Publications

high throughput screening for spores and vegetative forms of pathogenic b. anthracis by an internally controlled real-time pcr assay with automated dna preparation.	human infections with bacillus anthracis have become rare but in cases of intentional release, masses of samples would have to be expected. current pcr assays for anthrax are appropriate for use in single cases, but they have not been formulated for high throughput screening. this article describes a high throughput real-time pcr for anthrax, including automated sample preparation without the need for pre-culturing of samples. the assay detects single copies of target gene. an internal control m ...	2007	17093976
lethal and edema toxins in the pathogenesis of bacillus anthracis septic shock: implications for therapy.	recent research regarding the structure and function of bacillus anthracis lethal (letx) and edema (etx) toxins provides growing insights into the pathophysiology and treatment of shock with this lethal bacteria. these are both binary-type toxins composed of protective antigen necessary for their cellular uptake and either lethal or edema factors, the toxigenic moieties. the primary cellular receptors for protective antigen have been identified and constructed and key steps in the extracellular ...	2007	17095744
crystal structure of bacillus cereus hlyiir, a transcriptional regulator of the gene for pore-forming toxin hemolysin ii.	production of bacillus cereus and bacillus anthracis toxins is controlled by a number of transcriptional regulators. here we report the crystal structure of b. cereus hlyiir, a regulator of the gene encoding the pore-forming toxin hemolysin ii. we show that hlyiir forms a tight dimer with a fold and overall architecture similar to the tetr family of repressors. a remarkable feature of the structure is a large internal cavity with a volume of 550 a(3) suggesting that the activity of hlyiir is mod ...	2007	17097673
structure and molecular mechanism of bacillus anthracis cofactor-independent phosphoglycerate mutase: a crucial enzyme for spores and growing cells of bacillus species.	phosphoglycerate mutases (pgms) catalyze the isomerization of 2- and 3-phosphoglycerates and are essential for glucose metabolism in most organisms. this study reports the production, structure, and molecular dynamics analysis of bacillus anthracis cofactor-independent pgm (ipgm). the three-dimensional structure of b. anthracis pgm is composed of two structural and functional domains, the phosphatase and transferase. the structural relationship between these two domains is different than in the ...	2007	17085493
anthrax toxins inhibit immune cell chemotaxis by perturbing chemokine receptor signalling.	pathogenic strains of bacillus anthracis produce two potent toxins, lethal toxin (lt), a metalloprotease that cleaves mitogen-activated protein kinase kinases, and oedema toxin (et), a calcium/calmodulin-dependent adenylate cyclase. emerging evidence indicates a role for both toxins in suppressing the initiation of both innate and adaptive immune responses, which are essential to keep the infection under control. here we show that lt and et inhibit chemotaxis of t-cells and macrophages by subver ...	2007	17087730
inhibition of anthrax protective antigen outside and inside the cell.	in the course of bacillus anthracis infection, b. anthracis lethal factor (lf) and edema factor bind to a protective antigen (pa) associated with cellular receptors antxr1 (tem8) or antxr2 (cmg2), followed by internalization of the complex via receptor-mediated endocytosis. a new group of potential antianthrax drugs, beta-cyclodextrins, has recently been described. a member of this group, per-6-(3-aminopropylthio)-beta-cyclodextrin (amprbetacd), was shown to inhibit the toxicity of lf in vitro a ...	2007	17074791
poly-gamma-glutamate capsule-degrading enzyme treatment enhances phagocytosis and killing of encapsulated bacillus anthracis.	the poly-gamma-d-glutamic acid capsule confers antiphagocytic properties on bacillus anthracis and is essential for virulence. in this study, we showed that capd, a gamma-polyglutamic acid depolymerase encoded on the b. anthracis capsule plasmid, degraded purified capsule and removed the capsule from the surface of anthrax bacilli. treatment with capd induced macrophage phagocytosis of encapsulated b. anthracis and enabled human neutrophils to kill encapsulated organisms. a second glutamylase, p ...	2007	17074794
fully virulent bacillus anthracis does not require the immunodominant protein bcla for pathogenesis.	the bcla protein is the immunodominant epitope on the surface of bacillus anthracis spores; however, its roles in pathogenesis are unclear. we constructed a bcla deletion mutant (bcla) of the fully virulent ames strain. this derivative retained full virulence in several small-animal models of infection despite the bcla deletion.	2007	17074844
multicomponent anthrax toxin display and delivery using bacteriophage t4.	we describe a multicomponent antigen display and delivery system using bacteriophage t4. two dispensable outer capsid proteins, hoc (highly antigenic outer capsid protein, 155 copies) and soc (small outer capsid protein, 810 copies), decorate phage t4 capsid. these proteins bind to the symmetrically localized capsid sites, which appear following prohead assembly and expansion. we hypothesized that multiple antigens fused to hoc can be displayed on the same capsid and such particles can elicit br ...	2007	17069938
qualification and performance characteristics of a quantitative enzyme-linked immunosorbent assay for human lgg antibodies to anthrax lethal factor antigen.	the contribution of bacillus anthracis lethal factor (lf)-specific immune responses to protection against anthrax disease in humans remains incompletely defined due, in part, to a paucity of qualified reagents and a lack of standardized serological assays. toward this end, we have identified and characterized suitable positive quality control and standard reference sera and developed, optimized, and qualified an enzyme-linked immunosorbent assay (elisa) to measure lf-binding igg. herein we descr ...	2007	17071101
plasmid: a centralized repository for plasmid clone information and distribution.	the plasmid information database (plasmid; http://plasmid.hms.harvard.edu) was developed as a community-based resource portal to facilitate search and request of plasmid clones shared with the dana-farber/harvard cancer center (df/hcc) dna resource core. plasmid serves as a central data repository and enables researchers to search the collection online using common gene names and identifiers, keywords, vector features, author names and pubmed ids. as of october 2006, the repository contains >46 ...	2007	17132831
