Publications

tularaemia in europe: an epidemiological overview.	tularaemia exists endemically in most european countries. in some areas, such as finland and sweden, outbreaks comprising hundreds of cases are recorded at least once a decade. in other areas, outbreaks of such a magnitude occur only occasionally, except in times of war. between outbreaks, the natural reservoir of the causative agent, francisella tularensis, is unknown. the organism replicates intracellularly in protozoans. an association of tularaemia to natural water may be of significance in ...	2004	15287379
[tick borne zoonosis: selected clinical and diagnostic aspects].	tick-borne zoonotic infections are among the most diffuse vector borne diseases: these large group of infections is caused by different microorganisms: babesia spp., borrelia spp., rickettsia spp., ehrlichia spp., francisella tularensis, coxiella burnetii) and tick-borne encephalitis virus. babesiosis is caused by the protozoa (sporozoa) babesia microti and it is quite rare in humans in europe. the ixodids ticks are the competent vectors. a few symptomatic cases have been reported, mainly in spl ...	2004	15305697
mice sublethally infected with francisella novicida u112 develop only marginal protective immunity against systemic or aerosol challenge with virulent type a or b strains of f. tularensis.	the current study determined the ability of francisella novicida to act as a live vaccine against the much more virulent, but closely related pathogen, francisella tularensis. live attenuated strains of the latter are effective vaccines against human tularemia. however, the molecular cause of their attenuation remains unknown, and this is a regulatory barrier for licensing such vaccines. moreover, f. tularensis is exceptionally difficult to manipulate genetically. this is hampering the developme ...	2004	15312850
worldwide genetic relationships among francisella tularensis isolates determined by multiple-locus variable-number tandem repeat analysis.	the intracellular bacterium francisella tularensis is the causative agent of tularemia and poses a serious threat as an agent of bioterrorism. we have developed a highly effective molecular subtyping system from 25 variable-number tandem repeat (vntr) loci. in our study, multiple-locus vntr analysis (mlva) was used to analyze genetic relationships and potential population structure within a global collection of 192 f. tularensis isolates, including representatives from each of the four subspecie ...	2004	15317786
novel modification of lipid a of francisella tularensis.	we have investigated the lipid a of francisella tularensis subsp. holarctica strain 1547-57, a type b strain, by using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, nanoelectrospray quadrupole ion-trap mass spectrometry, and chemical methods. in accordance with the previously published structures of the lipid a from f. tularensis live vaccine strain (lvs) (atcc 29684) (e. vinogradov et al., eur. j. biochem. 269:6112-6118, 2002), all of the major lipid a forms from ...	2004	15322031
msba transporter-dependent lipid a 1-dephosphorylation on the periplasmic surface of the inner membrane: topography of francisella novicida lpxe expressed in escherichia coli.	the lipid a anchor of francisella tularensis lipopolysaccharide (lps) lacks both phosphate groups present in escherichia coli lipid a. membranes of francisella novicida (an environmental strain related to f. tularensis) contain enzymes that dephosphorylate lipid a and its precursors at the 1- and 4'-positions. we now report the cloning and characterization of a membrane-bound phosphatase of f. novicida that selectively dephosphorylates the 1-position. by transferring an f. novicida genomic dna l ...	2004	15339914
naturally occurring tularemia in a dog.	a 4-year-old spayed female irish setter was examined because of acute onset of lethargy, anorexia, and weakness. the dog had eaten an adult rabbit 36 hours earlier. tularemia was suspected because of the rabbit exposure; however, other common diseases characterized by fever, malaise, and lymphadenopathy of acute onset were also considered (ie, ehrlichiosis and rocky mountain spotted fever). the dog was treated with doxycycline (5 mg/kg [2.3 mg/lb], po, q 24 h) for 14 days as well as supportive t ...	2004	15344361
factors affecting the escape of francisella tularensis from the phagolysosome.	the highly virulent bacterium francisella tularensis is well adapted to the intracellular habitat but the mechanisms behind its intracellular survival have been elusive. recently, it was shown that the bacterium is capable of escaping from the phagosome of human and mouse monocytic cells. here it is shown that this escape is affected by gamma interferon (ifn-gamma) treatment of mouse peritoneal exudate cells since in treated cells the proportion that escaped was significantly lower (80%) than in ...	2004	15358816
a francisella tularensis pathogenicity island required for intramacrophage growth.	francisella tularensis is a gram-negative, facultative intracellular pathogen that causes the highly infectious zoonotic disease tularemia. we have discovered a ca. 30-kb pathogenicity island of f. tularensis (fpi) that includes four large open reading frames (orfs) of 2.5 to 3.9 kb and 13 orfs of 1.5 kb or smaller. previously, two small genes located near the center of the fpi were shown to be needed for intramacrophage growth. in this work we show that two of the large orfs, located toward the ...	2004	15375123
comparative proteome analysis of cellular proteins extracted from highly virulent francisella tularensis ssp. tularensis and less virulent f. tularensis ssp. holarctica and f. tularensis ssp. mediaasiatica.	francisella tularensis is the causative agent of the zoonotic disease tularemia. four subspecies of this pathogen, namely ssp. tularensis, mediaasiatica, holarctica, and novicida are spread throughout the northern hemisphere. although there are marked variations in their virulence to mammals, the subspecies are difficult to identify as they are closely genetically related. we carried out the comparative proteome analysis of cellular extracts from isolates representing the highly virulent subspec ...	2004	15378745
toll-like receptor 4 (tlr4) does not confer a resistance advantage on mice against low-dose aerosol infection with virulent type a francisella tularensis.	francisella tularensis, the causative agent of tularemia, is a gram-negative facultative intracellular bacterium. toll-like receptor (tlr) 4 is considered to be critical for inducing host innate immunity against many gram-negative bacteria including many respiratory pathogens. to determine the role of tlr4 in host defense against airborne f. tularensis infection, tlr4-defective c3h/hej (tlr4(d)) or wild-type c3h/heouj (wt) mice were challenged by low-dose aerosol with type a f. tularensis, and t ...	2004	15458779
