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long-range cooperative interactions modulate dimerization in sars 3clpro.severe acute respiratory syndrome (sars) is an infectious disease caused by the human coronavirus, sars-cov. the main viral protease, sars 3clpro, is a validated target for the development of antiviral therapies. since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. dimerization inhibitors are usually targeted to the dimerization interface a ...200617154528
augmentation of chemokine production by severe acute respiratory syndrome coronavirus 3a/x1 and 7a/x4 proteins through nf-kappab activation.severe acute respiratory syndrome (sars) is characterized by rapidly progressing respiratory failure resembling acute/adult respiratory distress syndrome (ards) associated with uncontrolled inflammatory responses. here, we demonstrated that, among five accessory proteins of sars coronavirus (sars-cov) tested, 3a/x1 and 7a/x4 were capable of activating nuclear factor kappa b (nf-kappab) and c-jun n-terminal kinase (jnk), and significantly enhanced interleukin 8 (il-8) promoter activity. furthermo ...200617141229
sars cov main proteinase: the monomer-dimer equilibrium dissociation constant.the sars coronavirus main proteinase (sars cov main proteinase) is required for the replication of the severe acute respiratory syndrome coronavirus (sars cov), the virus that causes sars. one function of the enzyme is to process viral polyproteins. the active form of the sars cov main proteinase is a homodimer. in the literature, estimates of the monomer-dimer equilibrium dissociation constant, kd, have varied more than 65,0000-fold, from <1 nm to more than 200 microm. because of these discrepa ...200617144656
progressive docking: a hybrid qsar/docking approach for accelerating in silico high throughput screening.a combination of protein-ligand docking and ligand-based qsar approaches has been elaborated, aiming to speed-up the process of virtual screening. in particular, this approach utilizes docking scores generated for already processed compounds to build predictive qsar models that, in turn, assess hypothetical target binding affinities for yet undocked entries. the "progressive docking" has been tested on drug-like substances from the nci database that have been docked into several unrelated target ...200617149875
anti-sars-cov activity of nucleoside analogs having 6-chloropurine as a nucleobase.nucleoside analogs that have 6-chloropurine as a nucleobase were synthesized and evaluated to develop novel anti-sars-cov agents. two compounds among them showed promising activity that was comparable to mizoribine and ribavirin. moreover, one of the compounds showed a structurally unique property.200617150843
porcine innate and adaptative immune responses to influenza and coronavirus infections.both innate and adaptative immune responses contribute to the control of infectious diseases, including by limiting the spreading of zoonotic diseases from animal reservoirs to humans. pigs represent an important animal reservoir for influenza virus infection of human populations and are also naturally infected by coronaviruses, an important group of viruses, which includes the recently emerged severe acute respiratory syndrome (sars) virus. studies on both innate and adaptative immune responses ...200617135502
generating vesicular stomatitis virus pseudotype bearing the severe acute respiratory syndrome coronavirus spike envelope glycoprotein for rapid and safe neutralization test or cell-entry assay.we generated a recombinant vesicular stomatitis virus (vsv) pseudotype (vsv delta g*sg) by replacing the envelope g gene with the gfp gene and complementing with spike glycoprotein (s) of sars-cov in trans. the neutralization and infection blocking tests showed that the vsv delta g*sg and sars-cov reacted similarly to sars-cov specific antiserum, suggesting the vsvdelta g*sg can be a safe replacement of the live sars-cov for neutralization test and cell-entry assay.200617135519
novel sars unique adomet-dependent methyltransferase. 200617102613
public health implications of using various case definitions in the netherlands during the worldwide sars outbreak.this study analysed the consequences of deviation from the who case definition for the assessment of patients with suspected severe acute respiratory syndrome (sars) in the netherlands during 2003. between 17 march and 7 july 2003, as a result of dilemmas in balancing sensitivity and specificity, five different case definitions were used. the patients referred for sars assessment were analysed from a public health perspective. none of the patients referred had sars, based on serological and viro ...200617121628
[the possible mechanism of lung injury induced by severe acute respiratory syndrome coronavirus spike glycoprotein].to investigate the role of severe acute respiratory syndrome coronavirus (sars-cov) in the induction of acute lung injury by promoting the synthesis of chemokine/cytokines in human endothelial cells.200617129463
the possible origin of recent human sars coronavirus isolate from china. 200617131942
[pharmacotherapy of severe acute respiratory syndrome (sars)].severe acute respiratory syndrome (sars) constitutes the first new infectious disease of the current millennium. it is caused by the novel sars-coronavirus (sars-cov). sars is related to a high morbidity and mortality and first appeared during an epidemic in 2002 - 2003. to date no specific therapy against the sars-cov is available. due to the rapid spread of sars during the epidemics in 2002 - 2003, randomised and controlled multicentre studies were not performed. therefore, general guidelines ...200617109267
radiology of severe acute respiratory syndrome (sars): the emerging pathologic-radiologic correlates of an emerging disease.severe acute respiratory distress syndrome (sars) caused by sars-associated coronavirus (sars-cov) is a systemic infection that clinically manifests as progressive pneumonia. during the initial phases of infection the virus causes pauci-inflammatory alveolar and interstitial edema that result in imaging abnormalities dominated by ground glass opacities (ggo). severe sars cases can develop radiologic and pathologic findings of diffuse alveolar damage. although radiologic evidence of acute bronchi ...200617110851
