Publications

the membrane protein of sars-cov suppresses nf-kappab activation.	severe acute respiratory syndrome coronavirus (sars-cov) infects many organs, such as lung, liver, and immune organs and causes life-threatening atypical pneumonia, sars causes high morbidity and mortality rates. the molecular mechanism of sars pathogenesis remains elusive. inflammatory stimuli can activate ikappab kinase (ikk) signalsome and subsequently the nuclear factor kappa b (nf-kappab), which influences gene expression of cyclooxygenase-2 (cox-2) along with other transcription factors. i ...	2007	17705188
priming with sars cov s dna and boosting with sars cov s epitopes specific for cd4+ and cd8+ t cells promote cellular immune responses.	cellular immune response plays an important role in antiviral immunity. in our previous study, immunization of mice with severe acute respiratory syndrome coronavirus (sars cov) spike (s) dna vaccine could induce both humoral and cellular immunity in response to a pool of entire overlapping s peptides. identification of functional dominant epitopes in sars cov s protein for t cells is crucial for further understanding of cellular immune responses elicited by sars cov s dna vaccine. in present st ...	2007	17709158
regulation of cell death during infection by the severe acute respiratory syndrome coronavirus and other coronaviruses.	both apoptosis and necrosis have been observed in cells infected by various coronaviruses, suggesting that the regulation of cell death is important for viral replication and/or pathogenesis. expeditious research on the severe acute respiratory syndrome (sars) coronavirus, one of the latest discovered coronaviruses that infect humans, has provided valuable insights into the molecular aspects of cell-death regulation during infection. apoptosis was observed in vitro, while both apoptosis and necr ...	2007	17714515
prevention and control of emerging infections: a challenge for the 3rd millennium.	in the last 30 years, several emerging infections due to novel viruses have been identified, from haemorrhagic fever viruses to hiv, from the sars-coronavirus to avian influenza viruses. ecological and genetic changes are important determinants of the emergence of new viral infections, driving to an increase of r0 (the basic reproductive number) through increasing the probability of transmission. the current h5n1 epidemic may be considered a prepandemic paradigm that needs thorough investigation ...	2007	17802926
severe acute respiratory syndrome coronavirus evades antiviral signaling: role of nsp1 and rational design of an attenuated strain.	the severe acute respiratory syndrome (sars) epidemic was caused by the spread of a previously unrecognized infectious agent, the sars-associated coronavirus (sars-cov). here we show that sars-cov could inhibit both virus- and interferon (ifn)-dependent signaling, two key steps of the antiviral response. we mapped a strong inhibitory activity to sars-cov nonstructural protein 1 (nsp1) and show that expression of nsp1 significantly inhibited the activation of all three virus-dependent signaling p ...	2007	17715225
structure-based design and synthesis of highly potent sars-cov 3cl protease inhibitors.		2007	17722121
specific plant terpenoids and lignoids possess potent antiviral activities against severe acute respiratory syndrome coronavirus.	in this study, 221 phytocompounds were evaluated for activity against anti-severe acute respiratory syndrome associated coronavirus (sars-cov) activities using a cell-based assay measuring sars-cov-induced cytopathogenic effect on vero e6 cells. ten diterpenoids (1-10), two sesquiterpenoids (11 and 12), two triterpenoids (13 and 14), five lignoids (15-19), curcumin (20), and reference controls niclosamide (21) and valinomycin (22) were potent inhibitors at concentrations between 3.3 and 10 micro ...	2007	17663539
optimization of a dna vaccine against sars.	severe acute respiratory syndrome coronavirus (sars-cov) first appeared in southern china in november 2002, and then quickly spread to 33 countries on five continents along international air travel routes. although the sars epidemic has been contained, there is a clear need for a safe and effective vaccine should an outbreak of a sars-cov infection reappear in human population. in this study, we tested four dna-vaccine constructs: (1) pll70, containing cdna for the sars-cov spike (s) gene; (2) p ...	2007	17665998
discovery of herpesviruses in bats.	seven novel gammaherpesviruses (ghv) and one novel betaherpesvirus were discovered in seven different european bat species (order chiroptera, family vespertilionidae) with a pan-herpesvirus pcr assay, targeting the dna polymerase (dpol) gene. the sequences of six bat ghv were similarly related to members of the gammaherpesvirus genera percavirus and rhadinovirus. the seventh ghv was related to the porcine lymphotropic herpesvirus 1 (genus macavirus). the betaherpesvirus appeared to be a distant ...	2007	17872515
enhancement of murine coronavirus replication by severe acute respiratory syndrome coronavirus protein 6 requires the n-terminal hydrophobic region but not c-terminal sorting motifs.	severe acute respiratory syndrome coronavirus encodes several accessory proteins of unknown function. we previously showed that one such protein, encoded by orf6, enhanced the growth of mouse hepatitis virus in tissue culture cells and in mice. protein 6 consists of an n-terminal hydrophobic peptide and a c-terminal region containing intracellular protein sorting motifs. herein, we show that mutation of the hydrophobic region but not the sorting motifs affected the ability of protein 6 to enhanc ...	2007	17670827
the coronavirus spike protein induces endoplasmic reticulum stress and upregulation of intracellular chemokine mrna concentrations.	murine hepatitis virus (mhv) and severe acute respiratory syndrome (sars) coronavirus (cov) are two of the best-studied representatives of the family coronaviridae. during cov infection, numerous cytokines and chemokines are induced in vitro and in vivo. human interleukin 8 and its mouse functional counterpart, cxcl2, are early-expressed chemokines. here we show that sars-cov and mhv induce endoplasmic reticulum (er) stress and cxcl2 mrna transcription during infection in vitro. expression of th ...	2007	17670839
structural proteomics of the sars coronavirus: a model response to emerging infectious diseases.	a number of structural genomics/proteomics initiatives are focused on bacterial or viral pathogens. in this article, we will review the progress of structural proteomics initiatives targeting the sars coronavirus (sars-cov), the etiological agent of the 2003 worldwide epidemic that culminated in approximately 8,000 cases and 800 deaths. the sars-cov genome encodes 28 proteins in three distinct classes, many of them with unknown function and sharing low similarity to other proteins. the structure ...	2007	17680348