identification of francisella tularensis genes encoding exported membrane-associated proteins using tnphoa mutagenesis of a genomic library.	francisella tularensis, the causative agent of tularemia, is a highly infectious pathogen of humans and animals, yet little is known about the surface proteins of this organism that mediate mechanisms of pathogenicity. lambdatnphoa was used to generate random alkaline phosphatase gene fusions in a f. tularensis subsp. tularensis (strain schu s4) genomic library to identify genes encoding exported extracytoplasmic proteins. eleven genes encoding membrane-associated proteins were identified by thi ...	2004	15458781
characterization of a wild-type strain of francisella tularensis isolated from a cat.	francisella tularensis type a is the primary cause of tularemia in animals and humans in north america. the majority of research on f. tularensis has been done with the attenuated live vaccine strain (lvs), which is a type b, but very few wild-type f. tularensis strains have been characterized. a gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the virginia-maryland regional college of v ...	2004	15460318
detection of staphylococcus aureus enterotoxin a and b genes with pcr-eia and a hand-held electrochemical sensor.	two electrochemical assays for detecting staphylococcus aureus enterotoxin a and b genes were developed. the assays are based on pcr amplification with biotinylated primers, hybridization to a fluorescein-labeled probe, and detection with horseradish peroxidase-conjugated anti-fluorescein antibody using a hand-held electrochemical detector. the limit of detection (lod) for both assays was approximately 16 copies of the sea and seb genes. the assays were evaluated in blinded studies, each with 81 ...	2004	15488376
did an epidemic of tularemia in ancient egypt affect the course of world history?	the dynamics of the spreading, and the identity of a virulent epidemic, similar to bubonic plague or typhus, which hit ancient egypt in the middle of the bronze age, are hereby presented. documented in medical papyri as well as archaeological findings, and re-echoed in biblical texts, a plague entered egypt's main harbor, avaris, around 1715 bc. as a result, the country was severely weakened at a time when it was already facing serious sociopolitical issues: crumbling central government, immigra ...	2004	15488667
[screening for antibodies against zoonotic agents among employees of the zoological garden of vienna, schönbrunn, austria].	the aim of this study was to investigate the prevalence of antibodies against zoonotic agents in employees of the zoological garden of vienna, schönbrunn, austria. sixty out of 120 employees participated in the study. in 97% of them antibodies to at least one zoonotic agent were identified. only two participants were free of antibodies to the zoonotic agents tested. the following seroprevalences (in brackets) were obtained: viral zoonotic (and potentially zoonotic) agents: influenzavirus a/h1n1 ...	2004	15495931
biochemical characterisation of francisella tularensis strains isolated in spain.		2004	14758833
target discrimination by surface-immobilized molecular beacons designed to detect francisella tularensis.	a molecular beacon (mb) array was designed based on unique regions of the 16s rrna of the bacterium francisella tularensis. nucleic acid molecular beacons undergo a spontaneous fluorogenic conformational change when they hybridize to specific complementary targets. the array was printed on aldehyde glass or hydrogel slides and evaluated for functioning in presence of complementary oligonucleotide sequences, single-nucleotide mismatch sequences and multiple nucleotide mismatch sequences. discrimi ...	2004	14709391
bichat guidelines for the clinical management of tularaemia and bioterrorism-related tularaemia.	francisella tularensis is one of the most infectious pathogenic bacteria known, requiring inoculation or inhalation of as few as 10 organisms to initiate human infection. inhalational tularaemia following intentional release of a virulent strain of f. tularensis would have great impact and cause high morbidity and mortality. another route of contamination in a deliberate release could be contamination of water. seven clinical forms, according to route of inoculation (skin, mucous membranes, gast ...	2004	15677845
a multiplexed pcr-coupled liquid bead array for the simultaneous detection of four biothreat agents.	we have developed a 10-plexed pcr assay coupled to a 12-plexed liquid bead array to rapidly screen environmental samples for b. anthracis, y. pestis, f. tularensis, and b. melitensis. highly validated species-specific primer sets were used to simultaneously amplify multiple diagnostic regions unique to each individual pathogen. resolution of the mix of amplified products was achieved by pcr product hybridization to corresponding probe sequences, attached to unique sets of fluorescent beads. the ...	2004	15680215
[francisella tularensis infection, or how to outsmart the certain death].	francisella tularensis is an intracellular bacterial pathogen and is the causative agent of tularemia. human become infected when manipulating with infected animals (usually rodents) or via arthropod vectors. in spite of the high virulence of the microbe still very little is known about its overall interaction with host cells and virulence factors identified to date show properties different from other bacterial species.	2004	15745053
aerosol-, but not intradermal-immunization with the live vaccine strain of francisella tularensis protects mice against subsequent aerosol challenge with a highly virulent type a strain of the pathogen by an alphabeta t cell- and interferon gamma- dependent mechanism.	francisella tularensis is an extremely virulent facultative intracellular bacterial pathogen of many mammalian species including mice and humans in which it causes a spectrum of disease collectively called tularemia. in humans, intradermal or inhaled inocula of 10cfu or less of the most virulent strains of the pathogen are sufficient to cause severe infection and possible death; in mice similar inocula are routinely lethal. an attenuated live vaccine strain, f. tularensis lvs, was developed almo ...	2005	15752834
the live vaccine strain of francisella tularensis replicates in human and murine macrophages but induces only the human cells to secrete proinflammatory cytokines.	francisella tularensis is the highly infectious agent of tularemia, a disease that can prove fatal in humans. an attenuated live vaccine strain (lvs) of this bacterium is avirulent in man but produces lethal illness in mice. as a step toward understanding the species specificity of the lvs, we compared its interactions with murine and human leukocytes. the bacterium replicated within murine bone marrow-derived macrophages (mubmdm), human monocyte-derived macrophages (humdm), and freshly isolated ...	2005	15758077