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
[a follow up study of total igm, igg, nucleoprotein and spike protein antibodies against severe acute respiratory syndrome (sars) coronavirus in patients with sars].to survey the dynamic changing and persistence of the special antibodies, including total igm, igg, nucleocapsid protein and spike protein antibodies, against severe acute respiratory syndrome coronavirus (sars-cov) in patients with sars.200617313874
[construction of genomic full-length cdna of sars coronavirus bj01 strain and identification of its biological characteristics].severe acute respiratory syndrome (sars) is an important emerging infectious disease which caused by sars coronavirus (sars-cov), and the study of its pathogenesis is needed for the treatment and prevention of this disease. to study the pathogenesis of sars-cov using reverse genetics technology, by in vitro ligation using 7 contiguous cdnas that span the entire genome of the sars-cov bj01 strain, a genomic full-length cdna was assembled, then using t7 ribomax large scale rna production systems w ...200617302155
indomethacin has a potent antiviral activity against sars coronavirus.severe acute respiratory syndrome (sars) is a newly emerging, highly transmissible and fatal disease caused by a previously unknown coronavirus (sars-cov). existing in non-identified animal reservoirs, sars-cov continues to represent a threat to humans because there is no effective specific antiviral therapy for coronavirus infections.200617302372
pathogenesis of avian flu h5n1 and sars.avian influenza a (h5n1) and severe acute respiratory syndrome (sars) coronavirus are infections that cause a severe viral pneumonia leading to acute respiratory dysfunction syndrome and carry a high case-fatality rate. we have investigated innate immune responses to both viruses using primary human macrophages and respiratory epithelial cells as in vitro models. in contrast to human influenza a h1ni viruses, the h5n1 viruses hyper-induce cytokines (tumour necrosis factor [tnf]alpha, interferon ...200617278385
[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
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
[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
viral load quantitation of sars-coronavirus rna using a one-step real-time rt-pcr.severe acute respiratory syndrome (sars) is an emerging infectious disease that first occurred in humans in the people's republic of china in november 2002 and has subsequently spread worldwide. a novel virus belonging to the coronaviridae family has been identified as the cause of this pulmonary disease. the severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays.200616023880
anti-sars-cov igg response in relation to disease severity of severe acute respiratory syndrome.the association between a robust or depressed antibody response and clinical severity of sars remains unknown.200616112612
using an integrated infection control strategy during outbreak control to minimize nosocomial infection of severe acute respiratory syndrome among healthcare workers.healthcare workers (hcws) are at risk of acquiring severe acute respiratory syndrome (sars) while caring for sars patients. personal protective equipment and negative pressure isolation rooms (npirs) have not been completely successful in protecting hcws. we introduced an innovative, integrated infection control strategy involving triaging patients using barriers, zones of risk, and extensive installation of alcohol dispensers for glove-on hand rubbing. this integrated infection control approach ...200616153744
bcl-2 inhibits the caspase-dependent apoptosis induced by sars-cov without affecting virus replication kinetics.vero cells transfected with either neo- or bcl-2-plasmid were infected with sars-cov at a high multiplicity of infection. apoptosis appeared after the onset of cpe and completion of virus replication, and could be prevented by bcl-2 expression. apoptosis is likely mediated by the mitochondrial pathway, as demonstrated by its inhibition using bcl-2, and by the activation of the caspase cascade, resulting in parp cleavage. prevention of apoptosis did not affect susceptibility to infection, kinetic ...200616155806
dissection of sars coronavirus spike protein into discrete folded fragments.the spike protein of the severe acute respiratory syndrome coronavirus (sars-cov) mediates cell fusion by binding to target cell surface receptors. this paper reports a simple method for dissecting the viral protein and for searching for foldable fragments in a random but systematic manner. the method involves digestion by dnase i to generate a pool of short dna segments, followed by an additional step of reassembly of these segments to produce a library of dna fragments with random ends but con ...200632288412
the renin-angiotensin system in acute respiratory distress syndrome.angiotensin-converting enzyme 2 (ace2) counterbalances with ace and functions as a negative regulator of the renin-angiotensin system (ras). the importance of ras in acute respiratory distress syndrome (ards) has recently re-emerged owing to the identification of ace2 as a receptor for the sars-coronavirus. recent studies have demonstrated that ace2 protects mice from acute lung injury as well as sars-mediated lung injury. we review the role of the ras, in particular ace2, in the pathogenesis of ...200632288774
absence of infection in asymptomatic contacts of index sars case in france.the first case of severe acute respiratory syndrome (sars) in france was diagnosed in march 2003. we conducted a serological survey to assess whether or not asymptomatic persons who had been in contact with this patient during his infectious stage had been infected. they were interviewed and asked to provide a blood sample for sars coronavirus immunoglobulin g antibody testing. despite the likely high infectivity of the sars patient, no asymptomatic sars infection was found in any of the 37 cont ...200629208086
