Publications

comparison of automated and nonautomated systems for identification of burkholderia pseudomallei.	the identification of burkholderia pseudomallei, the causative agent of melioidosis, is usually not difficult in laboratories in areas where it is endemic. with the increase in international travel and the threat of bioterrorism, it has become more likely that laboratories in areas where it is not endemic could encounter this organism. the increase in the use of and dependence upon automated identification systems makes accurate identification of uncommonly encountered organisms such as b. pseud ...	2002	12454163
analogous cytokine responses to burkholderia pseudomallei strains contrasting in virulence correlate with partial cross-protection in immunized mice.	cytokine mrna levels were assessed in burkholderia pseudomallei-susceptible balb/c mice and b. pseudomallei-resistant c57bl/6 mice following administration of a sublethal dose of less virulent (lv) b. pseudomallei, a candidate immunogen tested for protection against a highly virulent (hv) challenge. compared on the basis of the bacterial loads, the cytokine patterns induced by hv and lv b. pseudomallei were similar, involving gamma interferon, interleukin-10, and other cytokines. partial cross-p ...	2002	12065542
characterization of burkholderia pseudomallei infection and identification of novel virulence factors using a caenorhabditis elegans host system.	the environmental saphrophyte burkholderia pseudomallei is the causative agent of melioidosis, a systemic, potentially life-threatening condition endemic to many parts of south-east asia and northern australia. we have used the soil nematode caenorhabditis elegans as a model host to characterize the mechanisms by which this bacterium mounts a successful infection. we find that c. elegans is susceptible to a broad range of burkholderia species, and that the virulence mechanisms used by this patho ...	2002	12068805
burkholderia thailandensis e125 harbors a temperate bacteriophage specific for burkholderia mallei.	burkholderia thailandensis is a nonpathogenic gram-negative bacillus that is closely related to burkholderia mallei and burkholderia pseudomallei. we found that b. thailandensis e125 spontaneously produced a bacteriophage, termed phie125, which formed turbid plaques in top agar containing b. mallei atcc 23344. we examined the host range of phie125 and found that it formed plaques on b. mallei but not on any other bacterial species tested, including b. thailandensis and b. pseudomallei. examinati ...	2002	12081973
melioidosis--a report from pondicherry, south india.	melioidosis is an acute infectious disease caused by a safety-pin-shaped gram-negative bacteria called burkholderia pseudomallei. here, we report the first case of melioidosis in a middle aged male agricultural worker, from pondicherry. the isolation of this organism from subcutaneous nodules on the extensor aspect of his limbs underlines the diversity of its clinical presentation. difficulty in identifying the organism which mimics any other non-fementing gram-negative bacilli (nfgnb) on cursor ...	2002	12583480
primary melioidotic prostatic abscess: presentation, diagnosis and management.	in south-east asia and northern australia, melioidosis (infection with burkholderia pseudomallei) is a known cause of severe community-acquired sepsis. however, melioidosis presenting primarily as prostatic abscesses is very rare.	2002	12121159
characterization of a laboratory-generated variant of bps beta-lactamase from burkholderia pseudomallei that hydrolyses ceftazidime.	burkholderia pseudomallei produces an ambler class a beta-lactamase, known as bps-1. the beta-lactamase gene from a laboratory-derived, ceftazidime-resistant strain of b. pseudomallei (lh-1-2) was cloned and expressed in escherichia coli. the beta-lactamase, named bps-1m, had an identical isoelectric focusing point (pi 7.7) to that of bps-1, but differed in having a stronger hydrolytic activity against ceftazidime. susceptibility testing showed that bps-1m when expressed in e. coli conferred res ...	2002	12407130
the use of animal infection models to study the pathogenesis of melioidosis and glanders.	the use of animal infection models is central to the study of microbial pathogenesis. in combination with genetic, immunological and antigen purification techniques, much can be learned regarding the pathogenesis of diseases caused by microorganisms. this update focuses on the recent use of animal infection models to study the pathogenesis of melioidosis and glanders.	2002	12419603
characterisation of an acapsular mutant of burkholderia pseudomallei identified by signature tagged mutagenesis.	a burkholderia pseudomallei mutant which was attenuated in a mouse model of melioidosis was identified by a signature tagged mutagenesis approach. the transposon was shown to be inserted into a gene within the capsular biosynthetic operon. compared with the wild-type bacteria this mutant demonstrated a 10(5)-fold increase in the median lethal dose in a mouse model and it did not react with a monoclonal antibody against high mol. wt polysaccharide of b. pseudomallei. to determine the kinetics of ...	2002	12132769
a possible pitfall in the identification of burkholderia mallei using molecular identification systems based on the sequence of the flagellin flic gene.	amotile burkholderia mallei and motile burkholderia pseudomallei display a high similarity with regard to phenotype and clinical syndromes, glanders and melioidosis. the aim of this study was to establish a fast and reliable molecular method for identification and differentiation. despite amotility, the gene of the filament forming flagellin (flic) could be completely sequenced in two b. mallei strains. only one mutation was identified discriminating between b. mallei and b. pseudomallei. a poly ...	2002	12423776
demonstration of a cell-mediated immune response in melioidosis.	melioidosis is a bacterial infection caused by burkholderia pseudomallei. the aim of this study was to determine whether a cell-mediated adaptive immune response against b. pseudomallei developed in patients who had recovered from melioidosis. lymphocyte proliferation assays were done on peripheral blood mononuclear cells from patients (n=13) and control subjects (n=10) to determine the lymphocyte response to b. pseudomallei antigens. production of interferon-gamma and interleukin-10 was also de ...	2002	12134268
peripheral blood lymphocyte subsets in acute human melioidosis.		2002	12172754
