Publications

a human in vitro model system for investigating genome-wide host responses to sars coronavirus infection.	the molecular basis of severe acute respiratory syndrome (sars) coronavirus (cov) induced pathology is still largely unclear. many sars patients suffer respiratory distress brought on by interstitial infiltration and frequently show peripheral blood lymphopenia and occasional leucopenia. one possible cause of this could be interstitial inflammation, following a localized host response. in this study, we therefore examine the immune response of sars-cov in human peripheral blood mononuclear cells ...	2004	15357874
highly infectious sars-cov pseudotyped virus reveals the cell tropism and its correlation with receptor expression.	studies of sars coronavirus (sars-cov)-the causative agent of severe acute respiratory syndrome (sars)-have been hampered by its high transmission rate and the pathogenicity of this virus. to permit analysis of the host range and entry mechanism of sars-cov, we incorporated the humanized sars-cov spike (s) glycoprotein into hiv particles to generate a highly infectious sars-cov pseudotyped virus. the infection on vero e6-a permissive cell line to sars-cov-could be neutralized by sera from conval ...	2004	15358126
nucleocapsid protein of sars coronavirus tightly binds to human cyclophilin a.	severe acute respiratory syndrome coronavirus (sars-cov) is responsible for sars infection. nucleocapsid protein (np) of sars-cov (sars_np) functions in enveloping the entire genomic rna and interacts with viron structural proteins, thus playing important roles in the process of virus particle assembly and release. protein-protein interaction analysis using bioinformatics tools indicated that sars_np may bind to human cyclophilin a (hcypa), and surface plasmon resonance (spr) technology revealed ...	2004	15358143
"teaching old drugs to kill new bugs": structure-based discovery of anti-sars drugs.	severe acute respiratory syndrome (sars) main protease or 3c-like protease (3clpro) is essential for the propagation of the coronaviral life cycle and is regarded as one of the main targets for structure-based anti-sars drug design. it is an attractive approach to find new uses for old drugs as they have already been through extensive clinical testing and could easily be accelerated for clinical approval. briefly, we performed virtual screening of a database of small molecules against sars 3clpr ...	2004	15358186
molecular biology of severe acute respiratory syndrome coronavirus.	the worldwide epidemic of severe acute respiratory syndrome (sars) in 2003 was caused by a novel coronavirus called sars-cov. coronaviruses and their closest relatives possess extremely large plus-strand rna genomes and employ unique mechanisms and enzymes in rna synthesis that separate them from all other rna viruses. the sars epidemic prompted a variety of studies on multiple aspects of the coronavirus replication cycle, yielding both rapid identification of the entry mechanisms of sars-cov in ...	2004	15358261
evaluation of metal-conjugated compounds as inhibitors of 3cl protease of sars-cov.	3c-like (3cl) protease is essential for the life cycle of severe acute respiratory syndrome-coronavirus (sars-cov) and therefore represents a key anti-viral target. a compound library consisting of 960 commercially available drugs and biologically active substances was screened for inhibition of sars-cov 3cl protease. potent inhibition was achieved using the mercury-containing compounds thimerosal and phenylmercuric acetate, as well as hexachlorophene. as well, 1-10 microm of each compound inhib ...	2004	15358550
characterization of trans- and cis-cleavage activity of the sars coronavirus 3clpro protease: basis for the in vitro screening of anti-sars drugs.	severe acute respiratory syndrome (sars) has been globally reported. a novel coronavirus (cov), sars-cov, was identified as the etiological agent of the disease. sars-cov 3c-like protease (3clpro) mediates the proteolytic processing of replicase polypeptides 1a and 1ab into functional proteins, playing an important role in viral replication. in this study, we demonstrated the expression of the sars-cov 3clpro in escherichia coli and vero cells, and then characterized the in vitro trans-cleavage ...	2004	15358553
crystal structure of severe acute respiratory syndrome coronavirus spike protein fusion core.	severe acute respiratory syndrome coronavirus is a newly emergent virus responsible for a recent outbreak of an atypical pneumonia. the coronavirus spike protein, an enveloped glycoprotein essential for viral entry, belongs to the class i fusion proteins and is characterized by the presence of two heptad repeat (hr) regions, hr1 and hr2. these two regions are understood to form a fusion-active conformation similar to those of other typical viral fusion proteins. this hairpin structure likely jux ...	2004	15345712
mutational dynamics of the sars coronavirus in cell culture and human populations isolated in 2003.	the sars coronavirus is the etiologic agent for the epidemic of the severe acute respiratory syndrome. the recent emergence of this new pathogen, the careful tracing of its transmission patterns, and the ability to propagate in culture allows the exploration of the mutational dynamics of the sars-cov in human populations.	2004	15347429
emergency medical services utilization during an outbreak of severe acute respiratory syndrome (sars) and the incidence of sars-associated coronavirus infection among emergency medical technicians.	this was a study to evaluate the utilization of emergency medical services (ems) systems during the outbreak of severe acute respiratory syndrome (sars), and to assess the incidence of infection among emergency medical technicians (emts).	2004	15347538
determination of sars-coronavirus by a microfluidic chip system.	we have developed a new experimental system based on a microfluidic chip to determine severe acute respiratory syndrome coronavirus (sars-cov). the system includes a laser-induced fluorescence microfluidic chip analyzer, a glass microchip for both polymerase chain reaction (pcr) and capillary electrophoresis, a chip thermal cycler based on dual peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (rt-pcr) sars diagnostic kit, and a dna electrophoretic sizing kit. th ...	2004	15349945