dna microarray for detection of antibiotic resistance determinants in bacillus anthracis and closely related bacillus cereus.	we developed a multiplex pcr for amplification of ten genes involved in resistance to ciprofloxacin, doxycycline, rifampin, and vancomycin in bacillus anthracis and closely related bacillus cereus. enzymatic labelling of pcr products followed by hybridization to oligonucleotide probes on a dna microarray enabled simultaneous detection of resistance genes tetk, tetl, tetm, teto, vana, and vanb and resistance-mediating point mutations in genes gyra, gyrb, parc, and rpob. the presented assay allows ...	2007	17118627
operon prediction for sequenced bacterial genomes without experimental information.	various computational approaches have been proposed for operon prediction, but most algorithms rely on experimental or functional data that are only available for a small subset of sequenced genomes. in this study, we explored the possibility of using phylogenetic information to aid in operon prediction, and we constructed a bayesian hidden markov model that incorporates comparative genomic data with traditional predictors, such as intergenic distances. the prediction algorithm performs as well ...	2007	17122389
evaluation of a wipe surface sample method for collection of bacillus spores from nonporous surfaces.	polyester-rayon blend wipes were evaluated for efficiency of extraction and recovery of powdered bacillus atrophaeus spores from stainless steel and painted wallboard surfaces. method limits of detection were also estimated for both surfaces. the observed mean efficiency of polyester-rayon blend wipe recovery from stainless steel was 0.35 with a standard deviation of +/-0.12, and for painted wallboard it was 0.29 with a standard deviation of +/-0.15. evaluation of a sonication extraction method ...	2007	17122390
maasai perception of the impact and incidence of malignant catarrhal fever (mcf) in southern kenya.	we investigated the perceived impact of malignant catarrhal fever (mcf) to pastoralists in isinya division, a wildlife dispersal area of nairobi national park, and used a range of participatory epidemiology methodologies. we compared the relative importance, incidence and impact of mcf compared to other locally defined important diseases with a total of 158 respondents in 11 group meetings and 21 household meetings in july 2004. direct losses due to disease were investigated through lowered pric ...	2007	17123651
morphogenesis of the bacillus anthracis spore.	bacillus spp. and clostridium spp. form a specialized cell type, called a spore, during a multistep differentiation process that is initiated in response to starvation. spores are protected by a morphologically complex protein coat. the bacillus anthracis coat is of particular interest because the spore is the infective particle of anthrax. we determined the roles of several b. anthracis orthologues of bacillus subtilis coat protein genes in spore assembly and virulence. one of these, cote, has ...	2007	17114257
aluminum adjuvant linked to gulf war illness induces motor neuron death in mice.	gulf war illness (gwi) affects a significant percentage of veterans of the 1991 conflict, but its origin remains unknown. associated with some cases of gwi are increased incidences of amyotrophic lateral sclerosis and other neurological disorders. whereas many environmental factors have been linked to gwi, the role of the anthrax vaccine has come under increasing scrutiny. among the vaccine's potentially toxic components are the adjuvants aluminum hydroxide and squalene. to examine whether these ...	2007	17114826
insertion of anthrax protective antigen into liposomal membranes: effects of a receptor.	protective antigen (pa), the receptor-binding component of anthrax toxin, heptamerizes and inserts into the endosomal membrane at acidic ph, forming a pore that mediates translocation of the enzymic components of the toxin to the cytosol. when the heptameric pre-insertion form of pa (the prepore) is acidified in solution, it rapidly loses the ability to insert into membranes. to maximize insertion into model membranes, we examined two ways to bind the protein to large unilamellar vesicles (luv). ...	2007	17107945
n/c-4 substituted azetidin-2-ones: synthesis and preliminary evaluation as new class of antimicrobial agents.	a series of 3-chloro-4-(3-methoxy-4-acetyloxyphenyl)-1-[3-oxo-3-(phenylamino)propanamido] azetidin-2-ones 3a-g and 3-chloro-4-[2-hydroxy-5-(nitro substituted phenylazo)phenyl]-1-phenylazetidin-2-ones 6a-h were synthesized using appropriate synthetic route. structures of all the synthesized compounds were established on the basis of elemental analysis and spectroscopic data. the antimicrobial activity of the synthesized compounds was screened against several microbes. several of these molecules s ...	2007	17098426
structure of a carbohydrate esterase from bacillus anthracis.		2007	17063474
cloning and expression in e. coli of a functional fab fragment obtained from single human lymphocyte against anthrax toxin.	human lymphocytes derived from the blood of a donor immunized with anthrax vaccine were isolated and enriched for b-cells by nycoprep density centrifugation. individual anti-anthrax protective antigen (pa) b-cells were isolated by fluorescence activated cell sorting with fluorescence-labeled recombinant pa (rpa). the rna from sorted single b-cells was extracted using plant total rna as the carrier prior to purification by nanoprep rna isolation columns and then cdna was prepared. donor specific ...	2007	17045651
evaluation of the rapid analyte measurement platform (ramp) for the detection of bacillus anthracis at a crime scene.	the aim of this study was to evaluate the accuracy and reliability of the rapid analyte measurement platform (ramp) for presumptive identification of bacillus anthracis spores. test samples consisted of serial dilutions of spore preparations of several bacillus species, including b. anthracis, which were tested, using the ramp anthrax test cartridge, according to the manufacturer's instructions. the fluorescence labelled antibody-antigen complexes were detected in the portable reader after 15 mi ...	2007	17049777
plasmid exchanges among members of the bacillus cereus group in foodstuffs.	the bacillus cereus sensu lato group is genetically very close and possesses a remarkable plasmid gene pool that encodes a variety of functions such as virulence and self-transfer capabilities. the potential for horizontal transfer among the various subspecies of this group, which includes the human opportunistic pathogens b. cereus sensu stricto and b. anthracis as well as the biopesticide b. thuringiensis, has led to growing concerns regarding food safety and public health. in this study, the ...	2007	16996631
