Publications

internal and flanking sequence from aflp fragments using ligation-mediated suppression pcr.	amplification fragment-length polymorphism (aflp) analysis has proven to be a powerful tool for developing a large number of reliable genetic markers across a wide variety of organisms. often it is desirable to further characterize these markers by obtaining internal and flanking sequence information. here, we present a systematic approach for obtaining such information from aflp markers. aflp fragments can be isolated from dried polyacryamide sequencing gels (that have been stored for extended ...	1999	10337484
disruption of anthrax toxin binding with the use of human antibodies and competitive inhibitors.	the protective antigen (pa83) of bacillus anthracis is integral to the mechanism of anthrax toxicity. we have isolated a human single-chain fv antibody fragment (scfv) that blocks binding of a fluorescently tagged protective antigen (pa) moiety to cell surface receptors. several phage-displayed scfv were isolated from a naive library biopanned against pa83. soluble, monomeric scfv were characterized for affinity and screened for their capacity to disrupt receptor-mediated binding of pa. four uni ...	1999	10338505
anthrax toxin entry into polarized epithelial cells.	we examined the entry of anthrax edema toxin (edtx) into polarized human t84 epithelial cells using cyclic amp-regulated cl- secretion as an index of toxin entry. edtx is a binary a/b toxin which self assembles at the cell surface from anthrax edema factor and protective antigen (pa). pa binds to cell surface receptors and delivers ef, an adenylate cyclase, to the cytosol. edtx elicited a strong cl- secretory response when it was applied to the basolateral surface of t84 cells but no response wh ...	1999	10338515
proteasome activity is required for anthrax lethal toxin to kill macrophages.	anthrax lethal toxin (letx), consisting of protective antigen (pa) and lethal factor (lf), rapidly kills primary mouse macrophages and macrophage-like cell lines such as raw 264.7. lf is translocated by pa into the cytosol of target cells, where it acts as a metalloprotease to cleave mitogen-activated protein kinase kinase 1 (mek1) and possibly other proteins. in this study, we show that proteasome inhibitors such as acetyl-leu-leu-norleucinal, mg132, and lactacystin efficiently block letx cytot ...	1999	10338520
bctp sounds its battle cry.		1999	10348630
isolation of an asporogenic (spooa) protective antigen-producing strain of bacillus anthracis.	we found that congo red agar allows identification of sporulation-deficient bacillus anthracis. using congo red agar, we isolated an asporogenic derivative of the protective antigen-producing strain b. anthracis delta sterne-1(ppa102). polymerase chain reaction and southern hybridization analyses of dna from the asporogenic mutant revealed that a deletion was present in spooa, an essential gene for the initiation of sporulation. the deletion also encompassed the spoivb homologue and a portion of ...	1999	10349715
[adenylyl cyclase--isoforms, regulation and function].	since its discovery in 1956, cyclic amp (camp) has been shown to be a ubiquitous second messenger. it functions as one of many signaling molecules enabling cells to respond to external signals. camp is synthesized by adenylyl cyclases (acs), enzymes that convert adenosine triphosphate (atp) to camp. three classes of acs have been cloned based on the conservation of their catalytic domains; they include: class i-acs from enterobacteria, including escherichia coli; class ii-"toxic" acs, including ...	1999	10355282
plcr is a pleiotropic regulator of extracellular virulence factor gene expression in bacillus thuringiensis.	members of the bacillus cereus group (b. anthracis, b. cereus, b. mycoides and b. thuringiensis) are well-known pathogens of mammals (b. anthracis and b. cereus) and insects (b. thuringiensis). the specific diseases they cause depend on their capacity to produce specific virulence factors, such as the lethal toxin of b. anthracis and the cry toxins of b. thuringiensis. however, these bacillus spp. also produce a variety of proteins, such as phospholipases c, which are known to act as virulence f ...	1999	10361306
cytotoxic t-lymphocyte epitopes fused to anthrax toxin induce protective antiviral immunity.	we have investigated the use of the protective antigen (pa) and lethal factor (lf) components of anthrax toxin as a system for in vivo delivery of cytotoxic t-lymphocyte (ctl) epitopes. during intoxication, pa directs the translocation of lf into the cytoplasm of mammalian cells. here we demonstrate that antiviral immunity can be induced in balb/c mice immunized with pa plus a fusion protein containing the n-terminal 255 amino acids of lf (lfn) and an epitope from the nucleoprotein (np) of lymph ...	1999	10377103
understanding bacillus anthracis pathogenesis.		1999	10383221
bacillus anthracis: medical issues of biologic warfare.	recent world events refocused attention on the possibility of nations engaging in biologic warfare, including an attack with bacillus anthracis. the single available anthrax vaccine in the united states for human use, formerly known as mdph-pa, has decreased ability to protect laboratory animals against virulent b. anthracis strains, especially compared with new vaccines being developed. studies with these vaccines, however, have several shortcomings. the pathogenesis, diagnosis, treatment, and ...	1999	10391414
identification and characterization of a germination operon on the virulence plasmid pxo1 of bacillus anthracis.	the spores of bacillus anthracis, the agent of anthrax disease, germinate within professional phagocytes, such as murine macrophage-like raw264.7 cells and alveolar macrophages. we identified a cluster of germination genes extending for 3608 nucleotides between the pag and atxa genes on the b. anthracis virulence plasmid pxo1. the three predicted proteins (40, 55 and 37 kda in size) have significant sequence similarities to b. subtilis, b. cereus and b. megaterium germination proteins. northern ...	1999	10411756