[the interaction of dynamic speckles with suspensions of gram-negative cells].	the role of temporal coherence at photodynamic action of light on living cells was investigated. a mathematical model describing the interaction of low-coherent speckles with bacterial cells was suggested and its parameters were determined based on experimental data. the interrelation between the life time of dynamic optical speckles and the degree of photodestruction of illuminated cells was established by computer simulations. the conditions were determined under which the photoinactivation of ...	2005	16248164
biodefense labs. boston university under fire for pathogen mishap.		2005	15681355
oligo-chip based detection of tick-borne bacteria.	we have developed an oligonucleotide-chip based assay for detection of 16s ribosomal pcr products from tick-borne bacteria. this chip contains 14 specific probes, which target variable regions of 16s rdna of tick-borne bacteria including borrellia spp., rickettsia spp., anaplasma spp., coxiella burnetii and francisella tularensis. the specificity of these probes was tested by hybridization of the chip with fluorescently labeled pcr products amplified from the genomic dna of selected tick-borne b ...	2005	15686852
tularemia transmitted by insect bites--wyoming, 2001-2003.	tularemia is a zoonotic disease caused by francisella tularensis, a fastidious, gram-negative coccobacillus that infects vertebrates, especially rabbits and rodents. in humans, tularemia is classified into six major syndromes: ulceroglandular (the most common form), glandular, typhoidal, oculoglandular, oropharyngeal, and pneumonic. the case-fatality rate among humans can reach 30%-60% in untreated typhoidal cases. although bites from ticks and handling infected animals are considered the most c ...	2005	15729218
low dose aerosol infection of mice with virulent type a francisella tularensis induces severe thymus atrophy and cd4+cd8+ thymocyte depletion.	francisella tularensis is a gram-negative facultative intracellular bacterium and the causative agent of tularemia. two subspecies (type a and b strains) of the pathogen exist, the former being much more virulent than the latter for humans and other higher mammals. in this study, we examined the effect of virulent strains of f. tularensis infection on the thymus and thymocytes and the potential mechanisms involved. low-dose aerosol exposure of c57bl/6 mice with type a, but not type b, f. tularen ...	2005	16257504
discrimination between francisella tularensis and francisella-like endosymbionts when screening ticks by pcr.	the presence of francisella-like endosymbionts in tick species known to transmit tularemia poses a potential diagnostic problem for laboratories that screen tick samples by pcr for francisella tularensis. tick samples initially considered positive for f. tularensis based on standard 16s rrna gene pcr were found to be positive only for francisella-like endosymbionts using a multitarget f. tularensis taqman assay (isftu2, tul4, and iglc) and 16s rrna gene sequencing. specificity of pcr-based diagn ...	2005	16269811
francisella tularensis induces aberrant activation of pulmonary dendritic cells.	francisella tularensis is an obligate intracellular bacterium that induces severe, acute, often fatal disease when acquired by the respiratory route. despite the seriousness of this pathogen, very little is understood about its interaction with key target cells in the airways and lungs (alveolar macrophages and airway dendritic cells (dc)) after inhalation. in this study we demonstrate replication of f. tularensis in primary dc. early after infection, f. tularensis induced increased expression o ...	2005	16272336
[rodents as the vectors of infective agents in the natural foci of infections in the moscow megapolis].	this work deals with the results of 40-year observations on the circulation of infective agents in the natural foci of infections (tularemia, leptospirosis, hemorrhagic fever with renal syndrome) among rodents and other small mammals in the territory of the moscow. the monitoring of their frequency and the infection rates remains the main effective measure for the prophylaxis of dangerous infections among the population of the megapolis.	2005	16279545
[bioterrorism].	biological terrorism is intentionally to use infectious substances for developing diseases or death in animals or humans, leading to disaster and panic in our community. bioterrorism-associated diseases are mostly rare or eradicated infectious diseases, and recently, we do not have experience to make a clinical and laboratory diagnosis. in particular, these infectious diseases have incubation periods from infection to development of the disease. the staff working at public health institutions, i ...	2005	16296383
a 12-case outbreak of pharyngeal plague following the consumption of camel meat, in north-eastern jordan.	between late january and early february 1997, an outbreak of plague, associated with cervical lymphadenopathy and fever, occurred in the jordanian village of azraq ad-druze, which lies about 50 km west of the border with saudi arabia. the 12 cases who presented at hospital were initially assumed to have tularaemia, and all were successfully treated with gentamicin. when, however, their sera were tested for evidence of yersinia pestis or francisella tularensis infection (using haemagglutination, ...	2005	16297292
a mutant of francisella tularensis strain schu s4 lacking the ability to express a 58-kilodalton protein is attenuated for virulence and is an effective live vaccine.	francisella tularensis subsp. tularensis (type a) strain schu s4 is a prototypic strain of the pathogen that is highly virulent for humans and other mammals. its intradermal (i.d.) 50% lethal dose (ld50) for mice is <10 cfu. we discovered a spontaneous mutant, designated fsc043, of schu s4 with an i.d. ld50 of >10(8) cfu. fsc043 effectively vaccinated mice against challenge with a highly virulent type a strain, and the protective efficacy was at least as good as that of f. tularensis lvs, an emp ...	2005	16299332
clinical use of a diagnostic pcr for francisella tularensis in patients with suspected ulceroglandular tularaemia.	a retrospective analysis to evaluate the clinical use of a diagnostic pcr for francisella tularensis in patients with suspected ulceroglandular tularaemia was performed. 154 samples, 129 from patients with definitive tularaemia and 25 from patients where tularaemia could be ruled out, were analysed. the diagnostic pcr had a specificity of 96%, a sensitivity of 78.3%, and a positive predictive value of 99%. especially samples from encrusted lesions, even up to 4 weeks old, in patients with tulara ...	2005	16308216