a universal microarray for detection of sars coronavirus.	severe acute respiratory syndrome (sars) is caused by the sars coronavirus (sars-cov). there are many point mutations among sars-cov genome sequences. previous studies suggested that the mutations are correlated closely with the sars epidemic. it was found that the bases of six nucleotide positions (nt9404, nt9479, nt19838, nt21721, nt22222 and nt27827) with high-mutation rate have an important relationship with the sars epidemic. for viral detection as well as genotyping, a universal microarray ...	2004	15350733
inactivation of the coronavirus that induces severe acute respiratory syndrome, sars-cov.	severe acute respiratory syndrome (sars) is a life-threatening disease caused by a novel coronavirus termed sars-cov. due to the severity of this disease, the world health organization (who) recommends that manipulation of active viral cultures of sars-cov be performed in containment laboratories at biosafety level 3 (bsl3). the virus was inactivated by ultraviolet light (uv) at 254 nm, heat treatment of 65 degrees c or greater, alkaline (ph > 12) or acidic (ph < 3) conditions, formalin and glut ...	2004	15350737
importance of akt signaling pathway for apoptosis in sars-cov-infected vero e6 cells.	severe acute respiratory syndrome (sars) is an acute respiratory tract infectious disease that is associated with a new coronavirus (sars-cov). our recent study indicated that sars-cov infection induces activation of the p38 mitogen-activated protein kinase (mapk) signaling pathway and the p38 mapk inhibitor partially inhibited its cytopathic effect in vero e6 cells. the results of the present study indicated that before cell death, akt, which is an inhibitor of apoptosis, was also activated in ...	2004	15351204
characterization of protein-protein interactions between the nucleocapsid protein and membrane protein of the sars coronavirus.	the human coronavirus, associated with severe acute respiratory syndrome (sars-cov), was identified and molecularly characterized in 2003. sequence analysis of the virus indicates that there is only 20% amino acid (aa) identity with known coronaviruses. previous studies indicate that protein-protein interactions amongst various coronavirus proteins are critical for viral assembly. yet, little sequence homology between the newly identified sars-cov and those previously studied coronaviruses sugge ...	2004	15351485
in vitro inhibition of severe acute respiratory syndrome coronavirus by chloroquine.	we report on chloroquine, a 4-amino-quinoline, as an effective inhibitor of the replication of the severe acute respiratory syndrome coronavirus (sars-cov) in vitro. chloroquine is a clinically approved drug effective against malaria. we tested chloroquine phosphate for its antiviral potential against sars-cov-induced cytopathicity in vero e6 cell culture. results indicate that the ic50 of chloroquine for antiviral activity (8.8 +/- 1.2 microm) was significantly lower than its cytostatic activit ...	2004	15351731
emerging infectious disease issues in international adoptions: severe acute respiratory syndrome (sars), avian influenza and measles.	new emerging infections over the last few years demonstrate the potential for the introduction of epidemic illness through global migration. the increasing number of children adopted internationally (>20,000 in 2003, from the united states state department) provides a unique situation for the spread of emerging infections through the combination of international travel by parents through areas where such infections may be contracted and the nature of the living conditions for many of the orphans ...	2004	15353957
a review of viral gastroenteritis.	since kapakian first identified a virus in the stool of a patient with diarrhoea in 1972, many viruses have been described that cause diarrhoea directly or indirectly. it is now appreciated that viruses are the most common cause of diarrhoeal illness worldwide. although bacteria and other pathogens cause significant numbers of gastroenteritis, it is the viruses that are dealt with in this review. the viruses responsible will be discussed individually.	2004	15353966
osteonecrosis in children with severe acute respiratory syndrome.	osteonecrosis is a debilitating bone disease affecting adults who have recovered from severe acute respiratory syndrome in hong kong and china, but there are no data on its prevalence in children. we report 5 children with magnetic resonance imaging evidence of osteonecrosis. in view of the high prevalence and asymptomatic presentation of osteonecrosis, we suggest magnetic resonance imaging screening for osteonecrosis in children with severe acute respiratory syndrome.	2004	15361738
sars viral rna.		2004	15362378
[construction of phage antibody library for fab fragment from a convalescent patient infected with sars coronavirus].	to construct the phage antibody library for fab fragment from a convalescent patient infected with sars coronavirus.	2004	15367356
synthesis and characterization of a native, oligomeric form of recombinant severe acute respiratory syndrome coronavirus spike glycoprotein.	we have expressed and characterized the severe acute respiratory syndrome coronavirus (sars-cov) spike protein in cdna-transfected mammalian cells. the full-length spike protein (s) was newly synthesized as an endoglycosidase h (endo h)-sensitive glycoprotein (gp170) that is further modified into an endo h-resistant glycoprotein (gp180) in the golgi apparatus. no substantial proteolytic cleavage of s was observed, suggesting that s is not processed into head (s1) and stalk (s2) domains as observ ...	2004	15367599
retroviruses pseudotyped with the severe acute respiratory syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting enzyme 2.	infection of receptor-bearing cells by coronaviruses is mediated by their spike (s) proteins. the coronavirus (sars-cov) that causes severe acute respiratory syndrome (sars) infects cells expressing the receptor angiotensin-converting enzyme 2 (ace2). here we show that codon optimization of the sars-cov s-protein gene substantially enhanced s-protein expression. we also found that two retroviruses, simian immunodeficiency virus (siv) and murine leukemia virus, both expressing green fluorescent p ...	2004	15367630
