Publications

serum proteomic fingerprints of adult patients with severe acute respiratory syndrome.	severe acute respiratory syndrome (sars) is an emerging infectious disease caused by a new coronavirus strain, sars-cov. specific proteomic patterns might be present in serum in response to the infection and could be useful for early detection of the disease.	2006	16423906
nasopharyngeal detection of severe acute respiratory syndrome-associated coronavirus rna in health-care workers.		2006	16424407
colonization of severe acute respiratory syndrome-associated coronavirus among health-care workers screened by nasopharyngeal swab.	to report the efficacy and findings of a large-scale preventive screening program for severe acute respiratory syndrome-associated coronavirus (sars-cov) using amplification of the virus from a nasopharyngeal swab (nps) obtained from the health-care workers (hcws).	2006	16424418
the nucleocapsid protein of severe acute respiratory syndrome-coronavirus inhibits the activity of cyclin-cyclin-dependent kinase complex and blocks s phase progression in mammalian cells.	deregulation of the cell cycle is a common strategy employed by many dna and rna viruses to trap and exploit the host cell machinery toward their own benefit. in many coronaviruses, the nucleocapsid protein (n protein) has been shown to inhibit cell cycle progression although the mechanism behind this is poorly understood. the n protein of severe acute respiratory syndrome-coronavirus (sars-cov) bears signature motifs for binding to cyclin and phosphorylation by cyclin-dependent kinase (cdk) and ...	2006	16431923
amino terminus of the sars coronavirus protein 3a elicits strong, potentially protective humoral responses in infected patients.	the 3a protein of severe acute respiratory syndrome (sars)-associated coronavirus is expressed and transported to the plasma membrane in tissue cells of infected patients. its short n-terminal ectodomain was found to elicit strong humoral responses in half of the patients who had recovered from sars. the ectodomain-specific antibodies from the convalescent-phase plasma readily recognized and induced destruction of 3a-expressing cells in the presence of the human complement system, demonstrating ...	2006	16432024
antiviral applications of rnai for coronavirus.	until the appearance of severe acute respiratory syndrome (sars), caused by the sars coronavirus (sars-cov) in early 2003, coronavirus infection was not considered to be serious enough to be controlled by either vaccination or specific antiviral therapy. it is now believed that the availability of antiviral drugs effective against sars-cov will be crucial for the control of future sars outbreaks. recently, rna interference has been successfully used as a more specific and efficient method for ge ...	2006	16433589
china: open season.		2006	16437080
inhibitory effects of anti-sars traditional chinese medicines on the uv irradiation of lambda-lysogen.	by using lambda-lysogen as a model, the inhibitory effects of anti-severe acute respiratory syndrome (sars) traditional chinese medicines (tcms) prescription i on the uv irradiation were investigated in this present study. it was found that the prescription i possessed obvious inhibitory effects on the uv induction of lambda-lysogen, the inhibitory rate reaching 83.87%. among five medicinal herbs prescribed in that formula, herba patriniae, radix astragali and radix glycyrrhizae played important ...	2006	16437747
clinical and laboratory features in the early stage of severe acute respiratory syndrome.	to characterize the clinical and laboratory features of severe acute respiratory syndrome (sars) in the early stage and to compare them with those of patients initially suspected of having sars who were later determined to have other febrile diseases.	2006	16440123
development of human monoclonal antibodies against diseases caused by emerging and biodefense-related viruses.	polyclonal antibodies have a century-old history of being effective against some viruses; recently, monoclonal antibodies (mabs) have also shown success. the humanized mab synagis (palivizumab), which is still the only mab against a viral disease approved by the us fda, has been widely used as a prophylactic measure against respiratory syncytial virus infections in neonates and immunocompromised individuals. the first fully human mabs against two other paramyxoviruses, hendra and nipah virus, wh ...	2006	16441209
characterization of severe acute respiratory syndrome coronavirus membrane protein.	the coronavirus membrane protein (m) is the key player in the assembly of virions at intracellular membranes between endoplasmic-reticulum and golgi-complex. using a newly established human monoclonal anti-m antibody we detected glycosylated and nonglycosylated membrane-associated m in severe acute respiratory syndrome-associated coronavirus (sars-cov) infected cells and in purified virions. further analyses revealed that m contained a single n-glycosylation site at asparagine 4. recombinant m w ...	2006	16442106
patent pools and diagnostic testing.	there is increasing concern that overlapping patents in the field of genetics will create a costly and legally complex situation known as a patent thicket, which, along with the associated issues of accumulating royalty payments, can act as a disincentive for innovation. one potential means of preventing this is for the patent holders to enter into a so-called patent pool, such as those established in the electronics and telecommunications industries. precedents for these also exist in the field ...	2006	16443296
emerging viral diseases and infectious disease risks.	new pathogens and antimicrobial-resistant forms of older pathogens continue to emerge, some with the potential for rapid, global spread and high morbidity and mortality. pathogens can emerge either through introduction into a new population or when the interaction with the vector changes; emergence is also influenced by microbiological adaptation and change, global travel patterns, domestic and wild animal contact and other variants in human ecology and behaviour. quick, decisive action to detec ...	2006	16445811
synthesis of cyclopentenyl carbocyclic nucleosides as potential antiviral agents against orthopoxviruses and sars.	a practical and convenient methodology for the synthesis of chiral cyclopentenol derivative (+)-12a has been developed as the key intermediate that was utilized for the synthesis of biologically active carbocyclic nucleosides. the selective protection of allylic hydroxyl group followed by the ring-closing metathesis (rcm) reaction with grubbs catalysts provided (+)-12a on a 10 g scale with 52% overall yield from d-ribose (4). the key intermediate (+)-12a was utilized for the synthesis of unnatur ...	2006	16451078