the phagosomal transporter a couples threonine acquisition to differentiation and replication of legionella pneumophila in macrophages.	differentiation in response to environmental cues is integral to the success of many intracellular pathogens. by characterizing a legionella pneumophila mutant defective for differentiation in broth and replication in macrophages, we identified a subfamily of major facilitator superfamily transporters, here named pht (phagosomal transporter), that also is conserved in two other vacuolar pathogens, coxiella burnetii and francisella tularensis. biolog phenotype microarray analysis indicated that p ...	2005	15998735
cd4-cd8- t cells control intracellular bacterial infections both in vitro and in vivo.	memory t cells, including the well-known cd4(+) and cd8(+) t cells, are central components of the acquired immune system and are the basis for successful vaccination. after infection, cd4(+) and cd8(+) t cells expand into effector cells, and then differentiate into long-lived memory cells. we show that a rare population of cd4(-)cd8(-)cd3(+)alphabeta(+)gammadelta(-)nk1.1(-) t cells has similar functions. these cells potently and specifically inhibit the growth of the intracellular bacteria mycob ...	2005	16027239
[influence of cycloferon on the biological properties of bacterial intracellular pathogens].	the influence of cycloferon on some biological properties of pathogenic (shigella spp., salmonella spp., francisella tularensis, brucella spp.) and opportunistic (escherichia coli, staphylococcus spp.) microorganisms has been experimentally determined in vitro. as revealed in these experiments, the preparation used at concentrations under study considerably suppresses the antilysozyme and anticomplementary activity of gram-negative intracellular parasites (shigellae, salmonellae, f. tularensis, ...	2005	16028504
prevalence of infection with francisella tularensis, borrelia burgdorferi sensu lato and anaplasma phagocytophilum in rodents from an endemic focus of tularemia in bulgaria.	the prevalence of francisella tularensis, borrelia burgdorferi and anaplasma phagocytophilum in rodents is a determinant for their role in maintaining pathogens in the environment. a total of 169 rodents, trapped in an endemic focus of tularemia, were examined by pcr to asses the frequency of infection with the etiological agents of tularemia, lyme borreliosis and human granulocytic anaplasmosis. the overall prevalence of f. tularensis was 22 %. in 6 % of the black rats, f. tularensis was identi ...	2005	16028881
a novel screening elisa and a confirmatory western blot useful for diagnosis and epidemiological studies of tularemia.	a novel enzyme-linked immunosorbent assay (elisa) and a confirmatory western blot (wb) to detect human antibodies against francisella tularensis were evaluated. the elisa was based on partially purified lipopolysaccharide (lps), the wb on whole antigen of f. tularensis. positive wb showed a typical lps ladder. sensitivity and specificity of the elisa, as assessed in 104 positive sera and 1149 'normal' sera from healthy young adults, were 99.0% and 97.1% respectively. sensitivity of the wb was cl ...	2005	16050523
enteric fever-like illness caused by infection with citrobacter amalonaticus.	'enteric fever' is a potentially fatal, severe systemic disease, which is encountered worldwide. traditionally, enteric fever refers to a bacteremic illness caused by members of certain salmonella serotypes, notably: salmonella typhi, a gram-negative bacterium, and to a lesser extent, salmonella paratyphi a, b and c. in addition, other non-salmonella organisms may produce a syndrome clinically indistinguishable from "enteric fever". brucella sp., campylobacter sp., edwardsiella tarda, enterobact ...	2005	16083226
massive attack.		2005	16094740
dna assays for detection, identification, and individualization of select agent microorganisms.	the purpose of this article is to review the status of dna assays used for the detection, identification, and individualization of bacillus anthracis, yersinia pestis, francisella tularensis, burkholderia mallei, and brucella abortus. these select agent microorganisms are historically significant as they have either been used or experimented with as a bioweapon or as a terrorist agent and are the subject of intense research in the areas of biodefense and bioforensics. if the presence of a biolog ...	2005	16100754
[diagnostic tests: tularemia].		2005	16111241
francisella tularensis enters macrophages via a novel process involving pseudopod loops.	intracellular bacterial pathogens employ a variety of strategies to invade their eukaryotic host cells. from an ultrastructural standpoint, the processes that bacteria employ to invade their host cells include conventional phagocytosis, coiling phagocytosis, and ruffling/triggered macropinocytosis. in this paper, we describe a novel process by which francisella tularensis, the agent of tularemia, enters host macrophages. f. tularensis is a remarkably infectious facultative intracellular bacteria ...	2005	16113308
antimicrobial susceptibilities of austrian francisella tularensis holarctica biovar ii strains.	the antibiotic susceptibilities of 50 francisella tularensis subsp. holarctica biovar ii strains isolated from hares and human patients from the eastern part of austria were examined. minimum inhibitory concentrations (mics) of 24 antimicrobial agents were determined using eteststrade mark on cysteine heart agar plates supplemented with 10% sheep blood. all isolates were sensitive to tetracyclines, aminoglycosides, quinolones, chloramphenicol and rifampicin. resistance was observed in all isolat ...	2005	16143497
[phase variations of francisella tularensis lipopolysaccharide in human infection and immunization].	the comparative study of the specificity of antibodies in human sera after tularemia infection and immunization with live tularemia infection was carried out with the use of passive hemagglutination and immunoblotting techniques. the sera of tularemia patients contained two different types of immunoglobulins: strictly specific to the antigenic epitopes of f. tularensis iipopolysaccharide (lps) and strictly specific to f. tularensis subsp. novicida lps. such phenomenon may be due to phase variati ...	2005	16146218
detection of diverse new francisella-like bacteria in environmental samples.	following detection of putative francisella species in aerosol samples from houston, texas, we surveyed soil and water samples from the area for the agent of tularemia, francisella tularensis, and related species. the initial survey used 16s rrna gene primers to detect francisella species and related organisms by pcr amplification of dna extracts from environmental samples. this analysis indicated that sequences related to francisella were present in one water and seven soil samples. this is the ...	2005	16151142