phylogenetic analysis of the full-length sars-cov sequences: evidence for phylogenetic discordance in three genomic regions.	the origin of the severe acute respiratory syndrome-coronavirus (sars-cov) remains unclear. evidence based on bayesian scanning plots and phylogenetic analysis using maximum likelihood (ml) and bayesian methods indicates that sars-cov, for the largest part of the genome ( approximately 80%), is more closely related to group ii coronaviruses sequences, whereas in three regions in the orf1ab gene it shows no apparent similarity to any of the previously characterized groups of coronaviruses. there ...	2004	15368527
antivirals and antiviral strategies.	in recent years, the demand for new antiviral strategies has increased markedly. there are many contributing factors to this increased demand, including the ever-increasing prevalence of chronic viral infections such as hiv and hepatitis b and c, and the emergence of new viruses such as the sars coronavirus. the potential danger of haemorrhagic fever viruses and eradicated viruses such as variola virus being used as bioterrorist weapons has also increased the profile of antiviral drug discovery. ...	2004	15372081
identification of novel small-molecule inhibitors of severe acute respiratory syndrome-associated coronavirus by chemical genetics.	the severe acute respiratory syndrome-associated coronavirus (sars-cov) infected more than 8,000 people across 29 countries and caused more than 900 fatalities. based on the concept of chemical genetics, we screened 50,240 structurally diverse small molecules from which we identified 104 compounds with anti-sars-cov activity. of these 104 compounds, 2 target the sars-cov main protease (m(pro)), 7 target helicase (hel), and 18 target spike (s) protein-angiotensin-converting enzyme 2 (ace2)-mediat ...	2004	15380189
cellular entry of the sars coronavirus.		2004	15381196
[factors of avascular necrosis of femoral head and osteoporosis in sars patients' convalescence].	to explore the factors affecting the pathogenesis of avascular necrosis of femoral head and osteoporosis of sars patients during convalescent stage.	2004	15387943
immunogenicity of sars inactivated vaccine in balb/c mice.	severe acute respiratory syndrome (sars) is a serious infectious threat to public health. to create a novel trial vaccine and evaluate its potency, we attempted to generate a sars inactivated vaccine using sars coronavirus (sars-cov) strain f69 treated with formaldehyde and mixed with al(oh)3. three doses of the vaccine were used to challenge three groups of balb/c mice. we found that the mice exhibited specific igm on day 4 and igg on day 8. the peak titers of igg were at day 47 in low-dose gro ...	2004	15388253
protective humoral responses to severe acute respiratory syndrome-associated coronavirus: implications for the design of an effective protein-based vaccine.	some of the structural proteins of severe acute respiratory syndrome-associated coronavirus (sars-cov) carry major epitopes involved in virus neutralization and are essential for the induction of protective humoral responses and the development of an effective vaccine. rabbit antisera were prepared using full-length n and m proteins and eight expressed fragments covering the s protein. antisera to s and m proteins were found to have different neutralizing titres towards sars-cov infection in viv ...	2004	15448374
[current topics on sars coronavirus].	the serious respiratory disease, sars (severe acute respiratory syndrome), outbreaking in winter of 2003 to 2004 remained in a sporadic patient's generating at this winter. however, there is also a possibility that wild animals as the source of infection may not be specified and that it may be much in fashion again. the paper regarding sars and sars-cov is published at one per day now which has passed since fashion of sars in one or so year. there are many papers which the researchers of other v ...	2004	15449910
the spike protein of severe acute respiratory syndrome (sars) is cleaved in virus infected vero-e6 cells.	spike protein is one of the major structural proteins of severe acute respiratory syndrome-coronavirus. it is essential for the interaction of the virons with host cell receptors and subsequent fusion of the viral envelop with host cell membrane to allow infection. some spike proteins of coronavirus, such as mhv, hcov-oc43, aibv and bcov, are proteolytically cleaved into two subunits, s1 and s2. in contrast, tgv, fipv and hcov-229e are not. many studies have shown that the cleavage of spike prot ...	2004	15450134
application of real-time pcr for testing antiviral compounds against lassa virus, sars coronavirus and ebola virus in vitro.	this report describes the application of real-time pcr for testing antivirals against highly pathogenic viruses such as lassa virus, sars coronavirus and ebola virus. the test combines classical cell culture with a quantitative real-time pcr read-out. the assay for lassa virus was validated with ribavirin, which showed an ic(50) of 9 micrograms/ml. small-scale screening identified a class of imidazole nucleoside/nucleotide analogues with antiviral activity against lassa virus. the analogues cont ...	2004	15451189
detection of antibodies against sars-cov in serum from sars-infected donors with elisa and western blot.	recombinant fragments of s proteins from the severe acute respiratory syndrome (sars) coronavirus (sara-cov) were generated and used in a western blot (wb) assay that was compared to a commercial sars elisa method. in 85% of confirmed sars cases (n = 20), the s2 recombinant fragment based wb was positive and this was comparable to the commercial elisa using heat killed sars-cov. wb using the other four recombinant fragments in confirmed sars cases generated lower rates of detection (s1--75%, s1- ...	2004	15451470
detecting specific cytotoxic t lymphocytes against sars-coronavirus with dimerx hla-a2:ig fusion protein.	to assess specific cytotoxic t lymphocytes (ctls) against severe acute respiratory syndrome (sars)-coronavirus, a modified dimerx flow cytometry assay was performed with peripheral blood mononuclear cell (pbmc) from hla-a2+ sars-recovered donors at different time points post disease. cd8+dimerx-s1203+ ctls were detected in the pbmc from these donors up to 3 months after recovery. the percentages of cd8+dimerx-s1203+ cells paralleled the numbers of interferon-gamma-positive spots in an elispot as ...	2004	15451471
