Microb Drug Resist, 2004 Winter, 10(4), 359 - 63 Recurrent Klebsiella pneumoniae mycotic aneurysm in a diabetic patient and emergence of an extended-spectrum beta-lactamase (CTX-M-24)-containing Klebsiella pneumoniae strain after prolonged treatment with first-generation cephalosporins for mycotic aneurysm; Lee CH et al.; A 68-year-old diabetic woman suffered from mycotic aneurysm due to Klebsiella pneumoniae over her abdominal aorta; she received surgical intervention, followed by treatment with first-generation cephalosporins for 6 months . She was hospitalized again 11 months later because of another episode of mycotic aneurysm caused by K . pneumoniae on her thoracic aorta . Fingerprinting generated by pulsed-field gel electrophoresis and infrequent-restriction-site polymerase indicated K . pneumoniae isolates of the identical clonal strain were responsible for these two mycotic-aneurysm episodes . Unfortunately, nosocomial pneumonia developed at the second hospitalization; blood and purposefully sampled feces specimen cultures both grew CTX-M-24-producing K . pneumoniae, which were of the same strain and genetically nonrelated to the K . pneumoniae strain causing mycotic aneurysms earlier . This is the first report on infection due to CTX-M-24-producing K . pneumoniae . It is unclear whether the prolonged use of first-generation cephalosporins in this case selected a strain of enteric organism possessing the ESBL in question, which was capable of passing this ESBL plasmid to the K . pneumoniae strain causing the nosocomial infection . This report suggests that further observation is needed before one can draw a conclusion on the possibility of the selection of ESBL enteric organism by extensive exposure to first-generation cephalosporins. Microb Drug Resist, 2004 Winter, 10(4), 354 - 8 Outbreak of SHV-5 beta-lactamase-producing Klebsiella pneumoniae in a neonatal-pediatric intensive care unit in Spain; Brinas L et al.; The objective was to analyze the beta-lactamase genes and the clonal relationship in a series of 12 clinical Klebsiella pneumoniae strains resistant to ceftazidime or cefotaxime (MIC >/=16 microg/ml) recovered in the neonatalpediatric intensive care unit (ICU) ward of a Spanish hospital during a 1-year period . TEM, SHV, CTX-M, CMY, or FOX beta-lactamase genes were analyzed by PCR and sequencing . The clonal study was performed by pulsed-field gel electrophoresis (PFGE) using XbaI . All but one of the 12 K . pneumoniae strains harbored the bla (SHV-5) gene, and the bla TEM-1a gene was also detected in one of them . These 11 strains belonged to two different clonal types: A (9 strains) and B (2 strains) and were grouped in the subtypes A(1) (6 strains), A(2), A(3), A(4), B(1), and B(2) (1 strain each) . The clonal type A strains were isolated from 9 patients (in five cases from blood) during a 6-month period . The remaining K . pneumoniae strain harbored both the bla (SHV-11) + bla (CTX-M-14) genes and showed the clonal type C . A nosocomial outbreak by a SHV-5-producing multiresistant K . pneumoniae is reported in Spain in a neonatal-pediatric ICU ward . This is the first description of a K . pneumoniae harboring both the bla (SHV-11) and bla (CTX-M-14) genes in Spain. Clin Microbiol Infect, 2005 Jan, 11(1), 31 - 8 Activity of cefepime and carbapenems in experimental pneumonia caused by porin-deficient Klebsiella pneumoniae producing FOX-5 beta-lactamase; Pichardo C et al.; The in-vivo activities of cefepime, imipenem and meropenem against the porin-deficient strain Klebsiella pneumoniae C2 and its derivative K . pneumoniae C2(pMG252) coding for the AmpC-type beta-lactamase FOX-5 were determined . Bactericidal activities were determined with the kill-curve method . A pneumonia model in guinea-pigs was developed, and C(max), t((1/2)) and DeltaT/MIC were calculated for the three agents tested . Animals were treated for 72 h with sterile saline (control group) or with cefepime, imipenem or meropenem (240 mg/kg/day, intramuscularly, three times daily) . Bacterial counts in lungs (log(10) CFU/g tissue) were determined by serial dilution . MICs (mg/L) of cefepime, imipenem and meropenem against K . pneumoniae C2/K . pneumoniae C2(pMG252), determined by macrodilution, were: 0.5/4, 0.5/0.5 and 0.25/0.5, respectively . Bacterial counts in the lungs of animals infected with K . pneumoniae C2 and treated with antimicrobial agents were always lower than in the control group (cefepime, 4.4 +/- 0.5; imipenem, 4.6 +/- 0.4; meropenem, 4.7 +/- 0.5; control group, 5.6 +/- 0.8; p < 0.01) . No significant differences were observed among the groups receiving therapy (p > 0.05) . Bacterial lung clearance was higher in treated animals than in control animals following infection with K . pneumoniae C2(pMG252) (cefepime, 4.5 +/- 0.4; imipenem, 4.0 +/- 0.3; meropenem, 4.6 +/- 0.4; control group, 6.1 +/- 0.6; p < 0.01), with imipenem producing better clearance than either cefepime or meropenem (p < 0.05) . Thus, in the guinea-pig pneumonia model, cefepime, imipenem and meropenem were each effective against the porin-deficient K . pneumoniae strain C2 and its derivative expressing the plasmid-mediated AmpC type beta-lactamase FOX-5. Res Microbiol, 2005 Jan-Feb, 156(1), 76 - 81 Isolation and characterization of a high H(2)-producing strain Klebsiella oxytoca HP1 from a hot spring; Minnan L et al.; A hydrogen-producing bacterial strain was newly isolated from a hot spring and identified as Klebsiella oxytoca HP1 by 16S rRNA gene sequence analysis and detection by BioMerieux Vitek . Important parameters, including substrates, starting pH of culture, temperature and oxygen concentration for batch ferment hydrogen production, were investigated . Among different sugars, glucose and sucrose were the preferred substrates for hydrogen production . The optimal starting pH of culture was about 7.0 . The activity was drastically reduced in a prolonged fermentation due to the accumulation of organic acids . Increasing temperatures (from 25 to 35 degrees C) improved the hydrogen production activity . K . oxytoca HP1 produced hydrogen under different concentrations (1-10%) of oxygen in the gas phase, indicating that it is highly resistant to oxygen inhibition . Under batch ferment conditions, the maximal hydrogen production activity, rate and yield were obtained as 9.6 mmol/g dw h, 87.5 ml/l h and 1.0 mol/mol glucose (conversion 16.7%), respectively . In continuous hydrogen production, the maximum activity, rate and yield were 15.2 mmol/g dw h, 350.0 ml/l h and 3.6 mol/mol sucrose (conversion 32.5%), respectively . These results indicate that K . oxytoca HP1 is an ideal hydrogen producer. J Clin Microbiol, 2005 Jan, 43(1), 516 - 9 First isolation of metallo-beta-lactamase-producing multiresistant Klebsiella pneumoniae from a patient in Brazil; Lincopan N et al.; A multiresistant Klebsiella pneumoniae isolate was taken from the blood of a 75-year-old patient with nosocomial pneumonia who developed septic shock and failed therapy with imipenem . The isolate presented an MIC of imipenem of 128 microg/ml, and the production of a metallo-beta-lactamase was confirmed by phenotypic and genotypic techniques . We here report, for the first time, the detection of a metalloenzyme (IMP-1)-producing K . pneumoniae clinical strain in Latin America . The gene responsible for this phenotype was found to be bla(IMP-1), carried in a class 1 integron. J Clin Microbiol, 2005 Jan, 43(1), 494 - 6 Discrepancies and interpretation problems in susceptibility testing of VIM-1-producing Klebsiella pneumoniae isolates; Giakkoupi P et al.; Susceptibilities to beta-lactam antibiotics of five VIM-1-producing Klebsiella pneumoniae isolates were determined by broth microdilution, Etest, disk diffusion, and the automated systems Vitek 2, Phoenix, and MicroScan . Significant discrepancies were observed in the determination of susceptibility to imipenem and meropenem . Interpretation problems by the automated systems were also noted. Rev Med Chil, 2004 Oct, 132(10), 1173 - 8 {Resistance to gentamicin, amikacin and ciprofloxacin among nosocomial isolates of klebsiella pneumoniae subspecie pneumoniae producing extended spectrum beta-lactamases}; Diaz PQ et al.; BACKGROUND: Klebsiella pneumoniae is a pathogenic bacterium frequently isolated from nosocomial samples, specially the subspecie pneunonlae, with extensive antibiolic resistance profiles, including third generation cepbhalosporiis, aminoglycosides and quinolones . This is specially true for those strains producing extended spectrum beta lactamases (ESBL) . AIM: To investigate the susceptibility to gentamicin, amikacin and ciprofloxacin and the presence of some aminogloycoside modifying enzyme (AMEs) among nosocomial strains of K pneumoniae subspecie pneumoniae producing ESBL . MATERIAL AND METHODS: The antibiotic resistant patterns and the level of resistance (minimal inhibitory concentration, MIC) of 100 strains, isoklted from sel ,eal bospitals of dcifferent Chilean cities, were deterl,in,ed . Tbe presence of some aminoglycosides modifying enzyme (AMEs) was investigated by PCR . RESULTS: Sixty five percent of strains were resistant to gentamicin, 47% were resistant to amikacin, and 29% were resistant to ciprofloxacin . The most frequent AMEs genes detected were the aac(6')-Ib gene (6'N-Acetyltransferase type Ib enzyme) in 69% of strains, conferring resistance to amikacin, kanamycin, tobramycin, and nieoniycin, and the gene aac(3)-IIa (3-Acetyltransferase type 3-IIa enzyme), in 36% of strains, conferring resistance to gentamlicin . CONCLUSIONS: Among nosocomial strains of K pneumoniae subspecie pneumoniae isolaterd from Chilean hospitals, there is an association between the production of ESBL and the resistance to others antimicrobial agents, especially aminoglycosides . Nevertheless, 71% of isolates are susceptible to ciprofloxacin. Appl Microbiol Biotechnol . 