[cloning and expression of 7.55 kb kpni fragment of burkholderia pseudomallei ppm1 plasmid in escherichia coli].	bamhi, sali, psti, and kpni fragments of ppm1 (b. pseudomallei 12.95 kb plasmid) were cloned in e. coli. the recombinant clones carrying a 7.55 kb kpni fragment of ppm1 were highly resistant to several aminoglycosides (streptomycin, kanamycin, and gentamycin) and fluoroguinolones (perfloxacin, ofloxacin). two outer membrane proteins (23 and 27 kda) absent in e. coli and capable to form 120 kda oligomer complex were detected by the western blot method in the strain carrying recombinant ps19 plasm ...	2002	12180019
strategies for pcr based detection of burkholderia pseudomallei dna in paraffin wax embedded tissues.	recently, several cases of melioidosis imported to europe have been reported. the diagnosis of the acute or chronic infection remains challenging. this report describes an optimised protocol for fast and reliable dna preparation for use in two different polymerase chain reaction (pcr) assays, namely: (1) a seminested pcr assay targeting a genus specific sequence of the ribosomal protein subunit 21 (rpsu) gene and (2) a nested pcr assay targeting the gene encoding the filament forming flagellin ( ...	2002	12456780
single gene target bacterial identification. groel gene sequencing for discriminating clinical isolates of burkholderia pseudomallei and burkholderia thailandensis.	proper identification of burkholderia pseudomallei and burkholderia thailandensis is crucial in guiding clinical management of patients with suspected melioidosis, as more than 99% of cases of melioidosis are caused by b. pseudomallei, whereas b. thailandensis is only responsible for causing less than 1% of the cases. however, the difference between the 16s ribosomal rna gene sequences of b. pseudomallei and that of b. thailandensis is only 1%, and is therefore not discriminative enough for dist ...	2002	12458120
seroprevalence of melioidosis in dairy cattle in chiang mai province, northern thailand.	the seroprevalence of melioidosis in dairy cattle in chiang mai province was investigated using of the indirect hemagglutination antibody (iha) method. two hundred and fifty-three samples were tested for serum antibodies to burkholderia pseudomallei. the samples were from a total population of 8,688 dairy cattle in the province; random sampling, stratified by the location of cattle, was used. the seroprevalence was determined as 2% at 1:40 cut-off value, which was estimated to equate to 0.3% to ...	2002	12757220
passive protection against burkholderia pseudomallei infection in mice by monoclonal antibodies against capsular polysaccharide, lipopolysaccharide or proteins.	burkholderia pseudomallei, the aetiological agent of melioidosis, is endemic in south-east asia and northern australia, where it is an important cause of human disease. there is no vaccine available and antibiotic therapy is associated with high relapse rates. a panel of seven monoclonal antibodies (mabs) that recognise capsular polysaccharide, lipopolysaccharide or proteins was produced and their ability to protect mice passively against experimental melioidosis was evaluated. the mabs were cap ...	2002	12466403
burkholderia pseudomallei: abscess in an unusual site.	melioidosis is an infection caused by burkholderia pseudomallei. it is an important human pathogen in tropical area. the clinical manifestations are protean and multisystem involvement. we report an unusual case of melioidosis with abscess at root of mesentery in an elderly, non-insulin dependent diabetic thai women. she presented with prolonged fever and chronic abdominal pain. the early clinical diagnosis was carcinomatous mass with peritonitis. diagnosis of melioidosis arose from the surgical ...	2002	12215696
short report: a rapid method for the differentiation of burkholderia pseudomallei and burkholderia thailandensis.	a rapid method for the identification and differentiation of burkholderia pseudomallei and burkholderia thailandensis colonies is described. it consists of simultaneous use of 2 monoclonal antibody-based latex agglutination test systems. the anti-lipopolysaccharide test reacts with both species, whereas the anti-exopolysaccharide reacts only with b. pseudomallei. compared with classical biochemical tests, the method is highly reproducible and accurate. it is particularly useful for the identific ...	2002	12224587
biochemical-genetic analysis and distribution of des-1, an ambler class a extended-spectrum beta-lactamase from desulfovibrio desulfuricans.	desulfovibrio spp. are gram-negative anaerobes phylogenetically related to bacteroides spp., which are rarely isolated and which are mostly isolated from intra-abdominal abscesses. desulfovibrio desulfuricans clinical isolate d3 had a clavulanic acid-inhibited beta-lactam resistance profile and was resistant to some expanded-spectrum cephalosporins. a beta-lactamase gene, bla(des-1), was cloned from whole-cell dna of isolate d3 and expressed in escherichia coli. purified beta-lactamase des-1, wi ...	2002	12234847
cystic fibrosis and burkholderia pseudomallei infection: an emerging problem?	we recently managed 4 patients with cystic fibrosis who had acquired burkholderia pseudomallei infection after exposure in a region of endemicity. person-to-person transmission between 2 siblings may have occurred; otherwise, the evidence suggests that cystic fibrosis may increase the likelihood of infection with this organism, and patients should be warned of this possibility and cautioned to avoid high-risk activities.	2002	12471591
clinical features of community-acquired pneumonia treated at srinagarind hospital, khon kaen, thailand.	pneumonia is a serious illness associated with significant morbidity and mortality. the interpretation guidelines for pneumonia management requires knowledge of both the clinical presentation of the disease and local epidemiology. we studied the clinical features, initial laboratory results, antibiotic sensitivities, and outcomes of patients diagnosed with acute community-acquired pneumonia between january 1999 and december 2000 at srinagarind hospital. the causative organisms were identified in ...	2002	12236437
cloning of the class d beta-lactamase gene from burkholderia pseudomallei and studies on its expression in ceftazidime-susceptible and -resistant strains.	ceftazidime is the antibiotic of choice for the treatment of melioidosis. ceftazidime-resistant burkholderia pseudomallei have been identified and beta-lactamase production implicated in resistance. in this study, 25 strains of b. pseudomallei (15 clinical and 10 environmental strains) were examined for their ability to yield mutants that overexpress beta-lactamase. ceftazidime-resistant mutants were selected readily at high frequency and displayed four- to eight-fold increases in the mics of ce ...	2002	12356787
