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identification of a b-cell antigenic epitope at the n-terminus of sars-cov m protein and characterization of monoclonal antibody against the protein.to identify the potential b-cell antigenic epitopes within the n-terminus of sars-cov (sars-associated coronavirus, sars-cov) m protein and characterize monoclonal antibody (mab) against the protein as well as its recognizing region, we expressed and purified a portion of sars-cov m protein (amino acid 1-43) in escherichia coli (e. coli). by using western blot and enzyme-linked immunosorbent assay (elisa), we showed that the purified recombinant m protein could be recognized by four sars-cov-pos ...200616972028
three dimensional model of severe acute respiratory syndrome coronavirus helicase atpase catalytic domain and molecular design of severe acute respiratory syndrome coronavirus helicase inhibitors.the modeling of the severe acute respiratory syndrome coronavirus helicase atpase catalytic domain was performed using the protein structure prediction meta server and the 3d jury method for model selection, which resulted in the identification of 1jpr, 1uaa and 1w36 pdb structures as suitable templates for creating a full atom 3d model. this model was further utilized to design small molecules that are expected to block an atpase catalytic pocket thus inhibit the enzymatic activity. binding sit ...200616972168
a trimerizing gxxxg motif is uniquely inserted in the severe acute respiratory syndrome (sars) coronavirus spike protein transmembrane domain.in an attempt to understand what distinguishes severe acute respiratory syndrome (sars) coronavirus (scov) from other members of the coronaviridae, we searched for elements that are unique to its proteins and not present in any other family member. we identified an insertion of two glycine residues, forming the gxxxg motif, in the scov spike protein transmembrane domain (tmd), which is not found in any other coronavirus. this surprising finding raises an "oligomerization riddle": the gxxxg motif ...200616981695
antigenic and cellular localisation analysis of the severe acute respiratory syndrome coronavirus nucleocapsid protein using monoclonal antibodies.a member of the family of coronaviruses has previously been identified as the cause of the severe acute respiratory syndrome (sars). in this study, several monoclonal antibodies against the nucleocapsid protein have been generated to examine distribution of the nucleocapsid in virus-infected cells and to study antigenic regions of the protein. confocal microscopic analysis identified nucleocapsids packaged in vesicles in the perinuclear area indicating viral synthesis at the endoplasmic reticulu ...200616920216
putative cis-acting stem-loops in the 5' untranslated region of the severe acute respiratory syndrome coronavirus can substitute for their mouse hepatitis virus counterparts.consensus covariation-based secondary structural models for the 5' 140 nucleotides of the 5' untranslated regions (5'utrs) from mouse hepatitis virus (mhv) and severe acute respiratory syndrome coronavirus (scov) were developed and predicted three major helical stem-loop structures, designated stem-loop 1 (sl1), sl2, and sl4. the scov 5'utr was predicted to contain a fourth stem-loop, named sl3, in which the leader transcriptional regulatory sequence (trs) is folded into a hairpin loop. cdnas co ...200616920822
construction of a severe acute respiratory syndrome coronavirus infectious cdna clone and a replicon to study coronavirus rna synthesis.the engineering of a full-length infectious cdna clone and a functional replicon of the severe acute respiratory syndrome coronavirus (sars-cov) urbani strain as bacterial artificial chromosomes (bacs) is described in this study. in this system, the viral rna was expressed in the cell nucleus under the control of the cytomegalovirus promoter and further amplified in the cytoplasm by the viral replicase. both the infectious clone and the replicon were fully stable in escherichia coli. using the s ...200616928748
animal models and antibody assays for evaluating candidate sars vaccines: summary of a technical meeting 25-26 august 2005, london, uk.severe acute respiratory syndrome (sars) emerged in the guangdong province of china in late 2002 and spread to 29 countries. by the end of the outbreak in july 2003, the cdc and who reported 8437 cases with a 9.6% case fatality rate. the disease was caused by a previously unrecognized coronavirus, sars-cov. drawing on experience with animal coronavirus vaccines, several vaccine candidates have been developed and evaluated in pre-clinical trials. available data suggest that vaccines should be bas ...200616930781
experience of a global laboratory network in responding to infectious disease epidemics. 200616931402
extremely low exposure of a community to severe acute respiratory syndrome coronavirus: false seropositivity due to use of bacterially derived antigens.estimates of seropositivity to a new infectious agent in a community are useful to public health. for severe acute respiratory syndrome (sars), the figures are conflicting. herein, we screened 12,000 people in a community stricken by sars 10 months previously and found 53 individuals (0.44%) who had immunoglobulin g antibodies to the sars coronavirus (sars-cov) nucleocapsid (n) produced in bacteria. however, only seven of these (group 1) had sera which also reacted with the native n antigen expr ...200616940504
modulation of the unfolded protein response by the severe acute respiratory syndrome coronavirus spike protein.perturbation of the function of endoplasmic reticulum (er) causes stress leading to the activation of cell signaling pathways known as the unfolded protein response (upr). severe acute respiratory syndrome (sars) coronavirus (sars-cov) uses er as a site for synthesis and processing of viral proteins. in this report, we demonstrate that infection with sars-cov induces the upr in cultured cells. a comparison with m, e, and nsp6 proteins indicates that sars-cov spike (s) protein sufficiently induce ...200616940539
infectious diseases emerging from chinese wet-markets: zoonotic origins of severe respiratory viral infections.in china, close contacts between humans and food animals have resulted in the transmission of many microbes from animals to humans. the two most notable infectious diseases in recent years are severe acute respiratory syndrome and avian influenza. in this review, these two severe zoonotic viral infections transmitted by the respiratory route, with pandemic potential, are used as models to illustrate the role of chinese wet-markets in their emergence, amplification and dissemination.200616940861
