| using blood glucose data as an indicator for epidemic disease outbreaks. | in the future, transfer of vital sensor data from patients to the public health care system is likely to become commonplace. systems for automatic transfer of sensor data are now at the prototype stage. as electronic health record (ehr) systems adapt such functionality, widespread use may become an actuality in the foreseeable future.to prevent spreading of diseases, an early detection of infection is important. at the time an outbreak is diagnosed, many people may already be infected due to the ... | 2005 | 16160262 |
| decontamination assessment of bacillus anthracis, bacillus subtilis, and geobacillus stearothermophilus spores on indoor surfaces using a hydrogen peroxide gas generator. | to evaluate the decontamination of bacillus anthracis, bacillus subtilis, and geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. | 2005 | 16162224 |
| vaccines and the next pandemic. | | 2005 | 16162593 |
| t-cell response profiling to biological threat agents including the sars coronavirus. | the emergence of pathogens such as sars and the increased threat of bioterrorism has stimulated the development of novel diagnostic assays for differential diagnosis. rather than focusing on the detection of an individual pathogen component, we have developed a t cell profiling system to monitor responses to the pathogens in an array format. using a matrix of antigens specific for different pathogens, a specific t cell profile was generated for each individual by monitoring the intracellular pro ... | 2005 | 16164833 |
| identification of the bacillus anthracis (gamma) phage receptor. | bacillus anthracis, a gram-positive, spore-forming bacterium, is the etiological agent of anthrax. it belongs to the bacillus cereus group, which also contains bacillus cereus and bacillus thuringiensis. most b. anthracis strains are sensitive to phage gamma, but most b. cereus and b. thuringiensis strains are resistant to the lytic action of phage gamma. here, we report the identification of a protein involved in the bacterial receptor for the gamma phage, which we term gamr (gamma phage recept ... | 2005 | 16166537 |
| a comparative study of bacillus cereus, bacillus thuringiensis and bacillus anthracis extracellular proteomes. | bacillus cereus, bacillus thuringiensis and bacillus anthracis are closely related species that share a similar genetic background but occupy different ecological niches. virulence plasmids bearing genes coding for toxins, may explain, at least partly, this specialization. we have compared by 2-de in the early stationary phase of growth the extracellular proteomes of three strains of these species that have lost their virulence plasmids. proteins expected to be secreted or to belong to the cell ... | 2005 | 16167365 |
| pulmonary disease from biological agents: anthrax, plague, q fever, and tularemia. | anthrax, plague, q fever, and tularemia are all potential inhalational bioterrorism agents. the pulmonary manifestations of these agents can be readily confused with each other as well as other more common diseases such as influenza and atypical pneumonia. this article reviews the threat potential, microbiology, pathogenesis, clinical features, diagnosis, and treatment of each of these agents and highlights the similarities and differences between their pulmonary presentations. | 2005 | 16168313 |
| evidence for adjuvanticity of anthrax edema toxin. | bacillus anthracis edema factor (ef) is an adenylate cyclase that increases intracellular camp concentrations. since ef is present as a contaminant in the licensed protective antigen(pa)-based vaccines, we investigated its effect on anti-pa humoral immune response in balb/c mice. we observed a significant increase of anti-pa igg response in mice immunised with pa in association with ef as compared to pa alone. these results clearly show an adjuvant effect of ef, which is consistent with the data ... | 2006 | 16168529 |
| signaling pathways and genes that inhibit pathogen-induced macrophage apoptosis--creb and nf-kappab as key regulators. | certain microbes evade host innate immunity by killing activated macrophages with the help of virulence factors that target prosurvival pathways. for instance, infection of macrophages with the tlr4-activating bacterium bacillus anthracis triggers an apoptotic response due to inhibition of p38 map kinase activation by the bacterial-produced lethal toxin. other pathogens induce macrophage apoptosis by preventing activation of nf-kappab, which depends on ikappab kinase beta (ikkbeta). to better un ... | 2005 | 16169504 |
| piezoelectric-excited millimeter-sized cantilever (pemc) sensors detect bacillus anthracis at 300 spores/ml. | piezoelectric-excited millimeter-sized cantilever (pemc) sensors consisting of a piezoelectric and a borosilicate glass layer with a sensing area of 2.48 mm2 were fabricated. antibody specific to bacillus anthracis (ba, sterne strain 7702) spores was immobilized on pemc sensors, and exposed to spores (300 to 3x10(6) spores/ml). the resonant frequency decreased at a rate proportional to the spore concentration and reached a steady state frequency change of 5+/-5 hz (n=3), 92+/-7 hz (n=3), 500+/-1 ... | 2006 | 16169715 |
| beta-cyclodextrin derivatives that inhibit anthrax lethal toxin. | recently, we demonstrated that simultaneous blocking of bacterial growth by antibiotics and inhibition of anthrax toxin action with antibodies against protective antigen were beneficial for the treatment of anthrax. the present study examined the hypothesis that blocking the pore formed by protective antigen can inhibit the action of anthrax toxin. the potential inhibitors were chosen by a structure-based design using beta-cyclodextrin as the starting molecule. several beta-cyclodextrin derivati ... | 2006 | 16169738 |
| braf status and mitogen-activated protein/extracellular signal-regulated kinase kinase 1/2 activity indicate sensitivity of melanoma cells to anthrax lethal toxin. | anthrax lethal toxin, composed of protective antigen and lethal factor, was tested for cytotoxicity to human melanoma cell lines and normal human cells. eleven of 18 melanoma cell lines were sensitive to anthrax lethal toxin (ic(50) < 400 pmol/l) and 10 of these 11 sensitive cell lines carried the v599e braf mutation. most normal cell types (10 of 15) were not sensitive to anthrax lethal toxin and only 5 of 15 normal human cell types were sensitive to anthrax lethal toxin (ic(50) < 400 pmol/l). ... | 2005 | 16170021 |
| antiplatelet activities of anthrax lethal toxin are associated with suppressed p42/44 and p38 mitogen-activated protein kinase pathways in the platelets. | anthrax lethal toxin (lt) is the major virulence factor produced by bacillus anthracis, but the mechanism by which it induces high mortality remains unclear. we found that lt treatment could induce severe hemorrhage in mice and significantly suppress human whole-blood clotting and platelet aggregation in vitro. in addition, lt could inhibit agonist-induced platelet surface p-selectin expression, resulting in the inhibition of platelet-endothelial cell engagements. data from western blot analysis ... | 2005 | 16170766 |
| total synthesis of antigen bacillus anthracis tetrasaccharide--creation of an anthrax vaccine candidate. | | 2005 | 16172990 |