outbreak of tularaemia in golcuk, turkey in 2005: report of 5 cases and an overview of the literature from turkey.	tularaemia was diagnosed by taqman rt-pcr and microagglutination tests in 5 patients, all from a new settlement constructed after the earthquake of 1999. during the follow-up, 129 more cases were found in this settlement (data from the local health care authority). in this study, clinical features of 5 cases are presented briefly, and the turkish literature on past outbreaks of tularaemia is reviewed.	2005	16191887
use of shell-vial cell culture assay for isolation of bacteria from clinical specimens: 13 years of experience.	the shell-vial culture assay is performed routinely in our laboratory. recently we revisited our experience of using the shell-vial culture assay for the isolation of microorganisms from various clinical samples. over a 13-year period, we have isolated 580 bacterial strains (5%) from 11,083 clinical samples tested. over the same period, 285 isolates of rickettsiae, bartonellae, or coxiella burnetii were cultured from a total of 7,102 samples tested. these isolates include 55 rickettsia sp. isola ...	2005	16207953
tularemia in denmark: identification of a francisella tularensis subsp. holarctica strain by real-time pcr and high-resolution typing by multiple-locus variable-number tandem repeat analysis.	we report ulceroglandular tularemia affecting an 8-year-old boy and the first recovery of francisella tularensis in denmark. a novel real-time pcr assay was used to identify the strain as f. tularensis subsp. holarctica (type b). multiple-locus variable-number tandem repeat analysis demonstrated a close genetic relationship to strains from norway.	2005	16208017
[tularemia].		2005	16209241
francisella tularensis proteome: low levels of asb-14 facilitate the visualization of membrane proteins in total protein extracts.	proteomic analysis of bacterial pathogens isolated from in vivo sources, such as infected tissues, provides many challenges not the least of which is the limited quantity of sample available for analysis. it is, therefore, highly desirable to develop a one-step cellular lysis and protein solubilization method that minimizes protein losses and allows the maximum possible coverage of the proteome. here, we have used standard sample buffer constituents including urea, thiourea and dtt, but varied t ...	2005	16212441
diagnostic procedures in tularaemia with special focus on molecular and immunological techniques.	tularaemia is a severe bacterial zoonosis caused by the highly infectious agent francisella tularensis. it is endemic in countries of the northern hemisphere ranging from north america to europe, asia and japan. very recently, francisella-like strains causing disease in humans were described from tropical northern australia. in the last decade, efforts have been made to develop sensitive and specific immunological and molecular techniques for the laboratory diagnosis of tularaemia and also for t ...	2005	16219088
in vivo efficacy of fluoroquinolones against systemic tularaemia infection in mice.	the in vivo efficacy of ciprofloxacin, gatifloxacin and moxifloxacin were assessed in an experimental francisella tularensis schu s4 infection in the balb/c mouse model.	2005	16223941
innate immunity against francisella tularensis is dependent on the asc/caspase-1 axis.	francisella tularensis is a highly infectious gram-negative coccobacillus that causes the zoonosis tularemia. this bacterial pathogen causes a plague-like disease in humans after exposure to as few as 10 cells. many of the mechanisms by which the innate immune system fights francisella are unknown. here we show that wild-type francisella, which reach the cytosol, but not francisella mutants that remain localized to the vacuole, induced a host defense response in macrophages, which is dependent o ...	2005	16230474
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
seroprevalence of brucellosis, tularemia, and yersiniosis in wild boars (sus scrofa) from north-eastern germany.	brucellosis and tularemia are classical zoonotic diseases transmitted from an animal reservoir to humans. both, wildlife and domestic animals, contribute to the spreading of these zoonoses. the surveillance of the animal health status is strictly regulated for domestic animals, whereas systematic disease monitoring in wildlife does not exist. the aim of the present study was to provide data on the prevalence of anti-brucella, anti-francisella and anti-yersinia antibodies in wild boars from north ...	2005	16364020
natural killer and cd8 t cells dominate the response by human peripheral blood mononuclear cells to inactivated francisella tularensis live vaccine strain.	francisella tularensis is a category a biothreat agent, and as a result, it has recently generated much research interest. f. tularensis live vaccine strain (lvs) is an attenuated form of the virulent f. tularensis organism and has previously been used as a vaccine. however, because of safety concerns, it is no longer approved for this purpose. thus, the use of inactivated organisms is preferable for vaccine purposes. although many studies have been performed that examine human peripheral blood ...	2005	16386645
[incidence of zoonoses in petting zoos and evaluation of hygiene measures to prevent the transmission to humans].	in summer 2003, a study was performed in thirty swiss petting zoos with the objective to determine the prevalence of zoonotic agents, and to describe hygiene measures implemented to reduce the risk of human infection. fecal samples from different animal species were collected from the floor of pens to determine the prevalence of salmonella spp., campylobacter spp., verocytotoxin producing e. coli/ vtec and francisella tularensis. a questionnaire on hygiene measures, number of animals per species ...	2005	16398191
serologic survey of select infectious diseases in coyotes and raccoons in nebraska.	to obtain data about select zoonotic and other infectious diseases in free-ranging predators in five ecoregions in nebraska, sera were collected from 67 coyotes (canis latrans) and 63 raccoons (procyon lotor) from november 2002 through january 2003. for coyotes, antibodies were detected against canine distemper virus (cdv, 61%), francisella tularensis (32%), rickettsia rickettsi (13%), and flaviviruses (48%). none of the coyote sera had antibodies to borrelia burgdorferi, brucella canis, or six ...	2005	16456169
[rickettsia helvetica: an emerging tick-borne pathogen in hungary and europe].	rickettsia helvetica belonging to spotted fever group rickettsiae was recently detected by polymerase chain reaction followed by sequencing in european sheep ticks (ixodes ricinus) from hungary. current knowledge on these rickettsiae and the clinical and diagnostic aspects of r. helvetica infection is summarized. in acute cases, r. helvetica is generally responsible for flu-like symptoms. nevertheless, recent data indicate that in chronic cases, these rickettsiae can be responsible for perimyoca ...	2005	16440500
