Publications

first international quality assurance study on the rapid detection of viral agents of bioterrorism.	we have conducted an international quality assurance study of filovirus, lassa virus, and orthopox virus pcr with 24 participants. of the participating laboratories, 45.8 and 66.7% detected virus in all plasma samples, which contained > or = 5,000 and > or = 100,000 copies per ml, respectively. sensitivity levels were not significantly different between viruses. false-negative results were attributable to a lack of sensitivity.	2004	15071040
smallpox dna vaccine protects nonhuman primates against lethal monkeypox.	two decades after a worldwide vaccination campaign was used to successfully eradicate naturally occurring smallpox, the threat of bioterrorism has led to renewed vaccination programs. in addition, sporadic outbreaks of human monkeypox in africa and a recent outbreak of human monkeypox in the u.s. have made it clear that naturally occurring zoonotic orthopoxvirus diseases remain a public health concern. much of the threat posed by orthopoxviruses could be eliminated by vaccination; however, becau ...	2004	15078924
protective effect of exogenous recombinant mouse interferon-gamma and tumour necrosis factor-alpha on ectromelia virus infection in susceptible balb/c mice.	the resistance to mousepox is correlated with the production of type i cytokines: interleukin (il)-2, il-12, interferon (ifn)-gamma and tumour necrosis factor (tnf)-alpha. we intend to describe the modulation of generalized ectromelia virus (ev) infection with exogenous administration of mrifn-gamma and mrtnf-alpha separately and in combination using susceptible balb/c mice. the treatment schemes presented resulted in the localization of the generalized ev infection and its development into non- ...	2004	15086382
inhibitory activity of alkoxyalkyl and alkyl esters of cidofovir and cyclic cidofovir against orthopoxvirus replication in vitro.	a new series of ether lipid esters of cidofovir (cdv) were evaluated against vaccinia and cowpox viruses. activity was dependent on number of atoms in the alkyl or alkoxyalkyl chain, the linker moiety, and the presence of a double bond in the alkoxyalkyl chains linked to the phosphonate moiety of cdv.	2004	15105146
working safely with vaccinia virus: laboratory technique and the role of vaccinia vaccination.	vaccinia virus, the prototype orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology, virus-host interactions, and as a protein expression system and a vaccine vector. the ubiquitous use of vaccinia viruses in the laboratory raises certain safety concerns, because the virus can be a pathogen in individuals with immunological and dermatological abnormalities and, on occasion, can cause serious problems in normal hosts. this chapter reviews stan ...	2004	15114003
construction of recombinant vaccinia viruses using leporipoxvirus-catalyzed recombination and reactivation of orthopoxvirus dna.	poxvirus dna is not infectious because the initiation of the infective process requires proteins encapsidated along with the virus genome. however, infectious virus can be produced if purified poxvirus dna is transfected into cells previously infected with another poxvirus. this process is termed heterologous reactivation if the infecting virus is different from the transfected virus. we describe a method in which the high-frequency recombination and replication reactions catalyzed by the lepori ...	2004	15114007
orthopoxvirus diagnostics.	biologic and antigenic properties are often useful for identifying and differentiating orthopoxviruses (opv). however, polymerase chain reaction (pcr) amplification, with either restriction cleavage or sequencing of amplicons, has been gaining credibility as a more rapid, specific, sensitive, and often cost-saving technique for research and diagnostic laboratories. this chapter is consolidated using prior research papers from our laboratories with three different methods that should be suitable ...	2004	15114012
mouse models for studying orthopoxvirus respiratory infections.	concern regarding the use of variola and monkeypox viruses as bioterrorist agents has led to an increased study of orthopoxviruses to understand the molecular and cellular basis of pathogenesis and develop safe and effective antivirals and vaccines against smallpox. crucial to these efforts is the availability of animal models, which are inexpensive, genetically homogeneous, and recapitulate the human disease. the popular small-animal orthopoxvirus models employ the inbred mouse as the host, the ...	2004	15114022
real-time pcr system for detection of orthopoxviruses and simultaneous identification of smallpox virus.	a screening assay for real-time lightcycler (roche applied science, mannheim, germany) pcr identification of smallpox virus dna was developed and compiled in a kit system under good manufacturing practice conditions with standardized reagents. in search of a sequence region unique to smallpox virus, the nucleotide sequence of the 14-kda fusion protein gene of each of 14 variola virus isolates of the russian world health organization smallpox virus repository was determined and compared to publis ...	2004	15131152
polarized type 1 cytokine response and cell-mediated immunity determine genetic resistance to mousepox.	ectromelia virus (ectv), a natural mouse pathogen and an orthopoxvirus, has been used to investigate the correlation between polarized type 1 or type 2 immune responses and resistance to disease in poxvirus infections by using well defined resistant and susceptible mouse strains. our data show that distinct differences exist in the cytokine profiles expressed in resistant and susceptible mice infected with ectv. resistant c57bl/6 mice generate a type 1 cytokine response [ifn-gamma, il-2, and tum ...	2004	15184649
identification of residues in an orthopoxvirus interleukin-18 binding protein involved in ligand binding and species specificity.	interleukin-18 (il-18) is a critical cytokine in inflammation and adaptive immune responses. the il-18 binding proteins (il-18bp) are a family of proteins that bind to, and inhibit the activity of, il-18. using point mutagenesis, we analyzed the ectromelia virus il-18bp to identify residues involved in binding. because p13 can bind both human and murine il-18, and because it is highly homologous to the variola virus il-18bp, we set out to identify residues that may be involved in species specifi ...	2004	15193916
cutaneous infections of mice with vaccinia or cowpox viruses and efficacy of cidofovir.	orthopoxviruses, including smallpox, monkeypox and molluscipox, pose risks to human health through bioterrorist acts or natural transmission. there is no approved therapy for orthopoxvirus infections; however, cidofovir (cdv) has been approved as an investigational new drug for emergency treatment of adverse effects following smallpox vaccination. for evaluation of new therapies directed against orthopoxvirus infections, we have utilized immunocompetent, hairless mice (skh-1) inoculated by a cut ...	2004	15196818