recombinant vaccinia viruses protect against clostridium perfringens alpha-toxin.	recombinant vaccinia viruses that expressed the nontoxic c-domain of clostridium perfringens alpha-toxin were constructed. the j2r (thymidine kinase [tk] gene) and b13r (serpin 2 [spi-2] gene) loci were used as insertion sites for the clostridial dna, and expression of the foreign protein was measured in each case. a double recombinant that encoded the alpha-toxin truncate at the b13r locus and the protective antigen of bacillus anthracis at the j2r locus was also constructed. although differenc ...	1999	10413356
the respiratory burst-inhibiting acid phosphatase acpa is not essential for the intramacrophage growth or virulence of francisella novicida.	acid phosphatases capable of inhibiting the respiratory burst of neutrophils have been identified in certain intracellular pathogens. here we evaluate the role of acpa, a respiratory burst-inhibiting acid phosphatase of francisella, in the virulence and intracellular growth of this organism. an f. novicida acpa null mutant was created and found to exhibit wild-type growth kinetics in both cell-line and inflammatory mouse macrophages. the acpa mutant also shows wild-type replication in the spleen ...	1999	10418134
autogenous regulation of the bacillus anthracis pag operon.	protective antigen (pa) is an important component of the edema and lethal toxins produced by bacillus anthracis. pa is essential for binding the toxins to the target cell receptor and for facilitating translocation of the enzymatic toxin components, edema factor and lethal factor, across the target cell membrane. the structural gene for pa, paga (previously known as pag), is located on the 182-kb virulence plasmid pxo1 at a locus distinct from the edema factor and lethal factor genes. here we sh ...	1999	10419943
a case of anthrax sepsis: non-fatal course.		1999	10424807
principles for emergency response to bioterrorism.	the recent occurrence of a series of anthrax-related hoaxes illustrates the need to educate emergency services personnel about how to best ensure patient and worker safety in the case of suspected exposure to biological threat agents. there are very few data to support the methods being used or the variation in current care. emergency physicians, first responders, and hazardous materials response teams need a standardized approach to the management of patients who may have been exposed to biolog ...	1999	10424919
update on emerging infections from the centers for disease control and prevention. bioterrorism alleging use of anthrax and interim guidelines for management--united states, 1998.		1999	10424929
expression and purification of the recombinant protective antigen of bacillus anthracis.	protective antigen (pa) is a major component of the vaccine against anthrax. the structural gene for the 83-kda pa was expressed as fusion protein with 6x histidine residues in escherichia coli. expression of pa in e. coli under the transcriptional regulation of the t5 promoter yielded an insoluble protein aggregating to form inclusion bodies. the inclusion bodies were solubilized in 6 m guanidine-hcl and the protein was purified under denaturing conditions using nickel nitrilotriacetic acid (ni ...	1999	10425157
the effects of electron and chemical ionization modes on the ms profiling of whole bacteria.	free fatty acid profiling of whole bacteria [francisella tularensis, brucella melitensis, yersinia pestis, bacillus anthracis (vegetative and sporulated), and bacillus cereus] was carried out with direct probe mass spectrometry under 70-ev electron ionization (ei) and isobutane chemical ionization in both the positive (ci+) and negative modes (ci-). electron ionization produced spectra that contained molecular ions and fragment ions from various free fatty acids. spectra acquired with isobutane ...	1999	10439512
anthrax protective antigen: prepore-to-pore conversion.	pa(63), the active 63 kda form of anthrax protective antigen, forms a heptameric ring-shaped oligomer that is believed to represent a precursor of the membrane pore formed by this protein. when maintained at ph >/=8.0, this "prepore" dissociated to monomeric subunits upon treatment with sds at room temperature, but treatment at ph </=7 (or with beta-octylglucoside at ph 8.0) caused it to convert to an sds-resistant pore-like form. transition to this form involved major changes in the conformatio ...	1999	10441138
in vitro selection of dna aptamers to anthrax spores with electrochemiluminescence detection.	systematic evolution of ligands by exponential enrichment (selex) was used to select and pcr amplify dna sequences (aptamers) capable of binding to and detecting nonpathogenic sterne strain bacillus anthracis spores. a simplified affinity separation approach was employed, in which autoclaved anthrax spores were used as the separation matrix. an aptamer-magnetic bead-electrochemiluminescence (am-ecl) sandwich assay scheme was devised for detecting anthrax spores. using a low selex dna to spore ra ...	1999	10451913
cell surface-exposed tetanus toxin fragment c produced by recombinant bacillus anthracis protects against tetanus toxin.	bacillus anthracis, the causal agent of anthrax, synthesizes two surface layer (s-layer) proteins, ea1 and sap, which account for 5 to 10% of total protein and are expressed in vivo. a recombinant b. anthracis strain was constructed by integrating into the chromosome a translational fusion harboring the dna fragments encoding the cell wall-targeting domain of the s-layer protein ea1 and tetanus toxin fragment c (toxc). this construct was expressed under the control of the promoter of the s-layer ...	1999	10456940
vaccines in civilian defense against bioterrorism.		1999	10458959
vaccines, pharmaceutical products, and bioterrorism: challenges for the u.s. food and drug administration.		1999	10458960
clinical and epidemiologic principles of anthrax.		1999	10458964
anthrax: a possible case history.		1999	10458965