crystal structure of nonstructural protein 10 from the severe acute respiratory syndrome coronavirus reveals a novel fold with two zinc-binding motifs.the severe acute respiratory syndrome coronavirus (sars-cov) possesses a large 29.7-kb positive-stranded rna genome. the first open reading frame encodes replicase polyproteins 1a and 1ab, which are cleaved to generate 16 "nonstructural" proteins, nsp1 to nsp16, involved in viral replication and/or rna processing. among these, nsp10 plays a critical role in minus-strand rna synthesis in a related coronavirus, murine hepatitis virus. here, we report the crystal structure of sars-cov nsp10 at a re ...200616873246
dodecamer structure of severe acute respiratory syndrome coronavirus nonstructural protein nsp10.the severe acute respiratory syndrome coronavirus (sars-cov) nonstructural proteins nsp1 to nsp16 have been implicated by genetic analysis in the assembly of a functional replication/transcription complex. we report the crystal structure of nsp10 from sars-cov at 2.1-a resolution. the nsp10 structure has a novel fold, and 12 identical subunits assemble to form a unique spherical dodecameric architecture. two zinc fingers have been identified from the nsp10 monomer structure with the sequence mot ...200616873247
supramolecular architecture of severe acute respiratory syndrome coronavirus revealed by electron cryomicroscopy.coronavirus particles are enveloped and pleomorphic and are thus refractory to crystallization and symmetry-assisted reconstruction. a novel methodology of single-particle image analysis was applied to selected virus features to obtain a detailed model of the oligomeric state and spatial relationships among viral structural proteins. two-dimensional images of the s, m, and n structural proteins of severe acute respiratory syndrome coronavirus and two other coronaviruses were refined to a resolut ...200616873249
synthesis of stilbene derivatives with inhibition of sars coronavirus replication.stilbene derivatives have wide range of activities. in an effort to find other potential activities of this kind of compounds, 17 derivatives, including resveratrol, were synthesized. twelve of them were evaluated for their antiviral potential against severe acute respiratory syndrome (sars)-cov-induced cytopathicity in vero e6 cell culture. the result showed that sars virus was totally inhibited by compounds 17 and 19 (<or=0.5 mg ml(-1)) and no significant cytotoxic effects were observed in vit ...200616875760
the human severe acute respiratory syndrome coronavirus (sars-cov) 8b protein is distinct from its counterpart in animal sars-cov and down-regulates the expression of the envelope protein in infected cells.the severe acute respiratory syndrome coronavirus (sars-cov), isolated from humans infected during the peak of epidemic, encodes two accessory proteins termed as 8a and 8b. interestingly, the sars-cov isolated from animals contains an extra 29-nucleotide in this region such that these proteins are fused to become a single protein, 8ab. here, we compared the cellular properties of the 8a, 8b and 8ab proteins by examining their cellular localizations and their abilities to interact with other sars ...200616876844
crystal structure and mechanistic determinants of sars coronavirus nonstructural protein 15 define an endoribonuclease family.the approximately 30-kb coronavirus (+)rna genome is replicated and transcribed by a membrane-bound replicase complex made up of 16 viral nonstructural proteins (nsp) with multiple enzymatic activities. the complex includes an rna endonuclease, nendou, that is conserved among nidoviruses but no other rna virus, making it a genetic marker of this virus order. nendou (nsp15) is a mn(2+)-dependent, uridylate-specific enzyme, which leaves 2'-3'-cyclic phosphates 5' to the cleaved bond. neither bioch ...200616882730
use of human nasal cannulas during bronchoscopy procedures as a simple method for maintaining adequate oxygen saturation in pigtailed macaques (macaca nemestrina).rising concerns over respiratory illnesses caused by agents such as avian influenza viruses and sars coronavirus have prompted intensive research efforts and the resurgence of nonhuman primates as models for these human diseases. in the context of studying influenza infection and vaccine development, serial bronchoscopic procedures, including bronchial brush biopsies and bronchoalveolar lavage, were performed in pigtailed macaques (macaca nemestrina). the possible need for oxygen supplementation ...200616884179
use of human nasal cannulas during bronchoscopy procedures as a simple method for maintaining adequate oxygen saturation in pigtailed macaques (macaca nemestrina).rising concerns over respiratory illnesses caused by agents such as avian influenza viruses and sars coronavirus have prompted intensive research efforts and the resurgence of nonhuman primates as models for these human diseases. in the context of studying influenza infection and vaccine development, serial bronchoscopic procedures, including bronchial brush biopsies and bronchoalveolar lavage, were performed in pigtailed macaques (macaca nemestrina). the possible need for oxygen supplementation ...200616884179
synthesis, crystal structure, structure-activity relationships, and antiviral activity of a potent sars coronavirus 3cl protease inhibitor.a potent sars coronavirus (cov) 3cl protease inhibitor (tg-0205221, ki = 53 nm) has been developed. tg-0205221 showed remarkable activity against sars cov and human coronavirus (hcov) 229e replications by reducing the viral titer by 4.7 log (at 5 microm) for sars cov and 5.2 log (at 1.25 microm) for hcov 229e. the crystal structure of tg-0205221 (resolution = 1.93 a) has revealed a unique binding mode comprising a covalent bond, hydrogen bonds, and numerous hydrophobic interactions. structural c ...200616884309
can herbal medicine assist against avian flu? learning from the experience of using supplementary treatment with chinese medicine on sars or sars-like infectious disease in 2003. 200616884338
screening and identification of severe acute respiratory syndrome-associated coronavirus-specific ctl epitopes.severe acute respiratory syndrome (sars) is a highly contagious and life-threatening disease that emerged in china in november 2002. a novel sars-associated coronavirus was identified as its principal etiologic agent; however, the immunopathogenesis of sars and the role of special ctls in virus clearance are still largely uncharacterized. in this study, potential hla-a*0201-restricted spike (s) and nucleocapsid protein-derived peptides were selected from an online database and screened for poten ...200616887973
living donor renal transplantation using alemtuzumab induction and tacrolimus monotherapy.alemtuzumab was used as an induction agent in 205 renal transplant recipients undergoing 207 living donor renal transplants. all donor kidneys were recovered laparoscopically. postoperatively, patients were treated with tacrolimus monotherapy, and immunosuppression was weaned when possible. forty-seven recipients of living donor renal transplants prior to the induction era who received conventional triple drug immunosuppression without antibody induction served as historic controls. the mean fol ...200616889606