immune responses against severe acute respiratory syndrome coronavirus induced by virus-like particles in mice.	virus-like particles (vlps) represent a promising vaccine against severe acute respiratory syndrome coronavirus (sars cov). in this study, recombinant baculovirus vacs and vacme were constructed to express the s protein and the m and e proteins of sars cov, respectively. electron microscope analysis demonstrated the formation of vlps in vacme and vacs coinfected insect cells. mice immunized four times with vlps developed high antibody titres against sars cov. in addition, vlps elicited cell-medi ...	2007	17680799
selectivity in isg15 and ubiquitin recognition by the sars coronavirus papain-like protease.	the severe acute respiratory syndrome coronavirus papain-like protease (sars-cov plpro) carries out n-terminal processing of the viral replicase polyprotein, and also exhibits lys48-linked polyubiquitin chain debranching and isg15 precursor processing activities in vitro. here, we used sds-page and fluorescence-based assays to demonstrate that isg15 derivatives are the preferred substrates for the deubiquitinating activity of the plpro. with k(cat)/k(m) of 602,000 m(-1)s(-1), plpro hydrolyzes is ...	2007	17692280
human cd4(+) memory t-lymphocyte responses to sars coronavirus infection.	little is known about cd4(+) t-cell immunity to the severe acute respiratory syndrome (sars) coronavirus. in two sars patients, we examined the memory cd4(+) t-cell responses to peptides derived from sars coronavirus structural proteins. we generated cd4(+) t-cell lines to 3 peptides from the spike (s) protein and defined their hla restriction. in one patient, the predominant memory cd4(+) t-cell response was directed against an epitope outside the s protein receptor-binding domain. in both pati ...	2007	17692881
coronavirus non-structural protein 1 is a major pathogenicity factor: implications for the rational design of coronavirus vaccines.	attenuated viral vaccines can be generated by targeting essential pathogenicity factors. we report here the rational design of an attenuated recombinant coronavirus vaccine based on a deletion in the coding sequence of the non-structural protein 1 (nsp1). in cell culture, nsp1 of mouse hepatitis virus (mhv), like its sars-coronavirus homolog, strongly reduced cellular gene expression. the effect of nsp1 on mhv replication in vitro and in vivo was analyzed using a recombinant mhv encoding a delet ...	2007	17696607
biochemical and genetic analyses of murine hepatitis virus nsp15 endoribonuclease.	the goal of this project was to better define the relationship between the endoribonuclease activity of murine hepatitis virus (mhv) nsp15 (mnsp15) and its role in virus infection. molecular modeling demonstrated that the catalytic residues of mnsp15 are superimposable with its severe acute respiratory syndrome coronavirus ortholog. alanine substitutions at three key residues in the mnsp15 catalytic pocket (h262, h277, and g275) and a double-mutant version (h262p and h277a) generated proteins wi ...	2007	17898055
simultaneous detection and high-throughput identification of a panel of rna viruses causing respiratory tract infections.	clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, high-throughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. we evaluated two multiplex reverse transcription-pcr (rt-pcr) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens. the ngen respiratory virus analyte-specific assay (nanogen, san diego, ca) detects influenza a virus (flu-a) ...	2007	17507510
[loop-mediated isothermal amplification].	we have developed the loop-mediated isothermal amplification (lamp) method, an innovative gene amplification technique, which has the potential to become important in genetic testing. this method allows highly efficient amplification by a single enzyme at a constant temperature. it also has features such as high specificity, rapidity, and simplicity of detection. therefore, application to various life sciences, such as clinical medicine, is expected.	2007	17511240
the two faces of the tissue renin-angiotensin systems: implication in cardiovascular diseases.	the implication of the renin-angiotensin system (ras) in the regulation of the cardiovascular system has been well known for many years. accordingly, many pharmaceutical inhibitors have been developed to treat several pathologies, like hypertension and heart failure, and angiotensin converting enzyme (ace) became one of the major target in the treatment of these cardiovascular diseases. in the last decade however, it has become apparent that the classical view of the ras was not quite accurate. ...	2007	17504232
gxxxg motif of severe acute respiratory syndrome coronavirus spike glycoprotein transmembrane domain is not involved in trimerization and is not important for entry.	recently, a paper was published in which it was proposed that the gxxxg motif of the severe acute respiratory syndrome (sars) coronavirus spike (s) protein transmembrane domain plays a vital role in oligomerization of the protein (e. arbely, z. granot, i. kass, j. orly, and i. t. arkin, biochemistry 45:11349-11356, 2006). here, we show that the gxxxg motif is not involved in sars s oligomerization by trimerization analysis of s gxxxg mutant proteins. in addition, the capability of s to mediate e ...	2007	17507478
synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice.	the severe acute respiratory syndrome (sars) epidemic was characterized by high mortality rates in the elderly. the molecular mechanisms that govern enhanced susceptibility of elderly populations are not known, and robust animal models are needed that recapitulate the increased pathogenic phenotype noted with increasing age. using synthetic biology and reverse genetics, we describe the construction of a panel of isogenic sars coronavirus (sars-cov) strains bearing variant spike glycoproteins tha ...	2007	17507479
a retrospective serological study of severe acute respiratory syndrome cases in guangdong province, china.		2007	17517191
human immunodeficiency viral vector pseudotyped with the spike envelope of severe acute respiratory syndrome coronavirus transduces human airway epithelial cells and dendritic cells.	the human severe acute respiratory syndrome coronavirus (sars-cov) is a highly infectious virus that causes severe respiratory infections in humans. the spike envelope glycoprotein of sars-cov, the main determinant of sars-cov tropism, was isolated and used to pseudotype a human immunodeficiency virus (hiv)-based vector. spike-pseudotyped hiv vector was generated and evaluated in vitro on well-differentiated human airway epithelial cells and bronchial explants and in vivo in murine airways. the ...	2007	17518614
analysis of intraviral protein-protein interactions of the sars coronavirus orfeome.	the severe acute respiratory syndrome coronavirus (sars-cov) genome is predicted to encode 14 functional open reading frames, leading to the expression of up to 30 structural and non-structural protein products. the functions of a large number of viral orfs are poorly understood or unknown. in order to gain more insight into functions and modes of action and interaction of the different proteins, we cloned the viral orfeome and performed a genome-wide analysis for intraviral protein interactions ...	2007	17520018
