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rapid targeted gene disruption in bacillus anthracis.anthrax is a zoonotic disease recognized to affect herbivores since biblical times and has the widest range of susceptible host species of any known pathogen. the ease with which the bacterium can be weaponized and its recent deliberate use as an agent of terror, have highlighted the importance of gaining a deeper understanding and effective countermeasures for this important pathogen. high quality sequence data has opened the possibility of systematic dissection of how genes distributed on both ...201324047152
bacillus anthracis has two independent bottlenecks that are dependent on the portal of entry in an intranasal model of inhalational infection.bacillus anthracis can cause inhalational anthrax. murine inhalational b. anthracis infections have two portals of entry, the nasal mucosa-associated lymphoid tissue (nalt) and the lumen of the lungs. analysis of the dissemination from these sites is hindered because infections are asynchronous and asymptomatic until the hosts near death. to further understand and compare how b. anthracis disseminates from these two different environments, clonal analysis was employed using a library of equally ...201324042112
hollow-fiber pharmacodynamic studies and mathematical modeling to predict the efficacy of amoxicillin for anthrax postexposure prophylaxis in pregnant women and children.amoxicillin is considered an option for postexposure prophylaxis of bacillus anthracis in pregnant and postpartum women who are breastfeeding and in children because of the potential toxicities of ciprofloxacin and doxycycline to the fetus and child. the amoxicillin regimen that effectively kills b. anthracis and prevents resistance is unknown. fourteen-day dose range and dose fractionation studies were conducted in in vitro pharmacodynamic models to identify the exposure intensity and pharmacod ...201324041894
infectious diseases in poland in 2011.the aim of the study was assessment of the epidemiological situation of infectious and parasitic diseases in poland in 2011 materials and methods: the main source of data to develop the statistical overview was the annual bulletin "infectious diseases in poland in 2011," and "vaccinations in poland in 2011,"/niph-nih, csi, 2011 and information contained in the articles of epidemiological journal in which authors depth discussion of the epidemiological situation of 27 diseases or groups of diseas ...201324040711
evidence of local persistence of human anthrax in the country of georgia associated with environmental and anthropogenic factors.anthrax is a soil-borne disease caused by the bacterium bacillus anthracis and is considered a neglected zoonosis. in the country of georgia, recent reports have indicated an increase in the incidence of human anthrax. identifying sub-national areas of increased risk may help direct appropriate public health control measures. the purpose of this study was to evaluate the spatial distribution of human anthrax and identify environmental/anthropogenic factors associated with persistent clusters.201324040426
effect of delayed anthrax vaccine dose on bacillus anthracis protective antigen igg response and lethal toxin neutralization activity.we describe the bacillus anthracis protective antigen igg antibody response and the b. anthracis lethal toxin neutralization activity to a delayed dose of anthrax vaccine adsorbed (ava, biothrax(®)) using validated assays. 373 individuals received 1, 2, or 3 priming doses, 18-24 months afterward, they received a delayed dose of ava. overall, 23.6% of subjects showed detectable anti-pa igg before the boost, compared to 99.2% (p<0.0001) 28 days after the boost. geometric mean anti-pa igg concentra ...201324026013
[current situation of endemic status, prevention and control of neglected zoonotic diseases in china].neglected zoonotic diseases not only threaten the health of human, especially to the livestock keepers in poverty-stricken areas but also cause great economic losses to the animal husbandry. this paper reviews the current situation of the endemic status, prevention and control of neglected zoonotic diseases existing in china including rabies, bovine tuberculosis, brucellosis, anthrax, leptospirosis, echinococcosis, cysticercosis, leishmaniasis and fascioliasis, so as to provide the basic informa ...201324024457
beetroot-pigment-derived colorimetric sensor for detection of calcium dipicolinate in bacterial spores.in this proof-of-concept study, we describe the use of the main red beet pigment betanin for the quantification of calcium dipicolinate in bacterial spores, including bacillus anthracis. in the presence of europium(iii) ions, betanin is converted to a water-soluble, non-luminescent orange 1∶1 complex with a stability constant of 1.4 × 10(5) l mol(-1). the addition of calcium dipicolinate, largely found in bacterial spores, changes the color of the aqueous solution of [eu(bn)(+)] from orange to m ...201324019934
anthrax edema factor toxicity is strongly mediated by the n-end rule.anthrax edema factor (ef) is a calmodulin-dependent adenylate cyclase that converts adenosine triphosphate (atp) into 3'-5'-cyclic adenosine monophosphate (camp), contributing to the establishment of bacillus anthracis infections and the resulting pathophysiology. we show that ef adenylate cyclase toxin activity is strongly mediated by the n-end rule, and thus is dependent on the identity of the n-terminal amino acid. ef variants having different n-terminal residues varied by more than 100-fold ...201324015319
cyclodextrin derivatives as anti-infectives.cyclodextrin derivatives can be utilized as anti-infectives with pore-forming proteins as the targets. the highly efficient selection of potent inhibitors was achieved because per-substituted cyclodextrins have the same symmetry as the target pores. inhibitors of several bacterial toxins produced by bacillus anthracis, staphylococcus aureus, clostridium perfringens, clostridium botulinum, and clostridium difficile were identified from a library of ∼200 cd derivatives. it was demonstrated that mu ...201324011515
gastrointestinal anthrax: clinical experience in 5 cases.bacillus anthracis may usually cause three forms of anthrax: inhalation, gastrointestinal and cutaneous. the gastrointestinal (gi) anthrax develops after eating contaminated meat. thus, in this paper were report 5 cases of intestinal anthrax.201324009958
in silico and in vitro evaluation of pcr-based assays for the detection of bacillus anthracis chromosomal signature sequences.bacillus anthracis, the causative agent of anthrax, is a zoonotic pathogen that is relatively common throughout the world and may cause life threatening diseases in animals and humans. there are many pcr-based assays in use for the detection of b. anthracis. while most of the developed assays rely on unique markers present on virulence plasmids pxo1 and pxo2, relatively few assays incorporate chromosomal dna markers due to the close relatedness of b. anthracis to the b. cereus group strains. for ...201324005110