heat activation/shock temperatures for bacillus anthracis spores and the issue of spore plate counts versus true numbers of spores.	assessing true numbers of viable anthrax spores is complex. optimal heat activation conditions vary with species, media and germinants. published time/temperature combinations for bacillus anthracis spores range from 60 degrees c for <or=90 min to boiling for 1 min. results presented here indicate that temperatures are best kept to <or=70 degrees c and holding times need not exceed 15-30 min. under conditions of 60 degrees c for 90 min, 62-23 degrees c for 15 min and 70 degrees c for 15 or 30 mi ...	2007	17055602
the hc fragment of tetanus toxin forms stable, concentration-dependent dimers via an intermolecular disulphide bond.	protein oligomerisation is a prerequisite for the toxicity of a number of bacterial toxins. examples include the pore-forming cytotoxin streptolysin o, which oligomerises to form large pores in the membrane and the protective antigen of anthrax toxin, where a heptameric complex is essential for the delivery of lethal factor and edema factor to the cell cytosol. binding of the clostridial neurotoxins to receptors on neuronal cells is well characterised, but little is known regarding the quaternar ...	2007	17056064
protective and immunochemical activities of monoclonal antibodies reactive with the bacillus anthracis polypeptide capsule.	bacillus anthracis is surrounded by a polypeptide capsule composed of poly-gamma-d-glutamic acid (gammadpga). in a previous study, we reported that a monoclonal antibody (mab f26g3) reactive with the capsular polypeptide is protective in a murine model of pulmonary anthrax. the present study examined a library of six mabs generated from mice immunized with gammadpga. evaluation of mab binding to the capsule by a capsular "quellung" type reaction showed a striking diversity in capsular effects. m ...	2007	17060470
immunization against anthrax using bacillus subtilis spores expressing the anthrax protective antigen.	protective immunity to anthrax can be achieved by antibodies raised against the secreted protective antigen (pa) and this forms the basis of the current acellular vaccines for human use. bacillus subtilis spores have previously been used for delivery of heterologous antigens by the oral and nasal routes and their intrinsic heat-stability make them attractive vaccine vehicles. in this study we have expressed pa, or segments of pa, in b. subtilis using two strategies. first, display on the spore c ...	2007	17007969
bacillus anthracis: a multi-faceted role for anthrax lethal toxin in thwarting host immune defenses.	lethal factor (lf), along with its receptor-binding partner protective antigen (pa), forms lethal toxin (lt), a critical virulence factor for bacillus anthracis. lf is a zn(2+) protease that cleaves specific mitogen activated protein kinase kinases (mapkks), inactivating signal transduction intermediates required for normal immune function. initial research emphasized the role of lt in attenuating pro-inflammatory responses by macrophages, the primary targets of infection. more recent studies ha ...	2007	17008119
complete sequence analysis of novel plasmids from emetic and periodontal bacillus cereus isolates reveals a common evolutionary history among the b. cereus-group plasmids, including bacillus anthracis pxo1.	the plasmids of the members of the bacillus cereus sensu lato group of organisms are essential in defining the phenotypic traits associated with pathogenesis and ecology. for example, bacillus anthracis contains two plasmids, pxo1 and pxo2, encoding toxin production and encapsulation, respectively, that define this species pathogenic potential, whereas the presence of a bt toxin-encoding plasmid defines bacillus thuringiensis isolates. in this study the plasmids from b. cereus isolates that prod ...	2007	17041058
mechanistic differences between in vitro assays for hydrazone-based small molecule inhibitors of anthrax lethal factor.	a systematically generated series of hydrazones were analyzed as potential inhibitors of anthrax lethal factor. the hydrazones were screened using one uv-based and two fluorescence-based in vitro assays. the study identified several inhibitors with ic50 values in the micromolar range, and importantly, significant differences in the types of inhibition were observed with the different assays.	2007	16949126
combimatrix oligonucleotide arrays: genotyping and gene expression assays employing electrochemical detection.	electrochemical detection has been developed and assay performances studied for the combimatrix oligonucleotide microarray platform that contains 12,544 individually addressable microelectrodes (features) in a semiconductor matrix. the approach is based on the detection of redox active chemistries (such as horseradish peroxidase (hrp) and the associated substrate tmb) proximal to specific microarray electrodes. first, microarray probes are hybridized to biotin-labeled targets, second, the hrp-st ...	2007	16891109
increased potency of biothrax anthrax vaccine with the addition of the c-class cpg oligonucleotide adjuvant cpg 10109.	the inclusion of an adjuvant, in addition to the existing aluminum hydroxide, in the formulation of the licensed anthrax vaccine biothrax may have the potential to positively modify immune responses. some potential desirable outcomes from the inclusion of an additional adjuvant include increased immune response kinetics, increased response rates, more prolonged antibody decay rates, and the ability to use less antigen per dose or fewer doses to achieve immunity. one promising group of adjuvants ...	2007	16973247
murine splenocytes produce inflammatory cytokines in a myd88-dependent response to bacillus anthracis spores.	bacillus anthracis is a sporulating gram-positive bacterium that causes the disease anthrax. the highly stable spore is the infectious form of the bacterium that first interacts with the prospective host, and thus the interaction between the host and spore is vital to the development of disease. we focused our study on the response of murine splenocytes to the b. anthracis spore by using paraformaldehyde-inactivated spores (fis), a treatment that prevents germination and production of products a ...	2007	16978234
discernment between deliberate and natural infectious disease outbreaks.	public health authorities should be vigilant to the potential for outbreaks deliberately caused by biological agents (bioterrorism). such events require a rapid response and incorporation of non-traditional partners for disease investigation and outbreak control. the astute application of infectious disease epidemiological principles can promote an enhanced index of suspicion for such events. we discuss epidemiological indicators that should be considered during outbreak investigations, and also ...	2007	16893485