design and synthesis of dipeptidyl glutaminyl fluoromethyl ketones as potent severe acute respiratory syndrome coronovirus (sars-cov) inhibitors.	this paper describes the design and synthesis of dipeptidyl n,n-dimethyl glutaminyl fluoromethyl ketones (fmk) as severe acute respiratory syndrome coronovirus (sars-cov) inhibitors. the compounds were tested against sars-cov-induced cell death in vero or caco2 cells as a measurement of the inhibiting effects of the compounds on the replication of the virus. z-leu-gln(nme(2))-fmk (6a) was found to be a potent inhibitor with low toxicity in cells, protecting cells with an ec(50) value of 2.5 micr ...	2006	16451084
therapy with a severe acute respiratory syndrome-associated coronavirus-neutralizing human monoclonal antibody reduces disease severity and viral burden in golden syrian hamsters.	immunotherapy with monoclonal antibodies (mabs) offers safe interventions for the prevention of infection in patients after organ transplantation and for the treatment of cancers and autoimmune diseases. mab 201 is a severe acute respiratory syndrome-associated coronavirus (sars-cov)-specific mab that prevents establishment of viral replication in vitro and prevents viral replication in vivo when administered prophylactically. the efficacy of mab 201 in the treatment of sars was evaluated in gol ...	2006	16453264
epidemiological and genetic correlates of severe acute respiratory syndrome coronavirus infection in the hospital with the highest nosocomial infection rate in taiwan in 2003.	taiwan experienced a series of outbreaks of nosocomial severe acute respiratory syndrome (sars) infections in 2003. two months after the final outbreak, we recruited 658 employees from the hospital that suffered the first and most severe sars infections to help us investigate epidemiological and genetic factors associated with the sars coronavirus (sars-cov). sars-cov infections were detected by using enzyme immunoassays and confirmed by a combination of western blot assays, neutralizing antibod ...	2006	16455884
regulation of p90rsk phosphorylation by sars-cov infection in vero e6 cells.	the 90 kda ribosomal s6 kinases (p90rsks) are a family of broadly expressed serine/threonine kinases with two kinase domains activated by extracellular signal-regulated protein kinase in response to many growth factors. our recent study demonstrated that severe acute respiratory syndrome (sars)-coronavirus (cov) infection of monkey kidney vero e6 cells induces phosphorylation and dephosphorylation of signaling pathways, resulting in apoptosis. in the present study, we investigated the phosphoryl ...	2006	16458888
specific epitopes of the structural and hypothetical proteins elicit variable humoral responses in sars patients.	severe acute respiratory syndrome (sars) is an infectious disease which was caused by a novel coronavirus (sars-cov). sars has caused an outbreak in the world during 2003 and 2004, with 8098 individuals being infected and a death toll of 774 in 28 regions around the world. specific humoral responses to viral infection remain unclear. objective: to analyse the antigenicity of the sars-cov genome and identify potential antigenic epitopes in the structural proteins.	2006	16461566
[antiretroviral drugs in severe acute respiratory syndrome].	inhibition of viral assembly (protease inhibitors) and of fusion of viral and target membranes (fusion inhibitors) are general approaches to antiviral treatment, not specific for hiv. nonetheless, the agents that induce these phenomena are most often specific for a given virus or virus family. drugs developed for hiv treatment were reevaluated for use against severe acute respiratory syndrome-coronavirus (sars-cov) during and after the epidemic in 2003, in view of the absence of any other availa ...	2006	16462674
automated extraction protocol for quantification of sars-coronavirus rna in serum: an evaluation study.	we have previously developed a test for the diagnosis and prognostic assessment of the severe acute respiratory syndrome (sars) based on the detection of the sars-coronavirus rna in serum by real-time quantitative reverse transcriptase polymerase chain reaction (rt-pcr). in this study, we evaluated the feasibility of automating the serum rna extraction procedure in order to increase the throughput of the assay.	2006	16466582
lightup probes in clinical diagnostics.	the lightup probe technology has now matured and reached the phase where it has been implemented in commercial reagent kits, i.e. the ressq product line. several properties of the lightup probes make them particularly suitable for clinical settings. for instance, extraordinary shelf life and a chemical stability that allows convenient fridge storage. the origin of the higher stability of lightup probe kits compared to others, based on alternative probe technologies, is partly the relatively good ...	2006	16466783
promoter activity of sars coronavirus 5' utr sequence in eukaryotic cells.	to investigate 5'utr sequence in different sars-cov isolates, to identify the secondary structure, and to test the promoter activity of the cdna sequence corresponding to sars-cov 5'utr in eukaryotic cells.	2006	16468630
risk of ruling out severe acute respiratory syndrome by ruling in another diagnosis: variable incidence of atypical bacteria coinfection based on diagnostic assays.	severe acute respiratory syndrome (sars) caused the first epidemic of the 21st century and continues to threaten the global community.	2006	16470249
glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and m.	the severe acute respiratory syndrome coronavirus (sars-cov) open reading frame 3a protein has recently been shown to be a structural protein. the protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses. it does, however, have several similarities to the coronavirus m proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localiza ...	2006	16474139
characterization of the antiviral effects of interferon-alpha against a sars-like coronoavirus infection in vitro.	interferon (ifn)-alphas bind to and activate their cognate cell surface receptor to invoke an antiviral response in target cells. well-described receptor-mediated signaling events result in transcriptional regulation of ifn sensitive genes, effectors of this antiviral response. results from a pilot study to evaluate the clinical efficacy of ifn-alpha treatment of sars patients provided evidence for ifn-inducible resolution of disease. in this report we examined the contribution of ifn-inducible ...	2006	16474437
evaluation of measures to reduce international spread of sars.	mathematical models are used to quantify the effect of border control measures in reducing the international spread of sars. border screening is shown to play a relatively minor role in reducing disease spread. assuming detection rates similar to those reported for arrival screening in australia, screening can detect up to 10% (95% ci 3-23) of infected travellers, and reduce the probability of a large outbreak by up to 7% (95% ci 2-17). rapid reductions in the time to diagnosis and effective fac ...	2006	16476169