applying lessons learned from anthrax case history to other scenarios.		1999	10458966
polymorphism analysis and gene detection by minisequencing on an array of gel-immobilized primers.	two procedures, multibase and multiprimer, have been developed for single nucleotide extension of primers immobilized within polyacrylamide gel pads on a microchip. in the multibase assay, a primer is next to a polymorphic nucleotide; the nucleotide is identified by the specificity with which the primer incorporates fluorescently labeled dideoxyribo-nucleoside triphosphates. in the multiprimer assay, several primers containing different 3'-terminal nucleotides overlapping the variable nucleotide ...	1999	10471749
simulants, stimulants and diseases: the evolution of the united states biological warfare programme, 1945-60.	details about the us biological programme have largely been based on information in the open literature. more revealing aspects of the programme are now available through documents released under the freedom of information act. annual reports of the activities of the us army chemical corps from 1945 to 1959 have revealed significant increases in activity in biological warfare research. the corps research activity progressed from work on anthrax in 1941, through anti-crop agents in the mid-1940s, ...	1999	10472189
1996-97 global anthrax report.	while there is a general decrease in the number of anthrax outbreaks, and thus of human cases, worldwide this is still a disease that is extensively under-diagnosed and under-reported. however, it is now very infrequent to rare in canada, the united states, and many countries in europe. an increasing number of countries are now free. at the other extreme, it is a significant problem in west africa, spain, greece, turkey, albania, romania and in central asia. in spite of the textbooks, livestock ...	1999	10475945
a national register of historic and contemporary anthrax foci.	anthrax in russia has for a long time posed a serious problem for public health and veterinary services. at the beginning of the century, 40-60 thousand cases of this infection were annually reported in the country in agricultural animals and about 10-20 thousand cases in people where each fourth (25%) was dying. in the russian federation the registration of anthrax foci is obligatory for veterinary as well as for sanitary-epidemiological services. so our initial project, funded by the internati ...	1999	10475946
anthrax explodes in an australian summer.	anthrax occurred on 83 properties in an area of north central victoria between 26 january and 26 march in the summer of 1997. anthrax had not been recorded in the outbreak area since records were initiated in 1914, although anthrax did occur in the general area in the 1880s to 1890s. standard australian control measures were applied to the properties, including quarantine, tracing movements of animals on and off affected properties, secure disposal of carcases by burning, enhanced surveillance o ...	1999	10475947
identification of bacillus anthracis strains in china.	as part of a project to establish a reference strain collection, phenotypic, biochemical and genetic analyses were carried out on 84 strains of bacillus anthracis, 81 of them of chinese origin from various sources. particular differences from reports on isolates of other origins were the possession of fimbriae and single polar flagella with consequent motility in 77, self-agglutination by 64 and failure to ferment maltose in 60 of the chinese strains. the findings were considered to be of signif ...	1999	10475948
meso-scale ecology of anthrax in southern africa: a pilot study of diversity and clustering.	it has only recently been possible to detect sufficient genetic diversity among anthrax isolates to allow genotype grouping (keim et al. 1997). early results of such grouping suggest that the southern african subcontinent may be the geographical origin of bacillus anthracis. this report describes a pilot investigation of the genetic diversity of a study group of isolates from the kruger national park, south africa, and efforts to detect spatio-temporal clustering within the study group. this stu ...	1999	10475949
a review of anthrax in canada and implications for research on the disease in northern bison.	during the first half of the century, the majority of anthrax outbreaks in canada occurred in the southern portions of ontario and quebec and were often associated with pastures contaminated by effluent from textile industries dealing with imported animal materials. in 1952, introduction of federal regulations requiring disinfection of these materials greatly reduced the incidence of anthrax in eastern canada. since 1962, domestic outbreaks of the disease have been reported almost exclusively in ...	1999	10475950
antibody-based systems for the detection of bacillus anthracis in environmental samples	there is a necessity for rapid immunodiagnostic techniques for the detection and identification of bacillus anthracis in environmental specimens. the technology available for accomplishing this ranges in complexity from a simple dipstick type assay to complex biosensors. we have developed antigen capture dipstick assays for a series of infectious agents including an assay for b. anthracis protective antigen and one for b. anthracis spores. these immunochromatographic assays use colloidal gold to ...	1999	10475951
molecular diversity in bacillus anthracis.	molecular typing of bacillus anthracis has been extremely difficult due to the lack of polymorphic dna markers. we have identified nine novel variable number tandemly repeated loci from previously known amplified fragment length polymorphism markers or from the dna sequence. in combination with the previously known vrra locus, these markers provide discrimination power to genetically characterize b. anthracis isolates. the variable number tandem repeat (vntr) loci are found in both gene coding ( ...	1999	10475952