monitoring of elisa-reactive antibodies against anthrax protective antigen (pa), lethal factor (lf), and toxin-neutralising antibodies in serum of individuals vaccinated against anthrax with the pa-based uk anthrax vaccine.	the human anthrax vaccines currently licensed contain the protective antigen (pa) of bacillus anthracis as main antigen together with traces of some other bacillus components, e.g. lethal factor (lf). the present study aimed at monitoring the course of specific antibody titres against pa and lf by enzyme linked immunosorbent assays (elisa), as well as the levels of toxin-neutralising antibodies, in 11 volunteers vaccinated with the human anthrax vaccine uk. after an initial seroconversion in all ...	2007	17287051
turning biodefense dollars into products.	five years after the us anthrax attacks, and more than two years after bioshield legislation was ratified, a survey reveals that biodefense funding has thus far produced only a handful of products for clinical development.	2007	17287749
structural studies of thymidine kinases from bacillus anthracis and bacillus cereus provide insights into quaternary structure and conformational changes upon substrate binding.	thymidine kinase (tk) is the key enzyme in salvaging thymidine to produce thymidine monophosphate. owing to its ability to phosphorylate nucleoside analogue prodrugs, tk has gained attention as a rate-limiting drug activator. we describe the structures of two bacterial tks, one from the pathogen bacillus anthracis in complex with the substrate dt, and the second from the food-poison-associated bacillus cereus in complex with the feedback inhibitor dttp. interestingly, in contrast with previous s ...	2007	17288553
protection against anthrax by needle-free mucosal immunization with human anthrax vaccine.	human vaccination with biothrax requires six injections followed by annual boosters. this makes it difficult for the compliance of the immunization program and underscores the need for development of a new and optimized vaccination protocol. current research aims to demonstrate the proof of concept to develop a needle-free mucosal immunization protocol using a murine anthrax model. a/j mice were immunized with biothrax via an intranasal route. sera, saliva, vaginal, and nasal washes were evaluat ...	2007	17293013
reaerosolization of fluidized spores in ventilation systems.	this project examined dry, fluidized spore reaerosolization in a heating, ventilating, and air conditioning duct system. experiments using spores of bacillus atrophaeus, a nonpathogenic surrogate for bacillus anthracis, were conducted to delineate the extent of spore reaerosolization behavior under normal indoor airflow conditions. short-term (five air-volume exchanges), long-term (up to 21,000 air-volume exchanges), and cycled (on-off) reaerosolization tests were conducted using two common duct ...	2007	17293522
simultaneous real-time pcr detection of bacillus anthracis, francisella tularensis and yersinia pestis.	this report describes the development of in-house real-time pcr assays using minor groove binding probes for simultaneous detection of the bacillus anthracis pag and cap genes, the francisella tularensis 23 kda gene, as well as the yersinia pestis pla gene. the sensitivities of these assays were at least 1 fg, except for the assay targeting the bacillus anthracis cap gene, which showed a sensitivity of 10 fg when total dna was used as a template in a serial dilution. the clinical value of the ba ...	2007	17294160
identification of an in vivo inhibitor of bacillus anthracis spore germination.	germination of bacillus anthracis spores into the vegetative form is an essential step in anthrax pathogenicity. this process can be triggered in vitro by the common germinants inosine and alanine. kinetic analysis of b. anthracis spore germination revealed synergy and a sequential mechanism between inosine and alanine binding to their cognate receptors. because inosine is a critical germinant in vitro, we screened inosine analogs for the ability to block in vitro germination of b. anthracis spo ...	2007	17296608
discriminating inhalational anthrax from community-acquired pneumonia using chest radiograph findings and a clinical algorithm.	limiting the effects of a large-scale bioterrorist anthrax attack will require rapid and accurate detection of the earliest victims. we undertook this study to improve physicians' ability to rapidly detect inhalational anthrax victims.	2007	17296652
determination of antibiotic efficacy against bacillus anthracis in a mouse aerosol challenge model.	an anthrax spore aerosol infection mouse model was developed as a first test of in vivo efficacy of antibiotics identified as active against bacillus anthracis. whole-body, 50% lethal dose (ld50) aerosol challenge doses in a range of 1.9x10(3) to 3.4x10(4) cfu with spores of the fully virulent ames strain were established for three inbred and one outbred mouse strain (a/j, balb/c, c57bl, and swiss webster). the balb/c strain was further developed as a model for antibiotic efficacy. time course m ...	2007	17296745
infrared temperature control system for a completely noncontact polymerase chain reaction in microfluidic chips.	a completely noncontact temperature system is described for amplification of dna via the polymerase chain reaction (pcr) in glass microfluidic chips. an infrared (ir)-sensitive pyrometer was calibrated against a thermocouple inserted into a 550-nl pcr chamber and used to monitor the temperature of the glass surface above the pcr chamber during heating and cooling induced by a tungsten lamp and convective air source, respectively. a time lag of less than 1 s was observed between maximum heating r ...	2007	17297927
characteristics of spore germination in a mouse model of cutaneous anthrax.	cutaneous infection is the most common form of human anthrax, but little is known about bacillus anthracis spore germination in these infections.	2007	17299720
inactivation and purification of cowpea mosaic virus-like particles displaying peptide antigens from bacillus anthracis.	chimeric cowpea mosaic virus (cpmv) particles displaying foreign peptide antigens on the particle surface are suitable for development of peptide-based vaccines. however, commonly used peg precipitation-based purification methods are not sufficient for production of high quality vaccine candidates because they do not allow for separation of chimeric particles from cleaved contaminating species. moreover, the purified particles remain infectious to plants. to advance the cpmv technology further, ...	2007	17227681