rewiring the severe acute respiratory syndrome coronavirus (sars-cov) transcription circuit: engineering a recombination-resistant genome.live virus vaccines provide significant protection against many detrimental human and animal diseases, but reversion to virulence by mutation and recombination has reduced appeal. using severe acute respiratory syndrome coronavirus as a model, we engineered a different transcription regulatory circuit and isolated recombinant viruses. the transcription network allowed for efficient expression of the viral transcripts and proteins, and the recombinant viruses replicated to wt levels. recombinant ...200616891412
multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus rna in stool specimens.the emergence of a novel coronavirus (cov) as the cause of severe acute respiratory syndrome (sars) catalyzed the development of rapid diagnostic tests. stool samples have been shown to be appropriate for diagnostic testing for sars cov, although it has been recognized to be a heterogeneous and difficult sample that contains amplification inhibitors. limited information on the efficiency of extraction methods for the purification and concentration of sars cov rna from stool samples is available. ...200616891478
human neutralizing fab molecules against severe acute respiratory syndrome coronavirus generated by phage display.human recombinant fab fragments specific for the spike protein of severe acute respiratory syndrome coronavirus (sars-cov) were screened from a human fab library, which was generated from rnas from peripheral lymphocytes of convalescent sars patients. among 50 randomly picked clones, 12 fabs specially reacted with s protein by an enzyme-linked immunosorbent assay. the microneutralizing test showed that one clone, designated m1a, had neutralizing activity on vero e6 cells against sars-cov. dna se ...200616893997
severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release.fourteen orfs have been identified in the severe acute respiratory syndrome-associated coronavirus (sars-cov) genome. orf 3a of sars-cov codes for a recently identified transmembrane protein, but its function remains unknown. in this study we confirmed the 3a protein expression and investigated its localization at the surface of sars-cov-infected or 3a-cdna-transfected cells. our experiments showed that recombinant 3a protein can form a homotetramer complex through interprotein disulfide bridges ...200616894145
development of a red-shifted fluorescence-based assay for sars-coronavirus 3cl protease: identification of a novel class of anti-sars agents from the tropical marine sponge axinella corrugata.sars-coronavirus (sars-cov) encodes a main protease, 3clpro, which plays an essential role in the viral life cycle and is currently the prime target for discovering new anti-coronavirus agents. in this article, we report our success in developing a novel red-shifted (rs) fluorescence-based assay for 3clpro and its application for identifying small-molecule anti-sars agents from marine organisms. we have synthesised and characterised the first generation of a red-shifted internally quenched fluor ...200616895476
the structure of the endoribonuclease xendou: from small nucleolar rna processing to severe acute respiratory syndrome coronavirus replication.small nucleolar rnas (snornas) play a key role in eukaryotic ribosome biogenesis. in most cases, snornas are encoded in introns and are released through the splicing reaction. some snornas are, instead, produced by an alternative pathway consisting of endonucleolytic processing of pre-mrna. xendou, the endoribonuclease responsible for this activity, is a u-specific, metal-dependent enzyme that releases products with 2'-3' cyclic phosphate termini. xendou is broadly conserved among eukaryotes, an ...200616895992
congruent epidemic models for unstructured and structured populations: analytical reconstruction of a 2003 sars outbreak.both the threat of bioterrorism and the natural emergence of contagious diseases underscore the importance of quantitatively understanding disease transmission in structured human populations. over the last few years, researchers have advanced the mathematical theory of scale-free networks and used such theoretical advancements in pilot epidemic models. scale-free contact networks are particularly interesting in the realm of mathematical epidemiology, primarily because these networks may allow m ...200616904134
copious production of sars-cov nucleocapsid protein employing codon optimized synthetic gene.the severe acute respiratory syndrome coronavirus (sars-cov) nucleocapsid protein (np) is one of the predominant antigenic protein and the most abundant shed antigen throughout the sars-cov infection. this feature makes it a suitable molecular target for diagnostic applications. in this study the full length codon optimized np gene and its subfragment gene segment was cloned in a bacterial expression vector. the full length np could be expressed in e. coli at very high level within inclusion bod ...200616904198
factors associated with nosocomial sars-cov transmission among healthcare workers in hanoi, vietnam, 2003.in march of 2003, an outbreak of severe acute respiratory syndrome (sars) occurred in northern vietnam. this outbreak began when a traveler arriving from hong kong sought medical care at a small hospital (hospital a) in hanoi, initiating a serious and substantial transmission event within the hospital, and subsequent limited spread within the community.200616907978
severe acute respiratory syndrome coronavirus nsp1 protein suppresses host gene expression by promoting host mrna degradation.severe acute respiratory syndrome (sars) coronavirus (scov) causes a recently emerged human disease associated with pneumonia. the 5' end two-thirds of the single-stranded positive-sense viral genomic rna, gene 1, encodes 16 mature proteins. expression of nsp1, the most n-terminal gene 1 protein, prevented sendai virus-induced endogenous ifn-beta mrna accumulation without inhibiting dimerization of ifn regulatory factor 3, a protein that is essential for activation of the ifn-beta promoter. furt ...200616912115
structural and functional basis for adp-ribose and poly(adp-ribose) binding by viral macro domains.macro domains constitute a protein module family found associated with specific histones and proteins involved in chromatin metabolism. in addition, a small number of animal rna viruses, such as corona- and toroviruses, alphaviruses, and hepatitis e virus, encode macro domains for which, however, structural and functional information is extremely limited. here, we characterized the macro domains from hepatitis e virus, semliki forest virus, and severe acute respiratory syndrome coronavirus (sars ...200616912299