fluorescent detection techniques for real-time multiplex strand specific detection of bacillus anthracis using rapid pcr.	speed is a key area in our development of pcr assays for bacillus anthracis. we believe that the strand specific detection of amplicons within 10 min is a realistic goal and that this will be achieved through fluorescent in-tube assays. we have used the idaho lightcycler to study and develop candidate assays for b. anthracis. new strand specific fluorescent methods have been developed and a number of formats have been studied for speed and sensitivity. internal controls have been developed as a ...	1999	10475953
the ba813 chromosomal dna sequence effectively traces the whole bacillus anthracis community.	plasmid genes that are responsible for virulence of bacillus anthracis are important targets for the dna-based detection of anthrax. we evaluated the distribution of the ba813 chromosomal dna sequence (ba813) within closely related bacillus species. ba813 was systematically identified from 47 strains or isolates of b. anthracis tested, thus indicating its reliability as a tracer for that species. from the 60 strains of closely related bacillus spp. examined, three bona fide b. cereus and one bon ...	1999	10475954
polymerase chain reaction-elisa to detect bacillus anthracis from soil samples-limitations of present published primers.	a polymerase chain reaction (pcr)-elisa technique to detect bacillus anthracis from soil samples has been developed. the application of streptavidine-coated microtitre plates as well as plates covered with covalently linked oligonucleotides as catching probes led to a test sensitivity of about 100 fg pure genomic dna or of less than 10 spores seeded into 100 g soil material. some non-suspicious soil samples collected from different locations yielded positive results with presently published prim ...	1999	10475955
definitive identification of bacillus anthracis--a review.	the word 'problem' is seen with some frequency in relation to clear differentiation between bacillus anthracis and b. cereus. in fact, although the close relationship of these two species is undisputed, it is only in the case of a few borderline isolates, rarely encountered in practice, that any sort of identification problem exists. until recently this was only important to the taxonomist who found it unsatisfactory not to be able to identify definitively such isolates. to most others, if the i ...	1999	10475956
molecular recognition specificity of bacillus anthracis spore antibodies.	the sensitivity and specificity of polyclonal and monoclonal antibodies raised against anthrax spore preparations has been assessed by western blotting. none of the antibodies studied were completely specific in recognizing the anthrax spore surface. a polyclonal serum recognized a wide range of spore surface epitopes and demonstrated limited cross-reaction with the near-neighbour species bacillus cereus spore surface. two monoclonal antibodies studied demonstrated more extensive cross-reaction ...	1999	10475958
the capsule of bacillus anthracis, a review	the capsule of bacillus anthracis, composed of poly-d-glutamic acid, serves as one of the principal virulence factors during anthrax infection. by virtue of its negative charge, the capsule is purported to inhibit host defence through inhibition of phagocytosis of the vegetative cells by macrophages. in conjunction with lethal toxin and oedema toxin, whose target cells include macrophages and neutrophils, respectively, the capsule allows virulent anthrax bacilli to grow virtually unimpeded in th ...	1999	10475959
bacillus anthracis surface: capsule and s-layer.	two abundant surface proteins, ea1 and sap, are components of the bacillus anthracis surface layer (s-layer). their corresponding genes have been cloned, shown to be clustered on the chromosome and sequenced. ea1 and sap each possess three 's-layer homology' motifs. single and double disrupted mutants were constructed. ea1 and sap were co-localized at the cell surface of both the non-capsulated and capsulated bacilli. when present, the capsule is exterior to, and completely covers, the s-layer p ...	1999	10475960
the s-layer homology domain as a means for anchoring heterologous proteins on the cell surface of bacillus anthracis.	bacillus anthracis synthesizes two s-layer proteins, each containing three s-layer homology (slh) motifs towards their amino-terminus. in vitro experiments suggested that the three motifs of each protein were organized as a structural domain sufficient to bind purified cell walls. chimeric genes encoding the slh domains fused to the levansucrase of bacillus subtilis were constructed and integrated on the chromosome. cell fractionation and electron microscopy studies showed that both heterologous ...	1999	10475961
sequence, assembly and analysis of px01 and px02.	bacillus anthracis plasmids px01 and px02, harboured by the sterne and pasteur strains, respectively, have been sequenced by random 'shotgun' cloning and high throughout sequence analysis. these sequences have been assembled (sequencher) to generate a circulate px01 plasmid containing 181 656 bp and a single linear (gapped) px02 contig containing at least 93.479 bp. initial annotation suggests that the two plasmids combined contain at least 200 potential open reading frames (orfs) with < 40% hav ...	1999	10475962
genetic comparison of bacillus anthracis and its close relatives using amplified fragment length polymorphism and polymerase chain reaction analysis.	amplified fragment length polymorphism (aflp) analysis allows a rapid, relatively simple analysis of a large portion of a microbial genome, providing information about the species and its phylogenetic relationship to other microbes (vos et al. 1995). the method simply surveys the genome for length and sequence polymorphisms. the aflp pattern identified can be used for comparison to the genomes of other species. unlike other methods, it does not rely on analysis of a single genetic locus that may ...	1999	10475963