effect of nuclear motion on the absorption spectrum of dipicolinic acid.	using semiclassical electron-radiation-ion dynamics, the authors have examined the effect of nuclear motion, resulting from both finite temperature and the response to a radiation field, on the line broadening of the excitation profile of 2,6-pyridinedicarboxylic acid (dipicolinic acid). with nuclei fixed, there is a relatively small broadening associated with the finite time duration of an applied laser pulse. when the nuclei are allowed to move, the excitation spectrum exhibits a much larger b ...	2007	17228958
analysis of anti-protective antigen igg subclass distribution in recipients of anthrax vaccine adsorbed (ava) and patients with cutaneous and inhalation anthrax.	the anti-pa igg1, igg2, igg3, and igg4 subclass responses to clinical anthrax and to different numbers of anthrax vaccine adsorbed (ava, biothrax) injections were determined in a cross-sectional study of sera from 63 vaccinees and 13 clinical anthrax patients. the data show that both vaccination with three ava injections and clinical anthrax elicit anti-pa igg1, igg2, and igg3 subclass responses. an anti-pa igg4 response was detected in ava recipients after the fourth injection. the anthrax leth ...	2007	17229495
anthrax: from antiquity to answers.		2007	17230405
bacillus anthracis edema and lethal toxin have different hemodynamic effects but function together to worsen shock and outcome in a rat model.	to better define the contribution of edema toxins (etx) and lethal toxins (letx) to shock with bacillus anthracis, recombinant preparations of each were investigated alone or together in rats.	2007	17230417
results of a safety and immunogenicity study of an early-stage investigational rpa102 vaccine as compared to the fda licensed anthrax vaccine (anthrax vaccine adsorbed (ava. biothrax)).		2007	17234308
difference between the spore sizes of bacillus anthracis and other bacillus species.	to determine the size distribution of the spores of bacillus anthracis, and compare its size with other bacillus species grown and sporulated under similar conditions.	2007	17241334
murine innate immune response to virulent toxigenic and nontoxigenic bacillus anthracis strains.	effective treatment of anthrax is hampered by our limited understanding of the pathophysiology of bacillus anthracis infection. we used a genetically complete (pxo1(+) pxo2(+)) virulent b. anthracis strain and four isogenic toxin-null mutants to determine the effects of the anthrax edema toxin (et; edema factor [ef] plus protective antigen [pa]) and lethal toxin (lt; lethal factor [lf] plus pa) on the host innate response during systemic infection. using the spleen as an indicator for host respo ...	2007	17242059
cutaneous infection due to bacillus pumilus: report of 3 cases.	human infection due to bacillus pumilus is exceptional. we report 3 cases of cutaneous infection caused by b. pumilus that occurred in 3 shepherds, 2 of whom were members of the same family. the lesions appeared to have a morphology similar to that of cutaneous anthrax lesions. two patients were cured after treatment with amoxicillin-clavulanate, and the third patient was cured after prolonged treatment with ciprofloxacin. to our knowledge, primary cutaneous infection due to b. pumilus has not b ...	2007	17243047
label-free optical detection of anthrax-causing spores.		2007	17243788
experimental cutaneous bacillus anthracis infections in hairless hrs/j mice.	previous studies of experimental bacillus anthracis cutaneous infections in mice have implicated hair follicles as a likely entry site. hairless hrs/j mice were used to investigate this possibility because of their non-functional hair follicles that lack penetrating hair shafts. these mice also have diminished macrophage function, increased susceptibility to listeria, and enhanced neutrophil responses. hrs/j and balb/c mice were found to be resistant to epicutaneous inoculation with bacillus ant ...	2007	17244341
short-term reactogenicity and gender effect of anthrax vaccine: analysis of a 1967-1972 study and review of the 1955-2005 medical literature.	in the 1960s, the centers for disease control and prevention (cdc) held the investigational new drug (ind) application for the anthrax vaccine and collected short-term safety data from approximately 16,000 doses administered to almost 7000 individuals. while some recent anthrax vaccine safety studies have suggested that women experience more injection site reactions (isrs), to our knowledge the ind safety data were not previously examined for a gender-specific difference.	2007	17245803
selection of anthrax toxin protective antigen variants that discriminate between the cellular receptors tem8 and cmg2 and achieve targeting of tumor cells.	anthrax toxin, a three-component protein toxin secreted by bacillus anthracis, assembles into toxic complexes at the surface of receptor-bearing eukaryotic cells. the protective antigen (pa) protein binds to receptors, either tumor endothelial cell marker 8 (tem8) or cmg2 (capillary morphogenesis protein 2), and orchestrates the delivery of the lethal and edema factors into the cytosol. tem8 is reported to be overexpressed during tumor angiogenesis, whereas cmg2 is more widely expressed in norma ...	2007	17251181
dna-oligonucleotide encapsulating liposomes as a secondary signal amplification means.	a novel liposome-based signal amplification system was developed by encapsulating dna oligonucleotides within antibody-tagged liposomes and subsequently detecting the oligonucleotide with dye-encapsulating liposomes for double signal enhancement. in this sandwich immunoassay, the model analyte, protective antigen protein from b. anthracis, was captured by one set of antibodies immobilized in microtiter plate wells and detected using a second antibody conjugated to oligonucleotide-encapsulating l ...	2007	17253656
n-thiolated beta-lactams: studies on the mode of action and identification of a primary cellular target in staphylococcus aureus.	this study focuses on the mechanism of action of n-alkylthio beta-lactams, a new family of antibacterial compounds that show promising activity against staphylococcus and bacillus microbes. previous investigations have determined that these compounds are highly selective towards these bacteria, and possess completely unprecedented structure-activity profiles for a beta-lactam antibiotic. unlike penicillin, which inhibits cell wall crosslinking proteins and affords a broad spectrum of bacteriocid ...	2007	17258460
a comparative assessment of immunization records in the defense medical surveillance system and the vaccine adverse event reporting system.	we compared immunization data in the defense medical surveillance system (dmss) and immunization data for service members with an anthrax vaccine-associated adverse event reported to the vaccine adverse event reporting system (vaers) during january 1998 through december 2004. our main measure of agreement was sensitivity of the dmss conditional on an immunization record(s) occurring in vaers. the sensitivity of dmss was 73% for all vaccines and 74% for the anthrax vaccine on the vaers index immu ...	2007	17258846