highly conserved regions within the spike proteins of human coronaviruses 229e and nl63 determine recognition of their respective cellular receptors.we have recently demonstrated that the severe acute respiratory syndrome coronavirus (sars-cov) receptor angiotensin converting enzyme 2 (ace2) also mediates cellular entry of the newly discovered human coronavirus (hcov) nl63. here, we show that expression of dc-sign augments nl63 spike (s)-protein-driven infection of susceptible cells, while only expression of ace2 but not dc-sign is sufficient for entry into nonpermissive cells, indicating that ace2 fulfills the criteria of a bona fide hcov-n ...200616912312
structure-based drug design and structural biology study of novel nonpeptide inhibitors of severe acute respiratory syndrome coronavirus main protease.severe acute respiratory syndrome coronavirus (sars-cov) main protease (m(pro)), a protein required for the maturation of sars-cov, is vital for its life cycle, making it an attractive target for structure-based drug design of anti-sars drugs. the structure-based virtual screening of a chemical database containing 58,855 compounds followed by the testing of potential compounds for sars-cov m(pro) inhibition leads to two hit compounds. the core structures of these two hits, defined by the docking ...200616913704
molecular diagnosis of severe acute respiratory syndrome.the etiologic agent of severe acute respiratory syndrome (sars) has been identified as a new type of coronavirus, known as sars-coronavirus (sars-cov). although the sars epidemic has subsided, many authorities, including the world health organization (who) and the centers for disease control and prevention (cdc), have warned of the possible re-emergence of this highly infectious disease. although antibody-based diagnosis of sars has been demonstrated to be a reliable proof of sars infection, it ...200616916262
genomic sequencing of the severe acute respiratory syndrome-coronavirus.the polymerase chain reaction (pcr), which can exponentially replicate a target dna sequence, has formed the basis for the sensitive and direct examination of clinical samples for evidence of infection. during the epidemic of severe acute respiratory syndrome (sars) in 2003, pcr not only offered a rapid way to diagnose sars-coronavirus (sars-cov) infection, but also made the molecular analysis of its genomic sequence possible. sequence variations were observed in the sar-cov obtained from differ ...200616916263
a convenient cell fusion assay for the study of sars-cov entry and inhibition.sars-cov spike (s) protein-mediated cell fusion is important for the viral entry mechanism and identification of sars-cov entry inhibitors. in order to avoid the high risks involved in handling sars-cov and to facilitate the study of viral fusion mechanism, we established the cell lines: sr-cos7 cells that stably express both sars-cov s protein and red fluorescence protein, r-cos7 cells that stably express red fluorescence protein, and ag-cos7 cells that stably express both ace2 and green fluore ...200616916786
recombinant adeno-associated virus expressing the receptor-binding domain of severe acute respiratory syndrome coronavirus s protein elicits neutralizing antibodies: implication for developing sars vaccines.development of an effective vaccine for severe acute respiratory syndrome (sars) remains to be a priority to prevent possible re-emergence of sars coronavirus (sars-cov). we previously demonstrated that the receptor-binding domain (rbd) of sars-cov s protein is a major target of neutralizing antibodies. this suggests that the rbd may serve as an ideal vaccine candidate. recombinant adeno-associated virus (raav) has been proven to be an effective system for gene delivery and vaccine development. ...200616793110
generality of the final size formula for an epidemic of a newly invading infectious disease.the well-known formula for the final size of an epidemic was published by kermack and mckendrick in 1927. their analysis was based on a simple susceptible-infected-recovered (sir) model that assumes exponentially distributed infectious periods. more recent analyses have established that the standard final size formula is valid regardless of the distribution of infectious periods, but that it fails to be correct in the presence of certain kinds of heterogeneous mixing (e.g., if there is a core gr ...200616794950
human monoclonal antibody combination against sars coronavirus: synergy and coverage of escape mutants.experimental animal data show that protection against severe acute respiratory syndrome coronavirus (sars-cov) infection with human monoclonal antibodies (mabs) is feasible. for an effective immune prophylaxis in humans, broad coverage of different strains of sars-cov and control of potential neutralization escape variants will be required. combinations of virus-neutralizing, noncompeting mabs may have these properties.200616796401
accelerated induction of apoptosis in insect cells by baculovirus-expressed sars-cov membrane protein.it has been shown that severe acute respiratory syndrome-associated coronavirus (sars-cov) 3a and 7a proteins, but not membrane (m) protein, induce apoptosis in mammalian cells. upon expression of sars-cov m protein using the baculovirus/insect cell expression system, however, we found that the expressed m protein triggered accelerated apoptosis in insect cells, as characterized by rapid cell death, elevated cytotoxicity, cell shrinkage, nuclear condensation and dna fragmentation. conversely, th ...200616797548
preparation of a chimeric armored rna as a versatile calibrator for multiple virus assays. 200616798980
nmr assignment of the protein nsp3a from sars-cov. 200616799860
hosting the severe acute respiratory syndrome coronavirus: specific cell factors required for infection.as with all viruses, the severe acute respiratory syndrome coronavirus (sars-cov) utilizes specific host cell factors during its infection cycle. some of these factors have been identified and are now increasingly scrutinized as targets to intervene with infection. in this brief review, we describe the current understanding of how the sars-cov is able to use the cellular machinery for its replication.200616803585
severe acute respiratory syndrome (sars): development of diagnostics and antivirals.the previously unknown coronavirus that caused severe acute respiratory syndrome (sars-cov) affected more than 8,000 persons worldwide and was responsible for more than 700 deaths during the first outbreak in 2002-2003. for reasons unknown, the sars virus is less severe and the clinical progression a great deal milder in children younger than 12 years of age. in contrast, the mortality rate can exceed 50% for persons at or above the age of 60. as part of the sino-european project on sars diagnos ...200616804033