raccoon poxvirus as a mucosal vaccine vector for domestic cats.	in the present study, we evaluated both the immunogenicity and safety of recombinant raccoon poxvirus (rcn) as a mucosal vaccine vector for domestic cats. rcn is an orthopoxvirus that was isolated from healthy raccoons and has been used experimentally as a vaccine vector for rabies and other antigens in a variety of species, including raccoons, skunks, foxes, bobcats, rabbits, domestic cats, piglets, sheep and non-human primates. we evaluated the antibody response induced by a recombinant rcn va ...	2003	15203914
monkeypox virus detection in rodents using real-time 3'-minor groove binder taqman assays on the roche lightcycler.	during the summer of 2003, an outbreak of human monkeypox occurred in the midwest region of the united states. in all, 52 rodents suspected of being infected with monkeypox virus were collected from an exotic pet dealer and from private homes. the rodents were euthanized and submitted for testing to the united states army medical research institute of infectious diseases by the galesburg animal disease laboratory, illinois department of agriculture. the rodent tissue samples were appropriately p ...	2004	15208646
belo horizonte virus: a vaccinia-like virus lacking the a-type inclusion body gene isolated from infected mice.	here is described the isolation of a naturally occurring a-type inclusion body (ati)-negative vaccinia-like virus, belo horizonte virus (vbh), obtained from a mousepox-like outbreak in brazil. the isolated virus was identified and characterized as an orthopoxvirus by conventional methods. molecular characterization of the virus was done by dna cross-hybridization using vaccinia virus (vacv) dna. in addition, conserved orthopoxvirus genes such as vaccinia growth factor, thymidine kinase and haema ...	2004	15218187
differential efficacy of vaccinia virus envelope proteins administered by dna immunisation in protection of balb/c mice from a lethal intranasal poxvirus challenge.	dna vaccines might offer an alternative to the live smallpox vaccine in providing protective efficacy in an orthopoxvirus (opv) lethal respiratory challenge model. balb/c mice were immunised with dna vaccines coding for 10 different single vaccinia virus (vacv) membrane proteins. after an intranasal challenge with the vacv ihd strain, three gene candidates b5r, a33r and a27l produced > or =66% survival. the b5r dna vaccine consistently produced 100% protection and exhibited greatest efficacy aft ...	2004	15308360
anticipating smallpox and monkeypox outbreaks: complications of the smallpox vaccine.	the recent outbreak in the midwest of monkeypox, as well as the continued fears of a terrorist-induced epidemic of smallpox, prompted the authors' review of the literature regarding past and current experiences with smallpox vaccination. the smallpox vaccine, which is highly effective in preventing the spread of both these orthopoxvirus infectious illnesses, might be administered to numerous health care workers and, in the event of a smallpox attack, millions of other citizens. however, vaccinee ...	2004	15335444
in search of rnase p rna from microbial genomes.	a simple procedure has been developed to quickly retrieve and validate the dna sequence encoding the rna subunit of ribonuclease p (rnase p rna) from microbial genomes. rnase p rna sequences were identified from 94% of bacterial and archaeal complete genomes where previously no rnase p rna was annotated. a sequence was found in camelpox virus, highly conserved in all orthopoxviruses (including smallpox virus), which could fold into a putative rnase p rna in terms of conserved primary features an ...	2004	15337843
can we really learn from model pathogens?	cytolytic t (tc) lymphocytes are the first order response of the adaptive immune system in the recovery from primary viral infections. these effector cells execute their function either by direct cytotoxicity through the fas or perforin pathway and/or by the release of cytokines that either directly or indirectly exert antiviral activity. mice respond to infection by closely related viruses with a vigorous tc response, which is characterized by extensive crossreactivity on target cells infected ...	2004	15364054
[a study of neutralizing activity and cross-reactivity of monoclonal antibodies to ectromelia virus with orthopoxviruses pathogenic for man].	an extensive collection of 125 rat hybridomas secreting monoclonal antibodies (mabs) to ectromelia virus (ev) polypeptide (poxviridae family, orhtopoxvirus genus) was set up. a significant portion of mabs (37 types) recognized epitopes of the 14 kda polypeptide as well as the 37 and 35 kda polypeptides. however, a majority of mabs interacted with conformation-dependent epitopes, which were destroyed in immunoprecipitation. one hundred and thirteen of mabs cross-interacted with antigenic determin ...	2004	15455686
[human mini-antibodies to orthopoxviruses].	a library of human scfv antibodies displayed on the surface of bacteriophages (mrc, cambridge, england) was panned against the elstree strain of vaccinia virus (vacv), which resulted in the phage repertoire enriched with clones positive to the strain. individual clones from the repertoire were screened for binding, independently, to the vaccinia and ectromelia viruses; phage antibodies to the orthopoxviruses were selected. ten unique antibodies were identified after their vh- and vl-genes were s ...	2004	15455687
an oligonucleotide microarray for the detection of vaccinia virus.	vaccinia virus is a member of the orthopoxvirus group, to which also belongs variola virus, one of the most hazardous pathogens known to man. to establish a model system to detect orthopoxviruses, a vaccinia oligonucleotide microarray is designed, produced and tested. vaccinia virus is used to test the prepared microarrays. the virus dna samples in different propagation phases are extracted and hybridised with the oligonucleotide microarray. the results showed that the oligonucleotide microarray ...	2004	15462260
experimental infection of ground squirrels (spermophilus tridecemlineatus) with monkeypox virus.	a proposed new small-animal (rodent) model for studying the pathogenesis and treatment of severe orthopoxvirus infections is described. thirteen-lined ground squirrels (spermophilus tridecemlineatus) were infected intraperitoneally and intranasally with monkeypox virus (mpxv). a fulminant illness developed in all animals, and they died 6-9 days after infection. virus was cultured from the blood and oropharynx several days before death; at necropsy, all of the organs tested contained relatively h ...	2004	15498157