two anthrax cases with soft tissue infection, severe oedema and sepsis in danish heroin users.anthrax had become extremely rare in europe, but in 2010 an outbreak of anthrax among heroin users in scotland increased awareness of contaminated heroin as a source of anthrax. we present the first two danish cases of injectional anthrax and discuss the clinical presentations, which included both typical and more unusual manifestations.201324004900
discovery of inhibitors of bacillus anthracis primase dnag.primase dnag is an essential bacterial enzyme that synthesizes short ribonucleotide primers required for chromosomal dna replication. inhibitors of dnag can serve as leads for development of new antibacterials and biochemical probes. we recently developed a nonradioactive in vitro primase-pyrophosphatase assay to identify and analyze dnag inhibitors. application of this assay to dnag from bacillus anthracis (ba dnag), a dangerous pathogen, yielded several inhibitors, which include agents with dn ...201324004110
key tissue targets responsible for anthrax-toxin-induced lethality.bacillus anthracis, the causative agent of anthrax disease, is lethal owing to the actions of two exotoxins: anthrax lethal toxin (lt) and oedema toxin (et). the key tissue targets responsible for the lethal effects of these toxins are unknown. here we generated cell-type-specific anthrax toxin receptor capillary morphogenesis protein-2 (cmg2)-null mice and cell-type-specific cmg2-expressing mice and challenged them with the toxins. our results show that lethality induced by lt and et occurs thr ...201323995686
bacillus anthracis diagnostic detection and rapid antibiotic susceptibility determination using 'bioluminescent' reporter phage.genetically modified phages have the potential to detect pathogenic bacteria from clinical, environmental, or food-related sources. herein we assess an engineered 'bioluminescent' reporter phage (wß::luxab) as a clinical diagnostic tool for bacillus anthracis, the etiological agent of anthrax. wß::luxab is able to rapidly (within minutes) detect a panel of b. anthracis strains by transducing a bioluminescent phenotype. the reporter phage displays species specificity by its inability, or signific ...201323994352
structure of isochorismate synthase dhbc from bacillus anthracis.the isochorismate synthase dhbc from bacillus anthracis is essential for the biosynthesis of the siderophore bacillibactin by this pathogenic bacterium. the structure of the selenomethionine-substituted protein was determined to 2.4 å resolution using single-wavelength anomalous diffraction. b. anthracis dhbc bears the strongest resemblance to the escherichia coli isochorismate synthase entc, which is involved in the biosynthesis of another siderophore, namely enterobactin. both proteins adopt t ...201323989140
increasing the potency of an alhydrogel-formulated anthrax vaccine by minimizing antigen-adjuvant interactions.aluminum salts are the most widely used vaccine adjuvants, and phosphate is known to modulate antigen-adjuvant interactions. here we report an unexpected role for phosphate buffer in an anthrax vaccine (sparvax) containing recombinant protective antigen (rpa) and aluminum oxyhydroxide (aloh) adjuvant (alhydrogel). phosphate ions bind to aloh to produce an aluminum phosphate surface with a reduced rpa adsorption coefficient and binding capacity. however, these effects continued to increase as the ...201323986317
anthrax vaccines: present status and future prospects.the management of anthrax remains a top priority among the biowarfare/bioterror agents. it was the bacillus anthracis spore attack through the us mail system after the september 11, 2001, terrorist attacks in the usa that highlighted the potential of b. anthracis as a bioterrorism agent and the threat posed by its deliberate dissemination. these attacks invigorated the efforts toward understanding the anthrax pathogenesis and development of more comprehensive medical intervention strategies for ...201323984963
effect of anthrax immune globulin on response to biothrax (anthrax vaccine adsorbed) in new zealand white rabbits.development of anthrax countermeasures that may be used concomitantly in a postexposure setting requires an understanding of the interaction between these products. anthrax immune globulin intravenous (aigiv) is a candidate immunotherapeutic that contains neutralizing antibodies against protective antigen (pa), a component of anthrax toxins. we evaluated the interaction between aigiv and biothrax (anthrax vaccine adsorbed) in rabbits. while pharmacokinetics of aigiv were not altered by vaccinati ...201323979740
evaluation of intravenous anthrax immune globulin for treatment of inhalation anthrax.bacillus anthracis toxins can be neutralized by antibodies against protective antigen (pa), a component of anthrax toxins. anthrivig (human anthrax immunoglobulin), also known as aigiv, derived from plasma of humans immunized with biothrax (anthrax vaccine adsorbed), is under development for the treatment of toxemia following exposure to anthrax spores. the pharmacokinetics (pk) of aigiv was assessed in naive animals and healthy human volunteers, and the efficacy of aigiv was assessed in animals ...201323979731
the interdependence of antibody c and v regions on specificity and affinity: significant implications for the engineering of therapeutic antibodies. 201323979029
ecological niche modeling of bacillus anthracis on three continents: evidence for genetic-ecological divergence?we modeled the ecological niche of a globally successful bacillus anthracis sublineage in the united states, italy and kazakhstan to better understand the geographic distribution of anthrax and potential associations between regional populations and ecology. country-specific ecological-niche models were developed and reciprocally transferred to the other countries to determine if pathogen presence could be accurately predicted on novel landscapes. native models accurately predicted endemic areas ...201323977300
erythropoiesis suppression is associated with anthrax lethal toxin-mediated pathogenic progression.anthrax is a disease caused by the bacterium bacillus anthracis, which results in high mortality in animals and humans. although some of the mechanisms are already known such as asphyxia, extensive knowledge of molecular pathogenesis of this disease is deficient and remains to be further investigated. lethal toxin (lt) is a major virulence factor of b. anthracis and a specific inhibitor/protease of mitogen-activated protein kinase kinases (mapkks). anthrax lt causes lethality and induces certain ...201323977125
[four cases of cutaneous anthrax in diyarbakir, turkey].anthrax which is a rare disease in developed countries, is still a serious public health problem in countries like turkey where livestock is common. in this report, four cases of cutaneous anthrax detected in kirkira village of diyarbakir, southeast anatolia, turkey, were presented. three female and one male patients were admitted to our hospital with the complaints of skin lesions and high fever lasting for 10 days. their history indicated that they injured their fingers during slaughtering of ...201323971932