| use of in situ-generated dimethyldioxirane for inactivation of biological agents. | dimethyldioxirane (dmdo), generated in situ by adding acetone to an aqueous solution containing potassium peroxymonosulfate (oxone) at neutral ph, was investigated for inactivation of biological warfare agent simulants. the dmdo solution inactivated bacterial spores, fungal spores, vegetative bacterial cells, viruses, and protein by 7 orders of magnitude in less than 10 min. the kill rates of dmdo were more pronounced when compared to kill rates of buffered oxone alone. conditions for the use of ... | 2005 | 16173594 |
| an anthrax lethal factor-neutralizing monoclonal antibody protects rats before and after challenge with anthrax toxin. | lethal factor (lf) is a component of anthrax lethal toxin (letx). we generated anti-lf murine monoclonal antibodies (mabs) that show letx-neutralizing activity in vitro and in vivo. anti-lf mabs were generated by immunization with recombinant lf, and the mabs showing letx-neutralizing activity in vitro were selected. two mabs with the highest affinities, 5b13b1 (dissociation constant [k(d)], 2.62 nm) and 3c16c3 (k(d), 8.18 nm), were shown to recognize the same or closely overlapping epitopes on ... | 2005 | 16177329 |
| characterization of listeria monocytogenes expressing anthrolysin o and phosphatidylinositol-specific phospholipase c from bacillus anthracis. | two virulence factors of listeria monocytogenes, listeriolysin o (llo) and phosphatidylinositol-specific phospholipase c (pi-plc), mediate escape of this pathogen from the phagocytic vacuole of macrophages, thereby allowing the bacterium access to the host cell cytosol for growth and spread to neighboring cells. we characterized their orthologs from bacillus anthracis by expressing them in l. monocytogenes and characterizing their contribution to bacterial intracellular growth and cell-to-cell s ... | 2005 | 16177340 |
| induction of protective immunity to anthrax lethal toxin with a nonhuman primate adenovirus-based vaccine in the presence of preexisting anti-human adenovirus immunity. | prevention or therapy for bioterrorism-associated anthrax infections requires rapidly acting effective vaccines. we recently demonstrated (y. tan, n. r. hackett, j. l. boyer, and r. g. crystal, hum. gene ther. 14:1673-1682, 2003) that a single administration of a recombinant serotype 5 adenovirus (ad) vector expressing anthrax protective antigen (pa) provides rapid protection against anthrax lethal toxin challenge. however, approximately 35 to 50% of humans have preexisting neutralizing antibodi ... | 2005 | 16177368 |
| pathophysiological manifestations in mice exposed to anthrax lethal toxin. | pathophysiological changes associated with anthrax lethal toxin included loss of plasma proteins, decreased platelet count, slower clotting times, fibrin deposits in tissue sections, and gross and histopathological evidence of hemorrhage. these findings suggest that blood vessel leakage and hemorrhage lead to disseminating intravascular coagulation and/or circulatory shock as an underlying pathophysiological mechanism. | 2005 | 16177381 |
| bacillus anthracis edema toxin inhibits staphylococcus aureus enterotoxin b effects in vitro: a potential protein therapeutic? | various in vitro effects of staphylococcal enterotoxin b (seb) on human peripheral blood mononuclear cells were mitigated by bacillus anthracis edema toxin. in particular, levels of some seb-induced cytokines (tumor necrosis factor alpha, gamma interferon) and chemokines (monocyte chemoattractant protein 1, macrophage inflammatory protein 1 alpha [mip-1alpha], mip-1beta) were significantly diminished or even nonexistent, depending upon the timing of edema toxin administration. overall, these res ... | 2005 | 16177395 |
| simulation for assessing statistical methods of biologic terrorism surveillance. | multiple systems have been developed that use surveillance of health-care encounters to provide early warning of a terrorist attack. limited practical experience and the absence of adequate theoretical assessments have precluded determining which alarm-generating algorithms should be preferred. in the absence of practical and theoretical results, choosing a particular statistical algorithm can be difficult. one way to evaluate algorithms is through simulation. | 2005 | 16177700 |
| an evaluation model for syndromic surveillance: assessing the performance of a temporal algorithm. | syndromic surveillance offers the potential to rapidly detect outbreaks resulting from terrorism. despite considerable experience with implementing syndromic surveillance, limited evidence exists to describe the performance of syndromic surveillance systems in detecting outbreaks. | 2005 | 16177701 |
| emergency department visits for concern regarding anthrax--new jersey, 2001. | in october of 2001, after letters processed in trenton, new jersey, resulted in multiple cases of anthrax, emergency departments (eds) in new jersey experienced an increase in visits from patients concerned about possible exposure to agents of biologic terrorism. information about the effect of an actual biologic terrorism attack on the emergency department population might be useful in the design of biosurveillance systems, particularly with regard to their performance during the mitigation pha ... | 2005 | 16177709 |
| distribution of productive antigen-processing activity for mhc class ii presentation in macrophages. | we demonstrated that an epitope from the recombinant protective antigen (rpa) of bacillus anthracis was presented by mature major histocompatibility complex class ii (mhc-ii) molecules, whereas an epitope from the recombinant virulent (rv) antigen of yersinia pestis was presented by newly synthesized mhc-ii. we addressed which endosomal compartments were involved in the antigen processing of each epitope. bone-marrow-derived macrophages were subjected to subcellular fractionation; fractions were ... | 2005 | 16179011 |
| postal workers' perspectives on communication during the anthrax attack. | in 2001, the nation experienced its first bioterrorism attack, in the form of anthrax sent through the u.s. postal service, and public health professionals were challenged to communicate with a critical audience, u.s. postal workers. postal workers, the first cohort to receive public health messages during a bioterrorist crisis, offer a crucial viewpoint that can be used in the development of best practices in crisis and emergency risk communication. this article reports results of qualitative i ... | 2005 | 16181043 |
| whole-cell voltage clamp measurements of anthrax toxin pore current. | protective antigen (pa) of anthrax toxin binds cellular receptors and forms pores in target cell membranes, through which catalytic lethal factor (lf) and edema factor (ef) are believed to translocate to the cytoplasm. using patch clamp electrophysiological techniques, we assayed pore formation by pa in real time on the surface of cultured cells. the membranes of cho-k1 cells treated with activated pa had little to no electrical conductivity at neutral ph (7.3) but exhibited robust mixed ionic c ... | 2005 | 16183642 |
