 5-10-fold compared to conditional selection for expression plasmids. multicopy expression plasmids encoding essential mob1 or cdc28 genes are absolutely necessary for the viability of host cells with mob1 or cdc28 ...200717566670
cell entry by enveloped viruses: redox considerations for hiv and sars-coronavirus.for enveloped viruses, genome entry into the target cell involves two major steps: virion binding to the cell-surface receptor and fusion of the virion and cell membranes. virus-cell membrane fusion is mediated by the virus envelope complex, and its fusogenicity is the result of an active virus-cell interaction process that induces conformation changes within the envelope. for some viruses, such as influenza, exposure to an acidic milieu within the cell during the early steps of infection trigge ...200717567241
effectiveness of control measures during the sars epidemic in beijing: a comparison of the rt curve and the epidemic curve.one of the areas most affected by sars was beijing with 2521 reported cases. we estimate the effective reproductive number rt for the beijing sars epidemic, which represents the average number of secondary cases per primary case on each day of the epidemic and is therefore a measure of the underlying transmission dynamics. our results provide a quantitative assessment of the effectiveness of public health control measures. more generally, our results illustrate how changes in rt will reflect cha ...200817568476
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