anthrax of the lower lip.anthrax of the oral cavity and oropharynx is well known, whereas anthrax of the lips is very rare. we present a case of anthrax of the lower lip in a 57-year-old man. the infection was characterized by a wide, black eschar, surrounded by vesicles, crusts, and erythematous-edematous halo, with submandibular and laterocervical lymphadenopathy. the oral cavity, oropharynx, and tonsils were normal. laboratory examinations revealed leukocytosis and increased inflammatory markers. otolaryngologic, gas ...201324120906
zoonoses in rural veterinarians in the central region of argentina.to estimate the frequency of zoonoses in rural veterinarians and to search for risk factors.201324118152
acute infections in intravenous drug users. 201324115713
proteomic analysis of bacillus subtilis strains engineered for improved production of heterologous proteins.the use of bacterial systems for recombinant protein production has advantages of simplicity, time and cost over competing systems. however, widely used bacterial expression systems (e.g. escherichia coli, pseudomonas fluorescens) are not able to secrete soluble proteins directly into the culture medium. this limits yields and increases downstream processing time and costs. in contrast, bacillus spp. secrete native enzymes directly into the culture medium at grams-per-litre quantities, although ...201324115457
b. anthracis associated cardiovascular dysfunction and shock: the potential contribution of both non-toxin and toxin components.the development of cardiovascular dysfunction and shock in patients with invasive bacillus anthracis infection has a particularly poor prognosis. growing evidence indicates that several bacterial components likely play important pathogenic roles in this injury. as with other pathogenic gram-positive bacteria, the b. anthracis cell wall and its peptidoglycan constituent produce a robust inflammatory response with its attendant tissue injury, disseminated intravascular coagulation and shock. howev ...201324107194
quantitative models of the dose-response and time course of inhalational anthrax in humans.anthrax poses a community health risk due to accidental or intentional aerosol release. reliable quantitative dose-response analyses are required to estimate the magnitude and timeline of potential consequences and the effect of public health intervention strategies under specific scenarios. analyses of available data from exposures and infections of humans and non-human primates are often contradictory. we review existing quantitative inhalational anthrax dose-response models in light of criter ...201324058320
a sensitive & specific multiplex pcr assay for simultaneous detection of bacillus anthracis, yersinia pestis, burkholderia pseudomallei & brucella species.bacillus anthracis, yersinia pestis, burkholderia pseudomallei and brucella species are potential biowarfare agents. classical bacteriological methods for their identification are cumbersome, time consuming and of potential risk to the handler.201324056564
the two ccda proteins of bacillus anthracis differentially affect virulence gene expression and sporulation.the cytochrome c maturation system influences the expression of virulence factors in bacillus anthracis. b. anthracis carries two copies of the ccda gene, encoding predicted thiol-disulfide oxidoreductases that contribute to cytochrome c maturation, while the closely related organism bacillus subtilis carries only one copy of ccda. to investigate the roles of the two ccda gene copies in b. anthracis, strains were constructed without each ccda gene, and one strain was constructed without both cop ...201324056109
systemic use of fluoroquinolone in children.fluoroquinolones are an important class of antibiotics that are widely used in adult patients because of their broad spectrum of activity, good tissue penetration, and oral bioavailability. however, fluoroquinolone use in children is limited because juvenile animals developed arthropathy in previous experiments on fluoroquinolone use. indications for fluoroquinolone use in patients younger than 18 years, as stated by the u.s. food and drug administration, include treatment of complicated urinary ...201323741232
eco-genetic structure of bacillus cereus sensu lato populations from different environments in northeastern poland.the bacillus cereus group, which includes entomopathogens and etiologic agents of foodborne illness or anthrax, persists in various environments. the basis of their ecological diversification remains largely undescribed. here we present the genetic structure and phylogeny of 273 soil b. cereus s.l. isolates from diverse habitats in northeastern poland, with samplings acquired from the last european natural forest (białowieża national park), the largest marshes in europe (biebrza national park), ...201324312460
arsenic trioxide and other arsenical compounds inhibit the nlrp1, nlrp3, and naip5/nlrc4 inflammasomes.inflammasomes are large cytoplasmic multiprotein complexes that activate caspase-1 in response to diverse intracellular danger signals. inflammasome components termed nucleotide-binding oligomerization domain-like receptor (nlr) proteins act as sensors for pathogen-associated molecular patterns, stress, or danger stimuli. we discovered that arsenicals, including arsenic trioxide and sodium arsenite, inhibited activation of the nlrp1, nlrp3, and naip5/nlrc4 inflammasomes by their respective activ ...201324337744
direct proteolytic cleavage of nlrp1b is necessary and sufficient for inflammasome activation by anthrax lethal factor.inflammasomes are multimeric protein complexes that respond to infection by recruitment and activation of the caspase-1 (casp1) protease. activated casp1 initiates immune defense by processing inflammatory cytokines and by causing a rapid and lytic cell death called pyroptosis. inflammasome formation is orchestrated by members of the nucleotide-binding domain and leucine-rich repeat (nlr) or aim2-like receptor (alr) protein families. certain nlrs and alrs have been shown to function as direct re ...201323818853
draft genome sequences of pseudomonas fluorescens bs2 and pusillimonas noertemannii bs8, soil bacteria that cooperate to degrade the poly-γ-d-glutamic acid anthrax capsule.a mixed culture of pseudomonas fluorescens bs2 and pusillimonas noertemannii bs8 degraded poly-γ-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. here we report the draft genome sequences of both soil isolates.201323405351
anthracimycin, a potent anthrax antibiotic from a marine-derived actinomycete.licensed to kill: a new antibiotic, anthracimycin (see scheme), produced by a marine-derived actinomycete in saline culture, shows significant activity toward bacillus anthracis, the bacterial pathogen responsible for anthrax infections. chlorination of anthracimycin gives a dichloro derivative that retains activity against gram-positive bacteria, such as anthrax, but also shows activity against selected gram-negative bacteria.201323776159