2005 Jan 4; {Epub ahead of print} Application of cyanide hydrolase from Klebsiella sp . in a biosensor system for the detection of low-level cyanide; Mak KK et al.; A partially purified preparation of cyanide hydrolase (cyanidase) from a bacterium, Klebsiella sp., was applied as a biocatalyst in a biosensor system for low-level cyanide detection . In the biosensor system cyanide hydrolase converts cyanide into formate and ammonia . The formate produced in the cyanide degradation was detected with a formate biosensor, in which formate dehydrogenase (FDH; E.C . 1.2.1.2) was co-immobilized with salicylate hydroxylase (SHL; E.C . 1.14.13.1) on a Clark electrode . The principle of the formate sensor is that FDH converts formate into carbon dioxide using beta-nicotinamide adenine dinucleotide hydrate (NAD(+)) . The corresponding NADH produced is then oxidized to NAD(+) by SHL using salicylate and oxygen . The oxygen consumption is monitored with the Clark electrode . The optimum buffer pH and temperature for the enzymatic hydrolysis of potassium cyanide were studied . The preliminary experiments including the pretreatment of cyanide with cyanide hydrolase and then detection by the formate sensor gave a detection limit at 7.3 mumol l(-1) cyanide . The linear range of the calibration curve was between 30 mumol l(-1) and 300 mumol l(-1) cyanide. Diagn Microbiol Infect Dis, 2005 Jan, 51(1), 1 - 7 Evolution and dissemination of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae: Epidemiology and molecular report from the SENTRY Antimicrobial Surveillance Program (1997-2003); Dipersio JR et al.; During 2001, occurrences of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae isolates were detected in a single medical center (Hospital A) from the SENTRY Antimicrobial Surveillance Program that became endemic in long-term acute care areas and in the intensive care unit in 2002-2003 . Between 2001 and 2003, 123 patients were infected or colonized with ESBL-positive K . pneumoniae . Resistance profiles were determined by reference broth microdilution methods, and automated ribotyping and pulsed-field gel electrophoresis (PFGE) were performed . The ESBL-positive K . pneumoniae isolates were resistant to aztreonam, ceftazidime, aminoglycosides, and trimethoprim/sulfamethoxazole and susceptible to ciprofloxacin and tetracycline . In 1997, 1998, and 2000, 9 ESBL-producing K . pneumoniae strains from 2 New York City hospitals shared the same antibiograms and ribotype (204.2) as the strains from Hospital A . PFGE patterns divided Hospital A isolates into 2 subtypes (A and A(1)) and 3 New York City strains were similar to the Hospital A isolates (A(2), A(3), and A(4)) . Isoelectric focusing studies of 1 New York City isolate (A(4)) revealed pIs at 5.4, 7.7, and 8.2 . PCR and sequencing results from 1 strain of each Hospital A and 1 New York PFGE pattern determined that TEM-1 and SHV-5 (ESBL) were present in all strains . In addition, 2 New York isolates from 1998 (A(3) and A(4)) also had an OXA-2 enzyme . ESBL-producing K . pneumoniae isolates with ribotype 204.2 from SENTRY Program sites have been recognized in New York only since 1997 and in Hospital A beginning in 2001 . The similarities of the antibiogram and epidemiological patterns suggest that these isolates have persisted over time and may have evolved into different but genetically related endemic ESBL-positive K . pneumoniae clones that have the ability to cause sustained epidemic outbreaks in US medical centers. Salud Publica Mex, 2004 Nov-Dec, 46(6), 524 - 8 {Characterization of SHV-5 beta-lactamase-producing Klebsiella pneumoniae in an intensive care unit}; Andrade V et al.; OBJECTIVE: To perform the molecular characterization of Klebsiella pneumoniae isolates from pediatric patients and health care workers at the intensive care unit of a tertiary care hospital in Mexico City . MATERIAL AND METHODS: Fifteen Klebsiella pneumoniae isolates collected during an outbreak in June 1996 were analyzed; eight were from patients and seven from health care workers of Mexico's Children's Hospital . Characterization of isolates was carried out by pulsed field gel electrophoresis (PFGE), random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) and serotyping, beta-Lactamase isoelectric focusing (IEF), and nucleotide sequencing of PCR products . RESULTS: Serotype 61 was predominant and correlated with genomic fingerprints of RAPD and PFGE in 11 of 15 isolates . One SHV-5-producer predominant clone with a high case-fatality rate was identified . CONCLUSIONS: Molecular biology techniques are useful tools to characterize the K . pneumoniae clone isolated from patients and health care workers, suggesting potential cross-transmission . These data call for strengthening control programs to prevent dissemination of nosocomial infections in the studied hospital. Infect Immun, 2005 Jan, 73(1), 532 - 45 Central role of toll-like receptor 4 signaling and host defense in experimental pneumonia caused by Gram-negative bacteria; Schurr JR et al.; Toll-like receptor 4 (TLR4) has been identified as a receptor for lipopolysaccharide . However, the precise role of TLR4 in regulating gene expression in response to an infection caused by gram-negative bacteria has not been fully elucidated . The role of TLR4 signaling in coordinating gene expression was assessed by gene expression profiling in lung tissue in a mouse model of experimental pneumonia with a low-dose infection of Klebsiella pneumoniae . We analyzed four mouse strains: C57BL/6 mice, which are resistant to bacterial dissemination; 129/SvJ mice, which are susceptible; C3H/HeJ mice, which are susceptible and have defective TLR4 signaling; and their respective control strain, C3H/HeN (intermediate resistance) . At 4 h after infection, C57BL/6 and C3H/HeN mice demonstrated the greatest number of genes, with 67 shared induced genes which were TLR4 dependent and highly associated with the resistance phenotype . These genes included cytokine and chemokine genes required for neutrophil activation or recruitment, growth factor receptors, MyD88 (a critical adaptor protein for TLR signaling), and adhesion molecules . TLR4 signaling accounted for over 74% of the gene expression in the C3H background . These data suggest that early TLR4 signaling controls the vast majority of gene expression in the lung in response to an infection caused by gram-negative bacteria and that this subsequent gene expression determines survival of the host. Antimicrob Agents Chemother, 2005 Jan, 49(1), 276 - 80 Pharmacodynamic profile of ertapenem against Klebsiella pneumoniae and Escherichia coli in a murine thigh model; Maglio D et al.; The pharmacodynamic profile of ertapenem was evaluated in a neutropenic mouse thigh infection model . Extended-spectrum beta-lactamase (ESBL)-positive and ESBL-negative clinical strains of Escherichia coli and Klebsiella pneumoniae were studied . MICs ranged from 0.0078 to 0.06 microg/ml with standard inoculum tests . Ertapenem doses were administered once to five times daily to achieve various exposures, reported as the percentage of the dosing interval that the concentration of free ertapenem was in excess of the MIC (%T>MIC(free)) . Mean values for the static exposure and 80% maximally effective exposure (ED(80)) were 19% (range, 2 to 38%) and 33% (range, 13 to 65%) T>MIC(free), respectively . Differences in exposure requirements based on the presence of an ESBL resistance mechanism or bacterial species were not evident . In addition, experiments using a 100-fold higher inoculum did not decrease the magnitude of the reduction in bacterial density from baseline achieved compared to lower-inoculum studies . The pharmacodynamic parameter of %T>MIC(free) correlated well with bactericidal activity for all isolates, and the static and ED(80) exposures are consistent with those reported previously for carbapenems. Antimicrob Agents Chemother, 2005 Jan, 49(1), 256 - 63 bla(SHV) Genes in Klebsiella pneumoniae: different allele distributions are associated with different promoters within individual isolates; Hammond DS et al.; Extended-spectrum beta-lactamases (ESBLs) emerge by point mutation from non-extended-spectrum precursors . The aims of this study were to reveal the basis for variations in resistance levels found in a collection of 21 Klebsiella pneumoniae clinical isolates from Brisbane, Australia . Previous studies have shown that 20 