the native virulence plasmid combination affects the segregational stability of a theta-replicating shuttle vector in bacillus anthracis var. new hampshire.	the segregational stability of a small, theta-replicating, non-mobilizable shuttle plasmid (paex-5e) was determined in fully virulent (px01+/px02+), partially cured (px01+/px02- and px01-/px02+) and fully cured (px01-/px02-) derivatives of bacillus anthracis var. new hampshire. under the growth conditions used (l-broth, 37 degrees c, aerobic, batch culture), paex-5e remained segregationally stable in the px01-/px02+ and px01-/px02- derivatives for in excess of 100 culture generations, but was ex ...	1999	10475964
control of virulence gene expression in bacillus anthracis.	the atxa gene is an important regulator of virulence gene expression in bacillus anthracis. atxa positively regulates expression of the three genes encoding the anthrax toxin proteins and at least one gene is required for capsule production. here we report that an atxa-null mutant exhibits phenotypes unrelated to toxin and capsule synthesis. an atxa-null mutant grows poorly on minimal media and sporulates more efficiently than the parent strain. numerous transposon-generated promoter-lacz fusion ...	1999	10475965
crystallographic studies of the anthrax lethal toxin	anthrax lethal toxin comprises two proteins: protective antigen (pa; mw 83 kda) and lethal factor (lf; mw 87 kda). we have recently determined the crystal structure of the 735-residue pa in its monomeric and heptameric forms (petosa et al. 1997). it bears no resemblance to other bacterial toxins of known three-dimensional structure, and defines a new structural class which includes homologous toxins from other gram-positive bacteria. we have proposed a model of membrane insertion in which the wa ...	1999	10475966
mechanism of membrane translocation by anthrax toxin: insertion and pore formation by protective antigen	proteolytic activation of receptor-bound protective antigen (pa) at the cell surface removes pa20, allowing pa63 to oligomerize and form a ring-shaped heptameric prepore. the prepore binds edema factor (ef) and lethal factor (lf) and, after endocytosis and trafficking of the complex to an acidic, vesicular compartment, it undergoes membrane insertion and mediates translocation of ef/lf to the cytosol. data from membrane conductance experiments support a model of membrane insertion in which the 2 ...	1999	10475967
anthrax toxin fusion proteins for intracellular delivery of macromolecules	the dominant role played by the anthrax toxin in bacillus anthracis pathogenesis shows that the toxin has evolved to be an efficient system for delivering its two catalytic protein components, oedema factor and lethal factor (lf), into the cytosol of host cells. this system involves binding of the protective antigen (pa) toxin component to a ubiquitous (and still unidentified) receptor, proteolytic activation at the cell surface, internalization by endocytosis and translocation through an early ...	1999	10475968
lethal toxin actions and their consequences.	after entry of infectious anthrax spores into the body, host-specific signals induce spore germination, outgrowth of vegetative bacilli and the expression of lethal toxin and other virulence factors. anthrax lethal toxin (letx) is a virulence factor responsible for the major pathologies seen during systemic anthrax infections. injection of sterile letx into test animals mimics the shock and sudden death seen during active bacterial infections. once large levels of letx are produced within the bo ...	1999	10475969
anthrax lethal factor cleaves the n-terminus of mapkks and induces tyrosine/threonine phosphorylation of mapks in cultured macrophages	the lethal toxin (letx) of bacillus anthracis is the major virulence factor responsible for the death of infected animals and for cytolysis of cultured macrophages. its catalytic component, lf, contains the characteristic zinc-binding motif of metalloproteases, it binds zinc and indirect evidence suggests that this hydrolytic activity is essential for letx cytotoxicity (limpel et al. 1994; kochi et al. 1994). to identify substrates of lf, we have used the yeast two-hybrid system, employing an lf ...	1999	10475970
anthrax lethal factor causes proteolytic inactivation of mitogen-activated protein kinase kinase.	a search of the national cancer institute's anti-neoplastic drug screen for compounds with an inhibitory profile similar to that of the mitogen-activated protein kinase kinase (mapkk) inhibitor pd098059 yielded anthrax lethal toxin. anthrax lethal factor was found to inhibit progesterone-induced meiotic maturation of frog oocytes by preventing the phosphorylation and activation of mitogen-activated protein kinase (mapk). similarly, lethal toxin prevented the activation of mapk in serum stimulate ...	1999	10475971
experiences with vaccination and epidemiological investigations on an anthrax outbreak in australia in 1997.	between january and february 1997, there was a severe outbreak of anthrax on 83 properties in north-central victoria, australia. vaccination was used as a major tool to control the outbreak by establishing a vaccination buffer zone 30 km by 20 km. in all, 78, 649 cattle in 457 herds were vaccinated in a three week program. in the face of the outbreak, there was a delay before vaccination was able to stop deaths. in the 10 days following vaccination 144 cases of confirmed anthrax occurred and 38 ...	1999	10475972
antigen delivery by attenuated bacillus anthracis: new prospects in veterinary vaccines.	this report summarizes the recent investigations on the use of bacillus anthracis as a live vector for delivery of antigens. recombinant strains were constructed by engineering the current live sterne vaccine. this vaccine, used to prevent anthrax in cattle, causes side-effects due to anthrax toxin activities. bacteria producing a genetically detoxified toxin factor were devoid of lethal effects and were as protective as the sterne strain against experimental anthrax. moreover, b. anthracis expr ...	1999	10475973