simultaneous and rapid detection of salmonella typhi, bacillus anthracis, and yersinia pestis by using multiplex polymerase chain reaction (pcr).salmonella typhi, bacillus anthracis, and yersinia pestis are some serious human pathogens, which their early diagnosis is of great importance. salmonella typhi, bacillus anthracis, and yersinia pestis cause typhoid fever, anthrax, and plague respectively. these bacteria can be used to make biologic weapons.201324719692
reverse transcription-pcr-electrospray ionization mass spectrometry for rapid detection of biothreat and common respiratory pathogens.electrospray ionization mass spectrometry (esi-ms) analysis of reverse transcription (rt)-pcr amplicons from human respiratory samples allows for broad pathogen identification approximately 8 h after collection. we investigated the performance characteristics of a high-throughput rt-pcr-coupled esi-ms assay for distinguishing biothreat (bt) agents from common bacterial, fungal, and viral respiratory pathogens in bronchoalveolar lavage (bal) fluid specimens from subjects with suspected respirator ...201323903543
detection of bacillus anthracis dna in complex soil and air samples using next-generation sequencing.bacillus anthracis is the potentially lethal etiologic agent of anthrax disease, and is a significant concern in the realm of biodefense. one of the cornerstones of an effective biodefense strategy is the ability to detect infectious agents with a high degree of sensitivity and specificity in the context of a complex sample background. the nature of the b. anthracis genome, however, renders specific detection difficult, due to close homology with b. cereus and b. thuringiensis. we therefore elec ...201324039948
rapid identification of bacillus anthracis spores in suspicious powder samples by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (maldi-tof ms).rapid and reliable identification of bacillus anthracis spores in suspicious powders is important to mitigate the safety risks and economic burdens associated with such incidents. the aim of this study was to develop and validate a rapid and reliable laboratory-based matrix-assisted laser desorption ionization-time of flight mass spectrometry (maldi-tof ms) analysis method for identifying b. anthracis spores in suspicious powder samples. a reference library containing 22 different bacillus sp. s ...201323811517
live attenuated tularemia vaccines: recent developments and future goals.in the aftermath of the 2001 anthrax attacks in the u.s., numerous efforts were made to increase the level of preparedness against a biological attack both in the us and worldwide. as a result, there has been an increase in research interest in the development of vaccines and other countermeasures against a number of agents with the potential to be used as biological weapons. one such agent, francisella tularensis, has been the subject of a surge in the level of research being performed, leading ...201323764535
transient lipopolysaccharide-induced resistance to aerosolized bacillus anthracis in new zealand white rabbits.previous studies have demonstrated that prior infection by various bacterial pathogens induces nonspecific resistance to subsequent infection by other gram-negative and gram-positive bacterial pathogens. in the present study, we evaluated whether underlying inflammation enhanced host resistance to inhalational bacillus anthracis infection in new zealand white rabbits (spf; bordetella- and pasteurella-free). accordingly, rabbits were pretreated with either the inflammagen bacterial lps (60,000 eu ...201323759528
a systematic screen of fda-approved drugs for inhibitors of biological threat agents.the rapid development of effective medical countermeasures against potential biological threat agents is vital. repurposing existing drugs that may have unanticipated activities as potential countermeasures is one way to meet this important goal, since currently approved drugs already have well-established safety and pharmacokinetic profiles in patients, as well as manufacturing and distribution networks. therefore, approved drugs could rapidly be made available for a new indication in an emerge ...201323577127
development of a panel of recombinase polymerase amplification assays for detection of biothreat agents.syndromic panels for infectious disease have been suggested to be of value in point-of-care diagnostics for developing countries and for biodefense. to test the performance of isothermal recombinase polymerase amplification (rpa) assays, we developed a panel of 10 rpas for biothreat agents. the panel included rpas for francisella tularensis, yersinia pestis, bacillus anthracis, variola virus, and reverse transcriptase rpa (rt-rpa) assays for rift valley fever virus, ebola virus, sudan virus, and ...201323345286
evaluation of the filmarray® system for detection of bacillus anthracis, francisella tularensis and yersinia pestis.to evaluate the sensitivity and specificity of the biofire diagnostics filmarray(®) system in combination with their biothreat panel for the detection of bacillus anthracis (ba), francisella tularensis (ft) and yersinia pestis (yp) dna, and demonstrate the detection of ba spores.201323279070
recombinant vaccine displaying the loop-neutralizing determinant from protective antigen completely protects rabbits from experimental inhalation anthrax.we previously showed that a multiple antigenic peptide (map) vaccine displaying amino acids (aa) 304 to 319 from the 2β2-2β3 loop of protective antigen was capable of protecting rabbits from an aerosolized spore challenge with bacillus anthracis ames strain. antibodies to this sequence, referred to as the loop-neutralizing determinant (lnd), are highly potent at neutralizing lethal toxin yet are virtually absent in rabbit and human protective antigen (pa) antiserum. while the map vaccine was pro ...201323283638
stochastic humoral immunity to bacillus anthracis protective antigen: identification of anti-peptide igg correlating with seroconversion to lethal toxin neutralization.a substantial fraction of individuals vaccinated against anthrax have low to immeasurable levels of serum lethal toxin (letx)-neutralizing activity. the only known correlate of protection against bacillus anthracis in the currently licensed vaccine is magnitude of the igg response to protective antigen (pa); however, some individuals producing high serum levels of anti-pa igg fail to neutralize letx in vitro. this suggests that non-protective humoral responses to pa may be immunodominant in some ...201323415781
a preliminary study on the insect fauna of al-baha province, saudi arabia, with descriptions of two new species.a preliminary study was carried out on the insect fauna of al-baha province, south-western part of saudi arabia. a total number of 582 species and subspecies (few identified only to the genus level) belonging to 129 families and representing 17 orders were recorded. two of these species are described as new, namely: monomorium sarawatensis sharaf & aldawood, sp. n. [formicidae, hymenoptera] and anthrax alruqibi el-hawagry sp. n. [bombyliidae, diptera]. another eight species are recorded for the ...201323794807