severe acute respiratory syndrome in a naval diver.severe acute respiratory syndrome (sars) is a highly infectious, rapidly progressive, emerging disease. early diagnosis and preventive measures are key for treatment and minimization of secondary spread. in the context of the armed forces, aggressive containment measures are essential to prevent an outbreak. in this study, we present the first reported case, to our knowledge, of sars in a naval diver. the special physical requirements for divers and the potential complications associated with de ...200616808126
mechanisms of establishment of persistent sars-cov-infected cells.previously, we reported the establishment of cells with persistent sars-cov infection after apoptotic events and showed that both jnk and pi3k/akt signaling pathways are important for persistence by treatment with inhibitors at the early stages of sars-cov infection. however, the mechanisms of establishment of persistent infection are still unclear. in this study, we investigated which signaling pathways play important roles in escape from apoptosis in cells infected with sars-cov. in persistent ...200616808902
conformational states of the severe acute respiratory syndrome coronavirus spike protein ectodomain.the severe acute respiratory syndrome coronavirus enters cells through the activities of a spike protein (s) which has receptor-binding (s1) and membrane fusion (s2) regions. we have characterized four sequential states of a purified recombinant s ectodomain (s-e) comprising s1 and the ectodomain of s2. they are s-e monomers, uncleaved s-e trimers, cleaved s-e trimers, and dissociated s1 monomers and s2 trimer rosettes. lowered ph induces an irreversible transition from flexible, l-shaped s-e mo ...200616809285
enhanced induction of sars-cov nucleocapsid protein-specific immune response using dna vaccination followed by adenovirus boosting in balb/c mice.to investigate immunogenicity in the induction of humoral and cellular immune responses to genetic vaccines of the recombinant severe acute respiratory syndrome-associated coronavirus (sars-cov)-n gene expressing the same protein plasmid, pcdna3.1-n, and replication-defective adenoviral vector, rad-n, in a pcdna3.1-n prime-rad-n boost regimen and the reverse sequence in a rad-n prime-pcdna3.1-n boost regimen.200616809936
hexamethylene amiloride blocks e protein ion channels and inhibits coronavirus replication.all coronaviruses encode a small hydrophobic envelope (e) protein, which mediates viral assembly and morphogenesis by an unknown mechanism. we have previously shown that the e protein from severe acute respiratory syndrome coronavirus (sars-cov) forms cation-selective ion channels in planar lipid bilayers (wilson, l., mckinlay, c., gage, p., ewart, g., 2004. sars coronavirus e protein forms cation-selective ion channels. virology 330(1), 322-331). we now report that three other e proteins also f ...200616815524
[establishment of molecular differential diagnostic (mdd) technology for detection of common respiratory viruses].to provide rapid laboratory evidence for diagnosis of respiratory infection and help diagnose accurately and reduce the spread of disease, so that the patients can be diagnosed and treated early.200616816854
[follow up study on viruses associated with sars among the sars patients].to study the existence status of the sars-cov, retrovirus, and the poliovirus in the bodies of the patients with sars and the possible relationship between the three viruses and sars.200616816874
understanding the accessory viral proteins unique to the severe acute respiratory syndrome (sars) coronavirus.a novel coronavirus, termed the severe acute respiratory syndrome coronavirus (sars-cov), infected humans in guangdong, china, in november 2002 and the subsequent efficient human-to-human transmissions of this virus caused profound disturbances in over 30 countries worldwide in 2003. eventually, this epidemic was controlled by isolation and there has been no human infection reported since january 2004. however, research on different aspects of the sars-cov is not waning, as it is not known if th ...200616820226
nmr assignment of the sars-cov protein nsp1. 200616821128
substantial differences in preparedness for emergency infection control measures among major hospitals in japan: lessons from sars.emergency infection control measures are essential in hospitals. although japan was spared from the 2003 epidemic of severe acute respiratory syndrome (sars), hospitals were placed on high alert. the actual preparedness level of hospitals can be determined by examining individual perceptions among the hospital healthcare workers (hcws). the objective of this study was to assess the level of preparedness of emergency infection control measures in japan and to quantify the differences in preparedn ...200616826344
rna recognition and cleavage by the sars coronavirus endoribonuclease.the emerging disease sars is caused by a novel coronavirus that encodes several unusual rna-processing enzymes, including non-structural protein 15 (nsp15), a hexameric endoribonuclease that preferentially cleaves at uridine residues. how nsp15 recognizes and cleaves rna is not well understood and is the subject of this study. based on the analysis of rna products separated by denaturing gel electrophoresis, nsp15 has been reported to cleave both 5' and 3' of the uridine. we used several rnas, i ...200616828802
sars in singapore--predictors of disease severity.severe acute respiratory syndrome (sars) affected 8096 individuals in 29 countries, with 774 deaths. in singapore, there were 238 cases of sars with 33 deaths. a retrospective analysis was performed to identify predictors of poor outcome in patients with sars locally.200616829999
clinical and laboratory findings of sars in singapore.singapore was one of 29 countries worldwide affected by severe acute respiratory syndrome (sars) in 2003.200616830000
laboratory containment of sars virus.following the severe acute respiratory syndrome (sars) outbreak in 2003, a large number of clinical and environmental samples containing/potentially containing sars coronavirus (sarscov) as well as sars-cov stocks were retained in clinical and research laboratories. the importance of laboratory biosafety was demonstrated by the occurrence of laboratory incidents in singapore, taiwan and beijing. it is imperative that safe practice and techniques, safety equipment and appropriate facility design ...200616830004
[my greatest concern was decontaminating my family]. 200616836185
photochemical treatment of plasma with amotosalen and long-wavelength ultraviolet light inactivates pathogens while retaining coagulation function.the intercept blood system, a photochemical treatment (pct) process, has been developed to inactivate pathogens in platelet concentrates. these studies evaluated the efficacy of pct to inactivate pathogens in plasma and the effect of pct on plasma function.200616836564