comparative evaluation of two severe acute respiratory syndrome (sars) vaccine candidates in mice challenged with sars coronavirus.	two different severe acute respiratory syndrome (sars) vaccine strategies were evaluated for their ability to protect against live sars coronavirus (cov) challenge in a murine model of infection. a whole killed (inactivated by beta-propiolactone) sars-cov vaccine and a combination of two adenovirus-based vectors, one expressing the nucleocapsid (n) and the other expressing the spike (s) protein (collectively designated ad s/n), were evaluated for the induction of serum neutralizing antibodies an ...	2006	16476986
the catalysis of the sars 3c-like protease is under extensive regulation by its extra domain.	the 3c-like protease of the severe acute respiratory syndrome (sars) coronavirus has a c-terminal extra domain in addition to the chymotrypsin-fold adopted by picornavirus 3c proteases hosting the complete catalytic machinery. previously we identified the extra domain to be involved in enzyme dimerization which has been considered essential for the catalytic activity. in an initial attempt to map out the extra-domain residues critical for dimerization, we have systematically generated 15 point m ...	2006	16478476
two-year prospective study of the humoral immune response of patients with severe acute respiratory syndrome.	in a cohort study of 56 convalescent patients with severe acute respiratory syndrome (sars), titers of immunoglobulin g (igg) antibodies and neutralizing antibodies (nabs) against sars-associated coronavirus were assessed at regular intervals (at 1, 4, 7, 10, 16, and 24 months after the onset of disease) by use of enzyme-linked immunosorbent assay and neutralization assay. igg antibody and nab titers were highly correlated, peaking at month 4 after the onset of disease and decreasing thereafter. ...	2006	16479513
cytokine regulation in sars coronavirus infection compared to other respiratory virus infections.	the pathogenesis of severe acute respiratory syndrome (sars) is poorly understood and cytokine dysregulation has been suggested as one relevant mechanism to be explored. we compared the cytokine profile in caco2 cells after infection of sars coronavirus (sars-cov) with other respiratory viruses including respiratory syncytial virus (rsv), influenza a virus (fluav), and human parainfluenza virus type 2 (hpiv2). interferon (ifn) system (production and response) was not suppressed by sars-cov infec ...	2006	16482545
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 ...	2006	16484730
inhibition of cell proliferation by sars-cov infection in vero e6 cells.	severe acute respiratory syndrome (sars) is caused by sars-coronavirus (sars-cov). infection of vero e6 cells with sars-cov inhibits cell proliferation. our previous study indicated that akt, which is poorly phosphorylated in confluent cultures of vero e6 cells, is phosphorylated and then dephosphorylated upon infection by sars-cov. in the present study, we showed that a serine residue of akt was phosphorylated in vero e6 cells in subconfluent culture and that akt was dephosphorylated rapidly af ...	2006	16487305
seroprevalence of igg antibodies to sars-coronavirus in asymptomatic or subclinical population groups.	we systematically reviewed the current understanding of human population immunity against sars-cov in different groups, settings and geography. our meta-analysis, which included all identified studies except those on wild animal handlers, yielded an overall seroprevalence of 0.10% [95% confidence interval (ci) 0.02-0.18]. health-care workers and others who had close contact with sars patients had a slightly higher degree of seroconversion (0.23%, 95% ci 0.02-0.45) compared to healthy blood donor ...	2006	16490123
inactivation of sars coronavirus by means of povidone-iodine, physical conditions and chemical reagents.	the efficacy of several povidone-iodine (pvp-i) products, a number of other chemical agents and various physical conditions were evaluated for their ability to inactivate the severe acute respiratory syndrome coronavirus (sars-cov). treatment of sars-cov with pvp-i products for 2 min reduced the virus infectivity from 1.17 x 10(6) tcid(50)/ml to below the detectable level. the efficacy of 70% ethanol was equivalent to that of pvp-i products. fixation of sars-cov-infected vero e6 cells with a fix ...	2006	16490989
sars-associated coronavirus replication in cell lines.	given the potential for laboratory-associated severe acute respiratory syndrome-associated coronavirus (sars-cov) infections, we must know which cell lines are susceptible to the virus. we investigated 21 cell lines routinely used for virus isolation or research. after infection with sars-cov, cells were observed for cytopathic effects, and quantitative real-time polymerase chain reaction was used to measure ongoing viral replication. an indirect immunofluorescence assay was also used as a confi ...	2006	16494729
immunological characterizations of the nucleocapsid protein based sars vaccine candidates.	the recombinant nucleocapsid (rn) protein of the coronavirus (cov) responsible for severe acute respiratory syndrome (sars) was cloned and expressed in escherichia coli, extracted from cell lysates containing 6m urea, then purified by ni(2+)-affinity chromatography. in animal immunogenicity studies, we found that most anti-rn protein antibodies were igg2a in balb/c mice vaccinated with rn emulsified in montanide isa-51 containing the synthetic oligodeoxynucleotide, cpg. in contrast, anti-rn prot ...	2006	16494977
a recombinant baculovirus-expressed s glycoprotein vaccine elicits high titers of sars-associated coronavirus (sars-cov) neutralizing antibodies in mice.	a recombinant sars-cov spike (s) glycoprotein vaccine produced in insect cells in a pre-clinical development stage is described. a truncated version of s glycoprotein, containing only the ecto-domain, as well as a his-tagged full-length version were cloned and expressed in a serum-free insect cell line, expressf+. the proteins, purified to apparent homogeneity by liquid column chromatography, were formulated without adjuvant at 3, 9, 27, and 50 microg per dose in phosphate saline and used to imm ...	2006	16497416
linking bats to emerging diseases.		2006	16497913
voucher specimens for sars-linked bats.		2006	16497914