the diversity of viruses infecting humans.most human viruses have been discovered through the diseases they cause in animals, plants, bacteria or fungi. recent finds include human bocaviruses, which now seem to have a global distribution, and cause respiratory tract disease in infants, and several new pathogenic human coronaviruses. the sars coronavirus, genetically distinct from all previously known coronaviruses, caused a disease which was highly transmissible and very severe, eventually leading to 8000 cases worldwide with over 800 d ...200632309018
seroepidemiological study on sars-cov igg antibodies of different populations from several areas.in order to investigate the clinical and epidemiological rules of severe acute respiratory syndrome (sars), rates and levels sars coronavirus (sars-cov) igg antibodies of the patients and community populations from several areas were detected.200523674957
quality assurance for the diagnostics of viral diseases to enhance the emergency preparedness in europe.the threat posed by emerging and re-emerging communicable diseases and, more recently, by the intentional release of infectious agents in a susceptible population, has been receiving considerable attention at the national and international levels. public health efforts to strengthen disease detection, surveillance and control have been intensified. however, clinicians and clinical microbiology laboratories play an important role in the early detection of disease, the identification of the putati ...200529183474
control of severe acute respiratory syndrome in singapore.a severe acute respiratory syndrome (sars) outbreak occurred in singapore from february to may 2003. a high vigilance for the disease, frequent and regular temperature monitoring, early case identification and isolation of patients, as well as tracing and home quarantine of contacts, played major roles in controlling the outbreak. hospitals were dedicated to the screening and treatment of sars patients. within and between hospitals, movement by healthcare workers, patients and visitors were rest ...200521432128
an activated gops-poly-l-lysine- coated glass surface for the immobilization of 60mer oligonucleotides.to explore a method for enhancing the immobilization and hybridization efficiency of oligonucleotides on dna microarrays, conventional protocols of poly-l-lysine coating were modified by means of surface chemistry, namely, the slides were prepared by the covalently coupling of poly-l-lysine to a glycidoxy-modified glass surface. the modified slides were then used to print microarrays for the detection of the sars coronavirus by means of 60mer oligonucleotide probes. the characteristics of the mo ...200532313546
putative structure and function of orf3 in sars coronavirus.based on molecular modeling techniques we constructed a rational 3d model of orf3 in sars coronavirus (sars-cov). our studies suggest that the function of orf3 could be involved in fad/nad binding according to its predicted structure and comparison with other structure neighbors. furthermore, we identified three pairs of non-canonical n-h⋯π interactions in the structure of orf3, which can make contributions to the stability of protein structure. these results provide important clues for better u ...200532287548
influenza, respiratory syncytial virus and sars.acute lower respiratory tract infections (lrtis) are a major worldwide health problem, particularly in childhood, and are ranked first among the conditions contributing to the global burden of disease. about 30-50% of acute lrtis are viral in origin; of these, influenza and respiratory syncytial virus (rsv) are associated with the greatest disease burden in humans. vaccination against circulating human influenza strains and the use of neuraminidase inhibitor drugs have improved the options for c ...200532308534
infection of sars-cov on juvenile and adult brandt's volemicrotus brandtii.we studied the infectious effect of sars-cov virus on juvenile and adult brandt's vole (microtus brandtii) by nasal cavity spraying method (ccid50 is 105.7). sars virus caused serious deaths in adults. the death adults demonstrated hemorrhage from mouth, nasal cavity and intestine, hemorrhageious interstitial pneumonia and gore in liver, spleen and kidney. the survival adults demonstrated local hemorrhagic spot in lung and emphysema, but the other organs showed no pathological abnormality. sars ...200532214720
sensitive detection of sars coronavirus rna by a novel asymmetric multiplex nested rt-pcr amplification coupled with oligonucleotide microarray hybridization.we have developed a sensitive method for the detection of specific genes simultaneously. first, dna was amplified by a novel asymmetric multiplex pcr with universal primer(s). second, the 6-carboxytetramethylrhodamine (tamra)-labeled pcr products were hybridized specifically with oligonucleotide microarrays. finally, matched duplexes were detected by using a laser-induced fluorescence scanner. the usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (sars) cor ...200516156097
direct template matching reveals a host subcellular membrane gyroid cubic structure that is associated with sars virus.viral infection can result in alterations to the host subcellular membrane. this is often reported when using transmission electron microscopy (tem), resulting in a description of tubuloreticular membrane subcellular ultrastructure rather than a definition based on 3-d morphology. 2-d tem micrographs depicting subcellular membrane changes are associated with subcellular sars virion particles [goldsmith cs, tatti km, ksiazek tg et al. ultra-structural characterization of sars coronavirus. emerg i ...200516156956
severe acute respiratory syndrome: vaccine on the way. 200516157050
the interaction of the sars coronavirus non-structural protein 10 with the cellular oxido-reductase system causes an extensive cytopathic effect.the pathological mechanism of sars-cov infection was investigated. the gene for the sars-cov non-structural protein 10, which is located in the open reading frame of pp1a/pp1ab gene, was synthesized and used to screen for the specific cellular gene coding for the protein interacting with this nsp10 protein in a human embryo lung cdna library using a yeast trap method. the results indicated that apart from the two subunits of cellular rna polymerase complex, btf3 and atf5, this nsp10 protein was ...200516157265