coronaviruses and their therapy.coronaviruses may cause respiratory, enteric and central nervous system diseases in many species, including humans. until recently, the relatively low burden of disease in humans caused by few of these viruses hampered development of coronavirus specific therapeutics. however, the emergence of severe acute respiratory syndrome coronavirus (sars-cov) has prompted the discovery of such drugs. subsequent studies in animal models demonstrated the efficacy of sars-cov specific monoclonal antibodies, ...200616837072
severe acute respiratory syndrome coronavirus 7a accessory protein is a viral structural protein.severe acute respiratory syndrome coronavirus (scov) 7a protein is one of the viral accessory proteins. in expressing cells, 7a protein exhibits a variety of biological activities, including induction of apoptosis, activation of the mitogen-activated protein kinase signaling pathway, inhibition of host protein translation, and suppression of cell growth progression. analysis of scov particles that were purified by either sucrose gradient equilibrium centrifugation or a virus capture assay, in wh ...200616840309
antiviral drug discovery against sars-cov.severe acute respiratory syndrome (sars) is a life-threatening infectious disease caused by sars-cov. in the 2003 outbreak, it infected more than 8,000 people worldwide and claimed the lives of more than 900 victims. the high mortality rate resulted, at least in part, from the absence of definitive treatment protocols or therapeutic agents. although the virus spreading has been contained, due preparedness and planning, including the successful development of antiviral drugs against sars-cov, is ...200616842194
peptides derived from hiv-1, hiv-2, ebola virus, sars coronavirus and coronavirus 229e exhibit high affinity binding to the formyl peptide receptor.peptides derived from the membrane proximal region of fusion proteins of human immunodeficiency viruses 1 and 2, coronavirus 229 e, severe acute respiratory syndrome coronavirus and ebola virus were all potent antagonists of the formyl peptide receptor expressed in chinese hamster ovary cells. binding of viral peptides was affected by the naturally occurring polymorphisms at residues 190 and 192, which are located at second extracellular loop-transmembrane helix 5 interface. substitution of r190 ...200616842982
peptides derived from hiv-1, hiv-2, ebola virus, sars coronavirus and coronavirus 229e exhibit high affinity binding to the formyl peptide receptor.peptides derived from the membrane proximal region of fusion proteins of human immunodeficiency viruses 1 and 2, coronavirus 229 e, severe acute respiratory syndrome coronavirus and ebola virus were all potent antagonists of the formyl peptide receptor expressed in chinese hamster ovary cells. binding of viral peptides was affected by the naturally occurring polymorphisms at residues 190 and 192, which are located at second extracellular loop-transmembrane helix 5 interface. substitution of r190 ...200616842982
microwave-assisted tissue processing for same-day em-diagnosis of potential bioterrorism and clinical samples.the purpose of this study was to explore the turnaround times, section and image quality of a number of more "difficult" specimens destined for rapid diagnostic electron microscopy (em) after microwave-assisted processing. the results were assessed and compared with those of conventionally processed samples. a variety of infectious agents, some with a potential for bioterrorism, and liver biopsies serving as an example for routine histopathology samples were studied. the samples represented viru ...200616843832
common scaffolds, diverse recognition profiles. 200616843888
domain-swapped structure of the potent antiviral protein griffithsin and its mode of carbohydrate binding.the crystal structure of griffithsin, an antiviral lectin from the red alga griffithsia sp., was solved and refined at 1.3 a resolution for the free protein and 0.94 a for a complex with mannose. griffithsin molecules form a domain-swapped dimer, in which two beta strands of one molecule complete a beta prism consisting of three four-stranded sheets, with an approximate 3-fold axis, of another molecule. the structure of each monomer bears close resemblance to jacalin-related lectins, but its dim ...200616843894
the crystal structure of orf-9b, a lipid binding protein from the sars coronavirus.to achieve the greatest output from their limited genomes, viruses frequently make use of alternative open reading frames, in which translation is initiated from a start codon within an existing gene and, being out of frame, gives rise to a distinct protein product. these alternative protein products are, as yet, poorly characterized structurally. here we report the crystal structure of orf-9b, an alternative open reading frame within the nucleocapsid (n) gene from the sars coronavirus. the prot ...200616843897
human memory t cell responses to sars-cov e protein.e protein is a membrane component of severe acute respiratory syndrome coronavirus (sars-cov). disruption of e protein may reduce viral infectivity. thus, the sars-cov e protein is considered a potential target for the development of antiviral drugs. however, the cellular immune responses to e protein remain unclear in humans. in this study, we found that peripheral blood mononuclear cells (pbmcs) from fully recovered sars individuals rapidly produced ifn-gamma and il-2 following stimulation wit ...200616844400
sivirus: web-based antiviral sirna design software for highly divergent viral sequences.sivirus (http://sivirus.rnai.jp/) is a web-based online software system that provides efficient short interfering rna (sirna) design for antiviral rna interference (rnai). sivirus searches for functional, off-target minimized sirnas targeting highly conserved regions of divergent viral sequences. these sirnas are expected to resist viral mutational escape, since their highly conserved targets likely contain structurally/functionally constrained elements. sivirus will be a useful tool for designi ...200616845046
architecture of the sars coronavirus prefusion spike.the emergence in 2003 of a new coronavirus (cov) responsible for the atypical pneumonia termed severe acute respiratory syndrome (sars) was a stark reminder that hitherto unknown viruses have the potential to cross species barriers to become new human pathogens. here we describe the sars-cov 'spike' structure determined by single-particle cryo-em, along with the docked atomic structures of the receptor-binding domain and prefusion core.200616845391