activity of five fluoroquinolones against 71 isolates of burkholderia pseudomallei.		2002	12039903
melioidosis.	melioidosis is an important public health problem in some regions, and a potential bioweapon. recent reports confirm that it is endemic in china, taiwan and laos, but the true incidence in most countries is unknown, and the ecology poorly understood. potable water was the source of two recent outbreaks. the epidemiology and clinical manifestations of the disease in australia are similar to those in thailand, although prostatic abscesses and neurological manifestations are more common and parotid ...	2002	11964912
[burkholderia thailandensis: biological properties, identification and taxonomy].	burkholderia pseudomallei-like microorganisms have been isolated from soil and water in regions with endemic melioidosis. these strains have biochemical and antigenic profiles identical to melioidosis agents, except that they differ by virulence and l-arabinose (vir-, ara+). there are minor differences between these species by rrna sequence. dna hybridization and, more so, positive transformation of dna auxotrophic mutants of b. pseudomallei by cell lysates of b. thailandensis and b. mallei conf ...	2002	11904924
[melioidosis].	a 34-year-old woman presented two weeks after a visit to burma with fever peaking up to 39 degrees c, chills, non-productive cough, headache, muscle pain, shortness of breath and a painful swelling on the left lower leg. she was treated immediately with intravenous amoxycillin-clavulanic acid. the gram negative causative agent of melioidosis, burkholderia (previously pseudomonas) pseudomallei, was cultured from samples taken beforehand. the patient then received ceftazidime. she recovered. in vi ...	2002	11980374
sherlock holmes and a biological weapon.		2002	11823558
melioidotic osteomyelitis of the femur occurring in a traveler.		2002	11953265
a simple method to detect and differentiate burkholderia pseudomallei and burkholderia thailandensis using specific flagellin gene primers.	we have previously shown that burkholderia pseudomallei, the causative pathogen of melioidosis, may be discriminated from the closely related non-pathogenic species burkholderia thailandensis by the presence of a 15 base pair deletion in the flagellin gene of b. thailandensis. using specific flagellin gene primers flanking the distinctive region, pcr products of 191 and 176 bp in size were detected for b. pseudomallei and b. thailandensis, respectively. the sensitivity of detection is 20-80 colo ...	2002	12144773
cerebral melioidosis in singapore: a review of five cases.	a variety of neurological manifestations of infection with burkholderia pseudomallei have been described including cerebral abscesses, which are a well-recognized form of neurological melioidosis. the optimal antibiotic therapy for this condition has not been defined; however, combinations of intravenous antibiotics are frequently used in the early stages. five cases of melioidosis involving brain abscesses are described which presented in singapore over the past 3 years (1997-2000), 4 of which ...	2002	11926000
burkholderia pseudomallei infection, or melioidosis, and nephrotic syndrome.		2002	11773478
a mutant of burkholderia pseudomallei, auxotrophic in the branched chain amino acid biosynthetic pathway, is attenuated and protective in a murine model of melioidosis.	using a transposon mutagenesis approach, we have identified a mutant of burkholderia pseudomallei that is auxotrophic for branched chain amino acids. the transposon was shown to have interrupted the ilvi gene encoding the large subunit of the acetolactate synthase enzyme. compared to the wild type, this mutant was significantly attenuated in a murine model of disease. mice inoculated intraperitoneally with the auxotrophic mutant, 35 days prior to challenge, were protected against a challenge dos ...	2002	12183585
phage display of recombinant antibodies toward burkholderia pseudomallei exotoxin.	we have used the phagemid pcomb3h to construct recombinant phages displaying the single chain variable fragment (scfv) towards exotoxin of burkholderia pseudomallei. variable heavy and light chain fragments were amplified from the hybridoma 6e6a8f3b line, with a wide spectrum of primers specific to mouse antibody genes. through overlapping extension polymerase chain reaction, the heavy and light chain fragments were linked to form the scfv which was subsequently cloned into the phage display vec ...	2002	12186782
comparison of rapid, automated ribotyping and dna macrorestriction analysis of burkholderia pseudomallei.	an automated ribotyping device (riboprinter) was used to determine the ribotypes of a collection of burkholderia pseudomallei isolates. in a preliminary evaluation with the restriction enzymes bamhi and ecori, the protocol with ecori was more discriminating. the reproducibilities of the ribotypes obtained with ecori (ecori ribotypes) were determined by testing three levels of bacterial loads. the performance of the manufacturer's software was assessed by comparing the machine-optimized ribotypes ...	2002	12202553
model of differential susceptibility to mucosal burkholderia pseudomallei infection.	burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease with protean clinical manifestations. the major route of infection is thought to be through subcutaneous inoculation of contaminated soil and water, although ingestion and inhalation of contaminated aerosols are also possible. this study examines infection through the intranasal route in a murine model to mimic infection through inhalation. two strains of mice, c57bl/6 and balb/c, exhibit differential suscepti ...	2002	11796576
melioidosis presenting as an infected intrathoracic subclavian artery pseudoaneurysm treated with femoral vein interposition graft.	we present the first case of in situ replacement of an infected subclavian artery using superficial femoral vein and the fourth reported case of an infected arterial pseudoaneurysm caused by pseudomonas pseudomallei. sepsis and hoarseness developed in a 58-year-old man after recent travel to borneo, indonesia. indirect laryngoscopy revealed a paralyzed right vocal cord. computed tomography and arteriography revealed a 6.5-cm pseudoaneurysm of the proximal right subclavian artery. blood cultures ...	2002	11877708