francisella tularensis in the united states.	the causative agent of tularemia, francisella tularensis, is a formidable biologic agent that occurs naturally throughout north america. we examined genetic and spatial diversity patterns among 161 us f. tularensis isolates by using a 24-marker multiple-locus variable-number tandem repeat analysis (mlva) system. mlva identified 126 unique genotypes. phylogenetic analyses showed patterns similar to recently reported global-scale analyses. we observed clustering by subspecies, low genetic diversit ...	2005	16485467
[evaluation of the enzyme-linked immunosorbent assay for the serodiagnosis of tularemia].	the usefulness of the elisa using as antigen prepared in our laboratory supernatant obtained after centrifugation of sonicated f. tularensis cell suspension was compared with the tube agglutination test with commercial available antigen. paired serum specimens obtained from 6 patients with ulceroglandular syndrome of tularemia were tested in both tests. the cut-off limit of serum antibodies was set at mean antibody titre determined in the sera of 115 blood donors exceeded by three standard devia ...	2005	16773836
[pathogenicity of francisella].	the data of literature and the results of the author's research on the pathogenicity of the causative agent of tularemia and other francisella organisms are reviewed. the solution of the problem of their pathogenicity is based, as stated by the author, on the level of our knowledge of the genetics of francisella. the conclusion has been made that scientific achievements in the field of the genetics of francisella, obtained during the last 15 years, make it possible to find out the pathogenicity ...	2005	15773415
[tularemia acquired in denmark by an eight-year-old child].		2005	15779266
efficacy of the live attenuated francisella tularensis vaccine (lvs) in a murine model of disease.	a live attenuated vaccine francisella tularensis live vaccine strain (lvs), that confers protection against tularemia infection in a number of animal models including man was developed during the 1960s in the us. in this study, we have established the median lethal dose (mld) after intraperitoneal (i.p.) or intravenous (i.v.) delivery of ndbr lot 4 f. tularensis lvs to be 4 cfu and 2.24 x 10(4) cfu, respectively, in balb/c mice and less than 1 cfu and 1.29 x 10(4) cfu, respectively, in c57bl/6 m ...	2005	15780452
intranasal interleukin-12 treatment for protection against respiratory infection with the francisella tularensis live vaccine strain.	francisella tularensis is a gram-negative intracellular bacterium that can induce lethal respiratory infection in humans and rodents. however, little is known about the role of innate or adaptive immunity in protection from respiratory tularemia. in the present study, the role of interleukin-12 (il-12) in inducing protective immunity in the lungs against intranasal infection of mice with the live vaccine strain (lvs) of f. tularensis was investigated. it was found that gamma interferon (ifn-gamm ...	2005	15784575
the contribution of reactive nitrogen and oxygen species to the killing of francisella tularensis lvs by murine macrophages.	intracellular killing of francisella tularensis by macrophages depends on interferon-gamma (ifn-gamma)-induced activation of the cells. the importance of inducible nitric oxide synthase (inos) or nadph phagocyte oxidase (phox) for the cidal activity was studied. murine ifn-gamma-activated peritoneal exudate cells (pec) produced nitric oxide (no), measured as nitrite plus nitrate, and superoxide. when pec were infected with the live vaccine strain, lvs, of f. tularensis, the number of viable bact ...	2005	15788155
the role of mapk signal pathways during francisella tularensis lvs infection-induced apoptosis in murine macrophages.	francisella tularensis is a highly virulent intracellular pathogen responsible for tularemia. this bacterium is capable of infecting many mammalian species and various cell types, but little is known about the mechanisms of survival and interactions with host cells. we examined the number of infected host cells, cytotoxicity and the role of apoptosis or necrosis in infection-induced cell death. our results demonstrate that f. tularensis lvs induces apoptosis of infected macrophages within 10 h. ...	2005	15820149
epizootiologic investigations of selected infectious disease agents in free-ranging eurasian lynx from sweden.	serum samples from 106 eurasian lynx (lynx lynx) from across sweden, found dead or shot by hunters in 1993-99, were investigated for presence of antibodies to feline parvovirus (fpv), feline coronavirus, feline calicivirus, feline herpesvirus, feline immunodeficiency virus, francisella tularensis, and anaplasma phagocytophila, and for feline leukemia virus antigen. in addition, tissue samples from 22 lynx submitted in 1999 were analyzed by real-time polymerase chain reaction (pcr) to detect nucl ...	2005	15827211
tularemia: emergence/re-emergence.	francisella tularensis is a gram-negative coccobacillus and the etiologic agent of the zoonotic disease tularemia. first described in 1911 in tulare county, california, it has since been reported throughout the northern hemisphere, with natural infections reported among an unusually wide range of vertebrates and invertebrates. in recent years, tularemia has emerged in new geographic locations, populations, and settings. this review will serve to highlight mechanisms contributing to the recent em ...	2005	15845234
intranasal vaccination induces protective immunity against intranasal infection with virulent francisella tularensis biovar a.	the inhalation of francisella tularensis biovar a causes pneumonic tularemia associated with high morbidity and mortality rates in humans. exposure to f. tularensis usually occurs by accident, but there is increasing awareness that f. tularensis may be deliberately released in an act of bioterrorism or war. the development of a vaccine against pneumonic tularemia has been limited by a lack of information regarding the mechanisms required to protect against this disease. vaccine models for f. tul ...	2005	15845466
lipopolysaccharide microarrays for the detection of antibodies.	lipopolysaccharide (lps) is the major component of gram-negative bacterial outer membrane. lps are immunogenic and show species/strain specificity. the demonstration of anti-lps antibodies in clinical samples is of diagnostic value in certain gram-negative bacterial infections. in the present study we explored the possibility of immobilizing lps isolated from different bacteria in a microarray format for the detection of anti-lps antibodies. lps was successfully immobilized on nitrocellulose-coa ...	2005	15847798