small molecules blocking the entry of severe acute respiratory syndrome coronavirus into host cells.	severe acute respiratory syndrome coronavirus (sars-cov) is the pathogen of sars, which caused a global panic in 2003. we describe here the screening of chinese herbal medicine-based, novel small molecules that bind avidly with the surface spike protein of sars-cov and thus can interfere with the entry of the virus to its host cells. we achieved this by using a two-step screening method consisting of frontal affinity chromatography-mass spectrometry coupled with a viral infection assay based on ...	2004	15452254
macaque model for severe acute respiratory syndrome.	rhesus and cynomolgus macaques were challenged with 10(7) pfu of a clinical isolate of the severe acute respiratory syndrome (sars) coronavirus. some of the animals developed a mild self-limited respiratory infection very different from that observed in humans with sars. the macaque model as it currently exists will have limited utility in the study of sars and the evaluation of therapies.	2004	15452262
macaque model for severe acute respiratory syndrome.	rhesus and cynomolgus macaques were challenged with 10(7) pfu of a clinical isolate of the severe acute respiratory syndrome (sars) coronavirus. some of the animals developed a mild self-limited respiratory infection very different from that observed in humans with sars. the macaque model as it currently exists will have limited utility in the study of sars and the evaluation of therapies.	2004	15452262
resolution of primary severe acute respiratory syndrome-associated coronavirus infection requires stat1.	intranasal inhalation of the severe acute respiratory syndrome coronavirus (sars cov) in the immunocompetent mouse strain 129svev resulted in infection of conducting airway epithelial cells followed by rapid clearance of virus from the lungs and the development of self-limited bronchiolitis. animals resistant to the effects of interferons by virtue of a deficiency in stat1 demonstrated a markedly different course following intranasal inhalation of sars cov, one characterized by replication of vi ...	2004	15452265
efficient replication of severe acute respiratory syndrome coronavirus in mouse cells is limited by murine angiotensin-converting enzyme 2.	replication of viruses in species other than their natural hosts is frequently limited by entry and postentry barriers. the coronavirus that causes severe acute respiratory syndrome (sars-cov) utilizes the receptor angiotensin-converting enzyme 2 (ace2) to infect cells. here we compare human, mouse, and rat ace2 molecules for their ability to serve as receptors for sars-cov. we found that, compared to human ace2, murine ace2 less efficiently bound the s1 domain of sars-cov and supported less-eff ...	2004	15452268
globalization and risks to health.		2004	15459728
severe acute respiratory syndrome surveillance in australia.	in march 2003, the world health organization (who) issued a global alert recommending active worldwide surveillance for severe acute respiratory syndrome (sars). this paper describes the epidemiological features of cases reported by australian states and territories to the australian government department of health and ageing between 17 march and 31 july 2003. there were 138 people investigated for sars: 111 as suspect and 27 as probable. five probable cases were reported to who after review of ...	2004	15460954
american society for virology-23rd annual meeting. vaccines and antiviral agents 10-14 july 2004, montreal canada.		2004	15470595
antibody world summit 2004-sri conference 19-21 july 2004, philadelphia, pa, usa.	overall, the sessions were well attended and a lot of questions were asked. two-thirds of the speakers were from industry and the rest from academia. of particular note, the academics presented cutting-edge science in the antibody discovery and development area. business representatives from various therapeutic antibody companies discussed in detail various aspects and challenges of building a viable therapeutic antibody company.	2004	15470596
temporal relationship of viral load, ribavirin, interleukin (il)-6, il-8, and clinical progression in patients with severe acute respiratory syndrome.	although viral replication and overwhelming immune responses are believed to contribute to the progression of severe acute respiratory syndrome (sars), little is known about the temporal relationship between viral load, ribavirin, proinflammatory cytokines, and clinical progression. we report that ribavirin was not effective in reducing the sars coronavirus load in 3 of 8 probable cases studied and that elevated levels of interleukin (il)-6 and il-8 subsequent to the peak viral load were found i ...	2004	15472864
efficient assembly and release of sars coronavirus-like particles by a heterologous expression system.	virus-like particles (vlps) produced by recombinant expression of the major viral structural proteins could be an attractive method for severe acute respiratory syndrome (sars) control. in this study, using the baculovirus system, we generated recombinant viruses that expressed s, e, m and n structural proteins of sars-cov either individually or simultaneously. the expression level, size and authenticity of each recombinant sars-cov protein were determined. in addition, immunofluorescence and fa ...	2004	15474033
a novel auto-cleavage assay for studying mutational effects on the active site of severe acute respiratory syndrome coronavirus 3c-like protease.	the 3c-like protease (3clpro) of severe acute respiratory syndrome (sars) has been proposed as an attractive target for drug design. his41 and cys145 were essential for the active site as the principal catalytic residues. in this study, we mutated the two sites, expressed four resulting mutants in escherichia coli and characterized. all mutants showed undetectable activity in trans-cleavage assay. in addition, we introduced a 31-mer peptide containing an auto-cleavage site to the n-terminal of t ...	2004	15474466
immunological, structural, and preliminary x-ray diffraction characterizations of the fusion core of the sars-coronavirus spike protein.	the sars-cov spike protein, a glycoprotein essential for viral entry, is a primary target for vaccine and drug development. two peptides denoted hr-n(sn50) and hr-c(sc40), corresponding to the leu/ile/val-rich heptad-repeat regions from the n-terminal and c-terminal segments of the sars-cov spike s2 sequence, respectively, were synthesized and predicted to form trimeric assembly of hairpin-like structures. the polyclonal antibodies produced by recombinant s2 protein were tested for antigenicity ...	2004	15474492