human monkeypox infection: a family cluster in the midwestern united states.	the outbreak of monkeypox in the midwestern united states during june 2003 marks the first documented human infection in the western hemisphere. consistent with those in outbreaks in africa, most cases in this outbreak were associated with febrile rash illness. we describe a cluster of monkeypox in a family with a spectrum of clinical illness, including encephalitis, and outline the laboratory confirmation of monkeypox.	2004	15499541
new class of orthopoxvirus antiviral drugs that block viral maturation.	by using a homology-based bioinformatics approach, a structural model of the vaccinia virus (vv) i7l proteinase was developed. a unique chemical library of approximately 51,000 compounds was computationally queried to identify potential active site inhibitors. the resulting biased subset of compounds was assayed for both toxicity and the ability to inhibit the growth of vv in tissue culture cells. a family of chemotypically related compounds was found which exhibits selective activity against or ...	2004	15507601
modified vaccinia virus ankara as antigen delivery system: how can we best use its potential?	safety-tested modified vaccinia virus ankara (mva) has been established as a potent vector system for the development of candidate recombinant vaccines. the versatility of the vector system was recently demonstrated by the rapid production of experimental mva vaccines for immunization against severe acute respiratory syndrome associated coronavirus. promising results were also obtained in the delivery of epstein-barr virus or human cytomegalovirus antigens and from the clinical testing of mva ve ...	2004	15560976
5'-homoaristeromycin. synthesis and antiviral activity against orthopox viruses.	an efficient synthesis of 5'-homoaristeromycin has been developed. this permitted an extensive antiviral analysis, which found potent activity toward vaccinia, cowpox, and monkeypox viruses. for comparative purposes, 5'-homoadenosine was made available by a newly designed route and found to be inactive.	2005	15582429
[smallpox--in the past or not?].	smallpox is a potentially deadly illness caused by the variola virus, an orthopoxvirus. severe illness followed by blister-like body rash is the sign of smallpox. smallpox symptoms develop about 12 days after exposure. v. variole can spread very readily by aerosol, which may lead to explosive epidemics. for centuries, smallpox has been a worldwide cause of death, killing about 30% of the infected people. in 1972, the epidemic of smallpox in ex-yugoslavia was the largest postwar smallpox epidemic ...	2004	15615188
[characteristics of virus growth factor (vgf) encoded by vaccinia virus tian tan strain].	vaccinia virus growth factor (vgf) is encoded by an early gene located near the border between the terminal inverted repeats and the internal unique region of the viral genome. it has been suggested that secreted vgf may serve to stimulate the metabolism of neighboring uninfected cells, thereby increasing the capacity of these cells to support viral replication. in this study, we cloned and sequenced the vgf gene of vaccinia virus strain tian tap. the analytical results indicated that vgf of str ...	1997	15619809
multiplex pcr detection and species differentiation of orthopoxviruses pathogenic to humans.	a method for one-stage rapid identification of four orthopoxvirus species pathogenic to humans based on multiplex polymerase chain reaction (mpcr) was developed. five pairs of oligonucleotide primers--one, genus-specific; and the rest, species-specific for variola, monkeypox, cowpox, and vaccinia viruses, respectively--were used concurrently for mpcr assay of orthopoxvirus dnas. specificity and sensitivity of the method developed were evaluated using dnas of 57 orthopoxvirus strains, including t ...	2005	15652214
human vaccinia-like virus outbreaks in são paulo and goiás states, brazil: virus detection, isolation and identification.	since october 2001, the adolfo lutz institute has been receiving vesicular fluids and scab specimens of patients from paraíba valley region in the são paulo and minas gerais states and from são patricio valley, in the goiás state. epidemiological data suggested that the outbreaks were caused by cowpox virus or vaccinia virus. most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. virus particles with orthopoxvirus mo ...	2004	15654477
[orthopoxvirus immune evasion mechanism].		2004	15676243
[screening for orthopoxviruses by electron microscopy].		2004	15676244
[use of vaccinia immune globulin in the prophylaxis and the treatment of orthopoxvirus infections].		2004	15676246
[using antiviral drugs for orthopoxvirus infections].		2004	15676247
variola and camelpox virus-specific sequences are part of a single large open reading frame identified in two german cowpox virus strains.	a large open reading frame (orf) has been identified in two german cowpox virus strains. the orfs (5676 and 5679 nt, respectively) differ in 10 nucleotides, resulting in an amino acid homology of 99.8%. in searching genbank nucleotide sequences (>90% identity) were present in several small orfs in variola major, variola minor and camelpox virus genomes. alignments revealed that these small orfs are fragments of a large orf. however, sequences of the orf described here are entirely absent in the ...	2005	15681053
wiskott-aldrich syndrome protein is needed for vaccinia virus pathogenesis.	smallpox, caused by variola virus, was a devastating disease in humans, but how the virus evolved a strategy to spread to tissue remains unknown. through the use of microarrays, we identified the gene encoding the wiskott-aldrich syndrome protein (wasp), one of the five known wasp family members, which has been induced in the course of infection of human cells with different strains of vaccinia virus (vv) (s. guerra, l. a. lopez-fernandez, a. pascual-montano, m. munoz, k. harshman, and m. esteba ...	2005	15681416
electrostatic modeling predicts the activities of orthopoxvirus complement control proteins.	regulation of complement activation by pathogens and the host are critical for survival. using two highly related orthopoxvirus proteins, the vaccinia and variola (smallpox) virus complement control proteins, which differ by only 11 aa, but differ 1000-fold in their ability to regulate complement activation, we investigated the role of electrostatic potential in predicting functional activity. electrostatic modeling of the two proteins predicted that altering the vaccinia virus protein to contai ...	2005	15699145
cpg-dna protects against a lethal orthopoxvirus infection in a murine model.	cpg-dna has been described as a potent activator of the innate immune system, with potential to protect against infection caused by a range of pathogens in a non-specific manner. here two classes of cpg-dna (cpg-a and cpg-b) have been investigated for their abilities to protect mice from infection with an orthopoxvirus (vaccinia virus). dosing with either cpg-a or b by the intraperitonal or intranasal route protected mice against a subsequent intranasal challenge with vaccinia virus. to our know ...	2005	15708635