identification of hepta- and octo-uridine stretches as sole signals for programmed +1 and -1 ribosomal frameshifting during translation of sars-cov orf 3a variants.	programmed frameshifting is one of the translational recoding mechanisms that read the genetic code in alternative ways. this process is generally programmed by signals at defined locations in a specific mrna. in this study, we report the identification of hepta- and octo-uridine stretches as sole signals for programmed +1 and -1 ribosomal frameshifting during translation of severe acute respiratory syndrome coronavirus (sars-cov) orf 3a variants. sars-cov orf 3a encodes a minor structural prote ...	2006	16500894
modeling the early events of severe acute respiratory syndrome coronavirus infection in vitro.	the clinical picture of severe acute respiratory syndrome (sars) is characterized by pulmonary inflammation and respiratory failure, resembling that of acute respiratory distress syndrome. however, the events that lead to the recruitment of leukocytes are poorly understood. to study the cellular response in the acute phase of sars coronavirus (sars-cov)-host cell interaction, we investigated the induction of chemokines, adhesion molecules, and dc-sign (dendritic cell-specific icam-3-grabbing non ...	2006	16501078
going to bat.		2006	16502604
[some medical staff positive for serum sars coronavirus antibody igg have only mild symptoms].	to identify patients with sars coronavirus infection who have only mild symptoms.	2006	16503535
biochemical and functional characterization of the membrane association and membrane permeabilizing activity of the severe acute respiratory syndrome coronavirus envelope protein.	a diverse group of cytolytic animal viruses encodes small, hydrophobic proteins to modify host cell membrane permeability to ions and small molecules during their infection cycles. in this study, we show that expression of the sars-cov e protein in mammalian cells alters the membrane permeability of these cells. immunofluorescent staining and cell fractionation studies demonstrate that this protein is an integral membrane protein. it is mainly localized to the er and the golgi apparatus. the pro ...	2006	16507314
solution structure of the severe acute respiratory syndrome-coronavirus heptad repeat 2 domain in the prefusion state.	the envelope glycoprotein, termed the spike protein, of severe acute respiratory syndrome coronavirus (sars-cov) is known to mediate viral entry. similar to other class 1 viral fusion proteins, the heptad repeat regions of sars-cov spike are thought to undergo conformational changes from a prefusion form to a subsequent post-fusion form that enables fusion of the viral and host membranes. recently, the structure of a post-fusion form of sars-cov spike, which consists of isolated domains of hepta ...	2006	16507566
identification of critical determinants on ace2 for sars-cov entry and development of a potent entry inhibitor.	severe acute respiratory syndrome (sars) is caused by a novel coronavirus, sars-cov. virus entry into cells is mediated through interactions between spike (s) glycoprotein and angiotensin-converting enzyme 2 (ace2). alanine scanning mutagenesis analysis was performed to identify determinants on ace2 critical for sars-cov infection. results indicated that charged amino acids between residues 22 and 57 were important, k26 and d30, in particular. peptides representing various regions of ace2 critic ...	2006	16510163
nucleocapsid protein of sars-cov activates the expression of cyclooxygenase-2 by binding directly to regulatory elements for nuclear factor-kappa b and ccaat/enhancer binding protein.	sars-associated coronavirus (sars-cov) causes inflammation and damage to the lungs resulting in severe acute respiratory syndrome. to evaluate the molecular mechanisms behind this event, we investigated the roles of sars-cov proteins in regulation of the proinflammatory factor, cyclooxygenase-2 (cox-2). individual viral proteins were tested for their abilities to regulate cox-2 gene expression. results showed that the cox-2 promoter was activated by the nucleocapsid (n) protein in a concentratio ...	2006	16546436
[antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses sars-cov, 229e and oc43 with their monoclonal antibodies].	to prepare and characterize monoclonal antibodies (mabs) against the recombinant nucleocapsid (n) protein of 3 human coronaviruses sars-cov, 229e and oc43 and study the antigenic relationship between the 3 n proteins.	2006	16546729
discovery of an rna virus 3'->5' exoribonuclease that is critically involved in coronavirus rna synthesis.	replication of the giant rna genome of severe acute respiratory syndrome (sars) coronavirus (cov) and synthesis of as many as eight subgenomic (sg) mrnas are mediated by a viral replicase-transcriptase of outstanding complexity that includes an essential endoribonuclease activity. here, we show that the cov replicative machinery, unlike that of other rna viruses, also uses an exoribonuclease (exon) activity, which is associated with nonstructural protein (nsp) 14. bacterially expressed forms of ...	2006	16549795
sars: are we still at risk?		2006	16550673
high expression level of soluble sars spike protein mediated by adenovirus in hek293 cells.	to develop a highly efficacious method for preparation of soluble sars s-protein using adenovirus vector to meet the requirement for s-protein investigation.	2006	16552820
severe acute respiratory syndrome coronavirus 3c-like protease-induced apoptosis.	the pathogenesis of severe acute respiratory syndrome-associated coronavirus (sars-cov) is an important issue for the treatment and prevention of severe acute respiratory syndrome. recently, sars-cov has been demonstrated to induce cell apoptosis in vero-e6 cells. the possible role of sars-cov 3c-like protease (3clpro) in virus-induced apoptosis is characterized in this study. growth arrest and apoptosis via caspase-3 and caspase-9 activities were demonstrated in sars-cov 3clpro -expressing huma ...	2006	16553810
close relationship between sars-coronavirus and group 2 coronavirus.	the sudden appearance and potential lethality of severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) in humans has resulted in a focusing of new attention on the determination of both its origins and evolution. the relationship existing between sars-cov and other groups of coronaviruses was determined via analyses of phylogenetic trees and comparative genomic analyses of the coronavirus genes: polymerase (orf1ab), spike (s), envelope (e), membrane (m) and nucleocapsid (n). ...	2006	16554722
only one protomer is active in the dimer of sars 3c-like proteinase.	the severe acute respiratory syndrome coronavirus 3c-like protease has been proposed to be a key target for structurally based drug design against sars. the enzyme exists as a mixture of dimer and monomer, and only the dimer was considered to be active. in this report, we have investigated, using molecular dynamics simulation and mutational studies, the problems as to why only the dimer is active and whether both of the two protomers in the dimer are active. the molecular dynamics simulations sh ...	2006	16565086