m and n proteins of sars coronavirus induce apoptosis in hpf cells.sars-associated coronavirus (sars-cov) induced cell apoptosis and its structural proteins may play a role in this process.200616845612
interferon-gamma and interleukin-4 downregulate expression of the sars coronavirus receptor ace2 in vero e6 cells.interferons (ifns) inhibit severe acute respiratory syndrome coronavirus (sars-cov) replication and might be valuable for sars treatment. in this study, we demonstrate that treatment of vero e6 cells with interleukin-4 (il-4) decreased the susceptibility of these cells to sars-cov infection. in contrast to ifns, il-4 did not show antiviral activity when administered immediately after sars-cov infection, suggesting that il-4 acts early during the sars-cov replication cycle. indeed, binding of rec ...200616860835
[an epidemiological investigation of bats carrying sars-cov in guangzhou and its vicinity].to detect serve acute respiratory syndrome-associated coronavirus (sars-cov) and sars-like-cov in fruit bats captured in guangzhou and its vicinity.200616864084
rnai therapeutics: can sirnas conquer sars? 200617262907
comparative immunization in balb/c mice with recombinant replication-defective adenovirus vector and dna plasmid expressing a sars-cov nucleocapsid protein gene.in order to investigate immunogenicity in the induction of humoral and cellular immune responses, severe acute respiratory syndrome associated coronavirus (sars-cov)-n gene recombinant replication-defective adenoviral vector, rad-n, was generated and immunized balb/c mice in a pcdna3.1-n prime-rad-n boost regimen. after humoral and cellular immune response detection, different levels of sars-cov n protein specific antibodies and interferon-gamma (ifn-gamma) secretion are shown compared to contro ...200617257500
vaccine efficacy in senescent mice challenged with recombinant sars-cov bearing epidemic and zoonotic spike variants.in 2003, severe acute respiratory syndrome coronavirus (sars-cov) was identified as the etiological agent of severe acute respiratory syndrome, a disease characterized by severe pneumonia that sometimes results in death. sars-cov is a zoonotic virus that crossed the species barrier, most likely originating from bats or from other species including civets, raccoon dogs, domestic cats, swine, and rodents. a sars-cov vaccine should confer long-term protection, especially in vulnerable senescent pop ...200617194199
vaccine efficacy in senescent mice challenged with recombinant sars-cov bearing epidemic and zoonotic spike variants.in 2003, severe acute respiratory syndrome coronavirus (sars-cov) was identified as the etiological agent of severe acute respiratory syndrome, a disease characterized by severe pneumonia that sometimes results in death. sars-cov is a zoonotic virus that crossed the species barrier, most likely originating from bats or from other species including civets, raccoon dogs, domestic cats, swine, and rodents. a sars-cov vaccine should confer long-term protection, especially in vulnerable senescent pop ...200617194199
long-term persistence of robust antibody and cytotoxic t cell responses in recovered patients infected with sars coronavirus.most of the individuals infected with sars coronavirus (sars-cov) spontaneously recovered without clinical intervention. however, the immunological correlates associated with patients' recovery are currently unknown. in this report, we have sequentially monitored 30 recovered patients over a two-year period to characterize temporal changes in sars-cov-specific antibody responses as well as cytotoxic t cell (ctl) responses. we have found persistence of robust antibody and ctl responses in all of ...200617183651
baculovirus surface display of sars coronavirus (sars-cov) spike protein and immunogenicity of the displayed protein in mice models.the baculovirus surface display technique has provided an ideal tool to display foreign proteins with natural conformation, functions, and immunogenicity. in this work, we explored the application of this technique on sars-associated coronavirus (sars-cov) spike (s) protein, and further analyzed the immunogenicity of displayed s protein. the entire ectodomain of s protein was fused between the gp64 signal peptide and the vsv-g membrane anchor and successfully displayed on the baculovirus surface ...200617184168
public health awareness of emerging zoonotic viruses of bats: a european perspective.bats classified in the order chiroptera are the most abundant and widely distributed non-human mammalian species in the world. several bat species are reservoir hosts of zoonotic viruses and therefore can be a public health hazard. lyssaviruses of different genotypes have emerged from bats in america (genotype 1 rabies virus; rabv), europe (european bat lyssavirus; eblv), and australia (australian bat lyssavirus; ablv), whereas nipah virus is the most important recent zoonosis of bat origin in a ...200617187565
public health awareness of emerging zoonotic viruses of bats: a european perspective.bats classified in the order chiroptera are the most abundant and widely distributed non-human mammalian species in the world. several bat species are reservoir hosts of zoonotic viruses and therefore can be a public health hazard. lyssaviruses of different genotypes have emerged from bats in america (genotype 1 rabies virus; rabv), europe (european bat lyssavirus; eblv), and australia (australian bat lyssavirus; ablv), whereas nipah virus is the most important recent zoonosis of bat origin in a ...200617187565
development of a method for bacteria and virus recovery from heating, ventilation, and air conditioning (hvac) filters.the aim of the work presented here is to study the effectiveness of building air handling units (ahus) in serving as high volume sampling devices for airborne bacteria and viruses. an hvac test facility constructed according to ashrae standard 52.2-1999 was used for the controlled loading of hvac filter media with aerosolized bacteria and virus. nonpathogenic bacillus subtilis var. niger was chosen as a surrogate for bacillus anthracis. three animal viruses; transmissible gastroenteritis virus ( ...200617240906
an overall picture of sars coronavirus (sars-cov) genome-encoded major proteins: structures, functions and drug development.a severe atypical pneumonia designated as severe acute respiratory syndrome (sars) by the world health organization broke out in china and menaced to more than other 30 countries between the end of the year 2002 and june of the year 2003. a novel coronavirus called severe acute respiratory syndrome coronavirus (sars-cov) has been recently identified as the etiological agent responsible for the infectious sars disease. based on extensively scientific cooperation and almost two-year's studies, rem ...200617168760