identification of a novel repetitive dna element and its use as a molecular marker for strain typing and discrimination of ara- from ara+ burkholderia pseudomallei isolates.	a novel 10-bp repeat (5'-cgacgcaggc-3')34 was identified in a strain of burkholderia pseudomallei, the first repetitive element found in this species. a pair of primers, based on the flanking sequences of the repetitive region, was used in pcr and dna sequence analysis to determine the genomic structure and distribution of the repetitive element in 76 arabinose- (ara-) and 7 ara+ b. pseudomallei isolates. dna fragments of 400-700 bp were amplified in all ara- isolates. ara+ isolates were charact ...	2002	11800476
cloning and expression of class a beta-lactamase gene blaa(bps) in burkholderia pseudomallei.	the beta-lactamase gene blaa(bps) in burkholderia pseudomallei was cloned and expressed in escherichia coli. bps-1 is a cephalosporinase with an isoelectric point of 7.7. sequence analysis of bps-1 revealed conserved motifs typical of class a beta-lactamases and a relationship to the pena (in b. cepacia) and blai (in yersinia enterocolitica) lineages.	2002	11897607
melioidosis vaccines.	melioidosis is a disease caused by the facultative intracellular pathogen burkholderia pseudomallei iand is associated with a high mortality rate. melioidosis is endemic in the tropics of southeast asia and northern australia and is of worldwide concern, particularly as it is a potential agent of bioterrorism or biological warfare. also of concern is the lack of a fully effective antibiotic regime, as cases of bacteremia have unacceptably high mortality rates and relapse of melioidosis is common ...	2002	12901586
use of 16s rrna gene sequencing for rapid identification and differentiation of burkholderia pseudomallei and b. mallei.	burkholderia pseudomallei and b. mallei, the causative agents of melioidosis and glanders, respectively, are designated category b biothreat agents. current methods for identifying these organisms rely on their phenotypic characteristics and an extensive set of biochemical reactions. we evaluated the use of 16s rrna gene sequencing to rapidly identify these two species and differentiate them from each other as well as from closely related species and genera such as pandoraea spp., ralstonia spp. ...	2003	14532197
bacterial expression of the scfv fragment of a recombinant antibody specific for burkholderia pseudomallei exotoxin.	the scfv antibody towards the burkholderia pseudomallei exotoxin was previously constructed by phage display and exhibited good specificity towards the exotoxin. we report here the optimization of the scfv expression in an e. coli expression system. four different e. coli strains (er2537, tg1, hb2151, and xl1-blue) were examined for optimal expression of the scfv protein. two types of carbon source (i.e. 0.2% glucose and 0.2% glycerol) were also tested for their ability to induce the scfv expres ...	2003	14536033
a case of nonfatal cutaneous melioidosis.	melioidosis is a tropical infectious disease caused by the gram-negative bacterium burkholderia pseudomallei. it is endemic in many parts of the world, including southeast asia, and has a mortality rate of about 45%. we report a case of localized nonfatal cutaneous melioidosis presenting as a persistent ulcer in an otherwise healthy young woman.	2003	14604083
detection of burkholderia pseudomallei in rice fields with pcr-based technique.	burkholderia pseudomallei ara- in rice fields was detected using pcr-based techniques with 16s rna and flagella gene primer sets. the sensitivity of these pcrs was at least 1 cfu/ml of b. pseudomallei ara- preincubated into ashdown's medium for 6 h. b. pseudomallei ara- dna from watery soil were more detectable than from dry soil. the distribution of this dna was mainly found at a depth of 300-600 mm under crop-covered fields, but not detected in the location of soil close to the land surface. t ...	2003	14533484
compensatory increase in ahpc gene expression and its role in protecting burkholderia pseudomallei against reactive nitrogen intermediates.	in the human pathogen burkholderia pseudomallei, katg encodes the antioxidant defense enzyme catalase-peroxidase. interestingly, a b. pseudomallei mutant, disrupted in katg, is hyperresistant to organic hydroperoxide. this hyperresistance is due to the compensatory expression of the alkyl hydroperoxide reductase gene ( ahpc) and depends on a global regulator oxyr. the katg-deficient mutant is also highly resistant to reactive nitrogen intermediates (rni). when overproduced, the b. pseudomallei a ...	2003	14614594
burkholderia pseudomallei: another emerging pathogen in cystic fibrosis.	burkholderia pseudomallei is an important cause of acute fulminant pneumonia and septicaemia in tropical regions of northern australia and south east asia. subacute and chronic forms of the disease also occur. there have been three recent reports of adults with cystic fibrosis (cf) who presumably acquired b pseudomallei infection during extended vacations or residence in either thailand or northern australia.	2003	14645982
localized melioidosis.		2003	14558747
searching for virulence burkholderia pseudomallei genes by immunoscreening the lambda zapii expressed genomic library.	the lambdazap ii expressed genomic library of b. pseudomallei was screened with pooled melioidosis serum preabsorbed with e. coli host cell. the positive clones were detected by using protein a-cdp-star chemiluminescence. all of 14 positive clones reacted with only the pooled absorbed melioidosis serum and not the pooled absorbed normal serum when tested with the plaque dot blot analysis. the expressed genes were detected by using a combination of immunoscreening, bioinformatics and molecular bi ...	2003	15115093
severe community-acquired pneumonia and sepsis caused by burkholderia pseudomallei associated with flooding in puerto rico.	burkholderia pseudomallei (melioidosis) is usually found in endemic areas of southeast asia and northern australia. however, a few cases of confirmed melioidosis indigenous to puerto rico and the americas have been reported previously. we describe the occurrence of a b. pseudomallei infection in a female with insulin-dependent diabetes mellitus exposed to flood waters in puerto rico. we conclude that b. pseudomallei should be considered a potential pathogen in high-risk patients with severe comm ...	2003	15449787
one-step purification of chimeric green fluorescent protein providing metal-binding avidity and protease recognition sequence.	gene fusion technique was successfully applied as a potential approach to create a metal-binding site to assist one-step purification of green fluorescent protein (gfp). the chimeric gfp carrying hexapolyhistidine (h6gfpuv) was purified to homogeneous protein via the immobilized metal affinity chromatography charged with zinc ions. removal of metal tagger could readily be performed by using enterokinase enzyme. engineering of the hexahistidine and enterokinase cleavage sites (ddddk) onto the chi ...	2003	15198344