trafficking motifs in the sars-coronavirus nucleocapsid protein.	the severe acute respiratory syndrome-coronavirus nucleocapsid (n) protein is involved in virus replication and modulation of cell processes. in this latter respect control may in part be achieved through the sub-cellular localisation of the protein. n protein predominately localises in the cytoplasm (the site of virus replication and assembly) but also in the nucleus/nucleolus. using a combination of live-cell and confocal microscopy coupled to mutagenesis we identified a cryptic nucleolar loca ...	2007	17524366
[severe acute respiratory syndrome].		2007	17526201
angiotensin-converting enzyme 2 in acute respiratory distress syndrome.	angiotensin-converting enzyme (ace) and ace2 are highly homologous metalloproteases that provide essential catalytic functions in the renin-angiotensin system (ras). angiotensin ii is one key effector peptide of the ras, inducing vasoconstriction and exerting multiple biological functions. ace cleaves angiotensin i to generate angiotensin ii, whereas ace2 reduces angiotensin ii levels. accumulating evidence has demonstrated a physiological and pathological role of ace2 in the cardiovascular syst ...	2007	17558469
up-regulation of il-6 and tnf-alpha induced by sars-coronavirus spike protein in murine macrophages via nf-kappab pathway.	the clinical picture of severe acute respiratory syndrome (sars) is characterized by an over-exuberant immune response with lung lymphomononuclear cells infilteration and proliferation that may account for tissue damage more than the direct effect of viral replication. to understand how cells response in the early stage of virus-host cell interaction, in this study, a purified recombinant s protein was studied for stimulating murine macrophages (raw264.7) to produce proinflammatory cytokines (il ...	2007	17532082
cleavage of spike protein of sars coronavirus by protease factor xa is associated with viral infectivity.	the spike (s) protein of sars coronavirus (sars-cov) has been known to recognize and bind to host receptors, whose conformational changes then facilitate fusion between the viral envelope and host cell membrane, leading to viral entry into target cells. however, other functions of sars-cov s protein such as proteolytic cleavage and its implications to viral infection are incompletely understood. in this study, we demonstrated that the infection of sars-cov and a pseudovirus bearing the s protein ...	2007	17533109
lack of support for an association between clec4m homozygosity and protection against sars coronavirus infection.		2007	17534354
isolation and identification of an scfv antibody against nucleocapsid protein of sars-cov.	to develop reagents for early diagnosis and therapeutic drugs against sars-associated coronavirus (sars-cov), a large (3 x 10(9)) immunized human antibody library was constructed from peripheral blood mononuclear cells from six sars convalescent patients. a single chain variable fragment antibody (n18) with high affinity against n protein of sars-cov was isolated. sequence analysis revealed that the vl gene was composed of vl3h (v lambda subgroup) and jl2 regions and the vh gene was composed of ...	2007	17548223
the use of hepatitis c virus ns3/4a and secreted alkaline phosphatase to quantitate cell-cell membrane fusion mediated by severe acute respiratory syndrome coronavirus s protein and the receptor angiotensin-converting enzyme 2.	the membrane fusion process mediated by severe acute respiratory syndrome coronavirus (sars-cov) s protein and its cellular receptor angiotensin-converting enzyme 2 (ace2) had been reconstituted using two chinese hamster ovary (cho) cell lines that constitutively express these recombinant proteins separately. this system was applied to develop a quantitative measurement of cell-cell fusion using hepatitis c virus (hcv) ns3/4a protease and a secretion-blocked egfp-4a/4b-seap (egfp: enhanced green ...	2007	17553448
lack of support for an association between clec4m homozygosity and protection against sars coronavirus infection.		2007	17534355
newcastle disease virus, a host range-restricted virus, as a vaccine vector for intranasal immunization against emerging pathogens.	the international outbreak of the severe acute respiratory syndrome-associated coronavirus (sars-cov) in 2002-2003 highlighted the need to develop pretested human vaccine vectors that can be used in a rapid response against newly emerging pathogens. we evaluated newcastle disease virus (ndv), an avian paramyxovirus that is highly attenuated in primates, as a topical respiratory vaccine vector with sars-cov as a test pathogen. complete recombinant ndv was engineered to express the sars-cov spike ...	2007	17535926
the association of rantes polymorphism with severe acute respiratory syndrome in hong kong and beijing chinese.	chemokines play important roles in inflammation and antiviral action. we examined whether polymorphisms of rantes, ip-10 and mig affect the susceptibility to and outcome of severe acute respiratory syndrome (sars).	2007	17540042
adenovirus-based vaccine prevents pneumonia in ferrets challenged with the sars coronavirus and stimulates robust immune responses in macaques.	a ferret model of severe acute respiratory syndrome (sars)-cov infection was used to evaluate the efficacy of an adenovirus vaccine. animals were subjected to heterologous prime-boost using vectors from human serotype 5 and chimpanzee derived adenoviruses (human adhu5 and chimpanzee adc7) expressing spike protein followed by intranasal challenge with sars-cov. vaccination led to a substantial reduction in viral load and prevented the severe pneumonia seen in unvaccinated animals. the same prime- ...	2007	17559989
synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice.	the severe acute respiratory syndrome (sars) epidemic was characterized by high mortality rates in the elderly. the molecular mechanisms that govern enhanced susceptibility of elderly populations are not known, and robust animal models are needed that recapitulate the increased pathogenic phenotype noted with increasing age. using synthetic biology and reverse genetics, we describe the construction of a panel of isogenic sars coronavirus (sars-cov) strains bearing variant spike glycoproteins tha ...	2007	17507479
the use of hepatitis c virus ns3/4a and secreted alkaline phosphatase to quantitate cell-cell membrane fusion mediated by severe acute respiratory syndrome coronavirus s protein and the receptor angiotensin-converting enzyme 2.	the membrane fusion process mediated by severe acute respiratory syndrome coronavirus (sars-cov) s protein and its cellular receptor angiotensin-converting enzyme 2 (ace2) had been reconstituted using two chinese hamster ovary (cho) cell lines that constitutively express these recombinant proteins separately. this system was applied to develop a quantitative measurement of cell-cell fusion using hepatitis c virus (hcv) ns3/4a protease and a secretion-blocked egfp-4a/4b-seap (egfp: enhanced green ...	2007	17553448