evaluation of human-to-human transmission of monkeypox from infected patients to health care workers.	in 2003, human monkeypox was first identified in the united states. the outbreak was associated with exposure to infected prairie dogs, but the potential for person-to-person transmission was a concern. this study examines health care worker (hcw) exposure to 3 patients with confirmed monkeypox.	2005	15714414
analysis of the virus population present in equine faeces indicates the presence of hundreds of uncharacterized virus genomes.	virus dna was isolated from horse faeces and cloned in a sequence-independent fashion. 268 clones were sequenced and 178140 nucleotides of sequence obtained. statistical analysis suggests the library contains 17560 distinct clones derived from up to 233 different virus genomes. tblastx analysis showed that 32% of the clones had significant identity to genbank entries. of these 63% were viral; 20% bacterial; 7% archaeal; 6% eukarya; and 5% were related to mobile genetic elements. fifty-two percen ...	2005	15744573
nucleic acid test screening of blood donors for orthopoxviruses can potentially prevent dispersion of viral agents in case of bioterrorism.	microbiologic agents such as variola virus (var) are very attractive for terrorism. as a result of international collaboration under the who eradication campaign, smallpox was declared eradicated in 1980. therefore, the immunization programs were discontinued worldwide. because most people are now immunologically naive, var is considered to be a potential threat agent or bioterrorist weapon. real-time polymerase chain reaction (pcr) followed by melting analysis was developed for fast and safe an ...	2005	15752158
orthopoxvirus targets for the development of antiviral therapies.	the potential use of smallpox virus as a bioterror agent and the endemic presence of monkeypox virus in africa underscores the need for better therapies for orthopoxvirus infections. the only existing clinical experience treating vaccinia and smallpox infections has been with marboran, which suggested that antiviral therapies could be effective in treating and preventing smallpox infections, but this compound has not been pursued. drugs that have been approved for other indications, like cidofov ...	2005	15777195
preparation of monoclonal antibodies cross-reactive with orthopoxviruses and their application for direct immunofluorescence test.	variola virus (smallpox virus), vaccinia virus (vv), cowpox virus (cpv) and ectromelia virus (ev) belong to the genus orthopoxvirus of the family poxviridae. to establish the possible diagnosis for smallpox infection, monoclonal antibodies (mabs) against vv and cpv were produced. the cross-reactivity of seven mabs with cells infected with various strains of the orthopoxviruses (cpv, vv and ev) was confirmed by an immunofluorescence (if) test and other immunological analyses. four and three mabs ...	2005	15781995
an investigation of the therapeutic value of vaccinia-immune igg in a mouse pneumonia model.	vaccinia-immune globulin (vig) was used to treat severe complications of smallpox vaccination, but its use was controversial because it resolved disease in only some clinical cases. vig is a pool of hyperimmune sera collected from individuals with a high neutralizing titre against the intracellular mature form (imv) of vaccinia virus (vacv), but activity against the extracellular enveloped form (eev) was often not considered. here, the efficacy of anti-vacv antibodies (abs) in protecting mice fr ...	2005	15784892
combinatorial optimization of isatin-beta-thiosemicarbazones as anti-poxvirus agents.	novel strategies are required to combat pox virus infections, whether caused by escape of viruses such as monkeypox from indigenous areas or intentional release of smallpox. anti-smallpox drugs with a unique mode of antiviral action, inhibition of transcription termination, were known but not therapeutically useful. using a combinatorial method, variants of the basic isatin-beta-thiosemicarbazone structure were prepared and examined for cytotoxicity and antiviral activity in vaccinia virus- and ...	2005	15828843
the gene of the complement-binding protein, an important anti-inflammatory factor of orthopoxviruses, is deleted from the genome of western african strains of monkeypox virus.		2005	15846974
[expression of genes for orthopoxviral tnf-binding proteins and study resulted recombinant proteins].	genes for tnf-binding proteins (crmbs) of variola virus (varv), monkeypox virus (mpxv) or cowpox (cpxv) were isolated with pcr from viral genomes and expressed within baculovirus dnas in sf21 insect cell line. properties of resulted recombinant proteins were studied with physical-chemical and immunological methods. it was shown with solid phase enzyme-linked immunoassay that viral proteins inhibited htnf binding with polyclonal htnf-antibodies. the strongest inhibitor was varv-crmb, the less one ...	2005	15856948
development in lipid drugs.	lipopeptide lipid moieties induce dendritic cell (dc) internalization and epitopes are recognized by mhc, the major histocompatibility complex. hiv-1 (human immunodeficiency virus type 1) lipopeptide vaccine candidate elicits immune responses, and sustains hiv control after highly active antiretroviral therapy (haart). mp- and dp-mart (anti-melanoma lipopeptides) induce strong ctl (cytolytic t lymphocyte) response. new bgtc, bgda, tgkc lipoplexes mediate gene delivery, e.g., into mouse pancreati ...	2005	15892690
the 4',4'-difluoro analog of 5'-noraristeromycin: a new structural prototype for possible antiviral drug development toward orthopoxvirus and cytomegalovirus.	as a surrogate for 4'-hydroxy-5'-noraristeromycin and related carbocyclic nucleosides, an efficient, enantiodivergent synthetic route to both enantiomers of 5-(6-amino-9h-purin-9-yl)-3,3-difluorocyclopentane-1,2-diol (6 and ent-6) has been developed from a common starting material ((+)-(1r,4s)-4-hydroxy-2-cyclopenten-1-yl acetate, 10). both compounds were assayed versus a series of viruses. the only response found was for compound 6 toward vaccinia and cowpox (ec50 of 143 and 94 microm, respecti ...	2005	15908221
how similar are poxviruses?		2005	15919976
suppressors of a host range mutation in the rabbitpox virus serpin spi-1 map to proteins essential for viral dna replication.	the orthopoxvirus serpin spi-1 is an intracellular serine protease inhibitor that is active against cathepsin g in vitro. rabbitpox virus (rpv) mutants with deletions of the spi-1 gene grow on monkey kidney cells (cv-1) but do not plaque on normally permissive human lung carcinoma cells (a549). this reduced-host-range (hr) phenotype suggests that spi-1 may interact with cellular and/or other viral proteins. we devised a genetic screen for suppressors of spi-1 hr mutations by first introducing a ...	2005	15994811