innate immune protection against infectious diseases by pulmonary administration of a phospholipid-conjugated tlr7 ligand.pulmonary administration of toll-like receptor (tlr) ligands protects hosts from inhaled pathogens. however, systemic side effects induced by tlr stimulation limit clinical development. here, a small-molecule tlr7 ligand conjugated with phospholipid, 1v270 (also designated tmx201), was tested for innate immune activation and its ability to prevent pulmonary infection in mice. we hypothesized that phospholipid conjugation would increase internalization by immune cells and localize the compound in ...201324192551
rapid focused sequencing: a multiplexed assay for simultaneous detection and strain typing of bacillus anthracis, francisella tularensis, and yersinia pestis.the intentional release of bacillus anthracis in the united states in 2001 has heightened concern about the use of pathogenic microorganisms in bioterrorism attacks. many of the deadliest bacteria, including the class a select agents bacillus anthracis, francisella tularensis, and yersinia pestis, are highly infectious via the pulmonary route when released in aerosolized form. hence, rapid, sensitive, and reliable methods for detection of these biothreats and characterization of their potential ...201323418519
combinations of monoclonal antibodies to anthrax toxin manifest new properties in neutralization assays.monoclonal antibodies (mabs) are potential therapeutic agents against bacillus anthracis toxins, since there is no current treatment to counteract the detrimental effects of toxemia. in hopes of isolating new protective mabs to the toxin component lethal factor (lf), we used a strain of mice (c57bl/6) that had not been used in previous studies, generating mabs to lf. six lf-binding mabs were obtained, representing 3 igg isotypes and one igm. one mab (20c1) provided protection from lethal toxin ( ...201323509144
dna vaccines: roles against diseases.vaccination is the most successful application of immunological principles to human health. vaccine efficacy needs to be reviewed from time to time and its safety is an overriding consideration. dna vaccines offer simple yet effective means of inducing broad-based immunity. these vaccines work by allowing the expression of the microbial antigen inside host cells that take up the plasmid. these vaccines function by generating the desired antigen inside the cells, with the advantage that this may ...201324432284
identification of the immunogenic spore and vegetative proteins of bacillus anthracis vaccine strain a16r.immunoproteomics was used to screen the immunogenic spore and vegetative proteins of bacillus anthracis vaccine strain a16r. the spore and vegetative proteins were separated by 2d gel electrophoresis and transferred to polyvinylidene difluoride membranes, and then western blotting was performed with rabbit immune serum against b.anthracis live spores. immunogenic spots were cut and digested by trypsin. matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed to ...201323516421
menaquinone analogs inhibit growth of bacterial pathogens.gram-positive bacteria cause serious human illnesses through combinations of cell surface and secreted virulence factors. we initiated studies with four of these organisms to develop novel topical antibacterial agents that interfere with growth and exotoxin production, focusing on menaquinone analogs. menadione, 1,4-naphthoquinone, and coenzymes q1 to q3 but not menaquinone, phylloquinone, or coenzyme q10 inhibited the growth and to a greater extent exotoxin production of staphylococcus aureus, ...201323959313
surface display of recombinant proteins on escherichia coli by bcla exosporium of bacillus anthracis.the anchoring motif is one of the most important aspects of cell surface display as well as efficient and stable display of target proteins. thus, there is currently a need for the identification and isolation of novel anchoring motifs.201324053632
overcoming target-mediated quinolone resistance in topoisomerase iv by introducing metal-ion-independent drug-enzyme interactions.quinolones, which target gyrase and topoisomerase iv, are the most widely prescribed antibacterials worldwide. unfortunately, their use is threatened by the increasing prevalence of target-mediated drug resistance. greater than 90% of mutations that confer quinolone resistance act by disrupting enzyme-drug interactions coordinated by a critical water-metal ion bridge. quinazolinediones are quinolone-like drugs but lack the skeletal features necessary to support the bridge interaction. these comp ...201324047414
novel strategies for enhanced removal of persistent bacillus anthracis surrogates and clostridium difficile spores from skin.removing spores of clostridium difficile and bacillus anthracis from skin is challenging because they are resistant to commonly used antimicrobials and soap and water washing provides only modest efficacy. we hypothesized that hygiene interventions incorporating a sporicidal electrochemically generated hypochlorous acid solution (vashe(®)) would reduce the burden of spores on skin.201323844234
topoisomerase iv-quinolone interactions are mediated through a water-metal ion bridge: mechanistic basis of quinolone resistance.although quinolones are the most commonly prescribed antibacterials, their use is threatened by an increasing prevalence of resistance. the most common causes of quinolone resistance are mutations of a specific serine or acidic residue in the a subunit of gyrase or topoisomerase iv. these amino acids are proposed to serve as a critical enzyme-quinolone interaction site by anchoring a water-metal ion bridge that coordinates drug binding. to probe the role of the proposed water-metal ion bridge, w ...201323460203
anti-inflammatory and anti-superbacterial activity of polyphenols isolated from black raspberry.the fruit of the black raspberry (rubus coreanus miquel) has been employed in traditional medicine, and recent studies have demonstrated its measureable biological activities. however, the root of the black raspberry has not been studied. therefore, in this study, we evaluated the anti-inflammatory and antibacterial properties of the root and unripe fruit polyphenols of the black raspberry. both polyphenols proved to have anti-inflammatory activity as evidenced by the decreased nitric oxide (no) ...201323440625
storage effects on sample integrity of environmental surface sampling specimens with bacillus anthracis spores.the effect of packaging, shipping temperatures and storage times on recovery of bacillus anthracis. sterne spores from swabs was investigated. macrofoam swabs were pre-moistened, inoculated with bacillus anthracis spores, and packaged in primary containment or secondary containment before storage at -15°c, 5°c, 21°c, or 35°c for 0-7 days. swabs were processed according to validated centers for disease control/laboratory response network culture protocols, and the percent recovery relative to a r ...201327213119