clinical aspects, diagnosis and treatment of anthrax	there are three clinical presentations of anthrax in humans: cutaneous (>95% of cases), orogastric and inhalational. the infectious form, the spore, enters the body and is thought to germinate within macrophages either at the site of inoculation (cutaneous or orogastric) or in the regional lymph node (inhalational). the bacillus then synthesizes its antiphagocytic capsule and the lethal and oedema toxins which interfere with the non-specific host defences leading to the characteristic locally de ...	1999	10475974
in vitro correlate of immunity in an animal model of inhalational anthrax	the incidence of anthrax in humans is extremely low. human vaccine efficacy studies for inhalational anthrax cannot be conducted. the identification of a correlate of protection that predicts vaccine efficacy is crucial for determining the immune status of immunized humans. this surrogate marker of immunity can only be established by using an appropriate animal model. numerous studies showed that protective antigen (pa) is the principle protective antigen in naturally- or vaccine-induced immunit ...	1999	10475975
immune correlates of protection against anthrax	bacillus anthracis protective antigen (pa) has been produced from a recombinant b. subtilis and its efficacy, when combined with the ribi adjuvant (mpl-tdw-cws) or alhydrogel, has been compared with that of the licensed uk human vaccine, in guinea pigs challenged with aerosolized ames strain spores. recombinant pa combined with the ribi adjuvant performed as well as pa from b. anthracis cultures in previous reports (ivins & welkos 1986; ivins et al. 1990; turnbull et al. 1991; jones et al. 1996; ...	1999	10475976
human immune responses to the uk human anthrax vaccine.	the igg anti-protective antigen subclass antibody response of individuals who had been infected with anthrax was compared with that of healthy individuals immunized with the uk licensed anthrax vaccine. the predominant subclass in both groups was igg1. in addition, igg3 was seen in convalescent serum while vaccinees produced igg2, igg3 and igg4 subclass. the significance of these results is discussed. further work is required to determine the role of antibodies in mediating protective immunity i ...	1999	10475977
expression of the protective antigen of bacillus anthracis by lactobacillus casei: towards the development of an oral vaccine against anthrax.	bacillus anthracis is the causative organism of the disease anthrax. the ability of the organism to form resistant spores and infect via the aerosol route has led to it being considered as a potential biological warfare agent. the current available human vaccines are far from ideal, they are expensive to produce, require repeated doses and may invoke transient side-effects in some individuals. there is also evidence to suggest that they may not give full protection against all strains of b. anth ...	1999	10475978
presentation of protective antigen to the mouse immune system: immune sequelae.	protective antigen (pa), the major protective component of the existing vaccine, is a potent immunogen. protective antigen in alhydrogel induced a high serum igg titre (> log10 4) in both the c57b16 and balb/c mouse and the predominant subclass of antibody induced was igg1, indicating that the response to pa was predominantly th2 directed. when plasmid dna encoding pa was used to immunize the balb/c mouse, a low serum igg titre was detected (</=log10 1), which was slightly increased by boosting ...	1999	10475979
world health organization activities on anthrax surveillance and control.	the achievements of a world health organization anthrax working group, established in 1990, have been the production of two editions of guidelines on anthrax surveillance and control and the formulation of templates to assist countries in the construction of their surveillance and control programmes. the latter was made possible by the active participation of the department of animal production and health, ministry of agriculture, food and fisheries, zambia and the livestock development programm ...	1999	10475980
images in clinical medicine. bacillus anthracis meningitis.		1999	10477780
anthrax.		1999	10477781
a poly-gamma-glutamate synthetic system of bacillus subtilis ifo 3336: gene cloning and biochemical analysis of poly-gamma-glutamate produced by escherichia coli clone cells.	three genes encoding a poly-gamma-glutamate synthetic system of bacillus subtilis ifo 3336 (bacillus natto) were cloned and expressed in escherichia coli. the e. coli clone produced poly-gamma-glutamate extracellularly. the genes, newly designated as pgsbca, were homologous with capbca genes of bacillus anthracis. all of pgsb, pgsc, and pgsa genes were essential for the polymer production. addition of mn(2+), instead of mg(2+), to the polymer-synthesis medium resulted in an increase in the polym ...	1999	10486244
[aerosol vaccination against dangerous infectious diseases].	the paper summarizes the results of development of the aerosol method, one of the mass ways of human vaccination. analysis of materials suggests that russia has designed highly effective live plague, tularemia, and anthrax vaccines that can be used to immunize in different ways: by epicutaneous and subcutaneous, and inhalation routes. the advantages and disadvantages of aerosol vaccination are shown. the correct use of this method provides a substantial effect when the epidemic situation is comp ...	1999	10487124
toxins that are activated by hiv type-1 protease through removal of a signal for degradation by the n-end-rule pathway.	diphtheria toxin enters the cytosol of mammalian cells where it inhibits cellular protein synthesis, leading to cell death. recently we found that the addition of a signal for n-end-rule-mediated protein degradation to diphtheria toxin substantially reduced its intracellular stability and toxicity. these results prompted us to construct a toxin containing a degradation signal that is removable through the action of a viral protease. in principle, such a toxin would be preferentially stabilized, ...	1999	10493930