characterization of acute-phase humoral immunity to monkeypox: use of immunoglobulin m enzyme-linked immunosorbent assay for detection of monkeypox infection during the 2003 north american outbreak.	a monkeypox outbreak occurred in the united states in 2003. patient's sera were sent to the centers for disease control and prevention as a part of outbreak response measures. clinical and epidemiologic information was abstracted from the case investigation forms. serum samples from patients were tested by using an immunoglobulin m (igm)-capture and an igg enzyme-linked immunosorbent assay elisa against orthopoxvirus antigen. the detection of antiviral igg and igm antibodies and the kinetics of ...	2005	16002637
rapid, differential diagnosis of orthopox- and herpesviruses based upon real-time pcr product melting temperature and restriction enzyme analysis of amplicons.	orthopoxviruses tend to have non-specific early symptoms that cannot be differentiated readily from other infectious exanthemas, such as varicella-zoster virus (vzv) or disseminated herpes simplex virus (hsv) infections. a rapid assay was developed for the differential diagnosis of orthopoxviruses and herpesviruses based upon the melting temperatures of real-time pcr amplicons. a mean melting temperature difference of 8.7 degrees c was observed between the products amplified from the two virus f ...	2005	16005086
evolutionary history of orthopoxvirus proteins similar to human complement regulators.	orthopoxviruses include many important pathogens such as variola major virus, camelpox, buffalopox, monkeypox, cowpox, and variola minor viruses. this group of viruses also includes vaccinia virus, which is extensively used in human vaccine development. genomes of orthopoxviruses encode proteins with sequences similar to human regulators of complement activation (rca) that contain tandem short consensus repeats (scrs). we employed phylogenetic tree analysis to evaluate the structural relationshi ...	2005	16023794
two-color multiplex assay for the identification of orthopox viruses with real-time lux- pcr.	the lux [light upon extension] is a real-time detection system that can be used for the detection and quantification of pathogens nucleic acids. in this study we used a universal lux approach, a variation of the lux detection system, for identifying orthopoxvirus nucleic acids in real time. this approach enables the design of sequence-specific primer sets in high identity genome sequences. the assay described here is designed to allow simultaneous detection of variola and other orthopox viruses ...	2005	16023830
contact inactivation of orthopoxviruses by household disinfectants.	the aim of this study is to identify common household disinfectants that combine significant activity against the type orthopoxvirus, vaccinia virus with minimal impact in terms of potential toxicity and/or damage to household or personal items.	2005	16033458
nmr assignment of the vaccinia virus envelope protein a27l.		2005	16034674
in vitro activity of cyclosal-nucleoside monophosphates and polyhydroxycarboxylates against orthopoxviruses.	because variola virus might be used as a pathogen in biological attacks, there is an urgent need to provide effective antiviral drugs for the treatment of orthopoxvirus infections. thus, the aim of the present study was to test the antiviral activity of 3 pro-nucleotides of the acyclic nucleoside analogues aciclovir (acv), 3 of penciclovir (pcv) and 38 of the cyclic nucleoside analogue brivudin (bvdu), on the basis of cyclosaligenyl-nucleoside monophosphate approach against vaccinia virus and co ...	2005	16076502
multiple diagnostic techniques identify previously vaccinated individuals with protective immunity against monkeypox.	approximately 50% of the us population received smallpox vaccinations before routine immunization ceased in 1972 for civilians and in 1990 for military personnel. several studies have shown long-term immunity after smallpox vaccination, but skepticism remains as to whether this will translate into full protection against the onset of orthopoxvirus-induced disease. the us monkeypox outbreak of 2003 provided the opportunity to examine this issue. using independent and internally validated diagnost ...	2005	16086024
genomic multiple sequence alignments: refinement using a genetic algorithm.	genomic sequence data cannot be fully appreciated in isolation. comparative genomics--the practice of comparing genomic sequences from different species--plays an increasingly important role in understanding the genotypic differences between species that result in phenotypic differences as well as in revealing patterns of evolutionary relationships. one of the major challenges in comparative genomics is producing a high-quality alignment between two or more related genomic sequences. in recent y ...	2005	16086841
interferon function is not required for recovery from a secondary poxvirus infection.	ifn function is critical for recovery from most primary viral infections, including poxvirus infection. in contrast, very little is known about the requirement for ifn function in mediating recovery from a secondary virus infection. we have used ectromelia virus (ectv), an orthopoxvirus very closely related to variola virus, to investigate the importance of ifn function in recovery from a secondary infection. variola virus, the causative agent of smallpox in humans, and ectv, which causes mousep ...	2005	16123129
simultaneous identification of orthopoxviruses and alphaviruses by oligonucleotide macroarray with special emphasis on detection of variola and venezuelan equine encephalitis viruses.	the development of a method in macroarray format for the identification of alphaviruses and orthopoxviruses in samples of concern in biodefense is reported. capture oligonucleotides designed to bind generic members of the orthopox- or alphavirus families and a collection of additional oligonucleotides to bind specifically nucleic acids from five individual alphaviruses, including venezuelan equine encephalitis, or dna from each of four orthopoxviruses, including variola virus (var) were deposite ...	2006	16181687
ectromelia virus: the causative agent of mousepox.	ectromelia virus (ectv) is an orthopoxvirus whose natural host is the mouse; it is related closely to variola virus, the causative agent of smallpox, and monkeypox virus, the cause of an emerging zoonosis. the recent sequencing of its genome, along with an effective animal model, makes ectv an attractive model for the study of poxvirus pathogenesis, antiviral and vaccine testing and viral immune and inflammatory responses. this review discusses the pathogenesis of mousepox, modulation of the imm ...	2005	16186218