| ability of physicians to diagnose and manage illness due to category a bioterrorism agents. | early recognition of a terrorist attack with biologic agents will rely on physician diagnosis. physicians' ability to diagnose and care for patients presenting after a bioterror event is unknown. the role of online case-based didactics to measure and improve knowledge in the diagnosis and treatment of these patients is unknown. | 2005 | 16186470 |
| effects of spontaneous deamidation on the cytotoxic activity of the bacillus anthracis protective antigen. | protective antigen (pa) is a central virulence factor of bacillus anthracis and a key component in anthrax vaccines. pa binds to target cell receptors, is cleaved by the furin protease, self-aggregates to heptamers, and finally internalizes as a complex with either lethal or edema factors. under mild room temperature storage conditions, pa cytotoxicity decreased (t(1/2) approximately 7 days) concomitant with the generation of new acidic isoforms, probably through deamidation of asn residues. ran ... | 2005 | 16188881 |
| the solute-binding component of a putative mn(ii) abc transporter (mnta) is a novel bacillus anthracis virulence determinant. | here we describe the characterization of a lipoprotein previously proposed as a potential bacillus anthracis virulence determinant and vaccine candidate. this protein, designated mnta, is the solute-binding component of a manganese ion atp-binding cassette transporter. coupled proteomic-serological screen of a fully virulent wild-type b. anthracis vollum strain, confirmed that mnta is expressed both in vitro and during infection. expression of mnta is shown to be independent of the virulence pla ... | 2005 | 16194238 |
| cereulide-producing strains of bacillus cereus show diversity. | producers of cereulide, the emetic toxin of bacillus cereus, are known to constitute a specific subset within this species. we investigated physiological and genetic properties of 24 strains of b. cereus including two high cereulide producers (600-1,800 ng cereulide mg(-1) wet weight biomass), seven average producers (180-600 ng cereulide mg(-1) wet weight biomass), four low cereulide producers (20-160 ng cereulide mg(-1) wet weight biomass) and 11 non-producers representing isolates from food, ... | 2005 | 16195898 |
| functional characterization of peptide-based anthrax toxin inhibitors. | we have identified an optimized peptide inhibitor that can be used to develop potent anthrax toxin therapeutics. anthrax toxin, an essential virulence factor of bacillus anthracis, elicits many of the symptoms associated with the disease, and is responsible for death. the toxin is composed of a cell-binding component, protective antigen, and two enzymatic components, edema factor and lethal factor. the three proteins are secreted individually by the bacterium and then assemble into functional co ... | 2005 | 16196489 |
| an unusually extensive case of cutaneous anthrax in a patient with type ii diabetes mellitus. | naturally acquired anthrax infection remains an important public-health problem in developing countries. turkey is one of the countries in which the zoonotic form of anthrax may still be encountered. the most frequent portal of entry for anthrax spores is the skin. although cutaneous anthrax is usually self-limiting, complications may arise in untreated cases. underlying systemic disorders such as diabetes mellitus may confound the clinical picture and lead to atypical presentations. we present ... | 2005 | 16197380 |
| determination of the most closely related bacillus isolates to bacillus anthracis by multilocus sequence typing. | there have been many efforts to develop bacillus anthracis detection assays, but the problem of false-positive results has often been encountered. therefore, to validate an assay for b. anthracis detection, it is critical to examine its specificity with the most closely related bacillus isolates that are available. to define the most closely related bacillus isolates to b. anthracis in our bacillus collections, we analyzed by multilocus sequence typing (mlst) the phylogeny of 77 closely related ... | 2005 | 16197725 |
| comparison of the immunological memory after dna vaccination and protein vaccination against anthrax in sheep. | currently available live spore vaccines against anthrax in animals have many drawbacks, one of which is their presumed inability to induce a long lasting immunity. in the present study we compared the immunological memory after a protein vaccination with dna vaccinations in sheep. the antigen used was the protective antigen (pa83) of bacillus anthracis. sheep were vaccinated three times with either pa83 plus alhydrogel, or with one of four different plasmid dna formulations, which all encoded ei ... | 2006 | 16198452 |
| characterization of sporulation histidine kinases of bacillus anthracis. | the initiation of sporulation in bacillus species is regulated by the phosphorelay signal transduction pathway, which is activated by several histidine sensor kinases in response to cellular and metabolic signals. comparison of the protein components of the phosphorelay between bacillus subtilis and bacillus anthracis revealed high homology in the phosphorelay orthologs of spo0f, spo0b, and spo0a. the sensor domains of sensor histidine kinases are poorly conserved between species, making ortholo ... | 2005 | 16199567 |
| anthrax: a continuing concern in the era of bioterrorism. | anthrax, a potentially fatal infection, is a virulent and highly contagious disease. it is caused by a gram-positive, toxigenic, spore-forming bacillus: bacillus anthracis. for centuries, anthrax has caused disease in animals and, although uncommonly, in humans throughout the world. descriptions of this naturally occurring disease begin in antiquity. anthrax is primarily a disease of herbivores, which are infected by ingestion of spores from the soil. with the advent of modern microbiology, past ... | 2005 | 16200179 |
| pakistan's experience of a bioterrorism-related anthrax scare. | from november 2001 to march 2002, the national institute of health, islamabad, pakistan, received 230 samples from 194 different sources for analysis for anthrax spores. these samples were taken from letters/packages suspected of containing anthrax and from individuals exposed to them. when cultured on sheep blood agar, 141 samples yielded growth suggestive of bacillus species. on the basis of growth characteristics, absence of beta-haemolysis, absent or doubtful motility and morphological chara ... | 2004 | 16201705 |
| anthrax: pathological aspects in autopsy cases in shiraz, islamic republic of iran, 1960-2001. | anthrax, like tuberculosis, shows a new epidemic spread in industrialized countries, revealing some ambiguous aspects to the disease and providing new challenges to medicine. shiraz university of medical sciences has records of 7130 autopsies performed in the past 40 years, 33 of which are anthrax cases. we reviewed all the pathology slides of these cases and classified the organs involved in a search for unrecognized microscopic findings. the most common cause of death was sepsis, caused by org ... | 2004 | 16201706 |