intranasal protollin-formulated recombinant sars s-protein elicits respiratory and serum neutralizing antibodies and protection in mice.	the feasibility of developing a prophylactic vaccine against sars was assessed by comparing the immune responses elicited by immunizing mice with a recombinant sars spike glycoprotein (s-protein) formulated with different adjuvants, given by different routes. in both young and aged mice, an intranasal protollin-formulated s-protein vaccine elicited high levels of antigen-specific igg in serum, comparable to those elicited by an intramuscular alum-adsorbed s-protein vaccine. serum antibodies were ...	2007	17640780
the nonstructural protein 8 (nsp8) of the sars coronavirus interacts with its orf6 accessory protein.	severe acute respiratory syndrome (sars) coronavirus (sars-cov) caused a severe outbreak in several regions of the world in 2003. the sars-cov genome is predicted to contain 14 functional open reading frames (orfs). the first orf (1a and 1b) encodes a large polyprotein that is cleaved into nonstructural proteins (nsp). the other orfs encode for four structural proteins (spike, membrane, nucleocapsid and envelope) as well as eight sars-cov-specific accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b an ...	2007	17532020
[expression of severe acute respiratory syndrome coronavirus receptors, ace2 and cd209l in different organ derived microvascular endothelial cells].	to investigate the expression of the 2 severe acute respiratory syndrome coronavirus (sars-cov) receptors, angiotensin-converting enzyme 2 (ace2) and cd209l in different human organ/tissue derived microvascular endothelial cells.	2007	17565868
bismuth complexes inhibit the sars coronavirus.		2007	17645269
a saccharomyces cerevisiae autoselection system for optimised recombinant protein expression.	yeasts are attractive organisms for recombinant protein production. they combine highly developed genetic systems and ease of use with reductions in time and costs. we describe an autoselection system for recombinant protein expression in saccharomyces cerevisiae which increases yields 5-10-fold compared to conditional selection for expression plasmids. multicopy expression plasmids encoding essential mob1 or cdc28 genes are absolutely necessary for the viability of host cells with mob1 or cdc28 ...	2007	17566670
cell entry by enveloped viruses: redox considerations for hiv and sars-coronavirus.	for enveloped viruses, genome entry into the target cell involves two major steps: virion binding to the cell-surface receptor and fusion of the virion and cell membranes. virus-cell membrane fusion is mediated by the virus envelope complex, and its fusogenicity is the result of an active virus-cell interaction process that induces conformation changes within the envelope. for some viruses, such as influenza, exposure to an acidic milieu within the cell during the early steps of infection trigge ...	2007	17567241
estimating incubation period with multiple contact days.	multiple contacts with the infectious source may cause inconsistencies in determining the incubation period of a patient and hence the calculation of the mean and variance of the incubation period. a method based on probability distribution to deal with the problem is presented and an example is given for illustration.	2007	17577826
[ligand screening in affinity chromatography and its applications].	affinity chromatography is one of the most powerful techniques in selective purification and isolation of a great number of compounds. affinity ligand is the most important factor in affinity chromatography, therefore, the selection and the screening of affinity ligand become the first issue to be considered when a novel type of affinity packing or a novel system of affinity chromatography needs to be developed. in combination with the research work of the author' s group, this paper briefly sum ...	2007	17580675
heterologous mva-s prime ad5-s boost regimen induces high and persistent levels of neutralizing antibody response against sars coronavirus.	severe acute respiratory syndrome (sars) is caused by a novel coronavirus (cov), sars-cov. in previous studies, we showed that a sars-cov spike (s) glycoprotein-based modified vaccinia ankara (mva-s) vaccine could induce strong neutralizing antibody (nab) response which might have played a critical role in protecting chinese rhesus monkeys from the pathogenic viral challenge. to date, however, it remains unknown what the minimal level of nab is required to achieve sterile immunity in humans. it ...	2007	17581748
heterologous mva-s prime ad5-s boost regimen induces high and persistent levels of neutralizing antibody response against sars coronavirus.	severe acute respiratory syndrome (sars) is caused by a novel coronavirus (cov), sars-cov. in previous studies, we showed that a sars-cov spike (s) glycoprotein-based modified vaccinia ankara (mva-s) vaccine could induce strong neutralizing antibody (nab) response which might have played a critical role in protecting chinese rhesus monkeys from the pathogenic viral challenge. to date, however, it remains unknown what the minimal level of nab is required to achieve sterile immunity in humans. it ...	2007	17581748
sirnas targeting terminal sequences of the sars-associated coronavirus membrane gene inhibit m protein expression through degradation of m mrna.	sars-associated coronavirus (scov) m protein plays a key role in viral assembly and budding. recent studies revealed that m protein could interact with n protein in the golgi complex. in this study, we showed that scov m protein co-localized in the golgi apparatus with a golgi vector marker. to study m protein function, three candidate small interfering rnas (sirnas) corresponding to m gene sequences were designed, transcribed in vitro, and then tested for their ability to silence m protein expr ...	2007	17590445
severe acute respiratory syndrome coronavirus orf6 antagonizes stat1 function by sequestering nuclear import factors on the rough endoplasmic reticulum/golgi membrane.	the host innate immune response is an important deterrent of severe viral infection in humans and animals. nuclear import factors function as key gatekeepers that regulate the transport of innate immune regulatory cargo to the nucleus of cells to activate the antiviral response. using severe acute respiratory syndrome coronavirus (sars-cov) as a model, we demonstrate that sars-cov orf6 protein is localized to the endoplasmic reticulum (er)/golgi membrane in infected cells, where it binds to and ...	2007	17596301
severe acute respiratory syndrome coronavirus gene 7 products contribute to virus-induced apoptosis.	the proteins encoded by gene 7 of the severe acute respiratory syndrome coronavirus (sars-cov) have been demonstrated to have proapoptotic activity when expressed from cdna but appear to be dispensable for virus replication. recombinant sars-covs bearing deletions in gene 7 were used to assess the contribution of gene 7 to virus replication and apoptosis in several transformed cell lines, as well as to replication and pathogenesis in golden syrian hamsters. deletion of gene 7 had no effect on sa ...	2007	17686858