quaternary structure, substrate selectivity and inhibitor design for sars 3c-like proteinase.the sars coronavirus 3c-like proteinase is recognized as a potential drug design target for the treatment of severe acute respiratory syndrome. in the past few years, much work has been done to understand the catalytic mechanism of this target protein and to design its selective inhibitors. the protein exists as a dimer/monomer mixture in solution and the dimer was confirmed to be the active species for the enzyme reaction. quantitative dissociation constants have been reported for the dimer by ...200617168761
bioinformatics research on the sars coronavirus (sars_cov) in china.severe acute respiratory syndrome (sars) first appeared in 2002 in china, which fastly affected about 8000 patients over 29 countries and caused 774 fatalities. as its pathogen was identified as a new kind of coronavirus (sars_cov), its genome was quickly sequenced on several isolates. studies on its functional genomics were performed by combinatorial application of all the available bioinformatics tools and the development of new programs. in this way, it was found that the four proteins were a ...200617168762
drug design targeting the main protease, the achilles' heel of coronaviruses.coronaviruses (covs), a genus containing about 26 known species to date, cause highly prevalent diseases and are often severe or fatal in humans and animals. in 2003, a previously unknown coronavirus was identified to be the etiological agent of a global outbreak of a form of life-threatening pneumonia called severe acute respiratory syndrome (sars). no efficacious therapy is currently available, and vaccines and drugs are under development to prevent sars-cov infection in many countries. the co ...200617168763
progress in computational approach to drug development against sars.since the outbreak of sars (severe acute respiratory syndrome) in november 2002 in southern china's guangdong province, considerable progress has been made in the development of drugs for sars therapy. the present mini review is focused on the area of computer-aided drug discovery, i.e., the advances achieved mainly from the approaches of structural bioinformatics, pharmacophore modeling, molecular docking, peptide-cleavage site prediction, and other computational means. it is highlighted that t ...200617168850
[severe acute respiratory syndrome (sars)].the sudden appearance of the severe acute respiratory syndrome (sars) in 2003 demonstrated to the world at large that despite the high standard of medical care in affected countries, a highly contagious emerging infectious disease could spread rapidly worldwide. by application and improvement of stringent infection control measures, a further spread of sars could be stopped and the disease could so far be defeated. in the meantime, decisive progress in the knowledge about the structure and furth ...200617171319
core structure of s2 from the human coronavirus nl63 spike glycoprotein.human coronavirus nl63 (hcov-nl63) has recently been identified as a causative agent of acute respiratory tract illnesses in infants and young children. the hcov-nl63 spike (s) protein mediates virion attachment to cells and subsequent fusion of the viral and cellular membranes. this viral entry process is a primary target for vaccine and drug development. hcov-nl63 s is expressed as a single-chain glycoprotein and consists of an n-terminal receptor-binding domain (s1) and a c-terminal transmemb ...200617176042
evaluation of immunomodulators, interferons and known in vitro sars-cov inhibitors for inhibition of sars-cov replication in balb/c mice.compounds approved for therapeutic use and in vitro inhibitors of severe acute respiratory syndrome coronavirus (sars-cov) were evaluated for inhibition in the mouse sars-cov replication model. a hybrid interferon, interferon alpha (ifn-alpha) b/d, and a mismatched double-stranded (ds) rna interferon (ifn) inducer, ampligen (poly i:poly c124), were the only compounds that potently inhibited virus titres in the lungs of infected mice as assessed by cpe titration assays. when mice were dosed intra ...200617176632
antiviral activity of nucleoside analogues against sars-coronavirus (sars-cov).the recent outbreak of severe acute respiratory syndrome (sars), which is an acute respiratory illness, is caused by newly discovered sars coronavirus (sars-cov). herein we describe the antiviral activity of several classes of nucleoside analogues evaluated against sars-cov in vero 76 cells, some of which exhibited moderate activity.200617176633
coupling molecular beacons to barcoded metal nanowires for multiplexed, sealed chamber dna bioassays.we have combined molecular beacon (mb) probes with barcoded metal nanowires to enable no-wash, sealed chamber, multiplexed detection of nucleic acids. probe design and experimental parameters important in nanowire-based mb assays are discussed. loop regions of 24 bases and 5 base pair stem regions in the beacon probes gave optimal performance. our results suggest that thermodynamic predictions for secondary structure stability of solution-phase mb can guide probe design for nanowire-based assays ...200617177440
[severe acute respiratory syndrome].towards the end of 2002 it was the epidemic'outbreak of very severe atypical pneumonia of unknown cause. 32 countries were affected. during 7 months about 8000 cases and 900 fatalities were reported. world health organization defined the new disease as severe acute respiratory syndrome (sars). thanks to the international investigations the etiologic factor of the disease was quickly identified. new coronavirus was described. it allowed the introduction of new more effective methods of disease' i ...200617163178
the lesson of supplementary treatment with chinese medicine on severe laboratory-confirmed sars patients.chinese medicine (cm) has been used to control infectious diseases for thousands of years. in 2003 outbreaks of severe acute respiratory syndrome (sars) occurred in china, hong kong and taiwan. in view of the possible beneficial effect of cm on sars, we conducted this study to examine whether cm is of any benefit as a supplementary treatment of sars. four severe laboratory-confirmed sars patients received routine western-medicine treatment plus different supplementary treatment: cm a, cm b and c ...200617163582
symptom combinations associated with outcome and therapeutic effects in a cohort of cases with sars.severe acute respiratory syndrome (sars) is an infectious disease and some of its symptoms were clinically indistinguishable of those from similar diseases. this study aimed to find the symptom combinations associated with adverse outcome and the therapeutic effects in a cohort of patients with probable sars retrospectively. in 2003, 123 sars cases in beijing were subjected to a strictly western medicine (wm) treatment, or a combined treatment (wm plus herba houttuyniae injection, addition of in ...200617163583
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