plga-dendron nanoparticles enhance immunogenicity but not lethal antibody production of a dna vaccine against anthrax in mice.	dendriplexes, complexes of dendrons and condensed plasmids containing the gene for protective antigen (pa) of bacillus anthracis, were encapsulated in poly-lactide-co-glycolide (plga) particles using the double emulsion method. the two dendrons employed are a dendron with three c(18) chains (c(18) dendron) and one with no attached hydrocarbon chains (the c(0) dendron). three types of particles were examined, namely plga-c(18) dendriplexes, plga-c(0) dendriplexes and the control plga-naked dna sy ...	2007	17258876
negative regulation of bacillus anthracis sporulation by the spo0e family of phosphatases.	the initiation of sporulation in bacillus species is controlled by the phosphorelay signal transduction system. multiple regulatory elements act on the phosphorelay to modulate the level of protein phosphorylation in response to cellular, environmental, and metabolic signals. in bacillus anthracis nine possible histidine sensor kinases can positively activate the system, while two response regulator aspartyl phosphate phosphatases of the rap family negatively impact the pathway by dephosphorylat ...	2007	17259308
massive outbreak of anthrax in wildlife in the malilangwe wildlife reserve, zimbabwe.	a massive outbreak of anthrax in the wildlife of the malilangwe wildlife reserve in zimbabwe between august and november 2004 resulted in the death of almost all the reserve's estimated 500 kudu (tragelaphus strepsiceros). other species badly affected were nyala (tragelaphus angasi), bushbuck (tragelaphus scriptus), waterbuck (kobus ellipsiprymnus) and roan antelope (hippotragus equinus), which suffered losses of approximately 68 per cent, 48 per cent, 44 per cent and 42 per cent of their popula ...	2007	17259452
of spore opsonization and passive protection against anthrax.		2007	17259600
contributions of histamine, prostanoids, and neurokinins to edema elicited by edema toxin from bacillus anthracis.	bacillus anthracis edema toxin (et), composed of protective antigen and an adenylate cyclase edema factor (ef), elicits edema in host tissues, but the target cells and events leading from ef-mediated cyclic-amp production to edema are unknown. we evaluated the direct effect of et on several cell types in vitro and tested the possibility that mediators of vascular leakage, such as histamine, contribute to edema in rabbits given intradermal et. et increased the transendothelial electrical resistan ...	2007	17261611
method of measuring bacillus anthracis spores in the presence of copious amounts of bacillus thuringiensis and bacillus cereus.	a sensitive and reliable method for the detection of bacillus anthracis (ba; sterne strain 7702) spores in presence of large amounts of bacillus thuringiensis (bt) and bacillus cereus (bc) is presented based on a novel pzt-anchored piezoelectric excited millimeter-sized cantilever (papemc) sensor with a sensing area of 1.5 mm2. antibody (anti-ba) specific to ba spores was immobilized on the sensing area and exposed to various samples of ba, bt, and bc containing the same concentration of ba at 3 ...	2007	17263347
methyl bromide fumigant lethal to bacillus anthracis spores.	methyl bromide (mb), an agricultural fumigant used in the united states, is capable of reducing or eliminating bacillis anthracis spores. in the event of a bioterrorist attack, mb might serve as an excellent decontaminating agent because it leaves no residue and does not damage furnishings and commodities.	2007	17265727
tracing isolates of bacterial species by multilocus variable number of tandem repeat analysis (mlva).	all bacterial genomes contain multiple loci of repetitive dna. repeat unit sizes and repeat sequences may vary when multiple loci are considered for different isolates of an individual microbial species. moreover, it has been documented on many occasions that the number of repeat units per locus is a strain-defining parameter. consequently, there is isolate-specificity in the number of repeats per locus when different strains of a given bacterial species are compared. the experimental assessment ...	2007	17266711
large scale genotype-phenotype correlation analysis based on phylogenetic trees.	we provide two methods for identifying changes in genotype that are correlated with changes in a phenotype implied by phylogenetic trees. the first method, venn, works when the number of branches over which the change occurred are modest. venn looks for genetic changes that are completely penetrant with phenotype changes on a tree. the second method, cctsweep, allows for a partial matching between changes in phenotypes and genotypes and provides a score for each change using maddison's concentra ...	2007	17267431
microarray analysis of transposon insertion mutations in bacillus anthracis: global identification of genes required for sporulation and germination.	a transposon site hybridization (trash) assay was developed for functional analysis of the bacillus anthracis genome using a mini-tn10 transposon which permitted analysis of 82% of this pathogen's genes. the system, used to identify genes required for generation of infectious anthrax spores, spore germination, and optimal growth on rich medium, was predictive of the contributions of two conserved hypothetical genes for the phenotypes examined.	2007	17277068
universal sample preparation method for characterization of bacteria by matrix-assisted laser desorption ionization-time of flight mass spectrometry.	mass spectrometry has been a very useful method to rapidly identify microorganisms associated with infectious diseases, detect bioterrorism threats, and discriminate among different subtypes of a pathogen. in this study, we developed a universal method for bacterial identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry. the effects on the mass spectrum of different experimental conditions, including the amount of bacterial cells used and treatment procedu ...	2007	17277202
immunogenicity of a subunit vaccine against bacillus anthracis.	the current approved vaccine against anthrax is based on protective antigen (pa) of bacillus anthracis, requires six injections over an 18-month period and has a known history of side effects. therefore, there is significant effort towards developing an improved vaccine against b. anthracis. here we separately engineered and expressed domain 4 of pa (pad4) and domain 1 of lethal factor (lfd1) as fusions to lichenase (lickm), a thermostable enzyme from clostridium thermocellum, and transiently ex ...	2007	17280756
anthrax toxin receptors and infantile systemic hyalinosis.		2007	17280935