| clinical decision making during public health emergencies: ethical considerations. | recent public health emergencies involving anthrax, the severe acute respiratory syndrome (sars), and shortages of influenza vaccine have dramatized the need for restrictive public health measures such as quarantine, isolation, and rationing. front-line physicians will face ethical dilemmas during public health emergencies when patients disagree with these measures. patients might request interventions that are not recommended or for which they are not eligible, or they might object to intrusive ... | 2005 | 16204162 |
| recognizing biothreat diseases: realistic training using standardized patients and patient simulators. | the uofl center for the deterrence of biowarfare and bioterrorism's (cdbb's) training involving standardized patients, who can make convincingly accurate representations of clinical signs and symptoms, and patient simulators has proven to be an effective means of enhancing bioterrorism preparedness. in addition to providing ready access to formative measures of preparedness, both of these teaching and learning tools also offer the option of summative evaluation of skills and knowledge acquired d ... | 2005 | 16205536 |
| treatment of acute myeloid leukemia during the second and third trimesters of pregnancy. | fortunately, the occurrence of acute myeloid leukemia (aml) during pregnancy is rare. we report a case of successful fetal outcome with standard induction and consolidation treatment in the second and third trimesters, respectively. a 37-year-old woman in her second trimester (21 wks) of pregnancy was found to have acute myeloid leukemia. she elected to maintain the pregnancy and underwent induction with cytarabine and idarubicin. her hospital course was complicated by pseudomonas vesicularis an ... | 2005 | 16207105 |
| when bioterrorism strikes: communication issues for the local health department. | public health preparedness is a multifaceted planning process that becomes grounded in a response plan and in effective communications, internal and external. this article describes an incident when the presence of anthrax spores was detected in a postal facility within kansas city, missouri, and discusses the communications issues faced by the kansas city health department (kchd). this incident provided the kchd the first opportunity to operationalize its incident management system-based respon ... | 2005 | 16210684 |
| anthrax in new jersey: a health education experience in bioterrorism response and preparedness. | the anthrax attack in 2001 created new challenges to health educators working on the response effort in new jersey. never before had there been a need for educating a group of people who had been exposed to a biological weapon. coming on the heels of the catastrophic world trade center collapse on september 11, 2001, the new jersey department of health and senior services was entrenched in the response to, and management of, the anthrax attack that placed a heavy emphasis on educating the postal ... | 2005 | 16210685 |
| weapons of mass destruction: biological. | humans are susceptible to microbial infections from many sources. biological warfare is the use of microbial forms of life to diminish the capabilities, disrupt the organization, and terrorize the noncombatant population of an adversary. this form of warfare has been used throughout history and has gained renewed interest with the current use of asymmetrical warfare. the civilized world has condemned its use by the implementation of treaties specifically against it. this is a brief review of som ... | 2002 | 16212312 |
| blocking anthrax lethal toxin at the protective antigen channel by using structure-inspired drug design. | bacillus anthracis secretes three polypeptides: protective antigen (pa), lethal factor (lf), and edema factor (ef), which interact at the surface of mammalian cells to form toxic complexes. lf and ef are enzymes that target substrates within the cytosol; pa provides a heptameric pore to facilitate lf and ef transport into the cytosol. other than administration of antibiotics shortly after exposure, there is currently no approved effective treatment for inhalational anthrax. here we demonstrate a ... | 2005 | 16214885 |
| synthesis of the beta anomer of the spacer-equipped tetrasaccharide side chain of the major glycoprotein of the bacillus anthracis exosporium. | the beta glycoside of the tetrasaccharide sequence beta-ant-(1-->3)-alpha-l-rhap-(1-->3)-alpha-l-rhap-(1-->2)-l-rhap, whose aglycon allows conjugation to proteins, was synthesized for the first time. a stepwise synthetic approach was applied with thioglycosides as glycosyl donors, and the beta anomer of the compound was obtained equipped with a spacer group whose further transformation allows conjugation to suitable carriers. to synthesize the beta-anthrosyl linkage with high stereoselectivity, ... | 2005 | 16216230 |
| characterization of a complex genomic alteration on chromosome 2p that leads to four alternatively spliced fusion transcripts in the neuroblastoma cell lines imr-5, imr-5/75 and imr-32. | genetic aberrations in neuroblastoma (nb) have been extensively characterized over the last years. alterations of the short arm of chromosome 2 (2p) have been of particular interest, since amplification of the mycn oncogene on 2p24 is associated with an adverse outcome in nb patients. here, we report on the characterization of a novel genomic rearrangement involving genetic material from 2p13 and 2p24 in nb cell lines that was discovered based on a serial analysis of gene expression (sage) profi ... | 2005 | 16216448 |
| ii. cationic lipid-formulated plasmid dna-based bacillus anthracis vaccine: evaluation of plasmid dna persistence and integration potential. | several formulated plasmid dna (pdna)-based vaccines are being evaluated for safety and efficacy in healthy human subjects. a safety concern for any vaccine that contains genetic material, be it whole organism, live-attenuated, or gene-based, is the potential for integration into genomic dna (gdna). to address this concern, a preclinical pdna persistence/integration study was conducted in rabbits to determine the level of pdna in muscle 2, 28, and 64 days after intramuscular injection of dmrie:d ... | 2005 | 16218776 |
| cerebral diffusional changes in the early phase of anthrax: is cutaneous anthrax only limited to skin? | characteristics of cerebral diffusion in the acute period of the anthrax infection were investigated to understand the pathophysiology of the disease. | 2006 | 16219360 |
| bioterrorism and mass casualty preparedness in hospitals: united states, 2003. | this study examined the content of hospital terrorism preparedness emergency response plans; whether those plans had been updated since september 11, 2001; collaboration of hospitals with outside organizations; clinician training in the management of biological, chemical, explosive, and nuclear exposures; drills on the response plans; and equipment and bed capacity. | 2005 | 16220875 |
| [the explicative power of the rules: the case of the koch's postulates]. | this article traces the emergence of the standard for evidence of causation in infectious diseases, from the scientific discoveries on contagion by agostino bassi, in the first half of the nineteenth century. however, only after the work of robert koch on anthrax and tuberculosis, a set of practical guidelines (koch's postulates) were acknowledged, to establish rigorous criteria before a microbiologist could claim a strictly causal relationship between microbes and infectious diseases. the metho ... | 2004 | 16222849 |