receptor-binding domain of sars-cov spike protein induces highly potent neutralizing antibodies: implication for developing subunit vaccine.	the spike (s) protein of severe acute respiratory syndrome (sars) coronavirus (cov), a type i transmembrane envelope glycoprotein, consists of s1 and s2 domains responsible for virus binding and fusion, respectively. the s1 contains a receptor-binding domain (rbd) that can specifically bind to angiotensin-converting enzyme 2 (ace2), the receptor on target cells. here we show that a recombinant fusion protein (designated rbd-fc) containing 193-amino acid rbd (residues 318-510) and a human igg1 fc ...	2004	15474494
medical treatment of viral pneumonia including sars in immunocompetent adult.	since no randomized controlled trials have been conducted on the treatment of viral pneumonia by antivirals or immunomodulators in immunocompetent adults, a review of such anecdotal experience are needed for the more rational use of such agents. case reports (single or case series) with details on their treatment and outcome in the english literature can be reviewed for pneumonia caused by human or avian influenza a virus (50 patients), varicella zoster virus (120), adenovirus (29), hantavirus ( ...	2004	15474623
interferon-beta and interferon-gamma synergistically inhibit the replication of severe acute respiratory syndrome-associated coronavirus (sars-cov).	recent studies have shown that interferon-gamma (ifn-gamma) synergizes with ifn-alpha/beta to inhibit the replication of both rna and dna viruses. we investigated the effects of ifns on the replication of two strains of severe acute respiratory syndrome-associated coronavirus (sars-cov). while treatment of vero e6 cells with 100 u/ml of either ifn-beta or ifn-gamma marginally reduced viral replication, treatment with both ifn-beta and ifn-gamma inhibited sars-cov plaque formation by 30-fold and ...	2004	15476870
natural course of severe acute respiratory syndrome-associated coronavirus immunoglobulin after infection.		2004	15478079
dc-sign and dc-signr interact with the glycoprotein of marburg virus and the s protein of severe acute respiratory syndrome coronavirus.	the lectins dc-sign and dc-signr can augment viral infection; however, the range of pathogens interacting with these attachment factors is incompletely defined. here we show that dc-sign and dc-signr enhance infection mediated by the glycoprotein (gp) of marburg virus (marv) and the s protein of severe acute respiratory syndrome coronavirus and might promote viral dissemination. signr1, a murine dc-sign homologue, also enhanced infection driven by marv and ebola virus gp and could be targeted to ...	2004	15479853
a previously undescribed coronavirus associated with respiratory disease in humans.	the etiology of acute respiratory tract illnesses is sometimes unclear due to limitations of diagnostic tests or the existence of as-yet-unidentified pathogens. here we describe the identification and characterization of a not previously recognized coronavirus obtained from an 8-mo-old boy suffering from pneumonia. this coronavirus replicated efficiently in tertiary monkey kidney cells and vero cells, in contrast to human coronaviruses (hcov) 229e and oc43. the entire cdna genome sequence of the ...	2004	15073334
inhibition of severe acute respiratory syndrome-associated coronavirus (sarscov) by calpain inhibitors and beta-d-n4-hydroxycytidine.	we evaluated two types of compounds for efficacy in inhibiting sarscov replication in vitro: calpain inhibitors (a class of cellular cysteine proteinases) and a number of nucleoside analogues. cytopathic effect reduction assays visually determined with spectrophotometric verification by neutral red (nr) uptake assay were used to evaluate cytotoxicity and antiviral potency of the compounds. significantly inhibitory compounds were then evaluated in virus yield reduction assays. two calpain inhibit ...	2004	15074711
coronavirus infection in an aids patient.		2004	15075525
3c-like proteinase from sars coronavirus catalyzes substrate hydrolysis by a general base mechanism.	sars 3c-like proteinase has been proposed to be a key enzyme for drug design against sars. lack of a suitable assay has been a major hindrance for enzyme kinetic studies and a large-scale inhibitor screen for sars 3cl proteinase. since sars 3cl proteinase belongs to the cysteine protease family (family c3 in clan cb) with a chymotrypsin fold, it is important to understand the catalytic mechanism of sars 3cl proteinase to determine whether the proteolysis proceeds through a general base catalysis ...	2004	15078103
amino acids 270 to 510 of the severe acute respiratory syndrome coronavirus spike protein are required for interaction with receptor.	a novel coronavirus, severe acute respiratory syndrome coronavirus (sars-cov), has recently been identified as the causative agent of severe acute respiratory syndrome (sars). sars-cov appears similar to other coronaviruses in both virion structure and genome organization. it is known for other coronaviruses that the spike (s) glycoprotein is required for both viral attachment to permissive cells and for fusion of the viral envelope with the host cell membrane. here we describe the construction ...	2004	15078936
generation and characterization of dna vaccines targeting the nucleocapsid protein of severe acute respiratory syndrome coronavirus.	severe acute respiratory syndrome (sars) is a serious threat to public health and the economy on a global scale. the sars coronavirus (sars-cov) has been identified as the etiological agent for sars. thus, vaccination against sars-cov may represent an effective approach to controlling sars. dna vaccines are an attractive approach for sars vaccine development, as they offer many advantages over conventional vaccines, including stability, simplicity, and safety. our investigators have previously s ...	2004	15078946