severe acute respiratory syndrome coronavirus gene 7 products contribute to virus-induced apoptosis.	the proteins encoded by gene 7 of the severe acute respiratory syndrome coronavirus (sars-cov) have been demonstrated to have proapoptotic activity when expressed from cdna but appear to be dispensable for virus replication. recombinant sars-covs bearing deletions in gene 7 were used to assess the contribution of gene 7 to virus replication and apoptosis in several transformed cell lines, as well as to replication and pathogenesis in golden syrian hamsters. deletion of gene 7 had no effect on sa ...	2007	17686858
development of a respiratory virus panel test for detection of twenty human respiratory viruses by use of multiplex pcr and a fluid microbead-based assay.	virology laboratories historically have used direct fluorescent-antibody assay (dfa) and culture to detect six or seven respiratory viruses. following the discovery of five new human respiratory viruses since 2000, there is an increasing need for diagnostic tests to detect these emerging viruses. we have developed a new test that can detect 20 different respiratory virus types/subtypes in a single 5-h test. the assay employs multiplex pcr using 14 virus-specific primer pairs, followed by a multi ...	2007	17596360
type ivb pilus operon promoter controlling expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid gene in salmonella enterica serovar typhi elicits full immune response by intranasal vaccination.	attenuated salmonella enterica serovar typhi strains have been considered to be attractive as potential live oral delivery vector vaccines because of their ability to elicit the full array of immune responses in humans. in this study, we constructed an attenuated s. enterica serovar typhi strain stably expressing conserved nucleocapsid (n) protein of severe acute respiratory syndrome-associated coronavirus (sars-cov) by integrating the n gene into the pilv gene, which was under the control of th ...	2007	17596427
open reading frame 8a of the human severe acute respiratory syndrome coronavirus not only promotes viral replication but also induces apoptosis.	a unique genomic difference between human and civet severe acute respiratory syndrome coronaviruses (sars-covs) is that the former has a deletion of 29 nucleotides from open reading frame (orf) 8a' that results in the generation of orf8a and orf8b. the objectives of the present study were to analyze antibody reactivity to orf8a in patients with sars and to elucidate the function of orf8a.	2007	17597455
functional genomics highlights differential induction of antiviral pathways in the lungs of sars-cov-infected macaques.	the pathogenesis of severe acute respiratory syndrome coronavirus (sars-cov) is likely mediated by disproportional immune responses and the ability of the virus to circumvent innate immunity. using functional genomics, we analyzed early host responses to sars-cov infection in the lungs of adolescent cynomolgus macaques (macaca fascicularis) that show lung pathology similar to that observed in human adults with sars. analysis of gene signatures revealed induction of a strong innate immune respons ...	2007	17696609
functional genomics highlights differential induction of antiviral pathways in the lungs of sars-cov-infected macaques.	the pathogenesis of severe acute respiratory syndrome coronavirus (sars-cov) is likely mediated by disproportional immune responses and the ability of the virus to circumvent innate immunity. using functional genomics, we analyzed early host responses to sars-cov infection in the lungs of adolescent cynomolgus macaques (macaca fascicularis) that show lung pathology similar to that observed in human adults with sars. analysis of gene signatures revealed induction of a strong innate immune respons ...	2007	17696609
chicken single-chain variable fragments against the sars-cov spike protein.	the major concern for severe acute respiratory syndrome (sars), caused by the sars-associated coronavirus (sars-cov), is the lack of diagnostic and therapeutic agents. using a phage display technology in a chicken system, high-affinity monoclonal antibody fragments against the sars-cov spike protein were characterized. ten truncated spike protein gene fragments were expressed in escherichia coli cells. following the immunization of chickens with these recombinant spike proteins, two single-chain ...	2007	17643500
pharmacologic treatment of sars: current knowledge and recommendations.	the severe acute respiratory syndrome (sars) pandemic caught the world by surprise in 2003 and spread rapidly within a relatively short period of time. hence, randomised placebo-controlled clinical trials on the treatment of sars were not possible. our understanding was obtained from observational, cohort studies, case series and reports. nevertheless, such information is useful in providing clinical management guidelines and directing future research in case sars recurs. early in the pandemic, ...	2007	17597972
a mechanistic view of enzyme inhibition and peptide hydrolysis in the active site of the sars-cov 3c-like peptidase.	the 3c-like main peptidase 3cl(pro) is a viral polyprotein processing enzyme essential for the viability of the severe acute respiratory syndrome coronavirus (sars-cov). while it is generalized that 3cl(pro) and the structurally related 3c(pro) viral peptidases cleave their substrates via a mechanism similar to that underlying the peptide hydrolysis by chymotrypsin-like serine proteinases (clsps), some of the hypothesized key intermediates have not been structurally characterized. here, we prese ...	2007	17599357
quantitative temporal-spatial distribution of severe acute respiratory syndrome-associated coronavirus (sars-cov) in post-mortem tissues.	few post-mortem studies have been performed on patients who have died from severe acute respiratory syndrome (sars). no studies have examined how the sars-associated coronavirus (sars-cov) loads in different organs with respect to time, post-mortem. the aim of this study was to determine the quantitative temporal-spatial distribution of sars-cov in the post-mortem tissue samples of seven patients. quantitation of a house-keeping gene, glyceraldehyde-3-phosphate dehydrogenase (gapdh) was undertak ...	2007	17607787
potent cross-reactive neutralization of sars coronavirus isolates by human monoclonal antibodies.	the severe acute respiratory syndrome coronavirus (sars-cov) caused a worldwide epidemic in late 2002/early 2003 and a second outbreak in the winter of 2003/2004 by an independent animal-to-human transmission. the gd03 strain, which was isolated from an index patient of the second outbreak, was reported to resist neutralization by the human monoclonal antibodies (hmabs) 80r and s3.1, which can potently neutralize isolates from the first outbreak. here we report that two hmabs, m396 and s230.15, ...	2007	17620608