antigen-specific cd4+ t cells recognize epitopes of protective antigen following vaccination with an anthrax vaccine.	detection of antigen-specific cd4+ t cells is facilitated by the use of fluorescently labeled soluble peptide-major histocompatibility complex (mhc) multimers which mirror the antigen specificity of t-cell receptor recognition. we have used soluble peptide-mhc class ii tetramers containing peptides from the protective antigen (pa) of bacillus anthracis to detect circulating t cells in peripheral blood of subjects vaccinated with an anthrax vaccine. pa-specific hla class ii-restricted t lymphocyt ...	2007	17283103
mutagenesis and repair in bacillus anthracis: the effect of mutators.	we have generated mutator strains of bacillus anthracis sterne by using directed gene knockouts to investigate the effect of deleting genes involved in mismatch repair, oxidative repair, and maintaining triphosphate pools. the single-knockout strains are deleted for muts, muty, mutm, or ndk. we also made double-knockout strains that are muts ndk or muty mutm. we have measured the levels of mutations in the rpob gene that lead to the rif(r) phenotype and have examined the mutational specificity. ...	2007	17220233
contribution of toxins to the pathogenesis of inhalational anthrax.	inhalational anthrax is a life-threatening infectious disease of considerable concern, especially as a potential bioterrorism agent. progress is gradually being made towards understanding the mechanisms used by bacillus anthracis to escape the immune system and to induce severe septicaemia associated with toxaemia and leading to death. recent advances in fundamental research have revealed previously unsuspected roles for toxins in various cell types. we summarize here pathological data for anima ...	2007	17223930
gorse gj, et al. "immunogenicity and tolerance of ascending doses of a recombinant protective antigen (rpa102) anthrax vaccine: a randomized, double-blinded, controlled, multicenter trial" [vaccine 24 (2006) 5950-5959].	in the study reported by gorse et al. a unique, educational opportunity was lost. the vaccine and biodefense communities almost experienced the rare chance in a phase i study to scientifically compare head-to-head an early-stage, investigational recombinant anthrax vaccine (rpa102) with the safe, effective and already fda-licensed anthrax vaccine, ava (biothrax). the authors take a stab at making safety and immunogenicity comparisons between the candidate vaccine and ava (biothrax) but the study ...	2007	17224206
computer-aided diagnosis with potential application to rapid detection of disease outbreaks.	our objectives are to quickly interpret symptoms of emergency patients to identify likely syndromes and to improve population-wide disease outbreak detection. we constructed a database of 248 syndromes, each syndrome having an estimated probability of producing any of 85 symptoms, with some two-way, three-way, and five-way probabilities reflecting correlations among symptoms. using these multi-way probabilities in conjunction with an iterative proportional fitting algorithm allows estimation of ...	2007	17225213
learning from the 2001 anthrax attacks: immunological characteristics.		2007	17191158
the us capitol bioterrorism anthrax exposures: clinical epidemiological and immunological characteristics.	bioterrorism-related anthrax exposures occurred at the us capitol in 2001. exposed individuals received antibiotics and anthrax vaccine adsorbed immunization.	2007	17191162
anticipating demand for emergency health services due to medication-related adverse events after rapid mass prophylaxis campaigns.	mass prophylaxis against infectious disease outbreaks carries the risk of medication-related adverse events (mraes). the authors sought to define the relationship between the rapidity of mass prophylaxis dispensing and the subsequent demand for emergency health services due to predictable mraes.	2007	17192445
structural investigation of the glms ribozyme bound to its catalytic cofactor.	the glms riboswitch is located in the 5'-untranslated region of the gene encoding glucosamine-6-phosphate (glcn6p) synthetase. the glms riboswitch is a ribozyme with activity triggered by binding of the metabolite glcn6p. presented here is the structure of the glms ribozyme (2.5 a resolution) with glcn6p bound in the active site. the glms ribozyme adopts a compact double pseudoknot tertiary structure, with two closely packed helical stacks. recognition of glcn6p is achieved through coordination ...	2007	17196404
bacillus cereus strains fall into two clusters (one closely and one more distantly related) to bacillus anthracis according to amino acid substitutions in small acid-soluble proteins as determined by tandem mass spectrometry.	small acid-soluble proteins (sasps) are located in the core region of bacillus spores and have been previously demonstrated as reliable biomarkers for differentiating bacillus anthracis and bacillus cereus. using ms and ms-ms analysis of sasps further phylogenetic correlations among b. anthracis and b. cereus strains are described here. esi was demonstrated to be a more comprehensive method, allowing for the analysis of intact proteins in both ms and ms-ms mode, thus providing molecular weight ( ...	2007	17197155
oligonucleotide microarray identification of bacillus anthracis strains using support vector machines.	the capability of a custom microarray to discriminate between closely related dna samples is demonstrated using a set of bacillus anthracis strains. the microarray was developed as a universal fingerprint device consisting of 390 genome-independent 9mer probes. the genomes of b. anthracis strains are monomorphic and therefore, typically difficult to distinguish using conventional molecular biology tools or microarray data clustering techniques. using support vector machines (svms) as a supervise ...	2007	17204462
fatal pneumonia among metalworkers due to inhalation exposure to bacillus cereus containing bacillus anthracis toxin genes.	bacillus cereus pneumonia is unusual in nonimmunocompromised hosts. we describe fatal cases in 2 metalworkers and the associated investigation. anthrax toxin genes were identified in b. cereus isolates from both patients using polymerase chain reaction. finding anthrax toxin genes in non-bacillus anthracis isolates has, to our knowledge, only been reported once previously.	2007	17205450
photogenerated glycan arrays identify immunogenic sugar moieties of bacillus anthracis exosporium.	using photogenerated glycan arrays, we characterized a large panel of synthetic carbohydrates for their antigenic reactivities with pathogen-specific antibodies. we discovered that rabbit igg antibodies elicited by bacillus anthracis spores specifically recognize a tetrasaccharide chain that decorates the outermost surfaces of the b. anthracis exosporium. since this sugar moiety is highly specific for the spores of b. anthracis, it appears to be a key biomarker for detection of b. anthracis spor ...	2007	17205603