an orally bioavailable antipoxvirus compound (st-246) inhibits extracellular virus formation and protects mice from lethal orthopoxvirus challenge.	st-246 is a low-molecular-weight compound (molecular weight = 376), that is potent (concentration that inhibited virus replication by 50% = 0.010 microm), selective (concentration of compound that inhibited cell viability by 50% = >40 microm), and active against multiple orthopoxviruses, including vaccinia, monkeypox, camelpox, cowpox, ectromelia (mousepox), and variola viruses. cowpox virus variants selected in cell culture for resistance to st-246 were found to have a single amino acid change ...	2005	16189015
the longest micron; transporting poxviruses out of the cell.	the large size of poxvirus virions (approximately 250-300 microm) makes them dependent on active transport for intracellular movement during infection. several recent papers have reported the utilization of the microtubule network by poxviruses during viral egress and their use of conventional kinesin for intracellular transport. this review looks at recent reports of poxvirus intracellular transport for virion egress and their interaction with the microtubule network.	2005	16207240
characterization of the major capsid proteins of myxoma virus particles using maldi-tof mass spectrometry.	the protein composition of poxvirus particles remains uncertain because of their large size and structural complexity. this has complicated the characterization of even well-studied orthopoxviruses, like vaccinia virus, and little or nothing is known about the capsid composition of viruses belonging to other poxvirus genera. this paper describes methods that address this problem and have been used to identify 17 different leporipoxvirus capsid proteins. myxoma virus particles were purified using ...	2006	16226321
combinations of polyclonal or monoclonal antibodies to proteins of the outer membranes of the two infectious forms of vaccinia virus protect mice against a lethal respiratory challenge.	previous studies demonstrated that antibodies to live vaccinia virus infection are needed for optimal protection against orthopoxvirus infection. the present report is the first to compare the protective abilities of individual and combinations of specific polyclonal and monoclonal antibodies that target proteins of the intracellular (imv) and extracellular (ev) forms of vaccinia virus. the antibodies were directed to one imv membrane protein, l1, and to two outer ev membrane proteins, a33 and b ...	2005	16227266
analysis of putative rnase p rna from orthopoxviruses.	a putative rnase p rna gene in camelpox virus, one of the orthopoxviruses, was cloned and transcribed in vitro. no rnase p activity could be detected in vitro from camelpox virus rnase p rna alone, or by addition of the escherichia coli rnase p protein subunit to reaction mixtures. camelpox virus rnase p rna reconstituted in vitro with camel or hela cell extracts, which were pre-treated with micrococcal nuclease to degrade the endogenous rnase p rna, showed no rnase p activity. vaccinia virus, a ...	2005	16253270
immunization with a single extracellular enveloped virus protein produced in bacteria provides partial protection from a lethal orthopoxvirus infection in a natural host.	subunit vaccines that use the vaccinia virus extracellular envelope protein a33r alone or combined with other structural proteins are excellent candidates for a new smallpox vaccine. since a new smallpox vaccine would be used in humans, who are the natural hosts for the orthopoxvirus variola, the agent of smallpox, it would be important to determine whether a prospective smallpox vaccine can protect from a lethal orthopoxvirus infection in a natural host. we addressed this question using the mou ...	2006	16256161
synthesis and antiviral activity of 7-deazaneplanocin a against orthopoxviruses (vaccinia and cowpox virus).	an efficient method for the synthesis of 7-deazaneplanocin a (2) has been accomplished by the condensation of cyclopentenol 3 with 6-chloro-7-deazapurine followed by subsequent functional group manipulations. the synthesized 7-deazaneplanocin a (2) exhibited potent antiviral activity against cowpox and vaccinia viruses without cytotoxicity in hff cells.	2006	16275078
transmission of monkeypox among persons exposed to infected prairie dogs in indiana in 2003.	to describe a cluster of human monkeypox cases associated with exposure to ill prairie dogs in a home child care.	2005	16275790
clinical characteristics of human monkeypox, and risk factors for severe disease.	human monkeypox is an emerging smallpox-like illness that was identified for the first time in the united states during an outbreak in 2003. knowledge of the clinical manifestations of monkeypox in adults is limited, and clinical laboratory findings have been unknown.	2005	16288398
genetic identification of novel poxviruses of cetaceans and pinnipeds.	novel poxviruses were identified in skin lesions of several species of cetaceans and pinnipeds using polymerase chain reaction targeting dna polymerase and dna topoisomerase i genes of members of the subfamily chordopoxvirinae. with the exception of parapoxviruses, no molecular data of marine mammal poxviruses were available to infer genetic and evolutionary relatedness to terrestrial vertebrate poxviruses. viruses were assigned to a cetacean poxvirus 1 (cpv-1) group based on nucleotide and amin ...	2006	16328132
substrate specificity of vaccinia virus thymidylate kinase.	anti-poxvirus therapies are currently limited to cidofovir [(s)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine], but drug-resistant strains have already been characterized. in the aim of finding a new target, the thymidylate (tmp) kinase from vaccinia virus, the prototype of orthopoxvirus, has been overexpressed in escherichia coli after cloning the gene (a48r). specific inhibitors and alternative substrates of pox tmp kinase should contribute to virus replication inhibition. biochemical charac ...	2005	16336263
[fusion proteins encoded by orf 129l of ectromelia and orf a30l of smallpox viruses cross-react with neutralizing monoclonal antibodies].	open reading frame (orf) 129l of ectromelia (ev) and orf a30l of smallpox viruses (spv) encoding fusion proteins were cloned and expressed in e. coli cells. the recombinant polypeptides (pra30l h pr129l) were purified from cell lysates by ni-nta chromatography. recombinant polypeptides were able to form trimers in buffered saline and they destroyed under treatment with sds and 2-mercaptoethanol. reactivity of pra30l, pr129l and orthopoxvirus proteins was analyzed by elisa and western blotting wi ...	2005	16358742