specific asparagine-linked glycosylation sites are critical for dc-sign- and l-sign-mediated severe acute respiratory syndrome coronavirus entry.	severe acute respiratory syndrome (sars) is caused by a newly emerged coronavirus (cov) designated sars-cov. the virus utilizes angiotensin-converting enzyme 2 (ace2) as the primary receptor. although the idea is less clear and somewhat controversial, sars-cov is thought to use c-type lectins dc-sign and/or l-sign (collectively referred to as dc/l-sign) as alternative receptors or as enhancer factors that facilitate ace2-mediated virus infection. in this study, the function of dc/l-sign in sars- ...	2007	17715238
a massachusetts prototype like coronavirus isolated from wild peafowls is pathogenic to chickens.	coronavirus infection was investigated in apparently healthy wild peafowls in guangdong province of china in 2003, while severe acute respiratory syndrome (sars) broke out there. no sars-like coronavirus had been isolated but a novel avian coronavirus strain, peafowl/gd/kq6/2003 (kq6), was identified. sequence analysis revealed that kq6 was an avian coronavirus infectious bronchitis virus (ibv), a member of coronavirus in group 3. the genome sequence of kq6 had extremely high degree of identity ...	2007	17629993
the s proteins of human coronavirus nl63 and severe acute respiratory syndrome coronavirus bind overlapping regions of ace2.	the cellular receptor for human coronavirus nl63 (hcov-nl63), a group i coronavirus, is angiotensin-converting enzyme2 (ace2). ace2 is also the receptor for the sars coronavirus (sars-cov), a group ii coronavirus. here we describe the ability of hcov-nl63 to utilize a number of ace2 variants previously characterized as sars-cov receptors. several ace2 variants that reduced sars-cov s-protein association similarly reduced that of hcov-nl63, whereas alteration of a number of solvent-exposed ace2 r ...	2007	17631932
targeting the glycans of glycoproteins: a novel paradigm for antiviral therapy.	several chronic viral infections (such as hiv and hepatitis c virus) are highly prevalent and are a serious health risk. the adaptation of animal viruses to the human host, as recently exemplified by influenza viruses and the severe acute respiratory syndrome coronavirus, is also a continuous threat. there is a high demand, therefore, for new antiviral lead compounds and novel therapeutic concepts. in this review, an original therapeutic concept for suppressing enveloped viruses is presented tha ...	2007	17632570
towards our understanding of sars-cov, an emerging and devastating but quickly conquered virus.	severe acute respiratory syndrome (sars) is a newly emerging infectious disease caused by a novel coronavirus (sars-cov), which has overwhelmed more than 30 countries claiming nearly 8400 cases with over 800 fatalities. thanks to the unprecedented international collaboration, the whole-genomes of sars-covs were successfully deciphered shortly after the identification of the causative pathogen for outbreak of sars in southern china, in 2003. hitherto, the sars-cov, as a viral paradigm of emerging ...	2007	17640731
the immunity induced by recombinant spike proteins of sars coronavirus in balb/c mice.	the immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (sars cov) was investigated in balb/c mice. two partial spike gene fragments s1 (322 1464 bp) and s2 (2170 2814 bp) of sars coronavirus were amplified by rt-pcr, and cloned into pet-23a prokaryotic expression vector, then transformed into competent escherichia e. coli bl21 (de3)(plyss) respectively. recombinant proteins were expressed and purified by ni2+ immobilized metal ion ...	2007	17641827
substrate specificity profiling and identification of a new class of inhibitor for the major protease of the sars coronavirus.	severe acute respiratory syndrome (sars) is an emerging infectious disease associated with a high rate of mortality. the sars-associated coronavirus (sars-cov) has been identified as the etiological agent of the disease. although public health procedures have been effective in combating the spread of sars, concern remains about the possibility of a recurrence. various approaches are being pursued for the development of efficacious therapeutics. one promising approach is to develop small molecule ...	2007	17605471
induction of t-cell response by a dna vaccine encoding a novel hla-a*0201 severe acute respiratory syndrome coronavirus epitope.	the severe acute respiratory syndrome coronavirus nucleocapsid protein (sars-cov n) is one of the major targets for sars vaccine due to its high potency in triggering immune responses. in this study, we have identified a novel hla-a*0201 restricted epitope, n220 (lalllldrl), of the sars-cov n-protein through bioinformatics analysis. the n-protein peptide n220 shows a high binding affinity towards human mhc class i in t2-cells, and is capable of activating cytotoxic t-cells in human peripheral bl ...	2007	17629360
rapid multiplex nested pcr for detection of respiratory viruses.	respiratory tract infections can be caused by a heterogeneous group of viruses and bacteria that produce similar clinical presentations. specific diagnosis therefore relies on laboratory investigation. this study developed and evaluated five groups of multiplex nested pcr assays that could simultaneously detect 21 different respiratory pathogens: influenza a virus (h1n1, h3n2, and h5n1); influenza b virus; parainfluenza virus types 1, 2, 3, 4a, and 4b; respiratory syncytial virus a and b; human ...	2007	17804659
[three-dimensional structure prediction of the viral protein in the development of anti sars drugs].		2007	17824221
expression, post-translational modification and biochemical characterization of proteins encoded by subgenomic mrna8 of the severe acute respiratory syndrome coronavirus.	the most striking difference between the subgenomic mrna8 of severe acute respiratory syndrome coronavirus isolated from human and some animal species is the deletion of 29 nucleotides, resulting in splitting of a single orf (orf8) into two orfs (orf8a and orf8b). orf8a and orf8b are predicted to encode two small proteins, 8a and 8b, and orf8 a single protein, 8ab (a fusion form of 8a and 8b). to understand the functions of these proteins, we cloned cdna fragments covering these orfs into expres ...	2007	17645546
bats, civets and the emergence of sars.	severe acute respiratory syndrome (sars) was the first pandemic transmissible disease of previously unknown aetiology in the twenty-first century. early epidemiologic investigations suggested an animal origin for sars-cov. virological and serological studies indicated that masked palm civets ( paguma larvata), together with two other wildlife animals, sampled from a live animal market were infected with sars-cov or a closely related virus. recently, horseshoe bats in the genus rhinolophus have b ...	2007	17848070
amino acid substitutions in the s2 region enhance severe acute respiratory syndrome coronavirus infectivity in rat angiotensin-converting enzyme 2-expressing cells.	to clarify the molecular basis of severe acute respiratory syndrome coronavirus (sars-cov) adaptation to different host species, we serially passaged sars-cov in rat angiotensin-converting enzyme 2 (ace2)-expressing cells. after 15 passages, the virus (rat-p15) came to replicate effectively in rat ace2-expressing cells. two amino acid substitutions in the s2 region were found on the rat-p15 s gene. analyses of the infectivity of the pseudotype-bearing s protein indicated that the two substitutio ...	2007	17652383