nucleotidyl cyclase activity of particulate guanylyl cyclase a: comparison with particulate guanylyl cyclases e and f, soluble guanylyl cyclase and bacterial adenylyl cyclases cyaa and edema factor.guanylyl cyclases (gcs) regulate many physiological processes by catalyzing the synthesis of the second messenger cgmp. the gc family consists of seven particulate gcs (pgcs) and a nitric oxide-activated soluble gc (sgc). rat sgc α1β1 possesses much broader substrate specificity than previously assumed. moreover, the exotoxins cyaa from bordetella pertussis and edema factor (ef) from bacillus anthracis possess nucleotidyl cyclase (nc) activity. pgc-a is a natriuretic peptide-activated homodimer ...201323922959
increased camp in monocytes augments notch signaling mechanisms by elevating rbp-j and transducin-like enhancer of split (tle).in cells of the innate immune system, pathological increases in intracellular camp attenuate immune responses and contribute to infections by bacteria such as bacillus anthracis. in this work, camp from b. anthracis edema toxin (et) is found to activate the notch signaling pathway in both mouse macrophages and human monocytes. et as well as a cell-permeable activator of pka induce notch target genes (hes1, hey1, il2ra, and il7r) and are able to significantly enhance the induction of these notch ...201323775085
structural and functional insight into an unexpectedly selective n-methyltransferase involved in plantazolicin biosynthesis.plantazolicin (pzn), a polyheterocyclic, n(α),n(α)-dimethylarginine-containing antibiotic, harbors remarkably specific bactericidal activity toward strains of bacillus anthracis, the causative agent of anthrax. previous studies demonstrated that genetic deletion of the s-adenosyl-l-methionine-dependent methyltransferase from the pzn biosynthetic gene cluster results in the formation of desmethylpzn, which is devoid of antibiotic activity. here we describe the in vitro reconstitution, mutational ...201323878226
total synthesis of the ribosomally synthesized linear azole-containing peptide plantazolicin a from bacillus amyloliquefaciens. 201323761292
evaluation of ambulance decontamination using gaseous chlorine dioxide.we evaluated gaseous chlorine dioxide (clo2) decontamination of an ambulance using a variety of bacterial biological agents.201323734992
highly potent silver-organoalkoxysilane antimicrobial porous nanomembrane.we used a simple electrospinning technique to fabricate a highly potent silver-organoalkoxysilane antimicrobial composite from agno3-polyvinylpyrrolidone (pvp)/3-aminopropyltrimethoxysilane (aptms)/tetraethoxysilane (teos) solution. spectroscopic and microscopic analyses of the composite showed that the fibers contain an organoalkoxysilane 'skeleton,' 0.18 molecules/nm2 surface amino groups, and highly dispersed and uniformly distributed silver nanoparticles (5 nm in size). incorporation of orga ...201323574791
transcriptional response of bovine monocyte-derived macrophages after the infection with different argentinean mycobacterium bovis isolates.infection of bovines with mycobacterium bovis causes important financial hardship in many countries presenting also a risk for humans. m. bovis is known to be adapted to survive and thrive within the intramacrophage environment. in spite of its relevance, at present the information about macrophage expression patterns is scarce, particularly regarding the bovine host. in this study, transcriptomic analysis was used to detect genes differentially expressed in macrophages derived from peripheral b ...201323484118
bangladesh anthrax outbreaks are probably caused by contaminated livestock feed.in bangladesh from 1 july to 30 september 2010 there were 104 animal cases of anthrax and 607 associated human cases. this investigation was conducted in sirajganj district in december 2010, on eight farms where animal cases had occurred. bacillus anthracis was recovered from soil samples and turbinate bones on six farms. canonical single nucleotide polymorphism (snp) analysis showed that all the isolates belonged to the major lineage a, sublineage a.br.001/002 of china and south east asia while ...201322814512
host cell cytotoxicity and cytoskeleton disruption by ceradpr, an adp-ribosyltransferase of bacillus cereus g9241.bacillus cereus g9241 was isolated from a welder suffering from an anthrax-like inhalation illness. b. cereus g9241 encodes two megaplasmids, pbcxo1 and pbc210, which are analogous to the toxin- and capsule-encoding virulence plasmids of bacillus anthracis. protein modeling predicted that the pbc210 lf homologue contained an adp-ribosyltransferase (adpr) domain. this putative bacterial adp-ribosyltransferase domain was denoted ceradpr. iterative modeling showed that ceradpr possessed several con ...201322934824
brucella as a potential agent of bioterrorism.perception on bioterrorism has changed after the deliberate release of anthrax by the postal system in the united states of america in 2001. potential bioterrorism agents have been reclassified based on their dissemination, expected rate of mortality, availability, stability, and ability to lead a public panic. brucella species can be easily cultured from infected animals and human materials. also, it can be transferred, stored and disseminated easily. an intentional contamination of food with b ...201322934672
structure-activity relationship for enantiomers of potent inhibitors of b. anthracis dihydrofolate reductase.bacterial resistance to antibiotic therapies is increasing and new treatment options are badly needed. there is an overlap between these resistant bacteria and organisms classified as likely bioterror weapons. for example, bacillus anthracis is innately resistant to the anti-folate trimethoprim due to sequence changes found in the dihydrofolate reductase enzyme. development of new inhibitors provides an opportunity to enhance the current arsenal of anti-folate antibiotics while also expanding th ...201322999981
tumor therapy with a urokinase plasminogen activator-activated anthrax lethal toxin alone and in combination with paclitaxel.pa-u2, an engineered anthrax protective antigen that is activated by urokinase was combined with wildtype lethal factor in the treatment of colo205 colon adenocarcinoma in vitro and b16-bl6 mouse melanoma in vitro and in vivo. this therapy was also tested in combination with the small molecule paclitaxel, based on prior reports suggesting synergy between erk1/2 inhibition and chemotherapeutics. colo205 was sensitive to pa-u2/lf while b16-bl6 was not. for the combination treatment of b16-bl6, pac ...201322843210
multilocus sequence analysis of bacillus thuringiensis serovars navarrensis, bolivia and vazensis and bacillus weihenstephanensis reveals a common phylogeny.the bacillus cereus group sensu lato includes six closely-related bacterial species: bacillus cereus, bacillus anthracis, bacillus thuringiensis, bacillus mycoides, bacillus pseudomycoides and bacillus weihenstephanensis. b. thuringiensis is distinguished from the other species mainly by the appearance of an inclusion body upon sporulation. b. weihenstephanensis is distinguished based on its psychrotolerance and the presence of specific signature sequences in the 16s rrna gene and cspa genes. a ...201323073664