| identification of marker proteins for bacillus anthracis using maldi-tof ms and ion trap ms/ms after direct extraction or electrophoretic separation. | direct extraction of bacterial vegetative cells or spores followed by matrix-assisted laser desorption ionization/time of flight mass spectrometry (maldi tof ms) has become popular for bacterial identification, since it is simple to perform and mass spectra are readily interpreted. however, only high-abundance proteins that are of low mass and ionize readily are observed. in the case of b. anthracis spores, small acid-soluble spore proteins (sasps) have been the most widely studied. additional i ... | 2005 | 16224957 |
| genome sequence and gene expression of bacillus anthracis bacteriophage fah. | fah, a lytic bacteriophage of bacillus anthracis, is used widely in the former soviet union to identify anthrax bacteria. here, we present the analysis of a 37,974 bp sequence of the fah genome and examine gene expression of the phage in a model host, bacillus cereus. half of the fah genome contains genes coding for structural proteins and host lysis functions in an arrangement typical of syphoviridae. the other half of the genome contains genes coding for enzymes of viral genome replication and ... | 2005 | 16226766 |
| bio-terrorism, human security and public health: can international law bring them together in an age of globalization? | bio-terrorism, the use of a microorganism with the deliberate intent of causing infection, before and since the anthrax attacks in the united states in october 2001, has emerged as a real medical and public health threat. the link between bio-terrorism, human security and public health raises complex questions on the normative trajectories of international law, the mandates of international organizations, and global health governance. in may 2001, the world health assembly of the world health or ... | 2005 | 16229381 |
| biocidal activity of a light-absorbing fluorescent conjugated polyelectrolyte. | herein we describe studies that indicate a cationic conjugated polyelectrolyte shows biocidal activity against gram-negative bacteria (escherichia coli, e. coli, bl21, with plasmids for azurin and ampicillin resistance) and gram-positive bacterial spores (bacillus anthracis, sterne, b. anthracis, sterne). these studies were carried out with aqueous suspensions of the conjugated polyelectrolyte, with the polyelectrolyte in supported formats and with samples in which the conjugated polyelectrolyte ... | 2005 | 16229539 |
| simulated anthrax attacks and syndromic surveillance. | we measured sensitivity and timeliness of a syndromic surveillance system to detect bioterrorism events. a hypothetical anthrax release was modeled by using zip code population data, mall customer surveys, and membership information from healthpartners medical group, which covers 9% of a metropolitan area population in minnesota. for each infection level, 1,000 releases were simulated. timing of increases in use of medical care was based on data from the sverdlovsk, russia, anthrax release. case ... | 2005 | 16229768 |
| gram-positive rod surveillance for early anthrax detection. | connecticut established telephone-based gram-positive rod (gpr) reporting primarily to detect inhalational anthrax cases more quickly. from march to december 2003, annualized incidence of blood isolates was 21.3/100,000 persons; reports included 293 corynebacterium spp., 193 bacillus spp., 73 clostridium spp., 26 lactobacillus spp., and 49 other genera. around-the-clock gpr reporting has described gpr epidemiology and enhanced rapid communication with clinical laboratories. | 2005 | 16229790 |
| a conserved motif in transmembrane helix 1 of diphtheria toxin mediates catalytic domain delivery to the cytosol. | a 10-aa motif in transmembrane helix 1 of diphtheria toxin that is conserved in anthrax edema factor, anthrax lethal factor, and botulinum neurotoxin serotypes a, c, and d was identified by blast, clustal w, and meme computational analysis. using the diphtheria toxin-related fusion protein toxin dab(389)il-2, we demonstrate that introduction of the l221e mutation into a highly conserved residue within this motif results in a nontoxic catalytic domain translocation deficient phenotype. to further ... | 2005 | 16230620 |
| [syndromic surveillance for the detection of outbreaks of unusual infectious diseases]. | syndromic surveillance has been developed in order to detect outbreaks of unusual infectious diseases such as severe acute respiratory syndrome (sars) or anthrax at an early stage. whereas the usual surveillance systems are based on established diagnoses and emergency department discharge data, syndromic surveillance uses preliminary outcomes and derived data such as absenteeism, prescription medication and requests for laboratory tests. investigations abroad have indicated the potential ofsyndr ... | 2005 | 16235804 |
| advances in alfalfa mosaic virus-mediated expression of anthrax antigen in planta. | plant viruses show great potential for production of pharmaceuticals in plants. such viruses can harbor a small antigenic peptide(s) as a part of their coat proteins (cp) and elicit an antigen-specific immune response. here, we report the high yield and consistency in production of recombinant alfalfa mosaic virus (almv) particles for specific presentation of the small loop 15 amino acid epitope from domain-4 of the bacillus anthracis protective antigen (pa-d4s). the epitope was inserted immedia ... | 2005 | 16236249 |
| proximity ligation assays with peptide conjugate 'burrs' for the sensitive detection of spores. | the proximity ligation assay (pla) has previously been used for the sensitive and specific detection of single proteins. in order to adapt pla methods for the detection of cell surfaces, we have generated multivalent peptide-oligonucleotide-phycoerythrin conjugates ('burrs') that can bind adjacent to one another on a cell surface and be ligated together to form unique amplicons. real-time pcr detection of burr ligation events specifically identified as few as 100 bacillus anthracis, 10 bacillus ... | 2005 | 16237122 |
| a multiplex polymerase chain reaction microarray assay to detect bioterror pathogens in blood. | heightened concern about the dangers of bioterrorism requires that measures be developed to ensure the safety of the blood supply. multiplex detection of such agents using a blood-screening dna microarray is a sensitive and specific method to screen simultaneously for a number of suspected agents. we have developed and optimized a multiplex polymerase chain reaction microarray assay to screen blood for three potential bioterror bacterial pathogens and a human ribosomal rna gene internal control. ... | 2005 | 16237218 |
| clindamycin and quinolone therapy for bacillus anthracis sterne infection in 60co-gamma-photon-irradiated and sham-irradiated mice. | sublethal ionizing doses of radiation increase the susceptibility of mice to bacillus anthracis sterne infection. in this study, we investigated the efficacy of clindamycin in 60co-gamma-photon-irradiated and sham-irradiated mice after intratracheal challenge with b. anthracis sterne spores. clindamycin has in vitro activity against b. anthracis and inhibits the production of toxin from other species, although no direct evidence exists that production of b. anthracis toxin is inhibited. | 2005 | 16239289 |