the involvement of natural killer cells in the pathogenesis of severe acute respiratory syndrome.	by using peripheral blood samples from 221 cases of severe acute respiratory syndrome (sars), 34 of mycoplasma pneumoniae infection, and 44 healthy adults, we measured the total number of natural killer (nk) and cd158b+ nk cells (cd158b+ nk) using flow cytometric analysis and calculated the percentage of cd158b+ nk cells. the total number of nk and cd158b+ nk cells and the percentage of cd158b+ nk cells were significantly lower in patients with sars than in those with m. pneumoniae infection (p ...	2004	15080302
detection of severe acute respiratory syndrome-associated coronavirus in pneumocytes of the lung.	previous reports have indicated that patients with severe acute respiratory syndrome (sars)-associated coronavirus infection could develop atypical pneumonia with fulminant pulmonary edema. however, the target cells of sars viral infection have not been characterized in detail. we report the pathologic findings of the lung in 3 cases of sars. chest radiographs at 2 to 3 weeks of infection revealed an atypical pneumonia with pulmonary consolidation, a clinical characteristic of sars infection. th ...	2004	15080310
exploring the binding mechanism of the main proteinase in sars-associated coronavirus and its implication to anti-sars drug design.	the main proteinase of sars-associated coronavirus (sars-cov) plays an important role in viral transcription and replication, and is an attractive target for anti-sars drug development. the important thing is to understand its binding mechanism with possible ligands. in this study, we investigated possible noncanonical interactions, potential inhibitors, and binding pockets in the main proteinase of sars-cov based on its recently determined crystal structure. these findings provide a wide clue t ...	2004	15080921
induction of sars-nucleoprotein-specific immune response by use of dna vaccine.	induction of effective cytotoxic t lymphocyte (ctl) and/or a specific antibody against conserved viral proteins may be essential to the development of a safe and effective severe acute respiratory syndrome coronavirus (sars-cov) vaccine. dna vaccination represents a new strategy for induction of humoral and cellular immune response. to determine the ability of sars-cov nucleoprotein (n protein) to induce antiviral immunity, in this report, we established a stable c2c12 line expressing sars-cov n ...	2004	15081618
are there characteristics of infectious diseases that raise special ethical issues?	this paper examines the characteristics of infectious diseases that raise special medical and social ethical issues, and explores ways of integrating both current bioethical and classical public health ethics concerns. many of the ethical issues raised by infectious diseases are related to these diseases' powerful ability to engender fear in individuals and panic in populations. we address the association of some infectious diseases with high morbidity and mortality rates, the sense that infecti ...	2004	15086371
primary characterization of sars coronavirus strain frankfurt 1.		2004	15088406
the nucleocapsid protein of the sars coronavirus is capable of self-association through a c-terminal 209 amino acid interaction domain.	severe acute respiratory syndrome (sars) coronavirus (sars-cov) caused a severe outbreak in several regions of the world in 2003. the virus is a novel coronavirus isolated from patients exhibiting atypical pneumonia and may have originated from wild animals such as civet cats in southern china. the genome of sars-cov is a positive-sense, single-stranded rna whose sequence is distantly related to all known coronaviruses that infect humans and animals. like other known coronaviruses, sars-cov is a ...	2004	15094372
[sars].		2004	15095764
severe acute respiratory syndrome.	severe acute respiratory syndrome (sars) is a new respiratory illness due to a novel coronavirus called sars-cov. cases of sars were first identified from guangdong province in southern china in november 2002. over the next several months, international travel allowed for the rapid spread of >8,000 cases over three continents. the economic and psychological impact of this mysterious illness was profound. in order to better educate the public about this emerging infectious disease, we describe th ...	2004	15095827
the spectrum of severe acute respiratory syndrome-associated coronavirus infection.	whether subclinical or atypical presentations of severe acute respiratory syndrome (sars) occur and whether clinical judgment is accurate in detecting sars are unknown. objectives: to describe the spectrum of sars coronavirus infection in a large outbreak and to compare diagnoses based on clinical judgment with the sars coronavirus test.	2004	15096332
summaries for patients. do some patients with sars have mild disease?		2004	15096361
antibody response and viraemia during the course of severe acute respiratory syndrome (sars)-associated coronavirus infection.	to understand the time-course of viraemia and antibody responses to severe acute respiratory syndrome-associated coronavirus (sars-cov), rt-pcr and elisa were used to assay 376 blood samples from 135 sars patients at various stages of the illness, including samples from patients who were in their early convalescent phase. the results showed that igm antibodies decreased and became undetectable 11 weeks into the recovery phase. igg antibodies, however, remained detectable for a period beyond 11 w ...	2004	15096554
severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice.	the spike protein (s), a membrane component of severe acute respiratory syndrome coronavirus (sars-cov) is anticipated to be an important component of candidate vaccines. we constructed recombinant forms of the highly attenuated modified vaccinia virus ankara (mva) containing the gene encoding full-length sars-cov s with and without a c-terminal epitope tag called mva/s-ha and mva/s, respectively. cells infected with mva/sor mva/s-ha synthesized a 200-kda protein, which was recognized by antibod ...	2004	15096611
evidence of airborne transmission of the severe acute respiratory syndrome virus.	there is uncertainty about the mode of transmission of the severe acute respiratory syndrome (sars) virus. we analyzed the temporal and spatial distributions of cases in a large community outbreak of sars in hong kong and examined the correlation of these data with the three-dimensional spread of a virus-laden aerosol plume that was modeled using studies of airflow dynamics.	2004	15102999