interaction between respiratory syncytial virus and glycosaminoglycans, including heparan sulfate.	glycosaminoglycans (gags), including heparan sulfate (hs), are expressed on the surface of nearly all cells, linked to transmembrane proteins. these gags are sulfated to varying extents, lending a negative charge, and are used by a large number of viruses to initiate infection of immortalized cell lines. here we describe the rationale and methods for analyzing gag usage by one such virus, respiratory syncytial virus (rsv). the protocols presented allow the determination of which gag(s) is employ ...	2007	17502668
expression of the severe acute respiratory syndrome coronavirus 3a protein and the assembly of coronavirus-like particles in the baculovirus expression system.	the bac-to-bac baculovirus expression system was used to generate a recombinant baculovirus capable of expressing the severe acute respiratory syndrome (sars)-coronavirus (cov) 3a protein. using the same expression system, two structural proteins, membrane (m) and envelope (e), were co-expressed to form sars-cov virus-like particles (vlps) within an insect cell. expression of viral proteins was confirmed by western blot analysis and the formation of vlps was studied by transmission electron micr ...	2007	17502669
epidemic modeling in complex realities.	in our global world, the increasing complexity of social relations and transport infrastructures are key factors in the spread of epidemics. in recent years, the increasing availability of computer power has enabled both to obtain reliable data allowing one to quantify the complexity of the networks on which epidemics may propagate and to envision computational tools able to tackle the analysis of such propagation phenomena. these advances have put in evidence the limits of homogeneous assumptio ...	2007	17502293
signal transduction in sars-cov-infected cells.	severe acute respiratory syndrome (sars) is a newly found infectious disease that is caused by a novel human coronavirus, sars coronavirus (sars-cov). because the mortality rate of sars patients is very high, understanding the pathological mechanisms of sars not only in vivo but in vitro is important for the prevention of sars. activation of signaling pathways caused by sars-cov infection leads to the phosphorylation and activation of downstream molecules. two conflicting cellular programs, apop ...	2007	17470913
severe acute respiratory syndrome (sars) coronavirus.	the global severe acute respiratory syndrome (sars) outbreak was the first pandemic of the 21st century. although the outbreak was successfully controlled, evidence that sars emerged from an animal reservoir has raised concerns that another pandemic could occur. this review discusses the likelihood of another sars pandemic and reviews the epidemiological and clinical features of the disease with an emphasis on the clinical presentation, diagnosis, and management of sars.	2007	17458774
formalin-treated uv-inactivated sars coronavirus vaccine retains its immunogenicity and promotes th2-type immune responses.	the demand for rapid and simple development of a vaccine against a newly emerging infectious disease is increasing worldwide. we previously revealed that uv-inactivated severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) virions (uv-v) elicited high levels of humoral immunity and a weak th0 response in mice immunized subcutaneously. to ensure the safety of such a whole inactivated sars-cov vaccine, we additionally treated the uv-v vaccine with formalin, resulting in the uv ...	2007	17515642
in silico prediction of sars protease inhibitors by virtual high throughput screening.	a structure-based in silico virtual drug discovery procedure was assessed with severe acute respiratory syndrome coronavirus main protease serving as a case study. first, potential compounds were extracted from protein-ligand complexes selected from protein data bank database based on structural similarity to the target protein. later, the set of compounds was ranked by docking scores using a electronic high-throughput screening flexible docking procedure to select the most promising molecules. ...	2007	17461975
proceedings of a meeting entitled emerging viral infectious diseases, april 4-5, 2005, hanoi, vietnam.		2007	17470907
clathrin-dependent entry of severe acute respiratory syndrome coronavirus into target cells expressing ace2 with the cytoplasmic tail deleted.	the penetration of various viruses into host cells is accomplished by hijacking the host endocytosis machinery. in the case of severe acute respiratory syndrome coronavirus (sars-cov) infection, viral entry is reported to require a low ph in intracytoplasmic vesicles; however, little is known about how sars-cov invades such compartments. here we demonstrate that sars-cov mainly utilizes the clathrin-mediated endocytosis pathway for its entry to target cells by using infectious sars-cov, as well ...	2007	17522231
the molecular biology of sars coronavirus.	severe acute respiratory syndrome (sars) is the first emerging infectious disease of the 21st century that has been highly transmissible and fatal and was caused by a previously unknown coronavirus (sars-cov). the sars epidemic in 2003 resulted in more than 8400 sars cases and approximately 800 deaths. existing in non-identified animal reservoirs, sars-cov continues to represent a threat to humans although more than four years have passed since a large outbreak of sars, and no new cases have bee ...	2007	17470909
a fluorogenic peptide containing the processing site of human sars corona virus s-protein: kinetic evaluation and nmr structure elucidation.	human severe acute respiratory syndrome coronavirus (hsars-cov) is the causative agent for sars infection. its surface glycoprotein (spike protein) is considered to be one of the prime targets for sars therapeutics and intervention because its proteolytic maturation by a host protease is crucial for host-virus fusion. using intramolecularly quenched fluorogenic (iqf) peptides based on hsars-cov spike protein (abz-(755)glu-gln-asp-arg-asn-thr-arg-glu-val-phe-ala-gln(766)-tyx-nh(2)) and in vitro s ...	2007	17471479
comparison of immunoglobulin g responses to the spike and nucleocapsid proteins of severe acute respiratory syndrome (sars) coronavirus in patients with sars.	both the nucleocapsid (n) and the spike (s) proteins of severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) are able to induce strong humoral responses in humans following an infection. to compare the immunoglobulin g (igg) responses to the s and n proteins of sars-cov in sars patients during the manifestation/convalescent period with those during the postinfection period, serum samples were collected from hospitalized sars patients within 6 weeks after the onset of illnes ...	2007	17475765