| murine macrophages kill the vegetative form of bacillus anthracis. | anti-protective antigen antibody was reported to enhance macrophage killing of ingested bacillus anthracis spores, but it was unclear whether the antibody-mediated macrophage killing mechanism was directed against the spore itself or the vegetative form emerging from the ingested and germinating spore. to address this question, we compared the killing of germination-proficient (gp) and germination-deficient (deltagerh) sterne 34f2 strain spores by murine peritoneal macrophages. while macrophages ... | 2005 | 16239551 |
| myd88-dependent signaling contributes to protection following bacillus anthracis spore challenge of mice: implications for toll-like receptor signaling. | bacillus anthracis is a spore-forming, gram-positive organism that is the causative agent of the disease anthrax. recognition of bacillus anthracis by the host innate immune system likely plays a key protective role following infection. in the present study, we examined the role of tlr2, tlr4, and myd88 in the response to b. anthracis. heat-killed bacillus anthracis stimulated tlr2, but not tlr4, signaling in hek293 cells and stimulated tumor necrosis factor alpha (tnf-alpha) production in c3h/h ... | 2005 | 16239556 |
| metalloproteinase inhibitors, nonantimicrobial chemically modified tetracyclines, and ilomastat block bacillus anthracis lethal factor activity in viable cells. | lethal toxin, produced by the bacterium bacillus anthracis, is a major contributor to morbidity and mortality in animals and humans who have contracted anthrax. one component of this toxin, lethal factor (lf), proteolytically inactivates members of the mitogen-activated protein kinase kinase (mapkk or mek) family. in this study we show that cmt-300, cmt-308, and ilomastat, agents initially characterized as matrix metalloproteinase inhibitors which are in early stages of development as pharmaceut ... | 2005 | 16239558 |
| anthrax lethal toxin enhances cytokine-induced vcam-1 expression on human endothelial cells. | vascular endothelial dysfunction is thought to play a prominent role in systemic anthrax pathogenesis. we examined the effect of anthrax lethal toxin (ltx), a key virulence factor of bacillus anthracis, on the expression of vascular cell adhesion molecule-1 (vcam-1) on normal and cytokine-stimulated human lung microvascular endothelial cells. confluent endothelial monolayers were treated with lethal factor (lf), protective antigen (pa), or both (ltx) in the presence or absence of tumor necrosis ... | 2005 | 16242117 |
| the crystal structure of the bacillus anthracis spore surface protein bcla shows remarkable similarity to mammalian proteins. | the lethal disease anthrax is propagated by spores of bacillus anthracis, which can penetrate into the mammalian host by inhalation, causing a rapid progression of the disease and a mostly fatal outcome. we have solved the three-dimensional structure of the major surface protein bcla on b. anthracis spores. surprisingly, the structure resembles c1q, the first component of complement, despite there being no sequence homology. although most assays for c1q-like activity, including binding to c1q re ... | 2005 | 16249180 |
| a model of anthrax toxin lethal factor bound to protective antigen. | anthrax toxin is made up of three proteins: the edema factor (ef), lethal factor (lf) enzymes, and the multifunctional protective antigen (pa). proteolytically activated pa heptamerizes, binds the ef/lf enzymes, and forms a pore that allows for ef/lf passage into host cells. using directed mutagenesis, we identified three lf-pa contact points defined by a specific disulfide crosslink and two pairs of complementary charge-reversal mutations. these contact points were consistent with the lowest en ... | 2005 | 16251269 |
| bacillus anthracis edema toxin causes extensive tissue lesions and rapid lethality in mice. | bacillus anthracis edema toxin (et), an adenylyl cyclase, is an important virulence factor that contributes to anthrax disease. the role of et in anthrax pathogenesis is, however, poorly understood. previous studies using crude toxin preparations associated et with subcutaneous edema, and et-deficient strains of b. anthracis showed a reduction in virulence. we report the first comprehensive study of et-induced pathology in an animal model. highly purified et caused death in balb/cj mice at lower ... | 2005 | 16251415 |
| principal-components analysis of fluorescence cross-section spectra from pathogenic and simulant bacteria. | principal-components analysis of a new set of highly resolved (< 1 nm) fluorescence cross-section spectra excited at 354.7 nm over the 370-646 nm band has been used to demonstrate the potential ability of uv standoff lidars to discriminate among particular biological warfare agents and simulants over short ranges. the remapped spectra produced by this technique from bacillus globigii (bg) and bacillus anthracis (ba) spores were sufficiently different to allow them to be cleanly separated, and th ... | 2005 | 16252661 |
| anthrax lethal toxin-mediated killing of human and murine dendritic cells impairs the adaptive immune response. | many pathogens have acquired strategies to combat the immune response. bacillus anthracis interferes with host defenses by releasing anthrax lethal toxin (lt), which inactivates mitogen-activated protein kinase pathways, rendering dendritic cells (dcs) and t lymphocytes nonresponsive to immune stimulation. however, these cell types are considered resistant to killing by lt. here we show that lt kills primary human dcs in vitro, and murine dcs in vitro and in vivo. kinetics of lt-mediated killing ... | 2005 | 16254597 |
| bacteriophage therapy: a revitalized therapy against bacterial infectious diseases. | bacteriophage (phage) therapy involves using phages or their products as bioagents for the treatment or prophylaxis of bacterial infectious diseases. much evidence in support of the effectiveness of phage therapy against bacterial infectious diseases has accumulated since 1980 from animal model studies conducted in western countries. reports indicate that appropriate administration of living phages can be used to treat lethal infectious diseases caused by gram-negative bacteria, such as escheric ... | 2005 | 16258815 |
| bioterrorism and its aftermath: dealing individually and organizationally with the emotional reactions to an anthrax attack. | from september 2001 through april 2004, the united states postal service (usps) dealt, for the first time, with bioterrorism resulting in employee deaths and the closure of a large mail processing plant in washington, d.c. the employee assistance program (eap) partnered with the usps throughout this tumultuous time to meet the multiple and evolving behavioral health needs of the employees and facilitate the employees' emotional preparedness for their return to work at the closed facility. this p ... | 2005 | 16265976 |