of these isolates possess bla(SHV-11), bla(SHV-2a), and/or bla(SHV-12), and there is an association between the copy numbers of the ESBL-encoding genes and resistance levels . In this study, a real-time PCR method for interrogating the polymorphic sites at codons 238 and 240 was developed, and this confirmed the relationship between mutant gene copy numbers and resistance levels . The bla(SHV) promoter region was cloned from one of the ESBL-expressing isolates, and this showed that bla(SHV) genes exist downstream of two different promoters within this single isolate . These promoters have both been reported previously, and they differ by virtue of the presence or absence of an IS26 insertion . The bla(SHV) copy numbers in cis with the different promoters were measured, and the copy number of the IS26 promoter was correlated with resistance levels . Cloning and analysis of PCR products showed that different bla(SHV) variants existed in cis with individual promoters in individual isolates but that mutant genes were more abundant downstream of the IS26 promoter . There were no ESBL-positive isolates without this promoter . It was concluded that bla(SHV) in cis with the IS26 promoter is located on an amplifiable replicon, and the presence of the IS26 insertion may facilitate the acquisition of an ESBL-positive phenotype. Acta Pharm, 2004 Sep, 54(3), 231 - 42 Preparation and evaluation of a new erythromycin derivative -- erythromycin taurate; Manna PK et al.; Erythromycin taurate, a new derivative of erythromycin, was prepared by reacting erythromycin base with tauric acid and its physico-chemical and biological properties were evaluated . The derivative has reasonably good solubility in organic solvents . The partition coefficient values in chloroform/water 1.17 and octanol/water 1.16 systems indicate its good distribution in various tissues in vivo . The in vitro antimicrobial potency of the derivative (833.33 microg mg(-1)) is higher than that of the existing derivatives such as erythromycin estolate, erythromycin stearate, erythromycin ethyl succinate, erythromycin gluceptate, erythromycin lactobionate . The antimicrobial spectrum is comparable to that of the parent compound . Our results indicate that erythromycin taurate has a high potential for possible clinical application and is more efficient against Escherichia coli and Klebsiella pneumoniae than the parent base. Beijing Da Xue Xue Bao, 2004 Dec, 36(6), 637 - 41 {Genotype of extended-spectrum beta-lactamases produced by antibiotic-resistant Escherichia coli and Klebsiella pneumoniae in surgical intensive care unit.}; Wang PY et al.; OBJECTIVE:To describe the antibiotic resistant mode of extended-spectrum beta-lactamases(ESBLs) producing Escherichia coli (E . coli.) and Klebsiella pneumoniae (KPn) in surgical intensive care unit(SICU), and to implore the molecular epidemiology of ESBLs coding genes of these strains . METHODS: The minimal inhibitory concentrations (MICs) at antibiotics were examined by agar dilution method . The ESBLs coding genes were amplified by TEM, SHV and CTX-M specific primers . Amplicons of such genes with conjugates' plasmids as templates were sequenced . RESULTS: In vitro susceptibility tests of ESBLs producing strains showed a high level of resistance to most of the beta-lactam biotics, especially cefotaxime . 93.5% of these ESBLs positive strains contained CTX-M group genes,and 38.7% of the strains contained SHV genes . By sequencing, some genotype were determined: TEM-1, CTX-M-1,3,14,22 . CONCLUSION: ESBLs producing strains were resistant to most of the beta-lactam biotics . The most prevalent ESBLs genotype of ESBLs produced by E coli and KPn in SICU was CTX-M subgroup . The most probable reason might be the extensive use of cefotaxime. Protein Expr Purif, 2005 Jan, 39(1), 107 - 15 Purification and characterization of the recombinant arylsulfatase cloned from Pseudoalteromonas carrageenovora; Kim DE et al.; Arylsulfatase cloned from a marine aerobic Gram-negative bacterium, Pseudoalteromonas carrageenovora, was overexpressed in Escherichia coli with 10muM IPTG induction . The expressed recombinant arylsulfatase was purified to homogeneity from the harvested cells through osmotic disruption and column chromatography methods, such as DEAE-cellulose anion exchange chromatography and Heparin-Sepharose affinity chromatography . The purified arylsulfatase was kinetically characterized using the synthetic substrate of phenolic ester, p-nitrophenyl sulfate (pNPS) . One unit of arylsulfatase catalyzes the liberation of 1.0mumol p-nitrophenol from pNPS per minute . The purified enzyme has a specific activity of 468 U/mg with a purification yield of 27% from the cell lysate, and exhibited an estimated molecular mass of 33kDa in SDS-PAGE analysis . The precursor polypeptide of 36kDa was processed by releasing a putative signal peptide, and the mature arylsulfatase of 33.1kDa with a N-terminal sequence of S-E-T-K-N was trafficked to periplasmic space . The enzyme had optimum reaction conditions for activity at pH 7.0 and at a temperature range of 40-45 degrees C . The apparent K(M) and k(cat) of the enzyme for hydrolysis of pNPS at pH 7.0 and at 45 degrees C were determined to be 1.15mM and 1000s(-1), respectively . Based on inhibitor studies along with optimal pH values and preferential periplasmic location of the enzyme, we suggest that the recombinant arylsulfatase from P . carrageenovora is probably similar to the Klebsiella sulfatase with serine residue in the active site. Am J Respir Crit Care Med . 2004 Dec 10; {Epub ahead of print} A Role for Hydroxy-Methylglutaryl Coenzyme A Reductase in Pulmonary Inflammation and Host Defense; Fessler MB et al.; Rationale: A growing literature indicates that hydroxy-methylglutaryl coenzyme A reductase inhibitors ('statins') modulate pro-inflammatory cellular signaling and functions . No studies to date, however, have addressed whether statins modulate pulmonary inflammation triggered by aerogenic stimuli, nor whether they affect host defense . Objectives: To test whether lovastatin modulates lipopolysaccharide-induced pulmonary inflammation and anti-bacterial host defense . Methods: In order to address these questions, and to confirm any effect of statins as dependent upon inhibition of hydroxy-methylglutaryl coenzyme A reductase, we treated C57Bl/6 mice with three oral doses of 10 mg/kg lovastatin (or vehicle) and three intraperitoneal doses of 10 mg/kg mevalonic acid (or saline), and then exposed them to: 1) aerosolized lipopolysaccharide; 2) intratracheal CXC chemokine KC; or 3) intratracheal Klebsiella pneumoniae . Measurements and Main Results: Lipopolysaccharide- and KC-induced airspace neutrophils were reduced by lovastatin, an effect that was blocked by mevalonic acid co-treatment . Lovastatin was also associated with reduced parenchymal myeloperoxidase and microvascular permeability, and altered airspace and serum cytokines following lipopolysaccharide . Native pulmonary clearance of K . pneumoniae was inhibited by lovastatin, and extrapulmonary dissemination enhanced, both reversibly with mevalonic acid . Ex vivo studies of neutrophils isolated from lovastatin-treated mice confirmed inhibitory effects upon rac activation, actin polymerization, chemotaxis, and bacterial killing . Conclusions: Lovastatin attenuates pulmonary inflammation induced by aerosolized lipopolysaccharide, and impairs host defense. J Med Microbiol, 2005 Jan, 54(1), 7 - 13 Compensatory response of IL-1 gene knockout mice after pulmonary infection with Klebsiella pneumoniae; Tanabe M et al.; This study was designed to determine the role of interleukin (IL)-1 in the inflammatory response against experimentally induced pneumonia caused by Klebsiella pneumoniae . The host immune responses of IL-1 gene knockout (IL-1 KO) mice and immunocompetent wild-type (WT) mice were compared after pulmonary infection with K . pneumoniae . There were no significant differences between the survival rates and viable bacterial counts in lungs and blood of IL-1 KO and WT mice after pulmonary infections under different conditions . Histopathological analysis showed a similar inflammatory response in both groups of mice . However, in the early stage of infection, the level of tumour necrosis factor alpha (TNF-alpha) in homogenized lungs of IL-1 KO mice was significantly higher than in WT mice . To determine the role of endogenous TNF-alpha in the recovery of the defence mechanism in IL-1 KO mice, mice were treated with an anti-TNF-alpha mAb before infection with K . pneumoniae . The results revealed a significantly lower survival rate of anti-TNF-alpha mAb-treated IL-1 KO mice than BSA-treated IL-1 KO mice . The data suggest that compensatory production of TNF-alpha in IL-1 KO mice contributes to the host defence against K . pneumoniae infection. Acta Crystallogr D Biol Crystallogr, 2004 Dec, 60(Pt 12 Pt 2), 2352 - 4 Epub 2004 Dec. Expression, crystallization and preliminary X-ray crystallographic studies of Klebsiella pneumoniae maltohexaose-producing alpha-amylase; Momma M et al.; A recombinant form of Klebsiella pneumoniae maltohexaose-producing alpha-amylase has