orthopoxvirus infection transmitted by a domestic cat.	the variola virus was declared eradicated by the world health organization in 1980 but human infections by cowpox virus, another member of the genus orthopoxvirus, are still observed, mainly in european countries. we report a woman who presented with two umbilicated vesicles surrounded by an indurated erythematous edema within cat scratch injuries on her thigh. the diagnosis of an orthopoxvirus infection was based on the visualization of characteristic virus particles by electron microscopy and ...	2006	16427982
highly attenuated smallpox vaccine protects rabbits and mice against pathogenic orthopoxvirus challenge.	the possible reemergence of smallpox through bioterrorism requires the preparation of adequate stockpiles of vaccine. dryvax, the only us-licensed vaccinia virus smallpox vaccine, has an unacceptable safety profile in the pre-event setting. lc16m8 is a japanese-licensed attenuated vaccinia virus strain that has been safely used in over 50,000 persons. until now, efficacy of this vaccine was unproven. using two animal models, we show that lc16m8 and dryvax elicit comparable humoral immune respons ...	2006	16430997
synthesis of cyclopentenyl carbocyclic nucleosides as potential antiviral agents against orthopoxviruses and sars.	a practical and convenient methodology for the synthesis of chiral cyclopentenol derivative (+)-12a has been developed as the key intermediate that was utilized for the synthesis of biologically active carbocyclic nucleosides. the selective protection of allylic hydroxyl group followed by the ring-closing metathesis (rcm) reaction with grubbs catalysts provided (+)-12a on a 10 g scale with 52% overall yield from d-ribose (4). the key intermediate (+)-12a was utilized for the synthesis of unnatur ...	2006	16451078
poxvirus infection of steller sea lions (eumetopias jubatus) in alaska.	lesions suggestive of poxvirus infection were observed in two steller sea lions (eumetopias jubatus) in alaska during live capture-and-release studies during 2000 and 2001. both of these animals, female pups in poor body condition, were from prince william sound; this population is part of the declining western stock. umbilicated, typically ulcerated dermal nodules were present, primarily on the fore flippers in one case, and over most of the body in the second case. histologically, there were d ...	2005	16456163
recognition of vaccinia virus-infected cells by human natural killer cells depends on natural cytotoxicity receptors.	natural killer (nk) cells are important in the immune response to a number of viruses; however, the mechanisms used by nk cells to discriminate between healthy and virus-infected cells are only beginning to be understood. infection with vaccinia virus provokes a marked increase in the susceptibility of target cells to lysis by nk cells, and we show that recognition of the changes in the target cell induced by vaccinia virus infection depends on the natural cytotoxicity receptors nkp30, nkp44, an ...	2006	16474130
pox proteomics: mass spectrometry analysis and identification of vaccinia virion proteins.	although many vaccinia virus proteins have been identified and studied in detail, only a few studies have attempted a comprehensive survey of the protein composition of the vaccinia virion. these projects have identified the major proteins of the vaccinia virion, but little has been accomplished to identify the unknown or less abundant proteins. obtaining a detailed knowledge of the viral proteome of vaccinia virus will be important for advancing our understanding of orthopoxvirus biology, and s ...	2006	16509968
purification, crystallization and preliminary diffraction studies of an ectromelia virus glutaredoxin.	ectromelia, vaccinia, smallpox and other closely related viruses of the orthopoxvirus genus encode a glutaredoxin gene that is not present in poxviruses outside of this genus. the vaccinia glutaredoxin o2l has been implicated as the reducing agent for ribonucleotide reductase and may thus play an important role in viral deoxyribonucleotide synthesis. as part of an effort to understand nucleotide metabolism by poxviruses, evm053, the o2l ortholog of the ectromelia virus, has been crystallized. ev ...	2005	16511093
[the time scale in poxvirus evolution].	unlike vertebrates and rna-containing viruses, the objective estimate of molecular clock for dna-containing viruses was so far absent. an extended central conservative genomic region of orthopoxviruses (about 102 kbp) and the sequence of dna polymerase gene (about 3 kbp) of the viruses belonging to various genera from the family poxviridae were analyzed. during this analysis, the known dating of variola virus (varv) transfer from west africa to south america (xvi century) and our own data on clo ...	2006	16523687
distinct thymidine kinases encoded by cowpox virus and herpes simplex virus contribute significantly to the differential antiviral activity of nucleoside analogs.	orthopoxviruses and herpesviruses are both large enveloped dna viruses, yet these virus families exhibit very different susceptibilities to antiviral drugs. we investigated the activation of nucleoside analogs by the types i and ii thymidine kinase (tk) homologs expressed by herpes simplex virus type 1 (hsv-1) and cowpox virus (cv). antiviral activity against tk(-) and tk(+) strains of hsv-1 and cv was determined, and the ratio of the ec(50) values was used as a measurement of tk dependence. as ...	2006	16530858
fatal poxvirus outbreak in a colony of new world monkeys.	an epizootic infection was observed in a colony of 80 new world monkeys consisting of various species including a group of marmosets and saguinus species. during the summer and autumn of 2002, 30 animals died of unknown diseases. six animals were sent to the german primate center for investigation of the cause of death. a complete pathologic and histologic investigation was carried out. the animals exhibited erosive-ulcerative lesions of the oral mucous membranes. advanced stages of the disease ...	2006	16537943
synthesis and antiviral evaluation of alkoxyalkyl derivatives of 9-(s)-(3-hydroxy-2-phosphonomethoxypropyl)adenine against cytomegalovirus and orthopoxviruses.	9-(s)-(3-hydroxy-2-phosphonomethoxypropyl)adenine [(s)-hpmpa] was one of the first acyclic nucleoside phosphonates described and has been reported to have good antiviral activity against most double-stranded dna viruses, including the herpes group viruses and the orthopoxviruses. however, (s)-hpmpa is not orally bioavailable and has not been developed for clinical use. we have prepared orally bioavailable lipid esters of (s)-hpmpa and report their synthesis and antiviral evaluation against cytom ...	2006	16539388
inhibitory efficacy of cyclosal-nucleoside monophosphates of aciclovir and brivudin on dna synthesis of orthopoxvi ruses.	previous studies have shown that cyclosaligenyl-monophosphate (cyclosal-mp) derivatives of aciclovir (acv), penciclovir (pcv) and brivudin (bvdu) can act as inhibitors of vaccinia virus and cowpox virus replication in vitro. the aim of the present study was to evaluate the inhibatory efficacy on dna synthesis in vaccinia and cowpox viruses of several cyclosal-pro-nucleotides of acv and bvdu, which have proven activity against pox viruses. viral dna was quantified in treated and non-treated virus ...	2006	16542003