| genetic analysis of transfer-related regions of the vancomycin resistance enterococcus conjugative plasmid phtbeta: identification of orit and a putative relaxase gene. | the pht plasmids phtalpha (65.9 kbp), phtbeta (63.7 kbp), and phtgamma (66.5 kbp) are highly conjugative pheromone-independent pmg1-like plasmids that carry tn1546-like transposons encoding vancomycin resistance. phtbeta is the prototype plasmid, and the phtalpha and phtgamma plasmids are derivatives of the insertion into phtbeta of an is232-like (2.2 kbp) element and a group ii intron (2.8 kbp), respectively. the complete nucleotide sequence of the phtbeta plasmid was determined and, with the e ... | 2005 | 16267297 |
| cape, a 47-amino-acid peptide, is necessary for bacillus anthracis polyglutamate capsule synthesis. | polyglutamate is found in various bacteria, but displays different functions depending on the species and their environment. here, we describe a minimal polyglutamate synthesis system in bacillus anthracis. in addition to the three genes previously described as sufficient for polyglutamate synthesis, this system includes a small open reading frame, cape, belonging to the cap operon. the polyglutamate system's requirement for the five cap genes, for capsulation and anchoring, was assayed in nonpo ... | 2005 | 16267300 |
| comparison of minisatellite polymorphisms in the bacillus cereus complex: a simple assay for large-scale screening and identification of strains most closely related to bacillus anthracis. | polymorphism of five tandem repeats that are monomorphic in bacillus anthracis was investigated in 230 isolates of the b. cereus group and in 5 sequenced b. cereus genomes in search for markers allowing identification of b. cereus and b. thuringiensis strains most closely related to b. anthracis. using this multiple-locus variable number of tandem repeat analysis (mlva), a cluster of 30 strains was selected for further characterization. eventually, six of these were characterized by multilocus s ... | 2005 | 16269689 |
| photodynamic inactivation of bacillus spores, mediated by phenothiazinium dyes. | spore formation is a sophisticated mechanism by which some bacteria survive conditions of stress and starvation by producing a multilayered protective capsule enclosing their condensed dna. spores are highly resistant to damage by heat, radiation, and commonly employed antibacterial agents. previously, spores have also been shown to be resistant to photodynamic inactivation using dyes and light that easily destroy the corresponding vegetative bacteria. we have discovered that bacillus spores are ... | 2005 | 16269726 |
| inactivation of bacillus endospores in envelopes by electron beam irradiation. | the anthrax incidents in the united states in the fall of 2001 led to the use of electron beam (eb) processing to sanitize the mail for the u.s. postal service. this method of sanitization has prompted the need to further investigate the effect of eb irradiation on the destruction of bacillus endospores. in this study, endospores of an anthrax surrogate, b. atrophaeus, were destroyed to demonstrate the efficacy of eb treatment of such biohazard spores. eb exposures were performed to determine (i ... | 2005 | 16269738 |
| a recombinant 63-kda form of bacillus anthracis protective antigen produced in the yeast saccharomyces cerevisiae provides protection in rabbit and primate inhalational challenge models of anthrax infection. | infection by bacillus anthracis is preventable by prophylactic vaccination with several naturally derived and recombinant vaccine preparations. existing data suggests that protection is mediated by antibodies directed against the protective antigen (pa) component of the anthrax toxin complex. pa is an 83-kda protein cleaved in vivo to yield a biologically active 63-kda protein. in an effort to evaluate the potential of yeast as an expression system for the production of recombinant pa, and to de ... | 2006 | 16271808 |
| in vivo protective role of human group iia phospholipase a2 against experimental anthrax. | anthrax is an acute disease caused by bacillus anthracis. some animal species are relatively resistant to anthrax infection. this trait has been correlated to the extent of the local inflammatory reaction, suggesting innate immunity to be the first line of defense against b. anthracis infection in nonimmunized hosts. group iia secreted phospholipase a2 (spla2-iia) is produced in particular by macrophages and possesses potent antibacterial activity especially against gram-positive bacteria. we ha ... | 2005 | 16272335 |
| synthesis of the tetrasaccharide side chain of the major glycoprotein of the bacillus anthracis exosporium. | an alpha-glycoside of the tetrasaccharide sequence beta-ant-(1-->3)-alpha-l-rhap-(1-->3)-alpha-l-rhap-(1-->2)-alpha-l-rhap whose aglycon allows conjugation to suitable carriers was synthesized. the nmr characteristics of the compound are virtually identical with those of the alpha-anomer of the tetrasaccharide isolated from the major glycoprotein of the bacillus anthracis exosporium. thus, the correct structure of the natural product has been proven by chemical synthesis. | 2006 | 16275067 |
| comparison of clarithromycin and ciprofloxacin therapy for bacillus anthracis sterne infection in mice with or without (60)co gamma-photon irradiation. | biological agents and ionizing radiation lead to more severe clinical outcomes than either insult alone. this study investigated the survival of non-irradiated and (60)co-gamma-irradiated mice given therapy for inhalation anthrax with ciprofloxacin (cip) or a clinically relevant mixture of clarithromycin (clr) and its major human microbiologically important metabolite 14-hydroxy clarithromycin (14-oh clr). all b6d2f1/j 10-week-old female mice were inoculated intratracheally with 3 x 10(8) c.f.u. ... | 2005 | 16278429 |
| the cytotoxic effect of anthrax lethal toxin on human lung cells in vitro and the protective action of bovine antibodies to pa and lf. | the excretion of protein toxins by vegetative cells of bacillus anthracis is critical to the development of the lethal consequences of anthrax, particularly inhalational anthrax. whilst the lung macrophages and other phagocytic cells transfer the spores from the lung cavities into the lymphatic system, and provide an initial germination site for the proliferation of the vegetative cells, it appears that much of the tissue pathology at the time of the host's death could be due to the action of th ... | 2006 | 16278807 |
| capitol hill staff workers' experiences of bioterrorism: qualitative findings from focus groups. | little systematic information is available on mental health issues related to bioterrorism. five focus groups were conducted with capitol hill office staff (n = 28 total participants) to learn about their experience of the anthrax incident on october 15, 2001. more than 2,000 verbal passages were coded into categories and themes by using qualitative analysis software. issues emerging from the discussions included difficulties utilizing customary social supports, concerns over potential long-term ... | 2005 | 16281199 |