indirect detection of bacillus anthracis using real-time pcr to detect amplified gamma phage dna.	typical real-time pcr methods used to identify bacillus anthracis do not distinguish between viable and non-viable spores, which would be critical in any first response and remediation scenarios. this study combined both real-time pcr, using primers specifically designed for gamma phage, with the highly specific gamma phage amplification into one simple assay to indirectly detect bacillus anthracis. since the amplification of gamma phage only occurs in the presence of a suitable host, the detect ...	2007	17208322
selection of human antibodies against cell surface-associated oligomeric anthrax protective antigen.	the protective antigen (pa(83)) of bacillus anthracis is the dominant antigen in natural and vaccine-induced immunity to anthrax infection. three human single-chain variable fragments (scfvs) against cell bound pa were isolated from an antibody phage display library. specifically, the antibodies were evaluated for their ability to bind to cell bound heptameric pa and ultimately protect against the cytotoxicity of lethal toxin. in total, all three scfvs possessed neutralizing activity against the ...	2007	17210180
cutaneous anthrax in a remote tribal area--araku valley, visakhapatnam district, andhra pradesh, southern india.		2007	17214721
thymidyl biosynthesis enzymes as antibiotic targets.	the two long-known "classical" enzymes of uridyl-5-methylation, thymidylate synthase and ribothymidyl synthase, have been joined by two alternative methylation enzymes, flavin-dependent thymidylate synthase and folate-dependent ribothymidyl synthase. these two newly discovered enzymes have much in common: both contain flavin cofactors, utilize methylenetetrahydrofolate as a source of methyl group, and perform thymidylate synthesis via chemical pathways distinct from those of their classic counte ...	2007	17216455
virulent spores of bacillus anthracis and other bacillus species deposited on solid surfaces have similar sensitivity to chemical decontaminants.	to compare the relative sensitivity of bacillus anthracis and spores of other bacillus spp. deposited on different solid surfaces to inactivation by liquid chemical disinfecting agents.	2007	17184315
biosynthetic analysis of the petrobactin siderophore pathway from bacillus anthracis.	the asbabcdef gene cluster from bacillus anthracis is responsible for biosynthesis of petrobactin, a catecholate siderophore that functions in both iron acquisition and virulence in a murine model of anthrax. we initiated studies to determine the biosynthetic details of petrobactin assembly based on mutational analysis of the asb operon, identification of accumulated intermediates, and addition of exogenous siderophores to asb mutant strains. as a starting point, in-frame deletions of each of th ...	2007	17189355
the alternative sigma factor sigmah is required for toxin gene expression by bacillus anthracis.	expression of the structural genes for the anthrax toxin proteins is coordinately controlled by host-related signals, such as elevated co(2), and the trans-acting positive regulator atxa. in addition to these requirements, toxin gene expression is under growth phase regulation. the transition state regulator abrb represses atxa expression to influence toxin synthesis. during the late exponential phase of growth, when abrb levels begin to decrease, toxin synthesis increases. here we report that t ...	2007	17189374
silence: a new forward genetic technology.	silencing induced by long terminal repeat (ltr)-encoded cis-acting response element, termed silence, is a forward genetic system that allows for conditional, epigenetic control of host-gene transcription. this new research tool is independent of gene mutation or disruption, does not require complementation, and conditional gene repression appears complete at the level of protein function. silence functions in hypodiploid cells and is a platform technology with broad applications in gene discover ...	2007	17179935
ultrastructural features of lymphocyte suppression induced by anthrax lethal toxin and treated with chloroquine.	antibacterial therapy does not fully protect against anthrax because of severe systemic intoxication. lysosomal processing of anthrax lethal toxin (ltx) is a key event in the disease pathogenesis, and agents interfering with this process, like chloroquine (cq), may have practical applications. although ltx is known to induce t-cell suppression, precise mechanisms of this phenomenon are not completely characterized. in the present study, we investigated alterations of lymphocyte ultrastructure ca ...	2007	17179957
radiology of biological weapons--old and the new?		2007	17174174
the design of inhibitors for medicinally relevant metalloproteins.	a number of metalloproteins are important medicinal targets for conditions ranging from pathogenic infections to cancer. many but not all of these metalloproteins contain a zinc(ii) ion in the protein active site. small-molecule inhibitors of these metalloproteins are designed to bind directly to the active site metal ions. in this review several metalloproteins of interest are discussed, including matrix metalloproteinases (mmps), histone deacetylases (hdacs), anthrax lethal factor (lf), and ot ...	2007	17163561
adenovirus-based prime-boost immunization for rapid vaccination against anthrax.	prime-boost vaccination using plasmid dna and replication-defective adenovirus vectors has emerged as a highly effective strategy for vaccinating against viral pathogens. however, its ability to provide protection against bacterial disease has never been assessed. here we evaluate prime-boost vaccination approaches for immunizing against anthrax. we show that mice primed with dna and boosted with an adenovirus vector, both expressing domain four of bacillus anthracis protective antigen (pa), hav ...	2007	17164792
oral administration of a salmonella enterica-based vaccine expressing bacillus anthracis protective antigen confers protection against aerosolized b. anthracis.	bacillus anthracis is the causative agent of anthrax, a disease that affects wildlife, livestock, and humans. protection against anthrax is primarily afforded by immunity to the b. anthracis protective antigen (pa), particularly pa domains 4 and 1. to further the development of an orally delivered human vaccine for mass vaccination against anthrax, we produced salmonella enterica serovar typhimurium expressing full-length pa, pa domains 1 and 4, or pa domain 4 using codon-optimized pa dna fused ...	2007	17145938