activity and mechanism of action of n-methanocarbathymidine against herpesvirus and orthopoxvirus infections.	n-methanocarbathymidine [(n)-mct] is a conformationally locked nucleoside analog that is active against some herpesviruses and orthopoxviruses in vitro. the antiviral activity of this molecule is dependent on the type i thymidine kinase (tk) in herpes simplex virus and also appears to be dependent on the type ii tk expressed by cowpox and vaccinia viruses, suggesting that it is a substrate for both of these divergent forms of the enzyme. the drug is also a good inhibitor of viral dna synthesis i ...	2006	16569849
a chemokine-binding domain in the tumor necrosis factor receptor from variola (smallpox) virus.	variola virus (vav) is the causative agent of smallpox, one of the most devastating diseases encountered by man, that was eradicated in 1980. the deliberate release of vav would have catastrophic consequences on global public health. however, the mechanisms that contribute to smallpox pathogenesis are poorly understood at the molecular level. the ability of viruses to evade the host defense mechanisms is an important determinant of viral pathogenesis. here we show that the tumor necrosis factor ...	2006	16581912
smallpox antiviral drug development: satisfying the animal efficacy rule.	concerns over the potential use of variola virus as a biological weapon have prompted new interest in the development of small molecule therapeutics to prevent and treat smallpox infection. since smallpox is no longer endemic, human clinical trials designed to link antiviral efficacy to clinical outcome have been supplanted by antiviral efficacy evaluations in animal models of orthopoxvirus disease. this poses a unique challenge for drug development; how can animal efficacy data with a surrogate ...	2006	16597208
follow-up on diagnostic proficiency of laboratories equipped to perform orthopoxvirus detection and quantification by pcr: the second international external quality assurance study.	two years after the first external quality assurance study on bioterrorism-relevant viruses, we have conducted a follow-up study on orthopoxvirus detection by pcr. thirty-three laboratories (27 european, 4 austral-asian, and 2 american) participated. samples contained 0 to 40,000,000 dna copies of lyophilized monkeypox, cowpox, and vaccinia virus per ml. laboratories achieved a >80% detection chance above 56,234 copies per ml. global sensitivity was not significantly improved over that of the fi ...	2006	16597852
protection induced in mice against a lethal orthopox virus by the lister strain of vaccinia virus and modified vaccinia virus ankara (mva).	the lister (elstree) strain of vaccinia virus, used in israel for vaccination against smallpox, was studied in tissue cultures and in a mouse model. the virus failed to reach the brain of the mice when inoculated intranasally at a dose of 500,000 plaque forming units, but was lethal for 50% of them, when injected intracranially. lower doses of virus injected intracranially caused some weight loss initially, but later the mice completely recovered. modified vaccinia virus ankara (mva), when infec ...	2006	16603280
a pyrimidine-pyrazolone nucleoside chimera with potent in vitro anti-orthopoxvirus activity.	synthetic hybridization of two privileged drug scaffolds, pyrazolone on the one hand and pyrimidine nucleoside on the other, resulted in the generation of two novel 5-substituted pyrimidine nucleosides with potent in vitro antiviral activity against two representative orthopoxviruses, vaccinia virus and cowpox virus.	2006	16603351
in vitro and in vivo evaluation of isatin-beta-thiosemicarbazone and marboran against vaccinia and cowpox virus infections.	it has been reported previously that some thiosemicarbazone compounds have prophylactic activity against smallpox disease and therapeutic activity against vaccinia virus (vv) infections. in these studies, isatin-beta-thiosemicarbazone (ibt) and marboran were administered once daily by intraperitoneal (ip) injection to mice using 30, 10 or 3 mg/kg for 5 days beginning 24, 48 or 72 h after inoculation with vv or cowpox virus (cv). both compounds were highly effective (p < 0.01) at preventing morta ...	2006	16621041
a simple and rapid protein array based method for the simultaneous detection of biowarfare agents.	a protein chip has been developed that allows the simultaneous detection of a multitude of different biowarfare agents. the chip was developed for the arraytube platform providing a cheap and easy to handle technology solution that combines a microtube-integrated protein chip with the classical procedure of a sandwich-enzyme-linked immunosorbent assay and signal amplification by streptavidin-poly-horseradish peroxidase. specific immunoassays for staphylococcus enterotoxin b, ricin, venezuelan eq ...	2006	16622830
comparative sequence analysis of envelope protein genes of indian buffalopox virus isolates.	buffalopox virus (bpxv) is considered to be a close variant of vaccinia virus (vacv), the prototype member of the genus orthopoxvirus. in the present study, we have analyzed the sequences of h3l, a27l, and d8l gene-homologues of vacv in bpxv to elucidate its genetic relationship to vacv and other orthopoxviruses (opvs). products of these genes have been shown to be important in attachment of vacv to host cell surface receptors during viral entry. additionally, the a27l gene is also responsible f ...	2006	16625321
interferon-gamma-mediated antiviral immunity against orthopoxvirus infection is provided by gamma delta t cells.		2006	16652291
fk506, a secondary metabolite produced by streptomyces, presents a novel antiviral activity against orthopoxvirus infection in cell culture.	to investigate the antiviral potential of the macrolide fk506, produced by streptomyces tsukubaensis, against orthopoxvirus infection in cell culture, and determine the replicative stage of viral cycle affected by the treatment.	2006	16696686
cell line dependency for antiviral activity and in vivo efficacy of n-methanocarbathymidine against orthopoxvirus infections in mice.	a novel carbocyclic thymidine analog, n-methanocarbathymidine [(n)-mct], was evaluated for inhibition of orthopoxvirus infections. efficacy in vitro was assessed by plaque reduction assays against wild-type and cidofovir-resistant strains of cowpox and vaccinia viruses in nine different cell lines. minimal differences were seen in antiviral activity against wild-type and cidofovir-resistant viruses. (n)-mct's efficacy was affected by the cell line used for assay, with 50% poxvirus-inhibitory con ...	2007	16712967