sequence and organization of pxo1, the large bacillus anthracis plasmid harboring the anthrax toxin genes.	the bacillus anthracis sterne plasmid pxo1 was sequenced by random, "shotgun" cloning. a circular sequence of 181,654 bp was generated. one hundred forty-three open reading frames (orfs) were predicted using genemark and genemark.hmm, comprising only 61% (110,817 bp) of the pxo1 dna sequence. the overall guanine-plus-cytosine content of the plasmid is 32.5%. the most recognizable feature of the plasmid is a "pathogenicity island," defined by a 44.8-kb region that is bordered by inverted is1627 e ...	1999	10515943
a minisonicator to rapidly disrupt bacterial spores for dna analysis.	concerns about the use of anthrax spores as a weapon of mass destruction have motivated the development of portable instruments capable of detecting and monitoring a suspected release of the agent. optimal detection of bacterial spores by pcr requires that the spores be disrupted to make the endogenous dna available for amplification. the entire process of spore lysis, pcr, and detection can take several hours using conventional methods and instruments. in this report, a minisonicator and protot ...	1999	10517145
anthrax toxins.	though its lethal effects were ascribed to an exotoxin almost half a century ago, the pathogenesis of anthrax has yet to be satisfactorily explained. subsequent work has led to the molecular identification and enzymatic characterization of three proteins that constitute two anthrax toxins. protective antigen binds an as yet unknown cell receptor and mediates the entry of the other two components to the cytoplasm via the endosomal pathway. edema factor, so named for its ability to induce edema, i ...	1999	10526577
monitoring temperature-sensitive vaccines and immunologic drugs, including anthrax vaccine.	the experience of the u.s. army medical materiel center, europe (usammce), in monitoring temperature-sensitive vaccines and immunologic drugs, including anthrax vaccine, during storage and shipment is discussed. usammce uses an electronic monitoring device to monitor and archive the time-temperature history of shipments of various vaccines, immunoglobulins, and other drugs requiring refrigeration. using these monitors, usammce can track its carriers' performance, reduce product loss, and validat ...	1999	10541032
recognition and treatment of anthrax.		1999	10553786
a novel surfactant nanoemulsion with broad-spectrum sporicidal activity against bacillus species.	two nontoxic, antimicrobial nanoemulsions, bctp and bctp 401, have been developed. these emulsions are composed of detergents and oils in 80% water. bctp diluted up to 1:1000 inactivated>90% of bacillus anthracis spores in 4 h and was also sporicidal against three other bacillus species. this sporicidal activity is due to disruption of the spore coat after initiation of germination without complete outgrowth. bctp 401 diluted 1:1000 had greater activity than bctp against bacillus spores and had ...	1999	10558951
inhalational anthrax: epidemiology, diagnosis, and management.	anthrax, a disease of great historical interest, is once again making headlines as an agent of biological warfare. bacillus anthracis, a rod-shaped, spore-forming bacterium, primarily infects herbivores. humans can acquire anthrax by agricultural or industrial exposure to infected animals or animal products. more recently, the potential for intentional release of anthrax spores in the environment has caused much concern. the common clinical manifestations of anthrax are cutaneous disease, pulmon ...	1999	10559102
biological warfare: would you recognize an attack?		1998	10562136
a plague upon your cattle.		1998	9654895
deadly relic of the great war.		1998	9655389
how anthrax kills.		1998	9660700
[cutaneous anthrax in a child].		1998	9662859
the expression of the protective antigen of bacillus anthracis in bacillus subtilis.	the expression of bacillus anthracis protective antigen (pa) in b. subtilis from the pag gene in ppa101-1 was explored in different genetic backgrounds in an attempt to identify opportunities to maximize expression. introduction of atxa, which positively regulates pa expression in b. anthracis did not improve expression levels in the protease-deficient strain wb600. plasmid ppa101-1 was found to carry a deletion which created a new fusion point between vector and insert sequence, and which remov ...	1998	9674126
anthrax: a disease from antiquity visits the modern world.		1998	9676107
thucydides' syndrome.		1998	9679482
the effectiveness and safety of vaccines against human anthrax: a systematic review.	we report on the results of a systematic review of existing controlled clinical trials undertaken to assess the effectiveness and safety of vaccines against human anthrax in relation to disease incidence and side-effects. two articles retrieved by electronic and hand search fulfilling some of the inclusion criteria underwent a quality assessment by a group of reviewers. data synthesized from the two trials showed that estimates of overall effectiveness and safety favour treatment (overall odds r ...	1998	9682332
comparative efficacy of experimental anthrax vaccine candidates against inhalation anthrax in rhesus macaques.	the authors examined the efficacy of bacillus anthracis protective antigen (pa) combined with adjuvants as vaccines against an aerosol challenge of virulent anthrax spores in rhesus macaques. adjuvants tested included i) aluminum hydroxide (alhydrogel), ii) saponin qs-21 and iii) monophosphoryl lipid a (mpl) in squalene/lecithin/tween 80 emulsion (slt). animals were immunized once with either 50 micrograms of recombinant pa plus adjuvant, or with anthrax vaccine adsorbed (ava), the licensed huma ...	1998	9682372