laboratory-acquired severe acute respiratory syndrome.		2004	15103000
sars ctl vaccine candidates; hla supertype-, genome-wide scanning and biochemical validation.	an effective severe acute respiratory syndrome (sars) vaccine is likely to include components that can induce specific cytotoxic t-lymphocyte (ctl) responses. the specificities of such responses are governed by human leukocyte antigen (hla)-restricted presentation of sars-derived peptide epitopes. exact knowledge of how the immune system handles protein antigens would allow for the identification of such linear sequences directly from genomic/proteomic sequence information (lauemoller et al., re ...	2004	15104671
[severe acute respiratory syndrome].		2004	15108450
identification of novel inhibitors of the sars coronavirus main protease 3clpro.	sars (severe acute respiratory syndrome) is caused by a newly discovered coronavirus. a key enzyme for the maturation of this virus and, therefore, a target for drug development is the main protease 3cl(pro) (also termed sars-cov 3cl(pro)). we have cloned and expressed in escherichia coli the full-length sars-cov 3cl(pro) as well as a truncated form containing only the catalytic domains. the recombinant proteins have been characterized enzymatically using a fluorescently labeled substrate; their ...	2004	15109248
coronaviridae and sars-associated coronavirus strain hsr1.	during the recent severe acute respiratory (sars) outbreak, the etiologic agent was identified as a new coronavirus (cov). we have isolated a sars-associated cov (sars-cov) strain by injecting vero cells with a sputum specimen from an italian patient affected by a severe pneumonia; the patient traveled from vietnam to italy in march 2003. ultrastructural analysis of infected vero cells showed the virions within cell vesicles and around the cell membrane. the full-length viral genome sequence was ...	2004	15109406
development of taqman rt-nested pcr system for clinical sars-cov detection.	severe acute respiratory syndrome (sars) is an acute newly emerged infectious respiratory illness. the etiologic agent of sars was named 'sars-associated coronavirus' (sars-cov) that can be detected with reverse transcription-polymerase chain reaction (rt-pcr) assays. in this study, 12 sets of nested primers covering the sars-cov genome have been screened and showed sufficient sensitivity to detect sars-cov in rna isolated from virus cultured in vero 6 cells. to optimize further the reaction con ...	2004	15109816
microbicides and the environmental control of nosocomial viral infections.	viruses are important causes of acute and chronic diseases in humans. newer viruses are still being discovered and those that are already known are being incriminated in the aetiology of clinical conditions with hitherto unknown causes. apart from frequently causing infections in the general community, many types of viruses are also significant nosocomial pathogens. while it is generally agreed that we underestimate the proportion of nosocomial infections that are viral, due to a lack of routine ...	2004	15110126
old drugs as lead compounds for a new disease? binding analysis of sars coronavirus main proteinase with hiv, psychotic and parasite drugs.	the sars-associated coronavirus (sars-cov) main proteinase is a key enzyme in viral polyprotein processing. to allow structure-based design of drugs directed at sars-cov main proteinase, we predicted its binding pockets and affinities with existing hiv, psychotic and parasite drugs (lopinavir, ritonavir, niclosamide and promazine), which show signs of inhibiting the replication of sars-cov. our results suggest that these drugs and another two hiv inhibitors (pnu and uc2) could be used as templat ...	2004	15110833
severe acute respiratory syndrome: report of treatment and outcome after a major outbreak.	the outcome is reported of a prospective uncontrolled study based on a stepwise treatment protocol during an outbreak of severe acute respiratory syndrome (sars) in hong kong.	2004	15115870
monophyletic relationship between severe acute respiratory syndrome coronavirus and group 2 coronaviruses.	although primary genomic analysis has revealed that severe acute respiratory syndrome coronavirus (sars cov) is a new type of coronavirus, the different protein trees published in previous reports have provided no conclusive evidence indicating the phylogenetic position of sars cov. to clarify the phylogenetic relationship between sars cov and other coronaviruses, we compiled a large data set composed of 7 concatenated protein sequences and performed comprehensive analyses, using the maximum-lik ...	2004	15116304
severe acute respiratory syndrome (sars) and the gdp. part i : epidemiology, virology, pathology and general health issues.	the health profession faces a new challenge with the emergence of a novel viral disease severe acute respiratory syndrome (sars), a form of atypical pneumonia caused by a coronavirus termed sars-cov. this highly infectious disease has spread through 32 countries, infecting more than 8,400 patients with over 790 deaths in just over 6 months. over one quarter of those infected were unsuspecting healthcare workers. the major transmission mode of sars-coronavirus appears to be through droplet spread ...	2004	15272339
disease-specific b cell epitopes for serum antibodies from patients with severe acute respiratory syndrome (sars) and serologic detection of sars antibodies by epitope-based peptide antigens.	severe acute respiratory syndrome (sars) has emerged as a highly contagious, sometimes fatal disease. to find disease-specific b cell epitopes, phage-displayed random peptide libraries were panned on serum immunoglobulin (ig) g antibodies from patients with sars. forty-nine immunopositive phage clones that bound specifically to serum from patients with sars were selected. these phageborne peptides had 4 consensus motifs, of which 2 corresponded to amino acid sequences reported for sars-associate ...	2004	15272409
structural organization of the genome of sars-associated coronavirus (strain sod) isolated on the territory of the russian federation.	complete nucleotide cdna sequence (29715 nucleotides) of sars-associated coronavirus (strain sod) isolated for the first time in the territory of the russian federation was determined. phylogenetic analysis revealed maximum similarity between strain sod genome and frankfurt 1 strain genome. three nucleotide substitutions determining two amino acid substitutions were detected.	2004	15273774