seronegative bacteremic melioidosis caused by burkholderia pseudomallei with ambiguous biochemical profile: clinical importance of accurate identification by 16s rrna gene and groel gene sequencing.	an aerobic gram-negative bacterium was isolated from the blood and sputum of an 84-year-old, chair-bound nursing home resident with acute bacteremic pneumonia. although the phenotypic characteristics suggested that the bacterium could be burkholderia pseudomallei, the vitek 1 system (gni+), which can successfully identify 99% of b. pseudomallei strains, showed that the bacterium was "unidentified." immunoglobulin g against the lipopolysaccharide (lps) of b. pseudomallei, as detected by an lps-ba ...	2003	12904433
a proposed scoring system for predicting mortality in melioidosis.	melioidosis, due to infection with the environmental organism burkholderia pseudomallei, continues to be associated with high mortality despite improvements in antibiotic therapy. using simple clinical findings and baseline laboratory tests available at the time of admission, we attempted to define those patients with acute melioidosis who were at higher risk of death. using data, collected prospectively from the period october 1989 to june 2002, from patients with acute culture-confirmed melioi ...	2003	15307431
a comparison of antibiotic regimens in the treatment of acute melioidosis in a mouse model.	melioidosis is caused by the gram-negative bacillus burkholderia pseudomallei. most clinical reports of disease are from south-east asia and northern australia. the organism is intrinsically resistant to most commonly available antibiotics. standard therapy includes ceftazidime either alone or in combination with co-trimoxazole. the clinical advantage in adding co-trimoxazole has never been determined; nor has the activity of newer, fourth-generation cephalosporins, such as cefepime, been studie ...	2003	12493790
characterization of the catalase-peroxidase katg from burkholderia pseudomallei by mass spectrometry.	the electron density maps of the catalase-peroxidase from burkholderia pseudomallei (bpkatg) presented two unusual covalent modifications. a covalent structure linked the active site trp111 with tyr238 and tyr238 with met264, and the heme was modified, likely by a perhydroxy group added to the vinyl group on ring i. mass spectrometry analysis of tryptic digests of bpkatg revealed a cluster of ions at m/z 6585, consistent with the fusion of three peptides through trp111, tyr238, and met264, and a ...	2003	12832453
novel selective medium for isolation of burkholderia pseudomallei.	isolation of burkholderia pseudomallei currently relies on the use of ashdown's selective agar (asa). we designed a new selective agar (burkholderia pseudomallei selective agar [bpsa]) to improve recovery of the more easily inhibited strains of b. pseudomallei. b. pseudomallei, burkholderia cepacia, and pseudomonas aeruginosa were used to determine the selectivity and sensitivity of bpsa. bpsa was more inhibitory to p. aeruginosa and b. cepacia and should make recognition of burkholderia species ...	2003	12843080
actin-based motility of burkholderia pseudomallei involves the arp 2/3 complex, but not n-wasp and ena/vasp proteins.	the facultative intracellular bacterium burkholderia pseudomallei induces actin rearrangement within infected host cells leading to formation of actin tails and membrane protrusions. to investigate the underlying mechanism we analysed the contribution of cytoskeletal proteins to b. pseudomallei-induced actin tail assembly. by using green fluorescent protein (gfp)-fusion constructs, the recruitment of the arp2/3 complex, vasodilator-stimulated phosphoprotein (vasp), neural wiskott-aldrich syndrom ...	2003	12780776
flagella are virulence determinants of burkholderia pseudomallei.	burkholderia pseudomallei, a facultatively intracellular pathogen, is a flagellated and motile gram-negative bacterium and is the causative agent of melioidosis in humans. flagella are commonly recognized as important virulence determinants expressed by bacterial pathogens since the motility phenotype imparted by these organelles often correlates with the ability of an organism to cause disease. we used a virulent isolate of b. pseudomallei, khw, to construct an isogenic deletion mutant with a m ...	2003	12654773
flagellum-mediated adhesion by burkholderia pseudomallei precedes invasion of acanthamoeba astronyxis.	in this study we investigated the role of the bacterial flagellum in burkholderia pseudomallei entry to acanthamoeba astronyxis trophozoites. b. pseudomallei cells were tethered to the external amoebic surface via their flagella. mm35, the flagellum-lacking flic knockout derivative of b. pseudomallei nctc 1026b did not demonstrate flagellum-mediated endocytosis in timed coculture, confirming that an intact flagellar apparatus assists b. pseudomallei entry into a. astronyxis.	2003	12654857
[melioidosis: a disease that is advancing].		2003	12659091
relapsing melioidosis as cause of iliac mycotic aneurysm: an indigenous case in taiwan.	melioidosis, an infectious disease caused by burkholderia pseudomallei, an aerobic gram-negative bacillus, is normally transmitted through skin wounds and contact with infected human beings and animals. its primary source is rice paddy soil and stagnant water. melioidosis manifesting as an arterial mycotic aneurysm is rare, and, to our knowledge, infected true and false aneurysms of the iliac artery have never been reported. we report the case of a patient without contact with the normal sources ...	2003	12663992
biodefense-driven murine model of pneumonic melioidosis.	a whole-body mouse model of pneumonic melioidosis was established for future evaluation of biodefense vaccine candidates. the aerosol 50% lethal doses of burkholderia pseudomallei strain 1026b for balb/c and c57bl/6 mice and the times to death, dissemination in organs, and tissue loads after exposure of the mice to low- and high-dose aerosols are reported. in addition, rpsl mutant backgrounds were attenuated in this acute model of disease.	2003	12496217
ubiquity of putative type iii secretion genes among clinical and environmental burkholderia pseudomallei isolates in northern australia.	type iii secretion (ttsi) genes of an hrp (hypersensitivity response and pathogenicity)-like locus were present in all 116 northern australian burkholderia pseudomallei isolates tested but were not detected in other common environmental burkholderia species. pcr of tts1 genes may prove valuable as a diagnostic test [corrected].	2003	12574307