stem-loop iv in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering rna replication.the 210-nucleotide (nt) 5' untranslated region (utr) in the positive-strand bovine coronavirus (bcov) genome is predicted to contain four higher-order structures identified as stem-loops i to iv, which may function as cis-acting elements in genomic rna replication. here, we describe evidence that stem-loop iv, a bulged stem-loop mapping at nt 186 through 215, (i) is phylogenetically conserved among group 2 coronaviruses and may have a homolog in groups 1 and 3, (ii) exists as a higher-order stru ...200516160171
detection of severe acute respiratory syndrome coronavirus in the brain: potential role of the chemokine mig in pathogenesis.previous studies have shown that common human coronavirus might be neurotropic, although it was first isolated as a pathogen of the respiratory tract. we noticed that a few patients with severe acute respiratory syndrome (sars) experienced central nervous symptoms during the course of illness. in the present study, we isolated a sars coronavirus strain from a brain tissue specimen obtained from a patient with sars with significant central nervous symptoms.200516163626
t-cell response profiling to biological threat agents including the sars coronavirus.the emergence of pathogens such as sars and the increased threat of bioterrorism has stimulated the development of novel diagnostic assays for differential diagnosis. rather than focusing on the detection of an individual pathogen component, we have developed a t cell profiling system to monitor responses to the pathogens in an array format. using a matrix of antigens specific for different pathogens, a specific t cell profile was generated for each individual by monitoring the intracellular pro ...200516164833
structural biology. adaptation of sars coronavirus to humans. 200516166506
structure of sars coronavirus spike receptor-binding domain complexed with receptor.the spike protein (s) of sars coronavirus (sars-cov) attaches the virus to its cellular receptor, angiotensin-converting enzyme 2 (ace2). a defined receptor-binding domain (rbd) on s mediates this interaction. the crystal structure at 2.9 angstrom resolution of the rbd bound with the peptidase domain of human ace2 shows that the rbd presents a gently concave surface, which cradles the n-terminal lobe of the peptidase. the atomic details at the interface between the two proteins clarify the impor ...200516166518
severe acute respiratory syndrome coronavirus-like virus in chinese horseshoe bats.although the finding of severe acute respiratory syndrome coronavirus (sars-cov) in caged palm civets from live animal markets in china has provided evidence for interspecies transmission in the genesis of the sars epidemic, subsequent studies suggested that the civet may have served only as an amplification host for sars-cov. in a surveillance study for cov in noncaged animals from the wild areas of the hong kong special administration region, we identified a cov closely related to sars-cov (ba ...200516169905
severe acute respiratory syndrome coronavirus-like virus in chinese horseshoe bats.although the finding of severe acute respiratory syndrome coronavirus (sars-cov) in caged palm civets from live animal markets in china has provided evidence for interspecies transmission in the genesis of the sars epidemic, subsequent studies suggested that the civet may have served only as an amplification host for sars-cov. in a surveillance study for cov in noncaged animals from the wild areas of the hong kong special administration region, we identified a cov closely related to sars-cov (ba ...200516169905
association between mannose-binding lectin gene polymorphisms and susceptibility to severe acute respiratory syndrome coronavirus infection.genetic determinants of susceptibility to severe acute respiratory syndrome coronavirus (sars-cov) infection remain unknown. we assessed whether mannose-binding lectin (mbl) gene polymorphisms were associated with susceptibility to sars-cov infection or disease severity in an ethnically homogeneous population born in northern china.200516170752
generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (n) sars-cov protein using a phage display approach.severe acute respiratory syndrome (sars)-cov is a newly emerging virus that causes sars with high mortality rate in infected people. successful control of the global sars epidemic will require rapid and sensitive diagnostic tests to monitor its spread, as well as, the development of vaccines and new antiviral compounds including neutralizing antibodies that effectively prevent or treat this disease.200516171519
false positive antibody results against human t-cell lymphotropic virus in patients with severe acute respiratory syndrome.taiwan suffered from the outbreak of severe acute respiratory syndrome (sars) in 2003. our laboratory performed a series of virology and serology tests for sars patients admitted to our hospital. cross-reactivity was found when testing for antibody against human t-cell lymphotropic virus (htlv) in one patient with sars. therefore, antibodies against htlv were examined in paired-sera from 26 sars patients. elisa and a neutralization test were used to measure anti-sars antibodies. seroconversion f ...200516173022
properties of isonucleotide-incorporated oligodeoxynucleotides and inhibition of the expression of spike protein of sars-cov.antisense oligonucleotides are recognized to be very efficient tools for the inhibition of gene expression in a sequence specific way. for the discovery of a novel efficient way to modify oligonucleotides, a series of single isonucleotide-incorporated antisense oligodeoxynucleotides have been synthesized, in which an isonucleotide was introduced at different positions of the sequences. the binding behaviors of modified oligodeoxynucleotides to the complementary sequence were studied by uv, cd, a ...200516173783
molecular signature of clinical severity in recovering patients with severe acute respiratory syndrome coronavirus (sars-cov).severe acute respiratory syndrome (sars), a recent epidemic human disease, is caused by a novel coronavirus (sars-cov). first reported in asia, sars quickly spread worldwide through international travelling. as of july 2003, the world health organization reported a total of 8,437 people afflicted with sars with a 9.6% mortality rate. although immunopathological damages may account for the severity of respiratory distress, little is known about how the genome-wide gene expression of the host chan ...200516174304