screen for agents that induce autolysis in bacillus subtilis.the growing prevalence of antibiotic-resistant infections underscores the need to discover new antibiotics and to use them with maximum effectiveness. in response to these needs, we describe a screening protocol for the discovery of autolysis-inducing agents that uses two bacillus subtilis reporter strains, sh-536 and bau-102. to screen chemical libraries, autolysis-inducing agents were first identified with a bau-102-based screen and then subdivided with sh-536 into two major groups: those that ...201323089762
poly-γ-(d)-glutamic acid capsule interferes with lytic infection of bacillus anthracis by b. anthracis-specific bacteriophages.the poly-γ-d-glutamic acid capsule of bacillus anthracis is a barrier to infection by b. anthracis-specific bacteriophages. capsule expression was found to completely inhibit lytic infection by γ phage, an observation supported by the demonstration that this phage does not elaborate a hydrolase that would facilitate penetration through the protective capsule outer layer.201323124233
an adenovirus-vectored nasal vaccine confers rapid and sustained protection against anthrax in a single-dose regimen.bacillus anthracis is the causative agent of anthrax, and its spores have been developed into lethal bioweapons. to mitigate an onslaught from airborne anthrax spores that are maliciously disseminated, it is of paramount importance to develop a rapid-response anthrax vaccine that can be mass administered by nonmedical personnel during a crisis. we report here that intranasal instillation of a nonreplicating adenovirus vector encoding b. anthracis protective antigen could confer rapid and sustain ...201323100479
use of site-directed mutagenesis to model the effects of spontaneous deamidation on the immunogenicity of bacillus anthracis protective antigen.long-term stability is a desired characteristic of vaccines, especially anthrax vaccines, which must be stockpiled for large-scale use in an emergency situation; however, spontaneous deamidation of purified vaccine antigens has the potential to adversely affect vaccine immunogenicity over time. in order to explore whether spontaneous deamidation of recombinant protective antigen (rpa)--the major component of new-generation anthrax vaccines--affects vaccine immunogenicity, we created a "genetical ...201323115046
immunization of mice with formalin-inactivated spores from avirulent bacillus cereus strains provides significant protection from challenge with bacillus anthracis ames.bacillus anthracis spores are the infectious form of the organism for humans and animals. however, the approved human vaccine in the united states is derived from a vegetative culture filtrate of a toxigenic, nonencapsulated b. anthracis strain that primarily contains protective antigen (pa). immunization of mice with purified spore proteins and formalin-inactivated spores (fis) from a nonencapsulated, nontoxigenic b. anthracis strain confers protection against b. anthracis challenge when pa is ...201323114705
in vivo murine and in vitro m-like cell models of gastrointestinal anthrax.bacillus anthracis is the causative agent of anthrax and is acquired by three routes of infection: inhalational, gastrointestinal and cutaneous. gastrointestinal (gi) anthrax is rare, but can rapidly result in severe, systemic disease that is fatal in 25%-60% of cases. disease mechanisms of gi anthrax remain unclear due to limited numbers of clinical cases and the lack of experimental animal models. here, we developed an in vivo murine model of gi anthrax where spore survival was maximized throu ...201323108317
in vivo germination of bacillus anthracis spores during murine cutaneous infection.germination is a key step for successful bacillus anthracis colonization and systemic dissemination. few data are available on spore germination in vivo, and the necessity of spore and host cell interactions to initiate germination is unclear.201323148288
long genomic dna amplicons adsorption onto unmodified gold nanoparticles for colorimetric detection of bacillus anthracis.unmodified gold nanoparticles (gnps) can be wrapped with long genomic single- and double-stranded dna (ssdna and dsdna) molecules produced by asymmetric polymerase chain reaction (as-pcr). more importantly, the dna-au interaction can be utilized for colorimetric detection of a specific nucleic acid sequence in clinical samples.201323145437
estimated copy number of bacillus anthracis plasmids pxo1 and pxo2 using digital pcr.we evaluated digital pcr (dpcr) to directly enumerate plasmid and chromosome copies in three strains of bacillus anthracis. copy number estimates based on conventional quantitative pcr (qpcr) highlighted the variability of using qpcr to measure copy number whereas estimates based on direct sequencing are comparable to dpcr.201323142659
anthrax epizootic and migration: persistence or extinction.in this paper, we use an extension of the deterministic mathematical model of an anthrax epizootic of hahn and furniss to study the effects of anthrax transmission, carcass ingestion, carcass induced environmental contamination, and migration rates on the persistence or extinction of animal populations. we compute the basic reproduction number r(0) for the anthrax epizootic model with and without taking into account animal migration. we obtained conditions for an anthrax enzootic region. we demo ...201323137874
ability of elisa and a toxin neutralization assay to detect changes in immunogenicity of a recombinant bacillus anthracis protective antigen vaccine upon storage.we examined the capability of a mouse immunogenicity assay to detect improper storage of a recombinant protective antigen (rpa)-based anthrax vaccine formulated with an aluminum adjuvant, using elisa and a toxin neutralization assay (tna) to measure the antibody response to rpa. the vaccine was stored at 4 °c, room temperature (rt) or 37 °c for one, four and eight weeks and used for immunization, along with freshly prepared vaccine. results showed that, contrary to elisa, tna is suitable to dete ...201323137818
detection of multiple waterborne pathogens using microsequencing arrays.a microarray was developed to simultaneously detect cryptosporidium parvum, cryptosporidium hominis, enterococcus faecium, bacillus anthracis and francisella tularensis in water.201323167710
bacillus anthracis acetyltransferases pata1 and pata2 modify the secondary cell wall polysaccharide and affect the assembly of s-layer proteins.the envelope of bacillus anthracis encompasses a proteinaceous s-layer with two s-layer proteins (sap and ea1). protein assembly in the envelope of b. anthracis requires s-layer homology domains (slh) within s-layer proteins and s-layer-associated proteins (bsls), which associate with the secondary cell wall polysaccharide (scwp), an acetylated carbohydrate that is tethered to peptidoglycan. here, we investigated the contributions of two putative acetyltransferases, pata1 and pata2, on scwp acet ...201323243307