the wbia locus is required for the 2-o-acetylation of lipopolysaccharides expressed by burkholderia pseudomallei and burkholderia thailandensis.	burkholderia pseudomallei and burkholderia thailandensis express similar o-antigens (o-ps ii) in which their 6-deoxy-alpha-l-talopyranosyl (l-6dtalp) residues are variably substituted with o-acetyl groups at the o-2 or o-4 positions. in previous studies we demonstrated that the protective monoclonal antibody, pp-ps-w, reacted with o-ps ii expressed by wild-type b. pseudomallei strains but not by a b. pseudomallei wbia null mutant. in the present study we demonstrate that wbia activity is require ...	2003	12586411
the humoral immune response in melioidosis patients during therapy.	this study was undertaken to identify and quantify the class and subclass antibody responses to the culture filtrate antigen (cfa) of burkholderia pseudomallei in melioidosis patients under long-term maintenance or eradication therapy.	2003	12590329
a case of imported melioidosis presenting as prostatitis.	we report a case of melioidosis in a previously healthy belgian man. he presented with septicemia and prostatic abscesses 1 week after a trip to vietnam. burkholderia pseudomallei was isolated from multiple hemocultures. he was treated successfully with intravenous ceftazidime and trimethoprim-sulfamethoxazole, followed by a per-oral maintenance therapy of amoxicillin-clavulanate with supplementary amoxicillin. there was no need for surgical drainage. this is the second reported case of melioido ...	2003	12590337
catalase-peroxidase katg of burkholderia pseudomallei at 1.7a resolution.	the catalase-peroxidase encoded by katg of burkholderia pseudomallei (bpkatg) is 65% identical with katg of mycobacterium tuberculosis, the enzyme responsible for the activation of isoniazid as an antibiotic. the structure of a complex of bpkatg with an unidentified ligand, has been solved and refined at 1.7a resolution using x-ray synchrotron data collected from crystals flash-cooled with liquid nitrogen. the crystallographic agreement factors r and r(free) are 15.3% and 18.6%, respectively. th ...	2003	12628252
cpg odn enhances uptake of bacteria by mouse macrophages.	unmethylated cpg motif in synthetic oligodeoxynucleotide (cpg odn) or bacterial dna is well recognized for its role in innate immunity, including enhancing production of no and cytokines by macrophages. in the present study, we demonstrated the effect of cpg odn on the phagocytic uptake of bacteria by macrophages. flow cytometric analysis of mouse macrophages (raw 264.7) incubated with fluorescein isothiocyanate (fitc)-labelled burkholderia pseudomallei, salmonella enterica serovar typhi or esch ...	2003	12653838
automated identification systems and burkholderia pseudomallei.		2003	12682195
a transposase-like gene isolated from burkholderia pseudomallei by ddrt-pcr technique.		2003	12682867
structural and functional cellular changes induced by burkholderia pseudomallei rhamnolipid.	in this study we report that extracellular burkholderia pseudomallei rhamnolipid induced cytopathic changes characterized by retraction, rounding up, and, finally, detachment in phagocytic and nonphagocytic cell lines. these changes were due to a progressive reorganization of the f-actin network resulting in impaired cell cycle progression and a reduced phagocytic function of macrophages.	2003	12704181
[melioidosis].		2003	12722256
multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, burkholderia pseudomallei and burkholderia mallei.	a collection of 147 isolates of burkholderia pseudomallei, b. mallei, and b. thailandensis was characterized by multilocus sequence typing (mlst). the 128 isolates of b. pseudomallei, the causative agent of melioidosis, were obtained from diverse geographic locations, from humans and animals with disease, and from the environment and were resolved into 71 sequence types. the utility of the mlst scheme for epidemiological investigations was established by analyzing isolates from captive marine ma ...	2003	12734250
recombinant truncated flagellin of burkholderia pseudomallei as a molecular probe for diagnosis of melioidosis.	current serological tests for melioidosis, using impure or uncharacterized cell antigens from burkholderia pseudomallei, have problems in detection sensitivity and specificity. therefore, we designed and expressed the recombinant flagellin (truncated at both the n- and c-terminal ends), and used the antigen to develop an indirect enzyme-linked immunosorbent assay (elisa) to diagnose melioidosis. comparison of the immunoreactivities of the full-length and truncated flagellins reveals that the tru ...	2003	12738642
crystal structure of the potassium channel kirbac1.1 in the closed state.	the kirbac1.1 channel belongs to the inward-rectifier family of potassium channels. here we report the structure of the entire prokaryotic kir channel assembly, in the closed state, refined to a resolution of 3.65 angstroms. we identify the main activation gate and structural elements involved in gating. on the basis of structural evidence presented here, we suggest that gating involves coupling between the intracellular and membrane domains. this further suggests that initiation of gating by me ...	2003	12738871
regulation of the katg-dpsa operon and the importance of katg in survival of burkholderia pseudomallei exposed to oxidative stress.	homologues of the catalase-peroxidase gene katg and the gene for the non-specific dna binding protein dpsa were identified downstream of oxyr in burkholderia pseudomallei. northern experiments revealed that both katg and dpsa are co-transcribed during oxidative stress. under conditions where the katg promoter is not highly induced, dpsa is transcribed from a second promoter located within the katg-dpsa intergenic region. a katg insertion mutant was found to be hypersensitive to various oxidants. ...	2003	12729890
a family history of deoxyribonuclease ii: surprises from trichinella spiralis and burkholderia pseudomallei.	deoxyribonuclease iialpha (dnase iialpha) is an acidic endonuclease found in lysosomes and nuclei, and it is also secreted. though its caenorhabditis elegans homolog, nuc-1, is required for digesting dna of apoptotic cell corpses and dietary dna, it is not required for viability. however, dnase iialpha is required in mice for correct development and viability, because undigested cell corpses lead to lesions throughout the body. recently, we showed that, in contrast to previous reports, active dn ...	2003	12594037