inhibition of cytokine gene expression and induction of chemokine genes in non-lymphatic cells infected with sars coronavirus.	sars coronavirus (sars-cov) is the etiologic agent of the severe acute respiratory syndrome. sars-cov mainly infects tissues of non-lymphatic origin, and the cytokine profile of those cells can determine the course of disease. here, we investigated the cytokine response of two human non-lymphatic cell lines, caco-2 and hek 293, which are fully permissive for sars-cov.	2006	16571117
mucosal immunization with surface-displayed severe acute respiratory syndrome coronavirus spike protein on lactobacillus casei induces neutralizing antibodies in mice.	induction of mucosal immunity may be important for preventing sars-cov infections. for safe and effective delivery of viral antigens to the mucosal immune system, we have developed a novel surface antigen display system for lactic acid bacteria using the poly-gamma-glutamic acid synthetase a protein (pgsa) of bacillus subtilis as an anchoring matrix. recombinant fusion proteins comprised of pgsa and the spike (s) protein segments sa (residues 2 to 114) and sb (residues 264 to 596) were stably ex ...	2006	16571824
tertiary structure prediction of sars coronavirus helicase.	sars coronavirus, scv, has been recently responsible of a sudden and widespread infection which caused almost 800 victims. the limited amount of scv protein structural information is partially responsible of the lack of specific drugs against the virus. coronavirus helicases are very conserved and peculiar proteins which have been proposed as suitable targets for antiviral drugs, such as bananins, which have been recently shown to inhibit the scv helicase in vitro. here, the quaternary structure ...	2006	16579970
severe acute respiratory syndrome coronavirus protein 7a interacts with hsgt.	severe acute respiratory syndrome coronavirus (sars-cov) 7a is an accessory protein with no known homologues. in this study, we report the interaction of a sars-cov 7a and small glutamine-rich tetratricopeptide repeat-containing protein (sgt). sars-cov 7a and human sgt interaction was identified using a two-hybrid system screen and confirmed with interaction screens in cell culture and cellular co-localization studies. the sgt domain of interaction was mapped by deletion mutant analysis and resu ...	2006	16580632
angiotensin-converting enzyme 2 in lung diseases.	the renin-angiotensin system (ras) plays a key role in maintaining blood pressure homeostasis, as well as fluid and salt balance. angiotensin ii, a key effector peptide of the system, causes vasoconstriction and exerts multiple biological functions. angiotensin-converting enzyme (ace) plays a central role in generating angiotensin ii from angiotensin i, and capillary blood vessels in the lung are one of the major sites of ace expression and angiotensin ii production in the human body. the ras ha ...	2006	16581295
severe acute respiratory syndrome coronavirus papain-like protease: structure of a viral deubiquitinating enzyme.	replication of severe acute respiratory syndrome (sars) coronavirus (sars-cov) requires proteolytic processing of the replicase polyprotein by two viral cysteine proteases, a chymotrypsin-like protease (3clpro) and a papain-like protease (plpro). these proteases are important targets for development of antiviral drugs that would inhibit viral replication and reduce mortality associated with outbreaks of sars-cov. in this work, we describe the 1.85-a crystal structure of the catalytic core of sar ...	2006	16581910
expression, purification and crystallization of the sars-cov macro domain.	macro domains or x domains are found as modules of multidomain proteins, but can also constitute a protein on their own. recently, biochemical and structural studies of cellular macro domains have been performed, showing that they are active as adp-ribose-1''-phosphatases. macro domains are also present in a number of positive-stranded rna viruses, but their precise function in viral replication is still unknown. the major human pathogen severe acute respiratory syndrome coronavirus (sars-cov) e ...	2006	16582497
crystallization and preliminary x-ray diffraction analysis of nsp15 from sars coronavirus.	the non-structural protein nsp15 from the aetiological agent of sars (severe acute respiratory syndrome) has recently been characterized as a uridine-specific endoribonuclease. this enzyme plays an essential role in viral replication and transcription since a mutation in the related h229e human coronavirus nsp15 gene can abolish viral rna synthesis. sars full-length nsp15 (346 amino acids) has been cloned and expressed in escherichia coli with an n-terminal hexahistidine tag and has been purifie ...	2006	16582498
early diagnosis of sars: lessons from the toronto sars outbreak.	the clinical presentation of sars is nonspecific and diagnostic tests do not provide accurate results early in the disease course. initial diagnosis remains reliant on clinical assessment. to identify features of the clinical assessment that are useful in sars diagnosis, the exposure status and the prevalence and timing of symptoms, signs, laboratory and radiographic findings were determined for all adult patients admitted with suspected sars during the toronto sars outbreak. findings were compa ...	2006	16586072
anti-severe acute respiratory syndrome coronavirus immune responses: the role played by v gamma 9v delta 2 t cells.	severe acute respiratory syndrome (sars) is caused by a novel coronavirus (sars-cov) strain. analyses of t cell repertoires in health care workers who survived sars-cov infection during the 2003 outbreak revealed that their effector memory v gamma 9v delta 2 t cell populations were selectively expanded ~3 months after the onset of disease. no such expansion of their alpha beta t cell pools was detected. the expansion of the v gamma 9v delta 2 t cell population was associated with higher anti-sar ...	2006	16586361
oligonucleotide-based antiviral strategies.	in the age of extensive global traffic systems, the close neighborhood of man and livestock in some regions of the world, as well as inadequate prevention measures and medical care in poorer countries, greatly facilitates the emergence and dissemination of new virus strains. the appearance of avian influenza viruses that can infect humans, the spread of the severe acute respiratory syndrome (sars) virus, and the unprecedented raging of human immunodeficiency virus (hiv) illustrate the threat of ...	2006	16594620
potential antivirals and antiviral strategies against sars coronavirus infections.	there are a number of antivirals as well as antiviral strategies that could be envisaged to prevent or treat severe acute respiratory syndrome (sars) (or similar) coronavirus (cov) infections. targets for the prophylactic or therapeutic interventions include interaction of the spike (s) glycoprotein (s1 domain) with the host cell receptor, fusion of the s2 domain with the host cell membrane, processing of the replicase polyproteins by the virus-encoded proteases (3c-like cysteine protease [3clpr ...	2006	16597209