been overexpressed in Escherichia coli and purified to homogeneity . Crystals were obtained at 293 K by the microbatch technique using 80 mM sodium/potassium phosphate buffer pH 6.2 containing 8% polyethylene glycol 3000, 4% polyethylene glycol 3350 and 40 mM sodium thiocyanate . Crystals of the overexpressed recombinant enzyme diffracted to better than 2.5 A resolution at 95 K using a synchrotron-radiation source . The crystals belong to the primitive monoclinic space group P2(1), with unit-cell parameters a = 74.8, b = 107.6, c = 82.2 A, beta = 96.2 degrees . Assuming the presence of two molecules per asymmetric unit, the V(M) value for the crystal was 2.3 A(3) Da(-1), indicating a solvent content of 47%. J Clin Microbiol, 2004 Dec, 42(12), 5715 - 21 Phenotypic and molecular detection of CTX-M-beta-lactamases produced by Escherichia coli and Klebsiella spp; Pitout JD et al.; Organisms producing CTX-M-beta-lactamases are emerging around the world as a source of resistance to oxyiminocephalosporins such as cefotaxime (CTX) . However, the laboratory detection of these strains is not well defined . In this study, a molecular detection assay for the identification of CTX-M-beta-lactamase genes was developed and used to investigate the prevalence of these enzymes among clinical isolates of Escherichia coli and Klebsiella species in the Calgary Health Region during 2000 to 2002 . In addition, National Committee for Clinical Laboratory Standards (NCCLS) recommendations were evaluated for the ability to detect isolates with CTX-M extended-spectrum beta-lactamases (ESBLs) . The PCR assay consisted of four primer sets and demonstrated 100% specificity and sensitivity for detecting different groups of CTX-M-beta-lactamases in control strains producing well-characterized ESBLs . Using these primer sets, 175 clinical strains producing ESBLs were examined for the presence of CTX-M enzymes; 24 (14%) were positive for bla(CTX-M-1-like) genes, 95 (54%) were positive for bla(CTX-M-14-like) genes, and the remaining 56 (32%) were negative for bla(CTX-M) genes . Following the NCCLS recommendations for ESBL testing, all of the control and clinical strains were detected when screened with cefpodoxime and when both cefotaxime and ceftazidime with clavulanate were used as confirmation tests. Clin Infect Dis, 2004 Dec 1, 39(11), 1654 - 9 Epub 2004 Dec 1. Pyogenic liver abscess: recent trends in etiology and mortality; Rahimian J et al.; BACKGROUND: Pyogenic liver abscess, a potentially life-threatening disease, has undergone significant changes in epidemiology, management, and mortality over the past several decades . METHODS: We reviewed the data for patients admitted to Bellevue Hospital and New York University Downtown Hospital (New York, New York) over a 10-year period . RESULTS: Of 79 cases reviewed, 43% occurred in patients with underlying biliary disease . The most common symptoms were fever, chills, and right upper quadrant pain or tenderness . The most common laboratory abnormalities were an elevated white blood cell count (in 68% of cases), temperature >or=38.1 degrees C (90%), a low albumin level (70.2%), and an elevated alkaline phosphatase level (67%) . Seventy percent of the abscesses were in the right lobe, and 77% were solitary . Klebsiella pneumoniae was identified in 41% of cases in which a pathogen was recovered . Eighteen (50%) of 36 Asian patients had K . pneumoniae isolated, in contrast to 6 (27.3%) of 22 non-Asian patients (not statistically significant) . Fifty-six percent of cases involved treatment with percutaneous drainage . Although prior reports noted mortality of 11%-31%, we observed only 2 deaths (mortality, 2.5%) . CONCLUSIONS: The data suggest that K . pneumoniae has become the predominant etiology of pyogenic liver abscess and that mortality from this disease has decreased substantially. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4829 - 34 Molecular and kinetic comparison of the novel extended-spectrum beta-lactamases CTX-M-25 and CTX-M-26; Munday CJ et al.; CTX-M-25 is a novel extended-spectrum beta-lactamase isolated from a single Canadian Escherichia coli isolate . Susceptibility testing demonstrated that this enzyme confers resistance to both cefotaxime and ceftazidime, but the level of resistance was reduced with the addition of beta-lactamase inhibitors . The bla(CTX-M-25) gene was detected on a 111-kb plasmid . It is a member of the CTX-M-8 group and has the closest amino acid identity (99%; three amino acid substitutions) with CTX-M-26 . The bla(CTX-M-26) gene was detected on a 100-kb plasmid isolated from a Klebsiella pneumoniae strain from the United Kingdom, and plasmid profiling revealed that it showed some homology to the bla(CTX-M-25)-harboring plasmid . Both CTX-M genes were located downstream of ISEcp1, although the copy upstream of bla(CTX-M-25) was disrupted by IS50-A . Comparative kinetic studies of recombinant CTX-M-25 and CTX-M-26 enzymes showed that CTX-M-25 has a higher level of ceftazidime hydrolysis (kcat values, 33 and 0.005 s(-1) for CTX-M-25 and CTX-M-26, respectively). Antimicrob Agents Chemother, 2004 Dec, 48(12), 4574 - 81 Bloodstream infections due to extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae: risk factors for mortality and treatment outcome, with special emphasis on antimicrobial therapy; Kang CI et al.; This study was conducted to evaluate risk factors for mortality and treatment outcome of bloodstream infections due to extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae (ESBL-EK) . ESBL production in stored K . pneumoniae and E . coli blood isolates from Jan 1998 to Dec 2002 was phenotypically determined according to NCCLS guidelines and/or the double-disk synergy test . A total of 133 patients with ESBL-EK bacteremia, including 66 patients with ESBL-producing K . pneumoniae and 67 with ESBL-producing E . coli, were enrolled . The overall 30-day mortality rate was 25.6% (34 of 133) . Independent risk factors for mortality were severe sepsis, peritonitis, neutropenia, increasing Acute Physiology and Chronic Health Evaluation II score, and administration of broad-spectrum cephalosporin as definitive antimicrobial therapy (P < 0.05 for each of these risk factors) . In 117 of the 133 patients, excluding 16 patients who died within 3 days after blood culture sample acquisition, the 30-day mortality rates according to definitive antibiotics were as follows: carbapenem, 12.9% (8 of 62); ciprofloxacin, 10.3% (3 of 29); and others, such as cephalosporin or an aminoglycoside, 26.9% (7 of 26) . When patients who received appropriate definitive antibiotics, such as carbapenem or ciprofloxacin, were evaluated, mortality in patients receiving inappropriate empirical antimicrobial therapy was found not to be significantly higher than mortality in those receiving appropriate empirical antimicrobial therapy (18.9 versus 15.5%; P = 0.666) . Carbapenem and ciprofloxacin were the most effective antibiotics in antimicrobial therapy for ESBL-EK bacteremia . A delay in appropriate definitive antimicrobial therapy was not associated with higher mortality if antimicrobial therapy was adjusted appropriately according to the susceptibility results . Our data suggest that more prudent use of carbapenem as empirical antibiotic may be reasonable. Am J Kidney Dis, 2004 Dec, 44(6), e102 - 6 Expanded-spectrum beta-lactamase producing Klebsiella pneumoniae-related peritonitis in a patient on peritoneal dialysis; Yang CC et al.; While hospitalized for pneumonia with ventilator-dependent respiratory failure, a 45-year-old man on continuous ambulatory peritoneal dialysis (CAPD) had nosocomial peritonitis secondary to infection by expanded spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-Kp) . He was treated successfully with a 3-week course of intraperitoneal (IP) flomoxef therapy without subsequent relapse, loss of peritoneal catheter, ultrafiltration failure, or dialysis inadequacy . The International Consensus Panel recommends IP ceftazidime as the treatment of choice for CAPD patients suffering Klebsiella species-related peritonitis . However, the most appropriate form of IP antibiotic therapy and the outcomes for expanded-spectrum beta-lactamase (ESBL)-producing bacteria-related peritonitis for CAPD patients have not been established yet . Further, the ability to correctly report minimal inhibitory concentrations (MICs) of ceftazidime for ESBL bacteria in the resistant range varies between laboratories, making the diagnosis of ESBL-Kp-related CAPD peritonitis more complex and difficult . Thus, it appears reasonable to suggest that its incidence is probably underestimated and its significance ignored . The authors suggest that a 3-week IP treatment with flomoxef, a synthesized oxacephem, with loading and maintenance doses of 250 and 125 mg/L, respectively, is effective and safe for ESBL-Kp-related peritonitis in these patients . ESBL producing bacterial infection should be considered as a possible cause of overt CAPD-related peritonitis . Early detection of ESBLB pathogens and institution of effective antibiotic treatment may improve the prognosis. Infect Immun, 2004 Dec, 72(12), 7107 - 14 Capsule polysaccharide mediates bacterial resistance to antimicrobial peptides; Campos MA et al.; The innate immune system plays a critical role in the defense of areas exposed to microorganisms . There is an increasing body of evidence indicating that antimicrobial peptides and proteins (APs) are one of the most important weapons of this system and that they make up the protective front for the respiratory tract . On the other hand, it is known that pathogenic organisms have developed countermeasures to resist these agents such as reducing the net negative charge of the bacterial membranes . Here we report the characterization of a novel mechanism of resistance to APs that is dependent on the bacterial capsule polysaccharide (CPS) . Klebsiella pneumoniae CPS mutant was more sensitive than the wild type to human neutrophil defensin 1, beta-defensin 1, lactoferrin, protamine sulfate, and polymyxin B . K . pneumoniae lipopolysaccharide O antigen did not play an important role in AP resistance, and CPS was the only factor conferring protection against polymyxin B in strains lacking O antigen . In addition, we found a significant correlation between the amount of CPS expressed by a given strain and the resistance to polymyxin B . We also showed that K . pneumoniae CPS mutant bound more polymyxin B than the wild-type strain with a concomitant increased in the self-promoted pathway . Taken together, our results suggest that CPS protects bacteria by limiting the interaction of APs with the surface . Finally, we report that K . pneumoniae increased the amount of CPS and upregulated cps transcription when grown in the presence of polymyxin B and lactoferrin. Biomedica, 2004 Sep, 24(3), 252 - 61 {Molecular epidemiology of nosocomial infection by extended-spectrum beta-lactamases-producing Klebsiella pneumoniae}; Espinal PA et al.; Molecular epidemiology applied to the study of nosocomial infection has been fundamental in formulating and evaluating control methods . From patients in a level 3 Bogota hospital, Klebsiella pneumoniae samples were isolated that produced extended-spectrum beta-lactamases (ESBL) . Each of 15 isolates was characterized microbiologically and by molecular characters realized by pulsed field gel electrophoresis (PFGE) and by repetitive-DNA sequences amplification (REP-PCR) . Antimicrobial susceptibility and ESBL production was determined in accordance with NCCLS guidelines . The beta-lactamases were evaluated by isoelectric-focusing and PCR . Twelve (80%) of the isolates were associated with nosocomial infection; 11 of them were from intensive care units . The antibiotic susceptibility displayed 13 resistance patterns--87% presented co-resistance to amikacin, 53% to gentamicin, 33% to ciprofloxacin, 40% to cefepime, 67% to piperacillin/tazobactam, 60% to trimethoprim/sulfamethoxazole and 47% to chloranphenicol . All were sensitive to imipenem . Production of TEM and SHV beta-lactamases was detected simultaneously in most isolates by isoelectric focusing and 93.3% produced a ceftazidimase of pl 8.2 of the SHV-5 type . The 15 isolates were grouped into 11 and 12 electrophoretic patterns by PFGE and REP-PCR, respectively . The degree of genetic variability indicated an endogenous origin of the nosocomial infections. Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 2131 - 6 Klebsiella singaporensis sp . nov., a novel isomaltulose-producing bacterium; Li X et al.; Cells of strain LX3(T), isolated from soil, were Gram-negative, facultatively anaerobic, non-motile, capsulated and non-endospore-forming straight rods, able to grow at 10 degrees C, unable to produce gas from lactose at 45 degrees C and unable to produce indole . The isolate converted sucrose to isomaltulose and did not produce detectable glucose by-products . The G+C content of the DNA was 56.4 mol% . Furthermore, comparison of 16S rRNA and rpoB gene sequences showed that the isolate clearly belongs to the genus Klebsiella . The closest phylogenetic relative was Klebsiella pneumoniae, there being 99.3 and 97.5 % similarity in 16S rRNA and rpoB gene sequences, respectively . DNA-DNA hybridization analysis demonstrated a very low level of relatedness to other members of the genus Klebsiella, indicating that the isolated strain and other species in the genus Klebsiella were not related at the species level . The isolate could be differentiated from other previously described members of the genus Klebsiella on the basis of phenotypic differences and 16S rRNA and rpoB gene sequence divergence, together with DNA-DNA reassociation data . Therefore, it is proposed that strain LX3(T) (=DSM 16265(T)=JCM 12419(T)) should be classified as the type strain of a novel species of genus Klebsiella, Klebsiella singaporensis sp . nov. Med Dosw Mikrobiol, 2004, 56(2), 161 - 71 {Detection of expression of extended-spectrum beta-lactamases (ESBL) in clinical strains of Klebsiella pneumoniae and Escherichia coli; comparison of E-test and two disc methods}; Andrzejewska E et al.; The aim of this study was to compare the effectiveness of E-test and two disc methods applied for the detection of extended spectrum beta-lactamases . All strains were tested by E-test, by double-disc synergy test (DDST) according to Jarlier (cefotaxim, ceftazidim, aztreonam and clavulanic acid) and also by disc test according to Appleton (cefpodoxime and cefpodoxime with clavulanic acid, CPD and CD01 disc) . We tested 148 clinical strains of E . coli and 78 strains of K . Pneumoniae . In case of K . pneumoniae, the activity of the ESBLs was detected among 30 strains--both in E-test, Jarlier test and Appleton test . Among E . coli, four strains were found ESBL-positive in the test according to Jarlier but only three strain of these when E-test and Appleton test was used . The results of investigations performed suggest, that E-test and disc methods according both Jarlier and Appleton have the same effectiveness in detection ESBLs among K . pneumoniae strains . However, in case of E . coli, interpretation of results may present a problem. J Clin Microbiol, 2004 Nov, 42(11), 5337 - 40 Complexity of Klebsiella pneumoniae isolates resistant to both cephamycins and extended-spectrum cephalosporins at a teaching hospital in Taiwan; Yan JJ et al.; Among 99 clinical Klebsiella pneumoniae isolates resistant to cefoxitin and extended-spectrum cephalosporins, coexistence of AmpC (DHA-1, CMY-2, or CMY-8) and extended-spectrum beta-lactamases (CTX-M and/or SHV) was detected in a total of 35 . The remainder produced AmpC (n = 42), extended-spectrum beta-lactamases (n = 9), metallo-beta-lactamases (n = 2), or none of these enzymes (n = 11) . Phenotypic characteristics of these isolates were demonstrated. Appl Microbiol Biotechnol, 2005 Feb, 66(5), 589 - 96 Epub 2004 Nov 04. Klebsiella planticola strain DSZ mineralizes simazine: physiological adaptations involved in the process; Sanchez M et al.; We examined the ability of a soil bacterium, Klebsiella planticola strain DSZ, to degrade the herbicide simazine (SZ) . Strain DSZ is metabolically diverse and grows on a wide range of s-triazine and aromatic compounds . DSZ cells grown in liquid medium with SZ (in 10 mM ethanol) as carbon source mineralized 71.6+/-1.3% of 0.025 mM SZ with a yield of 4.6+/-0.3 mug cell dry weight mmol(-1) carbon . The metabolites produced by DSZ during SZ degradation included ammeline, cyanuric acid, N-formylurea and urea . We studied the physiological adaptations which allow strain DSZ to metabolize SZ . Using scanning electron microscopy, we detected DSZ cells covering the surfaces of SZ crystals when the herbicide was used at high concentrations (0.1 mM) . The membrane order observed by FTIR spectroscopy showed membrane activity at low temperature (4 degrees C) to assimilate the herbicide . Membrane fatty acid analysis demonstrated that strain DSZ adapted to grow on SZ by increasing the degree of saturation of membrane lipid fatty acid; and the opposite effect was detected when both SZ and ethanol were used as carbon sources . This confirms the modulator effect of ethanol on membrane fluidity. Int J Antimicrob Agents, 2004 Nov, 24(5), 508 - 10 Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in an urban hospital in Dhaka, Bangladesh; Rahman MM et al.; The prevalence of extended-spectrum beta-lactamases (ESBL)-producing organisms in an urban hospital in Dhaka City was assessed over a 10-month period . A double disk test was performed to detect ESBL-producing Escherichia coli and Klebsiella pneumoniae . 