melioidosis.	melioidosis, which is infection with the gram-negative bacterium burkholderia pseudomallei, is an important cause of sepsis in east asia and northern australia. in northeastern thailand, melioidosis accounts for 20% of all community-acquired septicaemias, and causes death in 40% of treated patients. b pseudomallei is an environmental saprophyte found in wet soils. it mostly infects adults with an underlying predisposing condition, mainly diabetes mellitus. melioidosis is characterised by formati ...	2003	12767750
effects of soil ph, temperature and water content on the growth of burkholderia pseudomallei.	optimum conditions were determined for the growth of burkholderia pseudomallei in natural soils or waters. it grows better in paddy soil, crop-covered and fallow field than in fresh and salty water. although the optimal temperature and ph for the growth were 37 or 42 degrees c, and 6.5 or 7.5 in an environmental-mimicking soil medium, this bacterium can still grow at 4 degrees c, which was suggested to be related with the occurrence of melioidosis in some cold areas. in soil media with water con ...	2003	12800512
septicaemic melioidosis in a tertiary care hospital in south india.	melioidosis and the causative organism burkholderia pseudomallei are being recognized gradually in various centres in india. in the septicaemic form, melioidosis is a serious and life threatening condition which requires early detection and specific treatment to avoid case fatality. a review of patients with septicaemic melioidosis at a tertiary care hospital in south india was carried out with a view to define the clinical features, predisposing conditions, if any, and the outcome.	2003	14575177
dna-dna hybridization study of burkholderia species using genomic dna macro-array analysis coupled to reverse genome probing.	the present study was aimed at simplifying procedures to delineate species and identify isolates based on dna-dna reassociation. dna macro-arrays harbouring genomic dna of reference strains of several burkholderia species were produced. labelled genomic dna, hybridized to such an array, allowed multiple relative pairwise comparisons. based on the relative dna-dna relatedness values, a complete data matrix was constructed and the ability of the method to discriminate strains belonging to differen ...	2003	12807195
burkholderia pseudomallei class a beta-lactamase mutations that confer selective resistance against ceftazidime or clavulanic acid inhibition.	burkholderia pseudomallei, the causative agent of melioidosis, is inherently resistant to a variety of antibiotics including aminoglycosides, macrolides, polymyxins, and beta-lactam antibiotics. despite resistance to many beta-lactams, ceftazidime and beta-lactamase inhibitor-beta-lactam combinations are commonly used for treatment of melioidosis. here, we examine the enzyme kinetics of beta-lactamase isolated from mutants resistant to ceftazidime and clavulanic acid inhibition and describe spec ...	2003	12821450
role of the stationary growth phase sigma factor rpos of burkholderia pseudomallei in response to physiological stress conditions.	the burkholderia pseudomallei rpos gene was identified, and an rpos null mutant was constructed. the mutant was shown to have an increased sensitivity to carbon starvation and oxidative stress. by using rpos-lacz fusions, transcription of rpos was shown to be growth phase regulated, reaching a peak upon entry into stationary phase.	2003	14617667
burkholderia pseudomallei mycotic aneurysm.		2003	12823682
splenic abscess: clinical features, microbiologic finding, treatment and outcome.	splenic abscess is a rare clinical entity but may be underreported. a retrospective study at srinagarind hospital revealed 60 cases of splenic abscess between 1992 and 2001. the causative organisms were identified in 41 cases (68.3%). gram negative bacilli were commonly isolated and burkholderia pseudomallei was the most predominant. diabetes mellitus and leukemia were common underlying diseases found in 46.3 per cent and 9.7 per cent of culture confirmed cases, respectively. the patients usuall ...	2003	12859100
indirect immunofluorescent antibody test for the rapid diagnosis of melioidosis.	culture is the only reliable method available at present for the diagnosis of melioidosis. though serological tests have been described, their value in routine diagnosis is controversial. all indirect immunofluorescent assay (ifa) was therefore evaluated to determine its use in the diagnosis of melioidosis. whole cell antigen prepared from a laboratory isolate of burkholderia pseudomallei was used to assay igg and igm antibodies. fourteen of the 22 (63.6%) culture proven cases had igm antibodies ...	2003	14680201
[effect of cultivation temperature on the extrachromosomal hereditary factors of the causative agent of melioidosis].	the cultivation temperature of burkholderia pseudomallei has been shown to determine both the direction of morphological dissociation and the prophage induction rate. inheriting plasmid replicons was found to depend on the temperature conditions during the growth of these bacteria. no influence of b. pseudomallei plasmids ppm1 and pcm2 on the lysogenic state of the bacterial cell and the formation of different b. pseudomallei variants was noted.	2003	12886599
cutaneous melioidosis and necrotizing fasciitis caused by burkholderia pseudomallei.	in areas where melioidosis is endemic, stress on the healthcare system is substantial. because clinical manifestations are protean, the illness is difficult to diagnose, and cutaneous burkholderia pseudomallei infections can progress to necrotizing fasciitis. while it is and uncommon complication of cutaneous melioidosis, necrotizing fasciitis is potentially fatal and requires aggressive management, including early diagnosis, appropriate antibiotics selection and operative débridement.	2003	14725266
involvement of beta interferon in enhancing inducible nitric oxide synthase production and antimicrobial activity of burkholderia pseudomallei-infected macrophages.	burkholderia pseudomallei is the causative agent of melioidosis, a life-threatening disease that affects both humans and animals. this bacterium is able to survive and multiply inside both phagocytic and nonphagocytic cells. we recently reported that mouse macrophages infected with b. pseudomallei fail to produce a significant level of inducible nitric oxide synthase (inos), a crucial enzyme needed for the cells to control the intracellular growth of this bacterium. in the present study, we exte ...	2003	12761082