[expression of the recombinant sars coronavirus nucleocapsid protein in pichia pastoris and identification of its bioactivity].the full length cdna of sars coronavirus nucleocapsid (n) protein was amplified by pcr and cloned into yeast expression vector ppic3.5k to generate expression vector ppic3.5k-scovn. the plasmid was linearized and then transformed into p. pastoris (his- mut+) by electroporation method. his+ mut+ recombinant strains were screened on g418-rdb and mm/md plates, and further confirmed by pcr. the influence of various inducing media, dissolved oxygen(do) and the different final concentration of methano ...200516176089
[expression of sars spike gene in shizomycete pombe].the viral spike protein is the main surface antigen of the coronavirus, and it could be useful in the research of clinical diagnosis, sars vaccine and the structure biology.according to the analysis of the main antigen of the sars spike protein, 5 fragments of the whole spike gene were cloned, and ligated to the vector pnmt1. through electroporation transformantion to tcp1, the recombinant s. pombe strains capable of expressing the 5 fragments were constructed. sds-page or western blot analysis ...200516176106
rhabdomyolysis associated with acute renal failure in patients with severe acute respiratory syndrome.an outbreak of severe acute respiratory syndrome (sars) occurred in taiwan in 2003. sars complicated with rhabdomyolysis has rarely been reported. this study reported three cases of rhabdomyolysis developing during the clinical course of sars. thirty probable sars patients were admitted to the isolation wards at linkou chang gung memorial hospital between 4 april and 4 june 2003. thirty patients, including four men and 26 women aged from 12 to 87 years (mean age 40). eleven (36.7%) patients had ...200516178983
post-sars sickness syndrome manifestations and endocrinopathy: how, why, and so what? 200516181227
experience of using convalescent plasma for severe acute respiratory syndrome among healthcare workers in a taiwan hospital.to describe the immunological responses and clinical outcome of coronavirus (sars) infected healthcare workers (hcw) who had been administered with convalescent plasma as a treatment.200516183666
influence of fcgammariia and mbl polymorphisms on severe acute respiratory syndrome.polymorphisms of human fc gamma-receptor iia (fcgammariia) and mannose-binding lectin (mbl) genes have been associated with susceptibility to or severity of some infectious diseases. in order to investigate whether these genetic factors might influence susceptibility to infection with the severe acute respiratory syndrome-associated coronavirus (sars-cov) as well as the course and severity of the infection, we evaluated polymorphisms of fcgammariia and mbl genes in dna samples from a group of ap ...200516185324
adp-ribose-1"-monophosphatase: a conserved coronavirus enzyme that is dispensable for viral replication in tissue culture.replication of the approximately 30-kb plus-strand rna genome of coronaviruses and synthesis of an extensive set of subgenome-length rnas are mediated by the replicase-transcriptase, a membrane-bound protein complex containing several cellular proteins and up to 16 viral nonstructural proteins (nsps) with multiple enzymatic activities, including protease, polymerase, helicase, methyltransferase, and rnase activities. to get further insight into the replicase gene-encoded functions, we characteri ...200516188975
structural genomics of the severe acute respiratory syndrome coronavirus: nuclear magnetic resonance structure of the protein nsp7.here, we report the three-dimensional structure of severe acute respiratory syndrome coronavirus (sars-cov) nsp7, a component of the sars-cov replicase polyprotein. the coronavirus replicase carries out regulatory tasks involved in the maintenance, transcription, and replication of the coronavirus genome. nsp7 was found to assume a compact architecture in solution, which is comprised primarily of helical secondary structures. three helices (alpha2 to alpha4) form a flat up-down-up antiparallel a ...200516188992
use of dual taqman probes to increase the sensitivity of 1-step quantitative reverse transcription-pcr: application to the detection of sars coronavirus. 200516189379
sars vaccine: progress and challenge.severe acute respiratory syndrome (sars) emerged in 2002 as a severe and highly contagious infectious disease that rapidly spread to a number of different countries. the collaborative efforts of the global scientific community have provided, within a short period of time, substantial insights into the molecular biology and immunology of sars-cov. although the outbreak has been contained, there is continuous concern that the virus may resurface into the human population through seasonal changes, ...200516191415
an atypical rna pseudoknot stimulator and an upstream attenuation signal for -1 ribosomal frameshifting of sars coronavirus.the -1 ribosomal frameshifting requires the existence of an in cis rna slippery sequence and is promoted by a downstream stimulator rna. an atypical rna pseudoknot with an extra stem formed by complementary sequences within loop 2 of an h-type pseudoknot is characterized in the severe acute respiratory syndrome coronavirus (sars cov) genome. this pseudoknot can serve as an efficient stimulator for -1 frameshifting in vitro. mutational analysis of the extra stem suggests frameshift efficiency can ...200516055920
quality assurance for the diagnostics of viral diseases to enhance the emergency preparedness in europe.the threat posed by emerging and re-emerging communicable diseases and, more recently, by the intentional release of infectious agents in a susceptible population, has been receiving considerable attention at the national and international levels. public health efforts to strengthen disease detection, surveillance and control have been intensified. however, clinicians and clinical microbiology laboratories play an important role in the early detection of disease, the identification of the putati ...200516077216