43.2% and 39.5% of E . coli and K . pneumoniae had ESBL phenotypes, respectively . The combination of augmentin with ceftazidime detected the most ESBL-producing E . coli (39.5%) while augmentin with ceftriaxone was the best combination for the detection of ESBL (31.6%) in K . pneumoniae. Immunobiology, 2004, 209(3), 277 - 82 Immunomodulating effects of cefodizime on Klebsiella pneumoniae-stimulated neutrophils; Song H et al.; To explore the immunoregulating effects of cefodizime on neutrophils and its mechanisms, we detected the expression of some cytokines secreted by neutrophils after heat-killed Klebsiella pneumoniae induced inflammation . We analyzed the changes of signal transduction in this process by detecting the mRNA expression of toll like receptor 4 (TLR4) and the inhibitor factor of kappaBalpha (I-kappaBalpha) expressed by neutrophils . The activity of nuclear factor kappaB (NF-kappaB) in neutrophils was also assayed by electrophoretic mobility shift assay (EMSA) . We found cefodizime increased neutrophil production of TNF-alpha, IL-1beta in the early stage of inflammation, which was in agreement with the enhanced mRNA expression of TLR4 and DNA-binding activity of NF-kappaB . Taken together, the immunomodulating effects of cefodizime on cytokine production of K . pneumoniae stimulated neutrophils is possibly due to its regulation of TLR4 mRNA expression and DNA binding activities of NF-kappaB through the LPS-TLR4-NF-kappaB pathway. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 883 - 5 Intrinsic Klebsiella pneumoniae contamination of liquid germicidal hand soap containing chlorhexidine; Brooks SE et al.; We describe intrinsic contamination with Klebsiella pneumoniae occurring during the manufacture of germicidal hand soap, labeled as containing 2% chlorhexidine, used throughout a 350-bed community medical center . A 3-year retrospective study failed to find evidence of increased incidence of clinical isolates of this strain. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 878 - 82 A cluster of nosocomial Klebsiella oxytoca bloodstream infections in a university hospital; Sardan YC et al.; BACKGROUND: On February 19, 2003, four patients (patients 1-4) in the neurology ward underwent cranial magnetic resonance angiography (MRA) and developed fever within 1 hour afterward . Klebsiella oxytoca was isolated from blood cultures of patients 1 through 3 . OBJECTIVE: To identify the source of this cluster of nosocomial K . oxytoca bloodstream infections . DESIGN: Outbreak investigation . SETTING: A 1,000-bed university hospital . METHODS: The infection control team reviewed patient charts and interviewed nursing staff about the preparation and administration of parenteral fluids . The procedure of cranial MRA was observed . Arbitrarily primed polymerase chain reaction (AP-PCR) was performed to show the clonal relationship among these three strains . RESULTS: AP-PCR revealed that three K . oxytoca isolates had the same molecular profile . Cranial MRA was found to be the only common source among these patients . During MRA, before injection of the contrast medium, normal saline solution was infused to check the functioning of the intravenous catheter . Use of the solution for multiple patients was routine, but the access diaphragm of the bottle was not cleansed . The bottle of normal saline solution used on February 19 had already been discarded and the culture sample taken from the solution on the day of observation was sterile . CONCLUSIONS: We speculate that normal saline solution became contaminated during manipulation and that successive uses might have been responsible for this cluster . Poor aseptic techniques employed during successive uses appear to be the most likely route of contamination . Use of parenteral solutions for multiple patients was discontinued. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 832 - 7 Reduced use of third-generation cephalosporins decreases the acquisition of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae; Lee SO et al.; OBJECTIVES: To identify risk factors for the respiratory acquisition of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae among patients admitted to a neurosurgical intensive care unit (NSICU) and to modify them without changing general infection control measures . DESIGN: Nested case-control and intervention study . SETTING: A 1,200-bed, tertiary-care teaching hospital with a 17-bed NSICU . METHODS: Sputa of all patients admitted to the NSICU were cultured weekly during the study . From October 2002 through February 2003, 29 case-patients from whose sputum ESBL-producing K . pneumoniae was isolated were detected and 59 controls-patients were randomly selected among patients without any positive isolate of ESBL-producing K . pneumoniae . After analyzing the risk factors, we intervened to modify them and compared the acquisition rate of ESBL-producing K . pneumoniae before (October 2002 to February 2003) and after (April to August 2003) the intervention . RESULTS: Multivariate analysis showed that prior exposure to third-generation cephalosporins (TGCs) (OR, 6.0; CI95, 1.9 to 18.6; P = .002) was an independent risk factor of ESBL-producing K . pneumoniae acquisition . The neurosurgical team was notified of the result, and the infectious diseases specialist visited the NSICU three times a week to regulate TGC use during the intervention period . Patients admitted before the intervention were older than patients admitted after . The respiratory acquisition of ESBL-producing K . pneumoniae per 1,000 patient-days (13.5 {CI95, 8.9 to 18.1} vs 2.7 {CI95, 0.9 to 4.6}) and the antimicrobial use density of TGCs (38.2 +/- 5.0 vs 17.3 +/- 2.6; P < .001) decreased significantly after the intervention . CONCLUSION: Prior exposure to TGCs was an independent risk factor for the respiratory acquisition of ESBL-producing K . pneumoniae, and less use of TGCs was associated with a decrease in acquisition. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 812 - 8 Surveillance in Taiwan using molecular epidemiology for extended-spectrum beta-lactamase-producing Klebsiella pneumoniae; Yu WL et al.; OBJECTIVE: To evaluate intrahospital and interhospital clonal dissemination of extended-spectrum beta-lactamase (ESBL)-producing strains of Klebsiella pneumoniae . SETTING: Eight tertiary-care university hospitals and 16 regional hospitals in Taiwan . METHODS: Two hundred eleven confirmed ESBL-producing isolates of K . pneumoniae were collected from January 1998 to June 2000 . The isolates were characterized by various typing methods, including antibiogram (9 antimicrobial agents), computer-based ribotyping, pulsed-field gel electrophoresis (PFGE), and isoelectric focusing of beta-lactamase . RESULTS: Ribotyping identified 70 distinct ribogroups among 200 isolates evaluated . Forty-three of these ribogroups were unique . Eleven ribogroups, comprising 115 isolates, were detected in more than one hospital (interhospital dissemination), whereas 16 groups (42 isolates) were detected in more than one patient within a hospital (intrahospital dissemination) . The combination of ribotyping and PFGE identified two large epidemic clones, which were called 691.5/PFGE-G and 595.7/PFGE-A . These epidemic clones were detected mainly in the hospitals located in the northern and central regions of Taiwan . However, variation of the profiles of antibiograms and isoelectric focusing was apparent within each clone . In addition, isolates with the same isoelectric focusing profile (isoelectric points 7.9, 8.2, and 8.4) and antibiogram (resistance to 9 compounds evaluated) were present among different molecular-typed clones . CONCLUSIONS: Our results showed that clonal dissemination (both interhospital and intrahospital dissemination) is occurring in several regions of Taiwan . Rapid computer-based ribotyping associated with PFGE demonstrated multiple epidemic clones of ESBL-producing K . pneumoniae in Taiwan . The combination of phenotypic and molecular methods has proved useful to characterize these epidemic clones. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4466 - 9 SHV-49, a novel inhibitor-resistant beta-lactamase in a clinical isolate of Klebsiella pneumoniae; Dubois V et al.; A clinical strain of Klebsiella pneumoniae carried the bla(SHV-49) gene, encoding a novel inhibitor-resistant beta-lactamase of pI 7.6, derived from SHV-1 by the single substitution M69I . It also harbored a gene differing from bla(SHV-11) by four silent mutations and coding for a penicillinase . Both genes were chromosome located and might represent either a species-specific gene or an acquired resistance gene. Zhonghua Yi Xue Za Zhi, 2004 Sep 2, 84(17), 1454 - 9 {Identification and expression of blaCTX-M-14 and blaCTX-M-24}; Xiong ZZ et al.; OBJECTIVE: To identify the ESBL gene and the prevalence in Escherichia coli and Klebsiella pneumoniae strain isolated from Huashan Hospital, Shanghai . METHODS: Isolates were confirmed as an ESBL producing strain by double-disk synergy test and NCCLS Confirmatory Test . Antibiotic susceptibilities were determined by standard agar dilution procedure on Mueller-Hinton agar . To determine whether the resistance was transferable, the conjugation experiment was performed; plasmids were isolated from clinical isolates and transcojugants . The partial bla(gene) of ESBL producing isolates and their transcojugants were detected by PCR using universal primers for TEM, SHV, CTX-M-1group, Toho-1group, CTX-M-13group respectively . The entire bla(CTX-M-13) group were amplified by PCR using the primers outside the Open Reading Frame (ORF) of CTX-M-13group beta-lactamases; the PCR products of entire bla(CTX-M-13)group were cloned into vector and the recombinant plasmids were transformed into the recipient strain for