melioidotic septic arthritis and its risk factors.	melioidotic septic arthritis is an infection caused by the gram-negative bacillus burkholderia pseudomallei. it is commonly found in northeast thailand. the goal of our study was to identify specific characteristics of patients with melioidotic septic arthritis by comparing them with patients with non-melioidotic septic arthritis and to describe the results of treatment of melioidotic septic arthritis.	2003	12784002
disseminated septicaemic melioidosis: an unusual presentation of masticator space infection.	melioidosis is an infectious disease caused by a saprophytic bacterium, burkholderia pseudomallei. it is endemic to southeast asia and northern australia. the spectrum of melioidosis in humans varies from sub-clinical to overwhelming protean manifestations resembling other acute and chronic bacterial infections. disseminated septicaemia melioidosis presenting as a masticator space infection is reported here. this is germane to those treating diabetic patients with deep neck infections living in, ...	2003	12803798
cellular fatty acid profile distinguishes burkholderia pseudomallei from avirulent burkholderia thailandensis.	burkholderia pseudomallei, the cause of melioidosis, can be distinguished from the closely related but nonpathogenic burkholderia thailandensis by gas chromatography (gc) analysis of fatty acid derivatives. a 2-hydroxymyristic acid derivative (14:0 2oh) was present in 95% of b. pseudomallei isolates and no b. thailandensis isolates. gc mass spectrophotometry confirmed that 2-hydroxymyristic acid was present in b. pseudomallei. gc-fatty acid methyl ester analysis may be useful in distinguishing t ...	2003	14532228
g-csf immunotherapy for treatment of acute disseminated murine melioidosis.	burkholderia pseudomallei, the causative agent of melioidosis, is an important pathogen in tropical regions of australia and southeast asia. antibiotic therapy can be ineffective in patients with acute septicaemic melioidosis. it has been proposed that adjunctive immunotherapy using granulocyte colony stimulating factor (g-csf) combined with antibiotics may provide an alternative approach to antibiotics alone. we have developed a murine model for melioidosis that allows novel treatment approache ...	2003	12892898
a burkholderia pseudomallei type iii secreted protein, bope, facilitates bacterial invasion of epithelial cells and exhibits guanine nucleotide exchange factor activity.	we report the characterization of bope, a type iii secreted protein that is encoded adjacent to the burkholderia pseudomallei bsa locus and is homologous to salmonella enterica sope/sope2. inactivation of bope impaired bacterial entry into hela cells, indicating that bope facilitates invasion. consistent with this notion, bope expressed in eukaryotic cells induced rearrangements in the subcortical actin cytoskeleton, and purified bope exhibited guanine nucleotide exchange factor activity for cdc ...	2003	12897019
the effect of free chlorine on burkholderia pseudomallei in potable water.	chlorine is widely used in public water supplies to provide a disinfection barrier. the effect of chlorine disinfection on the water-borne pathogen burkholderia pseudomallei was assessed using multiple techniques. after exposure to chlorine viable bacteria were undetectable by conventional plate count techniques; however, persistence of b. pseudomallei was verified by flow cytometry and bacteria were recoverable following a simple one-step broth procedure. the minimum residual chlorine concentra ...	2003	14511713
melioidosis: an important cause of pneumonia in residents of and travellers returned from endemic regions.	melioidosis is endemic in south east asia, asia and northern australia. infection usually follows percutaneous inoculation or inhalation of the causative bacterium, burkholderia pseudomallei, which is present in soil and surface water in the endemic region. while 20-36% of melioidosis cases have no evident predisposing risk factor, the vast majority of fatal cases have an identified risk factor, the most important of which are diabetes, alcoholism and chronic renal disease. half of all cases pre ...	2003	14516149
intensity of rainfall and severity of melioidosis, australia.	in a 12-year prospective study of 318 culture-confirmed cases of melioidosis from the top end of the northern territory of australia, rainfall data for individual patient locations were correlated with patient risk factors, clinical parameters, and outcomes. median rainfall in the 14 days before admission was highest (211 mm) for those dying with melioidosis, in comparison to 110 mm for those surviving (p=0.0002). median 14-day rainfall was also significantly higher for those with pneumonia. on ...	2003	14720392
an extracellular zinc metalloprotease gene of burkholderia cepacia.	burkholderia cepacia produces at least one extracellular zinc metalloprotease that may be involved in virulence. a b. cepacia zinc metalloprotease gene was cloned using a burkholderia pseudomallei zinc metalloprotease gene as a probe. the predicted amino acid sequences of these b. cepacia and a b. pseudomallei extracellular zinc metalloproteases indicate that they are similar to the thermolysin-like family of metalloproteases (m4 family of metalloendopeptidases) and they are likely to be secrete ...	2003	12904566
managing sars.		2003	12917310
melioidosis in northern australia, 2001-02.	melioidosis, caused by the gram negative bacterium burkholderia pseudomallei, is endemic in northern australia. using data collated from centres in western australia, the northern territory and queensland, this report describes the epidemiology of this disease between 1 november, 2001 and 31 october, 2002. there were 47 cases seen during this period with an average annual incidence of 5.8 cases per 100,000 population. in indigenous australians, an incidence of 25.5 cases per 100,000 population w ...	2003	12926743
[identification of the causative agents of glanders and melioidosis by polymerase chain reaction].	burkholderia mallei and b. pseudomallei are causative agents of glanders and melioidosis, respectively, i.e. severe and fatal infection diseases of man and animal. the computer-based analysis of the 23s rrna gene sites was used for selecting the primers. two pairs of primers were chosen for the identification of b. mallei and bpseudomallei. dnas from 48 b. pseudomallei and 15 strains of b. mallei, unlike from other geterological bacteria, were positively amplified. therefore, the method of polym ...	2003	12966921