anthrax lethal factor cleaves the n-terminus of mapkks and induces tyrosine/threonine phosphorylation of mapks in cultured macrophages.	lethal factor (lf) is the major virulence factor produced by bacillus anthracis. lf is sufficient to cause death in laboratory animals and cytolysis of peritoneal macrophages and macrophage cell lines. lf contains the characteristic zinc binding motif of metalloproteases and indirect evidence suggest that this hydrolytic activity is essential for its cytotoxicity. to identify the substrate(s) of lf, we have used the yeast two-hybrid system, employing a lf inactive mutant as bait. this approach h ...	1998	9703991
biological warfare.		1998	9708788
bioterrorism as a public health threat.	the threat of bioterrorism, long ignored and denied, has heightened over the past few years. recent events in iraq, japan, and russia cast an ominous shadow. two candidate agents are of special concern--smallpox and anthrax. the magnitude of the problems and the gravity of the scenarios associated with release of these organisms have been vividly portrayed by two epidemics of smallpox in europe during the 1970s and by an accidental release of aerosolized anthrax from a russian bioweapons facilit ...	1998	9716981
ltx1, a mouse locus that influences the susceptibility of macrophages to cytolysis caused by intoxication with bacillus anthracis lethal factor, maps to chromosome 11.	the lethal factor (lf) toxin that is produced by bacillus anthracis plays an important role in the pathogenesis of anthrax. lf has mononuclear phagocyte-specific intoxicating effects that are not well understood. we have identified genetic differences in inbred mouse strains that determine whether their cultured macrophages are susceptible to the cytolytic effect of lf intoxication. our identification of resistant and susceptible mouse strains enabled us to analyse crosses between these strains ...	1998	9720874
a novel dipstick developed for rapid bet v 1-specific ige detection: recombinant allergen immobilized via a monoclonal antibody to crystalline bacterial cell-surface layers.	the incidence of allergy to airborne proteins derived from tree and grass pollen, feces of mites, spores of molds, and pet dander has been increasing over the last decades. since precise diagnosis is a prerequisite for successful immunotherapy, there is a rising demand for rapid, reliable, and inexpensive screening methods such as dipstick assays. with the purified recombinant major birch-pollen allergen rbet v 1a as model protein, crystalline bacterial cell-surface layers (s-layers) were tested ...	1998	9722228
use of a photoactivatable lipid to probe the topology of pa63 of bacillus anthracis in lipid membranes.	the protective antigen of bacillus anthracis is a key protein that promotes the translocation of the enzymatic moieties of the two toxins of b. anthracis into the cell cytoplasm. the membrane topology of the active form of the protective antigen (pa63) was investigated by proteolysis of pa63 inserted into liposomes containing a photoactivatable, radioactive lipid, and characterization of the n-terminal moiety of the deeply-inserted (and therefore radiolabeled) peptides. a single sequence startin ...	1998	9746362
anthrax toxin as a molecular tool for stimulation of cytotoxic t lymphocytes: disulfide-linked epitopes, multiple injections, and role of cd4(+) cells.	we have previously demonstrated that anthrax toxin-derived proteins, protective antigen (pa) and the amino-terminal portion of lethal factor (lfn), can be used in combination to deliver heterologous molecules to the cytosol of mammalian cells. in this study we examined the ability of an lfn-peptide disulfide-linked heterodimer to prime cytotoxic t lymphocytes (ctl) in the presence of pa. a mutant of lfn that contains a carboxy-terminal reactive cysteine was generated. this form of lfn could be o ...	1998	9746566
first shots fired in biological warfare.		1998	9751039
[anthrax toxins].	bacillus anthracis, a gram positive bacterium, is the causative agent of anthrax. this organism is capsulogen and toxinogenic. it secretes two toxins which are composed of three proteins: the protective antigen (pa), the lethal factor (lf) and the edema factor (ef). the lethal toxin (pa + lf) provokes a subite death in animals, the edema toxin (pa + ef) induces edema. the edema and the lethal factors are internalised into the target cells via the protective antigen. ef and lf exert an adenylate ...	1998	9759382
[implications of bacterial protein toxins in infectious and food-borne diseases].	among the 315 protein toxins elicited by gram positive and gram negative bacteria so far characterized, about 50 toxins are currently considered as totally or partially, responsible of the pathological manifestations and/or lethality resulting from host infection or intoxication (contaminated food) by relevant toxinogenic bacteria. a certain number of criteria are required for the assessment of indisputable involvement of a toxin or an array of toxins (from the same bacteria) in infectious disea ...	1998	9759385
rapid pathogen detection using a microchip pcr array instrument.	an array of pcr microchips for rapid, parallel testing of samples for pathogenic microbes is described. the instrument, called the advanced nucleic acid analyzer (anaa), utilizes 10 silicon reaction chambers with thin-film resistive heaters and solid-state optics. features of the system include efficient heating and real-time monitoring, low power requirements for battery operation, and no moving parts for reliability and ruggedness. we analyzed cultures of erwinia herbicola vegetative cells, ba ...	1998	9761255
practicalities of warfare required service personnel to be vaccinated against anthrax.		1998	9774304
molecular characterization of bacillus strains involved in outbreaks of anthrax in france in 1997.	outbreaks of anthrax zoonose occurred in two regions of france in 1997. ninety-four animals died, and there were three nonfatal cases in humans. the diagnosis of anthrax was rapidly confirmed by bacteriological and molecular biological methods. the strains of bacillus anthracis in animal and soil samples were identified by a multiplex pcr assay. they all belonged to the variable-number tandem repeat (vntr) group (vntr)3. a penicillin-resistant strain was detected. nonvirulent bacilli related to ...	1998	9774609