structure of severe acute respiratory syndrome coronavirus receptor-binding domain complexed with neutralizing antibody.	the severe acute respiratory syndrome coronavirus (sars-cov, or scv), which caused a world-wide epidemic in 2002 and 2003, binds to a receptor, angiotensin-converting enzyme 2 (ace2), through the receptor-binding domain (rbd) of its envelope (spike, s) glycoprotein. the rbd is very immunogenic; it is a major scv neutralization determinant and can elicit potent neutralizing antibodies capable of out-competing ace2. however, the structural basis of rbd immunogenicity, rbd-mediated neutralization, ...	2006	16597622
rapid peptide-based screening on the substrate specificity of severe acute respiratory syndrome (sars) coronavirus 3c-like protease by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.	severe acute respiratory syndrome coronavirus (sars-cov) 3c-like protease (3cl(pro)) mediates extensive proteolytic processing of replicase polyproteins, and is considered a promising target for anti-sars drug development. here we present a rapid and high-throughput screening method to study the substrate specificity of sars-cov 3cl(pro). six target amino acid positions flanking the sars-cov 3cl(pro) cleavage site were investigated. each batch of mixed peptide substrates with defined amino acid ...	2006	16600962
cynomolgus macaque as an animal model for severe acute respiratory syndrome.	the emergence of severe acute respiratory syndrome (sars) in 2002 and 2003 affected global health and caused major economic disruption. adequate animal models are required to study the underlying pathogenesis of sars-associated coronavirus (sars-cov) infection and to develop effective vaccines and therapeutics. we report the first findings of measurable clinical disease in nonhuman primates (nhps) infected with sars-cov.	2006	16605302
obstacles and advances in sars vaccine development.	the emergence of the severe acute respiratory syndrome (sars) that resulted in a pandemic in 2003 spurred a flurry of interest in the development of vaccines to prevent and treat the potentially deadly viral infection. researchers around the world pooled their scientific resources and shared early data in an unprecedented manner in light of the impending public health crisis. there are still large gaps in knowledge about the pathogenesis of this virus. while significant advances have been made i ...	2006	16191455
influence of intron and exon splicing enhancers on mammalian cell expression of a truncated spike protein of sars-cov and its implication for subunit vaccine development.	the spike (s) protein of severe acute respiratory syndrome coronavirus (sars-cov) is important for vaccine development. a truncated s protein of the tw1 strain, str2 (88 kda), carrying three s fragments (s74-253, s294-739, and s1129-1255) was investigated to study the influences of intron and exon splicing enhancers to improve str2 protein expression in mammalian cells. our results showed that str2 protein expression with the use of an 138 base-pair intron addition increased by 1.9-, 2.5-, and 4 ...	2006	16194584
a double-inactivated whole virus candidate sars coronavirus vaccine stimulates neutralising and protective antibody responses.	a double-inactivated, candidate whole virus vaccine against severe acute respiratory syndrome associated coronavirus (sars-cov) was developed and manufactured at large scale using fermenter cultures of serum protein free vero cells. a two step inactivation procedure involving sequential formaldehyde and u.v. inactivation was utilised in order to ensure an extremely high safety margin with respect to residual infectivity. the immunogenicity of this double-inactivated vaccine was characterised in ...	2006	16214268
direct fmoc/tert-bu solid phase synthesis of octamannosyl polylysine dendrimer-peptide conjugates.	the mannose binding proteins on the surface of the dendritic cells are responsible for capture of pathogens in the early stages of immune response. conjugation to mannose dendrimers is a rarely explored but potentially powerful strategy for enhancing immunogenicity of synthetic peptides relying on direct delivery to dendritic cells. we describe a general protocol for preparation of pure, monodisperse third-generation mannosylated poly-l-lysine dendrimer-peptide conjugates using direct, machine-a ...	2006	16247759
modular organization of sars coronavirus nucleocapsid protein.	the sars-cov nucleocapsid (n) protein is a major antigen in severe acute respiratory syndrome. it binds to the viral rna genome and forms the ribonucleoprotein core. the sars-cov n protein has also been suggested to be involved in other important functions in the viral life cycle. here we show that the n protein consists of two non-interacting structural domains, the n-terminal rna-binding domain (rbd) (residues 45-181) and the c-terminal dimerization domain (residues 248-365) (dd), surrounded b ...	2006	16228284
orchitis: a complication of severe acute respiratory syndrome (sars).	severe acute respiratory syndrome (sars) coronavirus has been known to damage multiple organs; however, little is known about its impact on the reproductive system. in the present study, we analyzed the pathological changes of testes from six patients who died of sars. results suggested that sars caused orchitis. all sars testes displayed widespread germ cell destruction, few or no spermatozoon in the seminiferous tubule, thickened basement membrane, and leukocyte infiltration. the numbers of cd ...	2006	16237152
synthesis and activity of an octapeptide inhibitor designed for sars coronavirus main proteinase.	the outbreak of sars, a life-threatening disease, has spread over many countries around the world. so far there is no effective drug for the treatment of sars. stimulated by the binding mechanism of sars-cov mpro with the octapeptide avlqsgfr reported recently as well as the "chou's distorted key" theory, we synthesized the octapeptide avlqsgfr for conducting various biochemical experiments to investigate the antiviral potential of the octapeptide against sars coronavirus (bj-01). the results de ...	2006	16242214
antibody responses against sars coronavirus are correlated with disease outcome of infected individuals.	most of the sars-cov-infected patients spontaneously recovered without clinical intervention while a small percentage succumbed to the disease. here, we characterized temporal changes in n protein-specific and s glycoprotein-specific neutralizing antibody (nab) responses in infected patients who have either recovered from or succumbed to sars-cov infection. recovered patients were found to have higher and sustainable levels of both n protein-specific and s glycoprotein-specific nab responses, su ...	2006	16299724