tularemia in a kidney transplant recipient: an unsuspected case and literature review.	tularemia is a zoonotic infection that has rarely been reported in transplant recipients. the authors present a case of unsuspected tularemia in a kidney transplant patient that was diagnosed by isolation of francisella tularensis in the blood. the patient was treated successfully with antibiotics. during diagnostic workup, a laboratory technician was exposed to tularemia by inhalation of the culture plate and received postexposure prophylaxis. this report emphasizes the importance of exposure h ...	2005	15861359
insights into the oxidative stress response in francisella tularensis lvs and its mutant deltaiglc1+2 by proteomics analysis.	francisella tularensis is a facultative intracellular pathogen. its capacity to induce disease depends on the ability to invade and multiply within a wide range of eukaryotic cells, such as professional phagocytes. the comparative disinterest in tularemia in the past relative to other human bacterial pathogens is reflected in the paucity of information concerning the mechanisms of pathogenesis. only a few genes and gene products associated with francisella virulence are known to date. the aim of ...	2005	15869961
[tularemia. the disease and its epidemiology in france].	this study had for aim to determine whether the epidemiologic features of animal and human tularemia observed in france could explain its re-emergence since 1993.	2005	15878815
[cross-reacting antigens of pathogenic burkholderia and some dangerous causative agents of infectious diseases].	cross-reacting antigens in b. mallei, b. pseudomallei, b. thailandensis, francisella tularensis, yersinia pestis and mycobacterium tuberculosis were studied with the use of immuno- and electrophoretic techniques. the set of antigens was shown to be almost identical in the causative agents of glanders, melioidosis, as well as in b. thailandensis, though in the latter organism 200-kd glycoprotein was absent. the analysis of immuno- and proteinograms demonstrated the presence of cross-reactions in ...	2005	15881932
[investigation on mixed natural foci of zoonotic infections].		2005	15881957
proteomic analysis of anti-francisella tularensis lvs antibody response in murine model of tularemia.	francisella tularensis live vaccine strain infection of mice has been established as an experimental model of tularemia that is suitable for studies of immune mechanisms against the intracellular pathogen. in this study, the model was used to explore immunogenic repertoire of f. tularensis with the aim of identifying new molecules able to activate the host immune system, potential bacterial markers with vaccine, and diagnostic applications. immunoproteomic approach based on the combination of tw ...	2005	15892173
host-pathogen interactions: a proteomic view.	host-pathogen interactions reflect the balance of host defenses and pathogen virulence mechanisms. advances in proteomic technologies now afford opportunities to compare protein content between complex biologic systems ranging from cells to animals and clinical samples. thus, it is now possible to characterize host-pathogen interactions from a global proteomic view. most reports to date focus on cataloging protein content of pathogens and identifying virulence-associated proteins or proteomic al ...	2005	15892564
[case report: a case of tularemia with delayed diagnosis].	tularemia is a zoonotic disease caused by francisella tularensis. in this report, a tularemia case who was living in a non-endemic area, has been presented. a 24 years old female patient with multiple cervical lymphadenitis has been admitted to the outpatient clinic of ankara training and research hospital, turkey. her complaints started two months ago as signs and symptoms of an upper respiratory tract infection. as she had received non-specific treatment (amoxycillin clavulanate; 2x 1g/day) in ...	2005	15900842
evolution of subspecies of francisella tularensis.	analysis of unidirectional genomic deletion events and single nucleotide variations suggested that the four subspecies of francisella tularensis have evolved by vertical descent. the analysis indicated an evolutionary scenario where the highly virulent f. tularensis subsp. tularensis (type a) appeared before the less virulent f. tularensis subsp. holarctica (type b). compared to their virulent progenitors, attenuated strains of f. tularensis exhibited specific unidirectional gene losses.	2005	15901721
vaccination strategies for francisella tularensis.	francisella tularensis is the etiologic agent of tularemia, a severe debilitating disease of humans and animals. the low infectious dose of f. tularensis in humans and the relative ease of culture are probably the properties which originally attracted interest in this bacterium as a bioweapon. even today, f. tularensis is ranked as one of the pathogens most likely to be used as a biological warfare or bioterrorism agent. a live attenuated vaccine (lvs) has been available for over 50 years, but t ...	2005	15919131
francisella tularensis lvs initially activates but subsequently down-regulates intracellular signaling and cytokine secretion in mouse monocytic and human peripheral blood mononuclear cells.	monocytic cells constitute an important defense mechanism against invading pathogens by recognizing conserved pathogens components. the recognition leads to activation of intracellular pathways involving nuclear factor kappa b (nf-kappab) and mitogen-activated protein kinases (mapk), such as the c-jun nh2-terminal kinase (jnk), and p38. we show that in vitro infection with francisella tularensis results in activation of nf-kappab, phosphorylation of p38 and c-jun, and secretion of tnf-alpha in a ...	2005	15925273
tularemia vaccine: past, present and future.	francisella tularensis is a gram negative intracellular pathogen that causes the highly debilitating or fatal disease tularemia. f. tularensis can infect a wide range of animals and can be transmitted to humans in a variety of ways, the most common being by the bite of an infected insect or arthropod vector. the attenuated f. tularensis live vaccine strain (lvs) has been used previously under investigational new drug status to vaccinate at-risk individuals. however the history of the strain and ...	2005	15928980
simultaneous measurement of specific serum igg responses to five select agents.	select agents are defined by cdc and the usda animal and plant health inspection service (aphis) as biological agents or toxins deemed a threat to public, animal, or plant health, or to animal or plant products. they are classified on the basis of their ease of dissemination, mortality/morbidity rate, and potential for social disruption. a subset of these agents includes bacillus anthracis, yersinia pestis, francisella tularensis, ricin toxin (rt), and staphylococcal enterotoxin b (seb). infecti ...	2005	15931499