good ace, bad ace do battle in lung injury, sars. 200516079870
serine-scanning mutagenesis studies of the c-terminal heptad repeats in the sars coronavirus s glycoprotein highlight the important role of the short helical region.the fusion subunit of the sars-cov s glycoprotein contains two regions of hydrophobic heptad-repeat amino acid sequences that have been shown in biophysical studies to form a six-helix bundle structure typical of the fusion-active core found in class i viral fusion proteins. here, we have applied serine-scanning mutagenesis to the c-terminal-most heptad-repeat region in the sars-cov s glycoprotein to investigate the functional role of this region in membrane fusion. we show that hydrophobic side ...200516081124
inhibitors of cathepsin l prevent severe acute respiratory syndrome coronavirus entry.severe acute respiratory syndrome (sars) is caused by an emergent coronavirus (sars-cov), for which there is currently no effective treatment. sars-cov mediates receptor binding and entry by its spike (s) glycoprotein, and infection is sensitive to lysosomotropic agents that perturb endosomal ph. we demonstrate here that the lysosomotropic-agent-mediated block to sars-cov infection is overcome by protease treatment of target-cell-associated virus. in addition, sars-cov infection was blocked by s ...200516081529
identification of immunodominant epitopes on the membrane protein of the severe acute respiratory syndrome-associated coronavirus.similar to other coronaviruses, the membrane (m) protein of severe acute respiratory syndrome-associated coronavirus (sars-cov) is a major transmembrane glycoprotein with multiple biological functions. to date, limited information is available about its antigenic properties. in this study, we identified two major immunodominant epitopes on the m protein located in the extreme n-terminal region (residues 1 to 31) and the interior c-terminal region (residues 132 to 161), respectively, by pepscan a ...200516081901
performance of single-step gel-based reverse transcription-pcr (rt-pcr) assays equivalent to that of real-time rt-pcr assays for detection of the severe acute respiratory syndrome-associated coronavirus.simple gel-based one-step reverse transcription-pcr (rt-pcr) assays, used to investigate patients during the 2003 severe acute respiratory syndrome (sars) outbreak in singapore, were found to be as sensitive as commercial and in-house real-time rt-pcr assays. the detection limit was approximately 1 genome equivalent (ge) per 5 microl pcr mixture. one pfu of sars coronavirus was estimated to be 258 +/- 46 ge.200516081995
first external quality assurance of antibody diagnostic for sars-new coronavirus.to confirm an infection with the new coronavirus (sars-cov) causing the severe acute respiratory syndrome (sars) diagnostic assays for detection of sars-cov specific antibody are necessary. to evaluate the diagnostic performance of laboratories an external quality assurance (eqa) study was performed in 2004. participating laboratories (9/20) correctly detected anti-sars antibodies in serum samples without false positive results in an immunofluorescence assay. in contrast, only 4/13 laboratories ...200516087120
expression and purification of n and e proteins from severe acute respiratory syndrome (sars)-associated coronavirus: a comparative study.histidine-tagged n (rnh) and e (reh) proteins of severe acute respiratory syndrome (sars)-coronovirus were expressed in the baculovirus/insect cell system and purified by immobilized metal affinity chromatography. rnh and reh proteins differed markedly with respect to expression levels, cell death kinetics and subcellular localizations that led to different extraction and purification schemes. the features of both proteins are compared and the potential applications of purified rnh and reh are d ...200516091881
coordinate induction of ifn-alpha and -gamma by sars-cov also in the absence of virus replication.severe acute respiratory syndrome (sars) is an emerging infection caused by a novel coronavirus known as sars-cov, characterized by an over-exuberant immune response with lung lymphomononuclear cells infiltration and proliferation that may account for tissue damage more than the direct effect of viral replication. this study is aimed at investigating the capability of sars-cov to activate ifn-alpha and -gamma expression in lymphomonocytes (pbmc) from healthy donors, evaluating whether viral repl ...200516095648
effects of severe acute respiratory syndrome (sars) coronavirus infection on peripheral blood lymphocytes and their subsets.severe acute respiratory syndrome (sars) caused large outbreaks of atypical pneumonia in 2003, with the largest localized outbreak occurring in beijing, china. lymphopenia was prominent amongst the laboratory abnormalities reported in acute sars.200516095942
genetic analysis of the sars-coronavirus spike glycoprotein functional domains involved in cell-surface expression and cell-to-cell fusion.the sars-coronavirus (sars-cov) is the etiological agent of severe acute respiratory syndrome (sars). the sars-cov spike (s) glycoprotein mediates membrane fusion events during virus entry and virus-induced cell-to-cell fusion. to delineate functional domains of the sars-cov s glycoprotein, single point mutations, cluster-to-lysine and cluster-to-alanine mutations, as well as carboxyl-terminal truncations were investigated in transient expression experiments. mutagenesis of either the coiled-coi ...200516099010
development of a method for concentrating and purifying sars coronavirus rna by a magnetic bead capture system.severe acute respiratory syndrome (sars) is a recently emerged infectious disease caused by a novel coronavirus, which has been designated the sars coronavirus (sars-cov). to date, molecular assays for the detection of sars-cov has focused mainly on reverse transcriptase-pcr (rt-pcr) analysis of specimens. however, rt-pcr assays currently available have low sensitivity during the early stage of the disease in which the viral load in specimens is very low. a method for concentrating and purifying ...200516101344