the spectrum of pathological changes in severe acute respiratory syndrome (sars).	to analyse the lung pathology of severe acute respiratory syndrome (sars) and correlate the findings with the time sequence of the disease.	2004	15279629
antibody responses against sars-coronavirus and its nucleocaspid in sars patients.	sars-cov is the etiologic agent of severe acute respiratory syndrome. an understanding of the antibody responses to the viral components is very important for diagnosis and vaccine development.	2004	15288616
in vitro susceptibility of 10 clinical isolates of sars coronavirus to selected antiviral compounds.	effective antiviral agents are urgently needed to combat the possible return of severe acute respiratory syndrome (sars). commercial antiviral agents and pure chemical compounds extracted from traditional chinese medicinal herbs were screened against 10 clinical isolates of sars coronavirus by neutralisation tests with confirmation by plaque reduction assays. interferon-beta-1a, leukocytic interferon-alpha, ribavirin, lopinavir, rimantadine, baicalin and glycyrrhizin showed antiviral activity. t ...	2004	15288617
a highly unusual palindromic transmembrane helical hairpin formed by sars coronavirus e protein.	the agent responsible for the recent severe acute respiratory syndrome (sars) outbreak is a previously unidentified coronavirus. while there is a wealth of epidemiological studies, little if any molecular characterization of sars coronavirus (scov) proteins has been carried out. here we describe the molecular characterization of scov e protein, a critical component of the virus responsible for virion envelope morphogenesis. we conclusively show that scov e protein contains an unusually short, pa ...	2004	15288785
sars, emerging infections, and bioterrorism preparedness.		2004	15288816
the sars coronavirus nucleocapsid protein induces actin reorganization and apoptosis in cos-1 cells in the absence of growth factors.	in march 2003, a novel coronavirus was isolated from patients exhibiting atypical pneumonia, and was subsequently proven to be the causative agent of the disease now referred to as sars (severe acute respiratory syndrome). the complete genome of the sars-cov (sars coronavirus) has since been sequenced. the sars-cov nucleocapsid (sars-cov n) protein shares little homology with other members of the coronavirus family. in the present paper, we show that sars-cov n is capable of inducing apoptosis o ...	2004	15294014
pregnancy and perinatal outcomes of women with severe acute respiratory syndrome.	this study was undertaken to evaluate the pregnancy and perinatal outcomes of pregnant women with severe acute respiratory syndrome (sars).	2004	15295381
recombinant scfv antibodies against e protein and n protein of severe acute respiratory syndrome virus.	three single chain antibodies (scfv) against the proteins of severe acute respiratory syndrome coronavirus (sars-cov) were isolated by phage display from an scfv antibody library. bio-panning was carried out against immobilized purified envelope (e) and nucleocapsid (n) proteins of sars-cov. their binding activity and specificity to e or n protein of sars-cov were characterized by phage-elisa. two of them, b10 and c20, could recognize non-overlapping epitopes of the e protein according to the tw ...	2004	15295646
potent inhibition of sars-associated coronavirus (scov) infection and replication by type i interferons (ifn-alpha/beta) but not by type ii interferon (ifn-gamma).	we sought to investigate the anti-severe acute respiratory syndrome (sars)-associated coronavirus (scov) activities of type i (alpha and beta) and type ii (gamma) interferons (ifn) in vitro. type i ifns protected cells from cytopathic effects (cpe) induced by scov, and inhibited viral genomic rna replication in frhk-4 cells (measured by quantitative rt-pcr) in a dose-dependent manner. intracellular viral rna copies were reduced 50% by ifn-alpha at a concentration of 25 u/ml and by ifn-beta at a ...	2004	15296649
sars virus in tears?		2004	15298024
clinical and laboratory features of severe acute respiratory syndrome vis-a-vis onset of fever.	severe acute respiratory syndrome (sars) is a rapidly progressive disease caused by a novel coronavirus (cov) infection. however, the disease presentation is nonspecific. the aim of this study was to define clearly the presentation, clinical progression, and laboratory data in a group of patients who had sars.	2004	15302738
major genetic marker of nidoviruses encodes a replicative endoribonuclease.	coronaviruses are important pathogens that cause acute respiratory diseases in humans. replication of the approximately 30-kb positive-strand rna genome of coronaviruses and discontinuous synthesis of an extensive set of subgenome-length rnas (transcription) are mediated by the replicase-transcriptase, a barely characterized protein complex that comprises several cellular proteins and up to 16 viral subunits. the coronavirus replicase-transcriptase was recently predicted to contain rna-processin ...	2004	15304651
viral evolution and the emergence of sars coronavirus.	the recent appearance of severe acute respiratory syndrome coronavirus (sars-cov) highlights the continual threat to human health posed by emerging viruses. however, the central processes in the evolution of emerging viruses are unclear, particularly the selection pressures faced by viruses in new host species. we outline some of the key evolutionary genetic aspects of viral emergence. we emphasize that, although the high mutation rates of rna viruses provide them with great adaptability and exp ...	2004	15306390
influenza as a model system for studying the cross-species transfer and evolution of the sars coronavirus.	severe acute respiratory syndrome coronavirus (sars-cov) moved into humans from a reservoir species and subsequently caused an epidemic in its new host. we know little about the processes that allowed the cross-species transfer of this previously unknown virus. i discuss what we have learned about the movement of viruses into humans from studies of influenza a, both how it crossed from birds to humans and how it subsequently evolved within the human population. starting with a brief review of se ...	2004	15306391