age- and gender-related difference of ace2 expression in rat lung.	epidemiologic data suggested that there was an obvious predominance of young adult patients with a slight female proneness in severe acute respiratory syndrome (sars). the angiotensin-converting enzyme 2 (ace2) was very recently identified as a functional receptor for sars virus and is therefore a prime target for pathogenesis and pharmacological intervention. rats of both genders at three distinct ages (young-adult, 3 months; middle-aged, 12 months; old, 24 months) were evaluated to determine t ...	2006	16303146
sars coronavirus 7a protein blocks cell cycle progression at g0/g1 phase via the cyclin d3/prb pathway.	the genome of severe acute respiratory syndrome-associated coronavirus (sars-cov) contains four structural genes that are homologous to genes found in other coronaviruses, and also contains six subgroup-specific open reading frames (orfs). expression of one of these subgroup-specific genes, orf7a, resulted in apoptosis via a caspase-dependent pathway. here, we observed that transient expression of orf7a protein fused with myc or gfp tags at its n or c terminus inhibited cell growth and prevented ...	2006	16303160
a novel fingerprint map for detecting sars-cov.	spike (s) protein is the most important membrane protein on the surface of severe acute respiratory syndrome coronavirus (sars-cov). it associates with cellular receptors to mediate infection of their target cells. inspired by such a mechanism, an in-depth investigation into the genome sequences of s protein of sars-cov and its receptor are conducted thru a mathematical transformation and graphic approach. as an outcome, a novel method for visualizing the characteristic of sars-cov is suggested. ...	2006	16289934
why are hiv-1 fusion inhibitors not effective against sars-cov? biophysical evaluation of molecular interactions.	the envelope spike (s) glycoprotein of the severe acute respiratory syndrome associated coronavirus (sars-cov) mediates the entry of the virus into target cells. recent studies point out to a cell entry mechanism of this virus similar to other enveloped viruses, such as hiv-1. as it happens with other viruses peptidic fusion inhibitors, sars-cov s protein hr2-derived peptides are potential therapeutic drugs against the virus. it is believed that hr2 peptides block the six-helix bundle formation, ...	2006	16290276
programmed ribosomal frameshifting in hiv-1 and the sars-cov.	ribosomal frameshifting is a mechanism of gene expression used by several rna viruses to express replicase enzymes. this article focuses on frameshifting in two human pathogens, the retrovirus human immunodeficiency virus type 1 (hiv-1) and the coronavirus responsible for severe acute respiratory syndrome (sars). the nature of the frameshift signals of hiv-1 and the sars-cov will be described and the impact of this knowledge on models of frameshifting will be considered. the role of frameshiftin ...	2006	16310880
preparation and evaluation of anti-sars coronavirus igy from yolks of immunized spf chickens.	severe acute respiratory syndrome (sars) is a recently discovered viral disease, characterized by fever, cough, acute fibrinous pneumonia and high infectivity. specific pathogen-free (spf) chickens were immunized with inactivated sars coronavirus and their eggs were harvested at regular intervals. yolk immunoglobulin (igy) was extracted using the water dilution method, followed by further purification on a sephadex g-75 column. sds-polyacrylamide gel electrophoresis (sds-page), western blot and ...	2006	16325277
preparation and evaluation of anti-sars coronavirus igy from yolks of immunized spf chickens.	severe acute respiratory syndrome (sars) is a recently discovered viral disease, characterized by fever, cough, acute fibrinous pneumonia and high infectivity. specific pathogen-free (spf) chickens were immunized with inactivated sars coronavirus and their eggs were harvested at regular intervals. yolk immunoglobulin (igy) was extracted using the water dilution method, followed by further purification on a sephadex g-75 column. sds-polyacrylamide gel electrophoresis (sds-page), western blot and ...	2006	16325277
from genome to drug lead: identification of a small-molecule inhibitor of the sars virus.	virtual screening, a fast, computational approach to identify drug leads [perola, e.; xu, k.; kollmeyer, t. m.; kaufmann, s. h.; prendergast, f. g. j. med. chem.2000, 43, 401; miller, m. a. nat. rev. drug disc.2002, 1 220], is limited by a known challenge in crystallographically determining flexible regions of proteins. this approach has not been able to identify active inhibitors of the severe acute respiratory syndrome-associated coronavirus (sars-cov) using solely the crystal structures of a ...	2006	16325400
solution structure of the x4 protein coded by the sars related coronavirus reveals an immunoglobulin like fold and suggests a binding activity to integrin i domains.	the sars related coronavirus genome contains a variety of novel accessory genes. one of these, called orf7a or orf8, code for a protein, known as 7a, u122 or x4. we set out to determine the three-dimensional structure of the soluble ectodomain of this type-i transmembrane protein by nuclear magnetic resonance spectroscopy. the fold of the protein is the first member of a further variation of the immunoglobulin like beta-sandwich fold. because x4 does not reveal significant sequence homologies to ...	2006	16328780
the n-terminal octapeptide acts as a dimerization inhibitor of sars coronavirus 3c-like proteinase.	the 3c-like proteinase of severe acute respiratory syndrome (sars) coronavirus has been proposed to be a key target for structural-based drug design against sars. accurate determination of the dimer dissociation constant and the role of the n-finger (residues 1-7) will provide more insights into the enzyme catalytic mechanism of sars 3cl proteinase. the dimer dissociation constant of the wild-type protein was determined to be 14.0microm by analytical ultracentrifugation method. the n-finger frag ...	2006	16329994
the 2003 sars outbreak and its impact on infection control practices.	severe acute respiratory syndrome (sars) emerged recently as a new infectious disease that was transmitted efficiently in the healthcare setting and particularly affected healthcare workers (hcws), patients and visitors. the efficiency of transmission within healthcare facilities was recognised following significant hospital outbreaks of sars in canada, china, hong kong, singapore, taiwan and vietnam. the causative agent of sars was identified as a novel coronavirus, the sars coronavirus. this w ...	2006	16297415