| [carl adolph von basedow on the on the occasion of the 150th anniversary of the day he died]. | in 2004 we commemorate the 150th day of death of carl adolph von basedow, who takes a lasting place in the history of medicine for his description of the merseburg triad (exophthalmos, goitre, tachycardia) with hyperthyroidism. there has been a long-lasting dispute over who first described these diagnostic features. in germany, the dermatologically-relevant features of hyperthyroidism are known as basedow disease. in the anglo-american literature the name "graves' disease" is predominately used, ... | 2004 | 16281617 |
| bacillus anthracis: toxicology, epidemiology and current rapid-detection methods. | b. anthracis, the causative agent for anthrax, has been well studied for over 150 years. due to the genetic similarities among various bacillus species, as well as its existence in both a spore form and a vegetative state, the detection and specific identification of b. anthracis have been proven to require complex techniques and/or laborious methods. with the heightened interest in the organism as a potential biological threat agent, a large number of interesting detection technologies have rec ... | 2006 | 16283259 |
| a defensive strategy. | | 2005 | 16283702 |
| [expression of atr-fc fusion protein in cho cells]. | atr-fc is a fusion protein consisting of extracellular domain of human anthrax toxin receptor (atr) and a fragment (hinge, ch2, and ch3 domains) of the fc of human igg1. the aim of atr-fc expression is to get an antibody-like molecule binding to protective antigen (pa), a component of anthrax toxins, this fusion protein may compete with cell surface receptor for pa binding, and block the transport of lethal factor (lf) and edema factor (ef) into cells, thereby act as an antitoxin to prevent and ... | 2005 | 16285529 |
| robert koch: the grandfather of cloning? | this year marks the centenary of robert koch's nobel prize for discovering the cause of tuberculosis. koch was also the first scientist to isolate the anthrax and cholera microbes. yet perhaps one of his greatest contributions to biology is the least appreciated: his method for propagating individual colonies of bacteria on plates, a technique that came to be called cloning. | 2005 | 16286000 |
| crystal structure of dihydrodipicolinate synthase (ba3935) from bacillus anthracis at 1.94 a resolution. | | 2006 | 16287120 |
| husbandry practices and health in smallholder dairy farms near addis ababa, ethiopia. | our study was conducted from november 2001 to april 2002 in the debre-zeit area (near addis ababa), ethiopia to assess the husbandry practices and to identify health constraints in 100 market-oriented smallholder dairy farms. a questionnaire survey, farm visit and animal examination were conducted. thirty-eight percent of the smallholder farms were owned by women. women-owned farms had more cows (median=3) than men-owned ones. the median herd size (including young animals) was four animals per f ... | 2006 | 16289383 |
| are surgical residents prepared for mass casualty incidents? | we hypothesized that resident education is inadequate with respect to management of mass casualty incidents that may involve chemical, biological, and nuclear exposures. | 2006 | 16289591 |
| filtration methods for recovery of bacillus anthracis spores spiked into source and finished water. | spores of bacillus anthracis sterne strain were recovered from 100ml and 1l volumes of tap and source waters using filtration through a 0.45um filter, followed by overnight culture on agar plates. in a set of experiments comparing sheep red blood cell (srbc) plates with a chromogenic agar formulation designed by r & f laboratories, with a spiking dose of 47 plate-enumerated spores in 100 ml tap water, the mean spore recoveries were 34.0 and 30.8 spores, respectively. when a spiking dose of 100 f ... | 2005 | 16290183 |
| characterization of bacillus anthracis germinant receptors in vitro. | bacillus anthracis begins its infectious cycle as a metabolically dormant cell type, the endospore. upon entry into a host, endospores rapidly differentiate into vegetative bacilli through the process of germination, thus initiating anthrax. elucidation of the signals that trigger germination and the receptors that recognize them is critical to understanding the pathogenesis of b. anthracis. individual mutants deficient in each of the seven putative germinant receptor-encoding loci were construc ... | 2005 | 16291679 |
| human neutrophils kill bacillus anthracis. | bacillus anthracis spores cause natural infections and are used as biological weapons. inhalation infection with b. anthracis, the etiological agent of anthrax, is almost always lethal, yet cutaneous infections usually remain localized and resolve spontaneously. neutrophils are typically recruited to cutaneous but seldom to other forms of anthrax infections, raising the possibility that neutrophils kill b. anthracis. in this study we infected human neutrophils with either spores or vegetative ba ... | 2005 | 16292357 |
| inhibition of platelet aggregation by anthrax edema toxin. | edema toxin is a key virulence determinant in anthrax pathogenesis that causes augmentation of camp inside host cells. this exotoxin has been implicated in facilitating bacterial invasion by impairing host defenses. here, we report for the first time that edema toxin plays an important role in suppression of platelet aggregation; an effect that could be of vital significance in anthrax afflicted subjects. it was found that edema toxin induces a dose dependent and time dependent increase in camp ... | 2006 | 16293226 |
| structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen. | anthrax lethal toxin assembles at the surface of mammalian cells when the lethal factor (lf) binds via its amino-terminal domain, lf(n), to oligomeric forms of activated protective antigen (pa). lf x pa complexes are then trafficked to acidified endosomes, where pa forms heptameric pores in the bounding membrane and lf translocates through these pores to the cytosol. we used enhanced peptide amide hydrogen/deuterium exchange mass spectrometry and directed mutagenesis to define the surface on lf( ... | 2006 | 16293620 |
| immunogenicity and protective efficacy of bacillus anthracis poly-gamma-d-glutamic acid capsule covalently coupled to a protein carrier using a novel triazine-based conjugation strategy. | the capsular polypeptide of bacillus anthracis is composed of a unique polyglutamic acid polymer in which d-glutamate monomers are joined by gamma-peptidyl bonds. the capsule is poorly immunogenic, and efforts at exploiting the polymer for vaccine development have focused on increasing its inherent immunogenicity through chemical coupling to immune-stimulating protein carriers. the usual strategy has employed carbodiimide-based condensing reagents for activation of free alpha-carboxyl groups, de ... | 2006 | 16293624 |
| 2-d reference map of bacillus anthracis vaccine strain a16r proteins. | bacillus anthracis has always been an important pathogen because it can cause lethal inhalational anthrax, and may be used as a bioweapon or by bioterrorists. in this study, a 2-de reference map and database of b. anthracis a16r was constructed. in total, 534 spots were processed, and 406 spots representing 299 proteins were identified. gel-estimated pis and molecular masses mostly matched well with their theoretical predictions, but some discrepancies also existed. spot and protein correspondin ... | 2005 | 16294314 |