nucleocapsid protein of sars-cov activates interleukin-6 expression through cellular transcription factor nf-kappab.	high levels of interleukin-6 (il-6) in the acute stage associated with lung lesions were found in sars patients. to evaluate the mechanisms behind this event, we investigated the roles of sars-cov proteins in the regulation of il-6. results showed that the viral nucleocapsid (n) protein activated il-6 expression in a concentration-dependent manner. promoter analyses suggested that nf-kappab binding element was required for il-6 expression regulated by n protein. further studies demonstrated that ...	2007	17490702
the 3a accessory protein of sars coronavirus specifically interacts with the 5'utr of its genomic rna, using a unique 75 amino acid interaction domain.	more than four years have passed since the outbreak of the severe acute respiratory syndrome (sars) epidemic, and still very little is known about the molecular biology and pathogenesis of this deadly virus. among the accessory proteins of the sars coronavirus (sars-cov), the 3a protein has been shown to interact with the spike, envelope, and membrane glycoprotein and has recently been established to be a structural component of capsid. recent studies suggest that the 3a protein may function as ...	2007	17488094
induction of specific immune responses by severe acute respiratory syndrome coronavirus spike dna vaccine with or without interleukin-2 immunization using different vaccination routes in mice.	dna vaccines induce humoral and cellular immune responses in animal models and humans. to analyze the immunogenicity of the severe acute respiratory syndrome (sars) coronavirus (cov), sars-cov, spike dna vaccine and the immunoregulatory activity of interleukin-2 (il-2), dna vaccine plasmids pcdna-s and pcdna-il-2 were constructed and inoculated into balb/c mice with or without pcdna-il-2 by using three different immunization routes (the intramuscular route, electroporation, or the oral route wit ...	2007	17494640
antigenic characterization of severe acute respiratory syndrome-coronavirus nucleocapsid protein expressed in insect cells: the effect of phosphorylation on immunoreactivity and specificity.	the nucleocapsid (n) protein of severe acute respiratory syndrome-coronavirus (sars-cov) is involved in the pathological reaction to sars and is a key antigen for the development of a sensitive diagnostic assay. however, the antigenic properties of this n protein are largely unknown. to facilitate the studies on the function and antigenicity of the sars-cov n protein, 6x histidine-tagged recombinant sars-cov n (rsars-n) with a molecular mass of 46 and 48kda was successfully produced using the re ...	2007	17499376
new structure model for the packaging signal in the genome of group iia coronaviruses.	a 190-nucleotide (nt) packaging signal (ps) located in the 3' end of open reading frame 1b in the mouse hepatitis virus, a group iia coronavirus, was previously postulated to direct genome rna packaging. based on phylogenetic data and structure probing, we have identified a 95-nt hairpin within the 190-nt ps domain which is conserved in all group iia coronaviruses but not in the severe acute respiratory syndrome coronavirus (group iib), group i coronaviruses, or group iii coronaviruses. the hair ...	2007	17428856
sars-cov nucleocapsid protein induced apoptosis of cos-1 mediated by the mitochondrial pathway.	to investigate the apoptosis effect of sars coronavirus nucleocapsid protein on cultured cell lines and to explore the possible pathway of apoptosis. pcdna3.1(-)/his-myc vector containing the sars coronavirus nucleocapsid gene (n), matric gene (m), spike gene (s) were transfected into cos-1, huh-7 and hepg2 cells. apoptosis induced by sars coronavirus n protein under starvation of serum of cos-1 cells was monitored by annexin v and electron microscopy assays. intracellular reactive oxygen specie ...	2007	17453707
induction of apoptosis by the severe acute respiratory syndrome coronavirus 7a protein is dependent on its interaction with the bcl-xl protein.	the severe acute respiratory syndrome coronavirus (sars-cov) 7a protein, which is not expressed by other known coronaviruses, can induce apoptosis in various cell lines. in this study, we show that the overexpression of bcl-xl, a prosurvival member of the bcl-2 family, blocks 7a-induced apoptosis, suggesting that the mechanism for apoptosis induction by 7a is at the level of or upstream from the bcl-2 family. coimmunoprecipitation experiments showed that 7a interacts with bcl-xl and other prosur ...	2007	17428862
knowledge of and attitudes toward severe acute respiratory syndrome among a cohort of dental patients in hong kong following a major local outbreak.	to assess the knowledge of and attitudes toward severe acute respiratory syndrome (sars) among patients attending a teaching dental hospital and private dental practices in hong kong during a major local sars outbreak.	2007	17405470
the sars coronavirus spike glycoprotein is selectively recognized by lung surfactant protein d and activates macrophages.	the severe acute respiratory syndrome coronavirus (sars-cov) infects host cells with its surface glycosylated spike-protein (s-protein). here we expressed the sars-cov s-protein to investigate its interactions with innate immune mechanisms in the lung. the purified s-protein was detected as a 210 kda glycosylated protein. it was not secreted in the presence of tunicamycin and was detected as a 130 kda protein in the cell lysate. the purified s-protein bound to vero but not 293t cells and was its ...	2007	17412287
crystal structure of a monomeric form of severe acute respiratory syndrome coronavirus endonuclease nsp15 suggests a role for hexamerization as an allosteric switch.	mature nonstructural protein-15 (nsp15) from the severe acute respiratory syndrome coronavirus (sars-cov) contains a novel uridylate-specific mn2+-dependent endoribonuclease (nendou). structure studies of the full-length form of the obligate hexameric enzyme from two covs, sars-cov and murine hepatitis virus, and its monomeric homologue, xendou from xenopus laevis, combined with mutagenesis studies have implicated several residues in enzymatic activity and the n-terminal domain as the major dete ...	2007	17409150
g1 phase cell cycle arrest induced by sars-cov 3a protein via the cyclin d3/prb pathway.	sars-cov 3a is a structural protein, mainly localizing to golgi apparatus and co-localizing with sars-cov m in co-transfected cells. here we observed that transient expression of 3a inhibited cell growth and prevented 5-bromodeoxyuridine incorporation, suggesting that 3a deregulated cell cycle progression. cell cycle analysis demonstrated that 3a expression was associated with blockage of cell cycle progression at g1 phase in hek 293, cos-7, and vero cells 24-60 h after transfection. mutation an ...	2007	17413032
community acquired pneumonia in the tropics.	due to the exponential growth of international exchange, millions of travelers are exposed to respiratory pathogens in the tropics and may return ill. community-acquired pneumonia is one of the more prevalent infections.	2007	17414123