expression; the PCR products were also directly sequenced and analyzed; the clinical isolates of ESBL producers were detected by PFGE . RESULTS: ESBL producers were resistant to most beta-lactams and non-beta-lactams . Most transconjugants were obtained at frequency of 10(-4) approximately 10(-5) and resistance to non-beta-lactams was cotransferred with the ESBL activity to the transconjugant . A plasmid of about > 23.1 kb was obtained from each tansconjugant by plasmid extraction . Partial gene amplification products of CTX-M-13 group gene were obtained from isolates and their transconjugants . The bla(CTX-M-13)group from 4 transconjugants were identified as bla(CTX-M-14), and other six were bla(CTX-M-24); those ESBLs were mediated by plasmids (> 23.1 kb); the transformants producing CTX-M-14 or CTX-M-24 were resistant to most beta-lactams, which were much more resistant to cefotaxime than to ceftazidine; PFGE patterns of those isolates were different . CONCLUSION: clinical isolate of Escherichia coli and Klebsiella pneumoniae isolated from Huashan Hospital, Shanhai produced CTX-M-14 or CTX-M-24, which caused the isolate resistant to most beta-lactams; no clone spread in those isolates was found. Mol Plant Microbe Interact, 2004 Oct, 17(10), 1078 - 85 Nitrogen fixation in wheat provided by Klebsiella pneumoniae 342; Iniguez AL et al.; In this report, all of the criteria necessary for the demonstration of nitrogen fixation in wheat (Triticum aestivum L.), the world's most important crop, are shown upon inoculation with a nitrogen-fixing bacterium, Klebsiella pneumoniae 342 (Kp342) . Kp342 relieved nitrogen (N) deficiency symptoms and increased total N and N concentration in the plant . Nitrogen fixation was confirmed by 15N isotope dilution in the plant tissue and in a plant product, chlorophyll . All of these observations were in contrast to uninoculated plants, plants inoculated with a nitrogen-fixing mutant of Kp342, and plants inoculated with dead Kp342 cells . Nitrogenase reductase was produced by Kp342 in the intercellular space of the root cortex . Wild-type Kp342 and the nifH mutant colonized the interior of wheat roots in equal numbers on a fresh weight basis . The nitrogen fixation phenotype described here was specific to cv . Trenton . Inoculation of cvs . Russ or Stoa with Kp342 resulted in no relief of nitrogen deficiency symptoms. Mol Microbiol, 2004 Nov, 54(3), 647 - 64 Structure and assembly of the pseudopilin PulG; Kohler R et al.; The pseudopilin PulG is one of several essential components of the type II pullulanase secretion machinery (the Pul secreton) of the Gram-negative bacterium Klebsiella oxytoca . The sequence of the N-terminal 25 amino acids of the PulG precursor is hydrophobic and very similar to the corresponding region of type IV pilins . The structure of a truncated PulG (lacking the homologous region), as determined by X-ray crystallography, was found to include part of the long N-terminal alpha-helix and the four internal anti-parallel beta-strands that characterize type IV pilins, but PulG lacks the highly variable loop region with a disulphide bond that is found in the latter . When overproduced, PulG forms flexible pili whose structural features, as visualized by electron microscopy, are similar to those of bacterial type IV pili . The average helical repeat comprises 17 PulG subunits and four helical turns . Electron microscopy and molecular modelling show that PulG probably assembles into left-handed helical pili with the long N-terminal alpha-helix tightly packed in the centre of the pilus . As in the type IV pilins, the hydrophobic N-terminal part of the PulG alpha-helix is necessary for its assembly . Subtle sequence variations within this highly conserved segment seem to determine whether or not a type IV pilin can be assembled into pili by the Pul secreton. Arch Microbiol, 2004 Nov, 182(5), 414 - 20 Oxaloacetate decarboxylase of Archaeoglobus fulgidus: cloning of genes and expression in Escherichia coli; Dahinden P et al.; Archaeoglobus fulgidus harbors three consecutive and one distantly located gene with similarity to the oxaloacetate decarboxylase Na+ pump of Klebsiella pneumoniae (KpOadGAB) . The water-soluble carboxyl transferase (AfOadA) and the biotin protein (AfOadC) were readily synthesized in Escherichia coli, but the membrane-bound subunits AfOadB and AfOadG were not . AfOadA was affinity purified from inclusion bodies after refolding and AfOadC was affinity purified from the cytosol . Isolated AfOadA catalyzed the carboxyl transfer from {4-14C}-oxaloacetate to the prosthetic biotin group of AfOadC or the corresponding biotin domain of KpOadA . Conversely, the carboxyl transferase domain of KpOadA exhibited catalytic activity not only with its pertinent biotin domain but also withAfOadC. Am J Ophthalmol, 2004 Oct, 138(4), 662 - 3 Endophthalmitis caused by Klebsiella species; Scott IU et al.; PURPOSE: To investigate clinical settings, antibiotic sensitivities, and visual outcomes associated with endophthalmitis caused by Klebsiella species . DESIGN: Retrospective case series . METHODS: Record review of patients with endophthalmitis caused by Klebsiella (1984 through 2003) . RESULTS: Clinical settings included cataract surgery (one eye), trauma (two), perforated corneal ulcer (one), and endogenous associated with hepatic abscess (one) . Pretreatment vision was hand motions or better in four eyes (80%) . Initial treatment was enucleation (one eye), pars plana vitrectomy (two), and vitreous tap and injection (two); intravitreal antibiotics were administered to all nonenucleated eyes . Klebsiella was sensitive to one or more antibiotics administered initially in all cases . In nonenucleated eyes, final acuity was >or=20/400 in two, 1/200 in one, and light perception in one . CONCLUSION: Despite treatment with appropriate antibiotics, endophthalmitis caused by Klebsiella species is associated with generally poor visual outcomes. Microbes Infect, 2004 Nov, 6(13), 1191 - 8 High prevalence of phagocytic-resistant capsular serotypes of Klebsiella pneumoniae in liver abscess; Lin JC et al.; To better understand the role of capsular polysaccharide (CPS) K1 or K2 in Klebsiella pneumoniae liver abscess as well as the development of metastasis to eye, neutrophil phagocytosis of 70 CPS isolates including K1 (n = 23)/K2 (n = 10), non-K1/K2 (n = 37) was evaluated by flow cytometry, fluorescence imaging, and electron microscopy . K1/K2 isolates were significantly more resistant to phagocytosis (P < 0.0001) than non-K1/K2 isolates and displayed increased resistance to intracellular killing . Although mucoid phenotype (M-type) K1/K2 isolates were significantly more resistant to phagocytosis (P = 0.0029) than M-type non-K1/K2, no significant difference in the phagocytosis rate was observed between K1/K2 isolates with M-type and non-M-type (P = 0.0924) . Mucoidy is an associated factor that was predominant in K1/K2 isolates, but which itself is not an independent influence on phagocytic resistance . The K1/K2 CPS proved significantly more resistant to phagocytosis than non-K1/K2 CPS in liver abscess isolates (P < 0.0001) and non-abscess isolates (P = 0.0001), suggesting that K1/K2 isolates were generally more virulent in both liver abscess and in non-liver abscess conditions . These findings indicate that resistance of CPS K1 or K2 K . pneumoniae to phagocytosis and intracellular killing presumably contributes to their high prevalence in liver abscess and uniquely in endophthalmitis. J Antimicrob Chemother, 2004 Dec, 54(6), 1130 - 1133 Epub 2004 Oct 14. Cefepime and the inoculum effect in tests with Klebsiella pneumoniae producing plasmid-mediated AmpC-type {beta}-lactamase; Kang CI et al.; OBJECTIVE: In the past decade, a new problem in Klebsiella pneumoniae strains has emerged: plasmid-mediated AmpC enzymes . This study was conducted to investigate the activity of cefepime against clinical isolates by determining the activities of cefepime and three other parenteral beta-lactam agents in standard and high inoculum MIC tests . METHODS: A total of 61 K . pneumoniae blood isolates, including 28 isolates producing AmpC-type beta-lactamases (14 isolates of DHA-1 and 14 isolates of CMY-1-like) and 33 isolates producing extended-spectrum beta-lactamases (ESBLs) (32 isolates of TEM- or SHV-related and one isolate of CTX-M-14-like), were included in the study . Antimicrobial susceptibilities were determined using broth microdilution MIC tests with standard and 100-fold-higher inocula . The inoculum effect was defined as an eight-fold or greater MIC increase on testing with the higher inoculum . RESULTS: In tests with AmpC beta-lactamase-producing K . pneumoniae isolates and their transconjugants, the inoculum effect was most consistently detected with cefepime, cefotaxime and ceftazidime, as inoculum effects were consistently detected in ESBL-producing isolates . However, the inoculum effect was least frequently detected with imipenem . CONCLUSION: Although the inoculum effect is an in vitro laboratory phenomenon, these results suggest that cefepime may be a less than reliable agent for therapy in cases of high