anti-sars-cov immunity induced by a novel cpg oligodeoxynucleotide.	to develop cpg oligodeoxynucleotides (cpg odns) based therapy for prevention and treatment of severe acute respiratory syndrome (sars), we selected a novel cpg odn (bw001), which displays b-type cpg odn structure feature at the 5' and a-type cpg odn structure feature at the 3', and tested for its anti-sars-cov activity. we found that the supernatants of human pbmcs stimulated by bw001 significantly protected vero cells from sars-cov infection. bw001 could stimulate human pbmcs and pdcs to secret ...	2006	16298165
design and biological activities of novel inhibitory peptides for sars-cov spike protein and angiotensin-converting enzyme 2 interaction.	severe acute respiratory syndrome (sars) is an emerging infectious disease caused by a novel coronavirus (sars-cov). the binding of sars-cov spike (s) protein to cellular angiotensin-converting enzyme 2 (ace2) is the first step in sars-cov infection. therefore, we assayed the inhibitory effects of small peptides derived from s protein on the binding of s protein to ace2 and on the s-protein-pseudotyped retrovirus infectivity. sp-4 (residues 192-203), sp-8 (residues 483-494), and sp-10 (residues ...	2006	16337697
sars coronavirus, but not human coronavirus nl63, utilizes cathepsin l to infect ace2-expressing cells.	viruses require specific cellular receptors to infect their target cells. angiotensin-converting enzyme 2 (ace2) is a cellular receptor for two divergent coronaviruses, sars coronavirus (sars-cov) and human coronavirus nl63 (hcov-nl63). in addition to hostcell receptors, lysosomal cysteine proteases are required for productive infection by some viruses. here we show that sars-cov, but not hcov-nl63, utilizes the enzymatic activity of the cysteine protease cathepsin l to infect ace2-expressing ce ...	2006	16339146
severe acute respiratory syndrome coronavirus membrane protein interacts with nucleocapsid protein mostly through their carboxyl termini by electrostatic attraction.	the severe acute respiratory syndrome coronavirus (sars-cov) membrane protein is an abundant virion protein, and its interaction with the nucleocapsid protein is crucial for viral assembly and morphogenesis. although the interacting region in the nucleocapsid protein was mapped to residues 168-208, the interacting region in the membrane protein and the interaction nature are still unclear. in this work, by using yeast two-hybrid and surface plasmon resonance techniques, the residues 197-221 of t ...	2006	16343974
modelling sars data using threshold geometric process.	during the outbreak of an epidemic disease, for example, the severe acute respiratory syndrome (sars), the number of daily infected cases often exhibit multiple trends: monotone increasing during the growing stage, stationary during the stabilized stage and then decreasing during the declining stage. lam first proposed modelling a monotone trend by a geometric process (gp) [x(i), i=1,2,...] directly such that [a(i-1)x(i), i=1,2,...] forms a renewal process for some ratio a>0 which measures the d ...	2006	16345017
severe acute respiratory syndrome coronavirus 3a protein is released in membranous structures from 3a protein-expressing cells and infected cells.	severe acute respiratory syndrome coronavirus (scov) accessory protein 3a is a virus structural protein. we demonstrate here that 3a protein was released efficiently in membranous structures from various cell lines expressing 3a protein. a subpopulation of the released 3a protein is associated with detergent-resistant membranes. the presence of the yxxphi and diacidic motifs, located within the cytoplasmic tail of the 3a protein, was not required for its efficient release. analysis of supernatan ...	2006	16352545
binding site-based classification of coronaviral papain-like proteases.	the coronavirus replicase gene encodes one or two papain-like proteases (termed pl1pro and pl2pro) implicated in the n-terminal processing of the replicase polyprotein and thus contributing to the formation of the viral replicase complex that mediates genome replication. using consensus fold recognition with the 3d-jury meta-predictor followed by model building and refinement, we developed a structural model for the single plpro present in the severe acute respiratory syndrome coronavirus (scov) ...	2006	16358325
sars coronavirus e protein in phospholipid bilayers: an x-ray study.	we investigated the structure of the hydrophobic domain of the severe acute respiratory syndrome e protein in model lipid membranes by x-ray reflectivity and x-ray scattering. in particular, we used x-ray reflectivity to study the location of an iodine-labeled residue within the lipid bilayer. the label imposes spatial constraints on the protein topology. experimental data taken as a function of protein/lipid ratio p/l and different swelling states support the hairpin conformation of severe acut ...	2006	16361349
how the sars coronavirus causes disease: host or organism?	the previous epidemic of severe acute respiratory syndrome (sars) has ended. however, many questions concerning how the aetiological agent, the novel sars coronavirus (cov), causes illness in humans remain unanswered. the pathology of fatal cases of sars is dominated by diffuse alveolar damage. specific histological changes are not detected in other organs. these contrast remarkably with the clinical picture, in which there are apparent manifestations in multiple organs. both pathogen and host f ...	2006	16362992
7a protein of severe acute respiratory syndrome coronavirus inhibits cellular protein synthesis and activates p38 mitogen-activated protein kinase.	it was recently shown that the 7a protein of severe acute respiratory syndrome coronavirus induces biochemical changes associated with apoptosis. in this study, the mechanism by which the 7a protein induces apoptosis was examined. the 7a protein was tested for the ability to inhibit cellular gene expression because several proapoptotic viral proteins with this function have previously been identified. 7a protein inhibited expression of luciferase from an mrna construct that specifically measures ...	2006	16378980
monoclonal antibodies targeting the hr2 domain and the region immediately upstream of the hr2 of the s protein neutralize in vitro infection of severe acute respiratory syndrome coronavirus.	we have previously shown that an escherichia coli-expressed, denatured spike (s) protein fragment of the severe acute respiratory coronavirus, containing residues 1029 to 1192 which include the heptad repeat 2 (hr2) domain, was able to induce neutralizing polyclonal antibodies (c. t. keng, a. zhang, s. shen, k. m. lip, b. c. fielding, t. h. tan, c. f. chou, c. b. loh, s. wang, j. fu, x. yang, s. g. lim, w. hong, and y. j. tan, j. virol. 79:3289-3296, 2005). in this study, monoclonal antibodies ( ...	2006	16378996