rapid deamidation of recombinant protective antigen when adsorbed on aluminum hydroxide gel correlates with reduced potency of vaccine.deamidation of the recombinant protective antigen (rpa) correlates with decreased effectiveness of the vaccine in protecting against infection by bacillus anthracis. we present data demonstrating dramatic deamidation of amino acid positions 713 and 719 of rpa adsorbed onto aluminum hydroxide gel, an adjuvant, relative to rpa stored in solution without adjuvant. although deamidation did not impact total levels of rpa-specific antibodies in a mouse model, it did correlate with a decrease in toxin- ...201323242822
plasma bacterial and mitochondrial dna distinguish bacterial sepsis from sterile systemic inflammatory response syndrome and quantify inflammatory tissue injury in nonhuman primates.systemic inflammatory response syndrome (sirs) is a fundamental host response common to bacterial infection and sterile tissue injury. systemic inflammatory response syndrome can cause organ dysfunction and death, but its mechanisms are incompletely understood. moreover, sirs can progress to organ failure or death despite being sterile or after control of the inciting infection. biomarkers discriminating between sepsis, sterile sirs, and postinfective sirs would therefore help direct care. circu ...201323247122
dna sequence detection based on raman spectroscopy using single walled carbon nanotube.a biosensor for detection of dna sequence in bacterium bacillus anthracis has been developed using raman spectrum of single walled carbon nanotubes (swnts). we have utilized the fact that being hydrophobic in nature single strand of target dna (diseased dna) gets wrapped over swnt surface forming single stranded dna-swnt complex. the sensing ability of this sensor has been studied using the dependency of g peak intensity (in the raman spectrum of swnts) on the covered surface of swnts. when a dn ...201323207369
rapid detection of bacillus anthracis spores using a super-paramagnetic lateral-flow immunological detection system.there is an urgent need for convenient, sensitive, and specific methods to detect the spores of bacillus anthracis, the causative agent of anthrax, because of the bioterrorism threat posed by this bacterium. in this study, we firstly develop a super-paramagnetic lateral-flow immunological detection system for b. anthracis spores. this system involves the use of a portable magnetic assay reader, super-paramagnetic iron oxide particles, lateral-flow strips and two different monoclonal antibodies d ...201323206542
bacillus cereus g9241 s-layer assembly contributes to the pathogenesis of anthrax-like disease in mice.bacillus cereus g9241, the causative agent of anthrax-like disease, harbors virulence plasmids encoding anthrax toxins as well as hyaluronic acid (ha) and b. cereus exopolysaccharide (bps) capsules. b. cereus g9241 also harbors s-layer genes, including homologs of bacillus anthracis surface array protein (sap), extractable antigen 1 (ea1), and the s-layer-associated proteins (bsls). in b. anthracis, s-layer proteins and bsls attach via their s-layer homology domains (slh) to the secondary cell w ...201323204457
anthrax toxin receptor 2a controls mitotic spindle positioning.oriented mitosis is essential during tissue morphogenesis. the wnt/planar cell polarity (wnt/pcp) pathway orients mitosis in a number of developmental systems, including dorsal epiblast cell divisions along the animal-vegetal (a-v) axis during zebrafish gastrulation. how wnt signalling orients the mitotic plane is, however, unknown. here we show that, in dorsal epiblast cells, anthrax toxin receptor 2a (antxr2a) accumulates in a polarized cortical cap, which is aligned with the embryonic a-v axi ...201323201782
monoclonal antibody against the poly-gamma-d-glutamic acid capsule of bacillus anthracis protects mice from enhanced lethal toxin activity due to capsule and anthrax spore challenge.the poly-gamma-d-glutamic acid (pga) capsule, a major virulence factor of bacillus anthracis, protects bacilli from immune surveillance and allows its unimpeded growth in the host. recently, the importance of the pga in the pathogenesis of anthrax infection has been reported. the pga capsule is associated with lethal toxin (lt) in the blood of experimentally infected animals and enhances the cytotoxicity of lt.201323201204
recombinant expression and purification of a tumor-targeted toxin in bacillus anthracis.many recombinant therapeutic proteins are purified from escherichia coli. while expression in e. coli is easily achieved, some disadvantages such as protein aggregation, formation of inclusion bodies, and contamination of purified proteins with the lipopolysaccharides arise. lipopolysaccharides have to be removed to prevent inflammatory responses in patients. use of the gram-positive bacillus anthracis as an expression host offers a solution to circumvent these problems. using the multiple prote ...201323200832
role of mechanical vs. chemical action in the removal of adherent bacillus spores during cip procedures.this study was designed to evaluate the respective roles of mechanical and chemical effects on the removal of bacillus spores during cleaning-in-place. this analysis was performed on 12 strains belonging to the bacillus cereus group (b. cereus, bacillus anthracis, bacillus thuringiensis) or to less related bacillus species (bacillus pumilus, bacillus licheniformis, bacillus sporothermodurans, bacillus subtilis). adherent spores were subjected to rinsing-in-place (mechanical action) and cleaning- ...201323200646
perhydrolase-nanotube paint composites with sporicidal and antiviral activity.act (perhydrolase) containing paint composites were prepared leading to broad-spectrum decontamination. act was immobilized onto multi-walled carbon nanotubes (mwnts) and then incorporated into latex-based paints to form catalytic coatings. these act-based paint composites showed a 6-log reduction in the viability of spores of bacillus cereus and bacillus anthracis (sterne) within 60 min. the paint composites also showed >4-log reduction in the titer of influenza virus (x-31) within 10 min (init ...201323188457
bacillus anthracis-derived nitric oxide induces protein s-nitrosylation contributing to macrophage death.bacillus anthracis, a causative agent of anthrax, is able to germinate and survive within macrophages. a recent study suggested that b. anthracis-derived nitric oxide (bno) is a key aspect of bacterial defense that protects bacterial dna from oxidative burst in the macrophages. however, the virulent effect of bno in host cells has not been investigated. here, we report that bno contributes macrophage killing by s-nitrosylation of bioenergetic-relating proteins within mitochondria. toxigenic ster ...201323178574
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