crystallization of acpa, a respiratory burst-inhibiting acid phosphatase from francisella tularensis.	francisella tularensis is a highly infectious bacterial pathogen that is classified as a category a pathogen by the centers for disease control and prevention. here, we report crystallization of a recombinant form of f. tularensis acpa, a unique and highly expressed acid phosphatase that is thought to play a role in intracellular survival by inhibiting the host respiratory burst. three crystal forms have been obtained, with form iii being the most suitable for high-resolution structure determina ...	2005	15935744
francisella tularensis live vaccine strain: proteomic analysis of membrane proteins enriched fraction.	proteome analysis of gram-negative facultative intracellular pathogen francisella tularensis (f. tularensis) live vaccine strain has been performed only on whole-cell extracts so far. this is the first study dealing with the analysis of the membrane subproteome of this microorganism. a fraction enriched in membrane proteins obtained by carbonate extraction was separated using two-dimensional electrophoresis and all visualized spots were identified by mass spectrometry. the reference map is the b ...	2005	15937965
structural analysis of the o-antigen of francisella tularensis subspecies tularensis strain osu 10.		2005	15947437
multiplexed detection of pathogen dna with dna-based fluorescence nanobarcodes.	rapid, multiplexed, sensitive and specific molecular detection is of great demand in gene profiling, drug screening, clinical diagnostics and environmental analysis. one of the major challenges in multiplexed analysis is to identify each specific reaction with a distinct label or 'code'. two encoding strategies are currently used: positional encoding, in which every potential reaction is preassigned a particular position on a solid-phase support such as a dna microarray, and reaction encoding, w ...	2005	15951805
modulation of biogenesis of the francisella tularensis subsp. novicida-containing phagosome in quiescent human macrophages and its maturation into a phagolysosome upon activation by ifn-gamma.	francisella tularensis is a highly virulent facultative intracellular pathogen that has been categorized as a class a bioterrorism agent, and is classified into four subsp, tularensis, holarctica, mediasiatica and novicida. although the ability of f. tularensis subsp. novicida to cause tularemia in mice is similar to the virulent subsp. tularensis and holarctica, it is attenuated in humans. it is not known whether attenuation of f. tularensis subsp. novicida in humans is resulting from a differe ...	2005	15953028
the francisella tularensis pathogenicity island protein iglc and its regulator mgla are essential for modulating phagosome biogenesis and subsequent bacterial escape into the cytoplasm.	the francisella tularensis subsp. novicida-containing phagosome (fcp) matures into a late endosome-like stage that acquires the late endosomal marker lamp-2 but does not fuse to lysosomes, for the first few hours after bacterial entry. this modulation in phagosome biogenesis is followed by disruption of the phagosome and bacterial escape into the cytoplasm where they replicate. here we examined the role of the francisella pathogenicity island (fpi) protein iglc and its regulator mgla in the intr ...	2005	15953029
[possible mechanisms of implementation of toxic potential of lipopolysaccharides of pathogenic bacteria].	the significance of variability of biological properties of lipopolysaccharides (lps) is discussed in the paper within the pathogenesis of infectious process. on the basis of an analysis of published data and of results of independent research of two microorganisms (yersinia pestis and francisella tularensis) a conclusion is made on that a biologically inert lps form (with a weak cytokine-inducing ability, apirogenicity and non-toxicity etc.) is typical of highly pathogenic bacteria. it is sugge ...	2005	15954469
chips and snps, bugs and thugs: a molecular sleuthing perspective.	recent events both here and abroad have focused attention on the need for ensuring a safe and secure food supply. although much has been written about the potential of particular select agents in bioterrorism, we must consider seriously the more mundane pathogens, especially those that have been implicated previously in foodborne outbreaks of human disease, as possible agents of bioterrorism. given their evolutionary history, the enteric pathogens are more diverse than agents such as bacillus an ...	2005	15954721
a new francisella (beggiatiales: francisellaceae) inquiline within dermacentor variabilis say (acari: ixodidae).	while estimating the prevalence of the dermacentor variabilis (say) symbiont (dvs) in dog ticks on martha's vineyard, ma, we identified dna that may represent a heretofore unrecognized francisella sp. polymerase chain reaction targeting a portion of the 16s rdna specific for dvs yielded an amplicon that was only 96.6% similar to that of dvs accessioned in genbank. phylogenetic analysis of the 16s and 23s rdna genes suggests the presence of a distinct bacterium closely related to the other endosy ...	2005	15962806
discrimination of francisella tularensis subspecies using surface enhanced laser desorption ionization mass spectrometry and multivariate data analysis.	francisella tularensis causes the zoonotic disease tularemia, and is considered a potential bioterrorist agent due to its extremely low infection dose and potential for airborne transmission. presently, f. tularensis is divided into four subspecies; tularensis, holarctica, mediasiatica and novicida. phenotypic discrimination of the closely related subspecies with traditional methods is difficult and tedious. furthermore, the results may be vague and they often need to be complemented with virule ...	2005	15668033
infection scare inflames fight against biodefence network.		2005	15674255
the complete genome sequence of francisella tularensis, the causative agent of tularemia.	francisella tularensis is one of the most infectious human pathogens known. in the past, both the former soviet union and the us had programs to develop weapons containing the bacterium. we report the complete genome sequence of a highly virulent isolate of f. tularensis (1,892,819 bp). the sequence uncovers previously uncharacterized genes encoding type iv pili, a surface polysaccharide and iron-acquisition systems. several virulence-associated genes were located in a putative pathogenicity isl ...	2005	15640799
toll-like receptor 4 (tlr4) plays a relatively minor role in murine defense against primary intradermal infection with francisella tularensis lvs.		2005	15626487