apoptosis induced by the sars-associated coronavirus in vero cells is replication-dependent and involves caspase.the pathogenesis of the severe acute respiratory syndrome (sars), a newly emerging life-threatening disease in humans, remains unknown. it is believed that the modulation of apoptosis is relevant to diseases that are caused by various viruses. to examine potential apoptotic mechanisms related to sars, we investigated features of apoptosis induced by the sars-associated coronavirus (sars-cov) in host cells. the results indicated that the sars-cov-induced apoptosis in vero cells in a virus replica ...200516101347
identification and antigenic epitope mapping of immunodominant region amino residues 510 to 672 on the spike protein of the severe acute respiratory syndrome coronavirus.the severe acute respiratory syndrome (sars) is a newly emerging human infectious disease caused by the severe acute respiratory syndrome coronavirus (sars-cov). the spike (s) protein of sars-cov is a major virion structural protein. it plays an important role in the interaction with receptors and neutralizing antibodies. in this study, the s1 domain of the spike protein and three truncated fragments were expressed by fusion with gst in a pgex-6p-1 vector. western blot results demonstrated that ...200516101348
elicitation of immunity in mice after immunization with the s2 subunit of the severe acute respiratory syndrome coronavirus.the s2 domain of the severe acute respiratory syndrome coronavirus (sars-cov) spike (s) protein is responsible for fusion between virus and target cell membranes, and is expected to be immungenic. in this study, we investigated the immune responses against the s2 subunit in balb/c mice, which were vaccinated either with plasmid dna encoding the s2 domain (residues 681-1120), the recombinant s2 fragment (residues 681-980) in incomplete freund's adjuvant, or with inactivated sars-cov. the increase ...200516101349
construction of a eukaryotic expression plasmid encoding partial s gene fragments of the sars-cov and its potential utility as a dna vaccine.the spike (s) protein, a main surface antigen of the sars coronavirus (sars-cov), is considered to be one of the most important protective antigen candidates for targets for vaccine design against the virus. in this study, a secreted recombinant expression plasmid, pvax-s1, encoding the partial s protein with a signal peptide, was constructed and used to immunize balb/c mice through electroporation. it was demonstrated that the eukaryotic expression vector pvax-s1 was successfully constructed by ...200516101350
profiles of igg antibodies to nucleocapsid and spike proteins of the sars-associated coronavirus in sars patients.to evaluate humoral immunity against the sars-associated coronavirus (sars-cov), we studied the profiles of igg antibodies to the nucleocapsid (n) and spike (s) proteins of sars-cov. serum specimens from 10 sars patients were analyzed by western blotting and an enzyme-linked immunosorbent assay (elisa) using purified recombinant n and truncated s (s1, s2, and s3) proteins as antigens. western blotting results demonstrated that 100% of the sars patients tested positive for n protein-specific anti ...200516101351
synthesis of novel test compounds for antiviral chemotherapy of severe acute respiratory syndrome (sars).this contribution reviews the synthesis of a range of experimental drugs designed for and aiming at antiviral chemotherapy of severe acute respiratory syndrome (sars) coronavirus (sars-cov)-induced human disease conditions. the selection of 25 test materials includes eleven trioxa-adamantane-triols (tats) [bn, ibnca, abnca, vanba, ethylvanba, eubn, euvanba, ansabn, ehrlich bn, [6]prismanebn, nitrodibn], trivially termed bananins, one trioxa-adamantan-ol (tao) thymoba, one bis-bananin pi-bananin ...200516101496
amotosalen photochemical inactivation of severe acute respiratory syndrome coronavirus in human platelet concentrates.a novel human coronavirus causing severe acute respiratory syndrome (sars) emerged in epidemic form in early 2003 in china and spread worldwide in a few months. every newly emerging human pathogen is of concern for the safety of the blood supply during and after an epidemic crisis. for this purpose, we have evaluated the inactivation of sars-coronavirus (cov) in platelet concentrates using an approved pathogen inactivation device, the intercept blood system. apheresis platelet concentrates (apcs ...200516101804
posttraumatic stress after sars.posttraumatic stress disorder (ptsd) can arise in patients with medical illness. we used 2 chinese self-report measures to examine features of ptsd, anxiety, and depression in 131 survivors of severe acute respiratory syndrome at 1 month and 3 months after discharge from the hospital. risk factors associated with psychological distress were identified.200516102324
a severe acute respiratory syndrome-associated coronavirus-specific protein enhances virulence of an attenuated murine coronavirus.most animal species that can be infected with the severe acute respiratory syndrome-associated coronavirus (sars-cov) do not reproducibly develop clinical disease, hindering studies of pathogenesis. to develop an alternative system for the study of sars-cov, we introduced individual sars-cov genes (open reading frames [orfs]) into the genome of an attenuated murine coronavirus. one protein, the product of sars-cov orf6, converted a sublethal infection to a uniformly lethal encephalitis and enhan ...200516103185
the severe acute respiratory syndrome coronavirus nucleocapsid protein is phosphorylated and localizes in the cytoplasm by 14-3-3-mediated translocation.the severe acute respiratory syndrome coronavirus(sars-cov) nucleocapsid (n) protein is one of the four structural proteins of the virus and is predicted to be a 46-kda phosphoprotein. our in silico analysis predicted n to be heavily phosphorylated at multiple residues. experimentally, we have shown in this report that the n protein of the sars-cov gets serine-phosphorylated by multiple kinases, in both the cytoplasm and the nucleus. the phosphoprotein is stable and localizes in the cytoplasm an ...200516103198
intracellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid protein: absence of nucleolar accumulation during infection and after expression as a recombinant protein in vero cells.the nucleocapsid (n) protein of several members within the order nidovirales localizes to the nucleolus during infection and after transfection of cells with n genes. however, confocal microscopy of n protein localization in vero cells infected with the severe acute respiratory syndrome coronavirus (sars-cov) or transfected with the sars-cov n gene failed to show the presence of n in the nucleoplasm or nucleolus. amino acids 369 to 389, which contain putative nuclear localization signal (nls) an ...200516103202
outbreak of severe acute respiratory syndrome in southern taiwan, 2003.this study describes the epidemiologic features of the severe acute respiratory syndrome (sars) outbreak in southern taiwan in 2003. according to the official files of reported cases of sars from february 21 to june 19, 2003, there were 586 cases in southern taiwan. symptom onset occurred between february 21 and june 19 in reported cases, between march 13 and may 30 in probable cases, and between march 17 and may 23 in polymerase chain reaction (pcr)-positive probable cases. dates of symptom ons ...200516103615
g0/g1 arrest and apoptosis induced by sars-cov 3b protein in transfected cells.severe acute respiratory syndrome coronavirus (sars-cov), cause of the life-threatening atypical pneumonia, infects many organs, such as lung, liver and immune organ, and induces parenchyma cells apoptosis and necrosis. the genome of sars-cov, not closely related to any of the previously characterized coronavirus, encodes replicase and four major structural proteins and a number of non-structural proteins. published studies suggest that some non-structural proteins may play important roles in th ...200516107218
biosafety level 3 laboratory for autopsies of patients with severe acute respiratory syndrome: principles, practices, and prospects.during the outbreak of the emergent severe acute respiratory syndrome (sars) infection, >30% of the approximately 8000 infected persons were health care workers. the highly infectious nature of sars coronavirus (sars-cov) compelled our pathologists to consider biosafety issues in the autopsy room and for tissue processing procedures.200516107979
sars vaccine protective in mice. 200516110580
[diagnostic tests: sars-corona virus]. 200516111269
characterization and application of monoclonal antibodies against n protein of sars-coronavirus.severe acute respiratory syndrome-coronavirus (sars-cov) causes an infectious disease through respiratory route. diagnosing the disease effectively and accurately at early stage is essential for preventing the disease transmission and performing antiviral treatment. in this study, we raised monoclonal antibodies (mabs) against the nucleocapsid (n) protein of sars-cov and mapped epitopes by using different truncated n protein fragments. the mapping of those epitopes was valuable for constructing ...200516112641
molecular evolution and multilocus sequence typing of 145 strains of sars-cov.in this study, we have identified 876 polymorphism sites in 145 complete or partial genomes of sars-cov available in the ncbi genbank. one hundred and seventy-four of these sites existed in two or more sars-cov genome sequences. according to the sequence polymorphism, all sars-covs can be divided into three groups: (i) group 1, animal-origin viruses (such as sars-cov sz1, sz3, sz13 and sz16); (ii) group 2, all viruses with clinical origin during first epidemic; and (iii) group 3, sars-cov gd03t0 ...200516112670
chloroquine is a potent inhibitor of sars coronavirus infection and spread.severe acute respiratory syndrome (sars) is caused by a newly discovered coronavirus (sars-cov). no effective prophylactic or post-exposure therapy is currently available.200516115318
anti-sars coronavirus 3c-like protease effects of isatis indigotica root and plant-derived phenolic compounds.the 3c-like protease (3clpro) of sars-coronavirus mediates the proteolytic processing of replicase polypeptides 1a and 1ab into functional proteins, becoming an important target for the drug development. in this study, isatis indigotica root extract, five major compounds of i. indigotica root, and seven plant-derived phenolic compounds were tested for anti-sars-cov 3clpro effects using cell-free and cell-based cleavage assays. cleavage assays with the 3clpro demonstrated that ic50 values were in ...200516115693
protease-mediated enhancement of severe acute respiratory syndrome coronavirus infection.a unique coronavirus severe acute respiratory syndrome-coronavirus (sars-cov) was revealed to be a causative agent of a life-threatening sars. although this virus grows in a variety of tissues that express its receptor, the mechanism of the severe respiratory illness caused by this virus is not well understood. here, we report a possible mechanism for the extensive damage seen in the major target organs for this disease. a recent study of the cell entry mechanism of sars-cov reveals that it take ...200516116101
using sirna in prophylactic and therapeutic regimens against sars coronavirus in rhesus macaque.development of therapeutic agents for severe acute respiratory syndrome (sars) viral infection using short interfering rna (sirna) inhibitors exemplifies a powerful new means to combat emerging infectious diseases. potent sirna inhibitors of sars coronavirus (scv) in vitro were further evaluated for efficacy and safety in a rhesus macaque (macaca mulatta) sars model using clinically viable delivery while comparing three dosing regimens. observations of sars-like symptoms, measurements of scv rna ...200516116432
using sirna in prophylactic and therapeutic regimens against sars coronavirus in rhesus macaque.development of therapeutic agents for severe acute respiratory syndrome (sars) viral infection using short interfering rna (sirna) inhibitors exemplifies a powerful new means to combat emerging infectious diseases. potent sirna inhibitors of sars coronavirus (scv) in vitro were further evaluated for efficacy and safety in a rhesus macaque (macaca mulatta) sars model using clinically viable delivery while comparing three dosing regimens. observations of sars-like symptoms, measurements of scv rna ...200516116432
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