inoculum infections caused by AmpC beta-lactamase-producing K . pneumoniae. Biotechnol Lett, 2004 Aug, 26(16), 1301 - 6 Improved process for exopolysaccharide production by Klebsiella pneumoniae sp . pneumoniae by a fed-batch strategy; Ramirez-Castillo ML et al.; Exopolysaccharide (EPS) was produced by Klebsiella pneumoniae K63 grown in fed-batch cultures using different procedures of the supply of carbon or nitrogen (N) source, or both . Cultures grown with excess of glucose and limitation or exhaustion of N produced 54.8 and 47.4 g(EPS) l(-1), respectively . These cultures also led to an accumulation of 'overflow' metabolites representing more than 16% of carbon conversion . The consistency indexes ( K ) obtained to the end of the cultures, characteristic of the rheological property of the biopolymer, were 16.4 Pa s(n) for N deficiency and 5.2 Pa s(n) for N limitation conditions . The simultaneous limitation of glucose and N decreased the excretion of co-metabolites (6.4% of carbon conversion) and the EPS production (18.1 g(EPS) l(-1)), while improving the quality of the polysaccharide, characterized by the highest K of 126.2 Pa s(n) and the highest pseudoplasticity degree (flow behaviour index, n=0.2). Zh Mikrobiol Epidemiol Immunobiol, 2004 Jul-Aug, (4), 7 - 11 {Proteinase activity of Klebsiella pneumoniae of different virulence}; Acute exacerbation of chronic hepatitis B during thalidomide therapy for multiple myeloma: a case report; Department of Internal Medicine, Gachon Medical School, Incheon, KoreaWe report a case of acute fatal exacerbation of chronic hepatitis B in a 50-year-old man with multiple myeloma being treated with thalidomide . The patient had a medical history of chronic hepatitis B and was diagnosed with stage IIIA multiple myeloma . He suffered two episodes of transient transaminitis of unknown origin after successive autologous stem cell transplantations . Spontaneous resolutions of the transaminitis were observed without special management . At that time, PCR of hepatitis B virus (HBV) were all-negative . After 5-months' administration of thalidomide for the second relapse of the multiple myeloma, he suddenly experienced dizziness and jaundice . The level of HBV DNA was 1,641 pg/mL and the serologic tests for other viruses were negative . Despite conventional supportive care, he expired due to septic shock caused by Klebsiella pneumonia . Based on the stable disease status of the multiple myeloma and exclusion of other hepatotoxic agents, it was assumed that the exacerbation of the hepatitis B virus during the thalidomide therapy preceded the bacterial sepsis . With the increased use of thalidomide in cancer treatment, cautious monitoring of the viral burden should be performed in patients with chronic hepatitis B. Eur J Clin Microbiol Infect Dis, 2004 Nov, 23(11), 813 - 7 Epub 2004 Nov. Evaluation of the current National Committee for Clinical Laboratory Standards guidelines for screening and confirming extended-spectrum beta-lactamase production in isolates of Escherichia coli and Klebsiella species from bacteremic patients; Katz OT et al.; The National Committee for Clinical Laboratory Standards (NCCLS) recommendations for screening and confirming the production of extended-spectrum beta-lactamases (ESBLs) were evaluated in 115 isolates of Escherichia coli and 157 isolates of Klebsiella spp . from Israeli patients with bacteremia . All isolates were screened using cefotaxime, ceftazidime, and cefpodoxime discs . Confirmatory tests using pairs of discs containing ceftazidime, cefotaxime, or cefpodoxime in which clavulanic acid was added to one of the discs in each pair {inhibitor-potentiated disc diffusion test (IPDDT)} and two double-sided E test strips containing ceftazidime or cefotaxime with and without clavulanic acid were performed on all isolates regardless of the results of screening tests . Isolates that tested positive by one or more confirmatory tests were considered ESBL producers . Overall, 69 (25.4%) strains were found to be ESBL producers . The sensitivity of the NCCLS screening criteria ranged between 98.6% for cefotaxime and 92.8% for ceftazidime, and the specificity ranged between 100% for cefotaxime and cefpodoxime and 99.0% for ceftazidime . The sensitivity of the confirmatory tests ranged between 97.1% for the cefotaxime E test and only 75.4% for the ceftazidime IPDDT discs . All 64 isolates that fell in the intermediate and resistant categories for cefotaxime, as well as all 41 in the same categories for ceftazidime and 68 of 69 in these categories for cefpodoxime, were confirmed as ESBL producers . The use of multiple antimicrobial discs for screening isolates and combinations of IPPDT discs is needed to improve the sensitivity of confirmatory testing . It is recommended that isolates falling in the intermediate and resistant categories in disc diffusion testing be reported as ESBL producers . The use of confirmatory tests should be limited to organisms with inhibition zone diameters ranging between the NCCLS recommendations for ESBL screening and the intermediate category breakpoints. Nucleic Acids Res . 2004 Oct 08;32(18):e138. KpnBI is the prototype of a new family (IE) of bacterial type I restriction-modification system; Chin V et al.; KpnBI is a restriction-modification (R-M) system recognized in the GM236 strain of Klebsiella pneumoniae . Here, the KpnBI modification genes were cloned into a plasmid using a modification expression screening method . The modification genes that consist of both hsdM (2631 bp) and hsdS (1344 bp) genes were identified on an 8.2 kb EcoRI chromosomal fragment . These two genes overlap by one base and share the same promoter located upstream of the hsdM gene . Using recently developed plasmid R-M tests and a computer program RM Search, the DNA recognition sequence for the KpnBI enzymes was identified as a new 8 nt sequence containing one degenerate base with a 6 nt spacer, CAAANNNNNNRTCA . From Dam methylation and HindIII sensitivity tests, the methylation loci were predicted to be the italicized third adenine in the 5' specific region and the adenine opposite the italicized thymine in the 3' specific region . Combined with previous sequence data for hsdR, we concluded that the KpnBI system is a typical type I R-M system . The deduced amino acid sequences of the three subunits of the KpnBI system show only limited homologies (25 to 33% identity) at best, to the four previously categorized type I families (IA, IB, IC, and ID) . Furthermore, their identity scores to other uncharacterized putative genome type I sequences were 53% at maximum . Therefore, we propose that KpnBI is the prototype of a new 'type IE' family. J Clin Microbiol, 2004 Oct, 42(10), 4799 - 802 New method for laboratory detection of AmpC beta-lactamases in Escherichia coli and Klebsiella pneumoniae; Nasim K et al.; A new cefoxitin-agar medium (CAM)-based assay was compared to the previously published modified three-dimensional (M3D) assay for the detection of AmpC production in Escherichia coli and Klebsiella pneumoniae . Clinical isolates of cefoxitin-resistant E . coli (n = 5) and K . pneumoniae (n = 7) and multiple control strains with and without AmpC enzymes were tested by both methods . The CAM method with 4 microg of cefoxitin/ml was equivalent to the M3D method for detecting AmpC production in E . coli and K . pneumoniae . This new method is easier to perform and interpret and allows for testing of multiple isolates on a single plate. J Antimicrob Chemother, 2004 Nov, 54(5), 881 - 888 Epub 2004 Oct 7. Outbreak of multi-resistant Klebsiella oxytoca involving strains with extended-spectrum {beta}-lactamases and strains with extended-spectrum activity of the chromosomal {beta}-lactamase; Decre D et al.; OBJECTIVES: This study was conducted to analyse broad-spectrum cephalosporin-resistant Klebsiella oxytoca strains . METHODS: The 57 isolates studied were recovered from clinical specimens (n=23) or from rectal swabs (n=34) during a 26-month period . Antibiotic susceptibility patterns were determined using standard agar diffusion and dilution methods including the synergy test between extended-spectrum cephalosporins and clavulanic acid . ERIC-2 PCR and pulsed-field gel electrophoresis (PFGE) methods were used to study the clonal relatedness of the strains . Plasmid-mediated and chromosomal beta-lactamases were characterized by mating and specific bla gene amplification and sequencing . RESULTS: Four different antibiotic resistance patterns were identified whereas ERIC-2 PCR and PFGE revealed six main profiles . Extended-spectrum beta-lactamases (ESBLs) were found in 32 strains: TEM-7 (n=26), TEM-129 (n=1), TEM-3 (n=4), SHV-2 (n=1) . The new TEM-type beta-lactamase, TEM-129, differed from TEM-7 by one mutation (Glu-104-->Lys) . All TEM-7 or TEM-129 producers were genetically related . Twenty-five other strains with identical ERIC-2 PCR and PFGE profiles harboured a bla(OXY-2) gene different from the reference gene: 24 strains displayed one substitution (Ala-237-->Ser) in the KTG motif and one strain, highly resistant to ceftazidime, showed an additional substitution (Pro-167-->Ser) . CONCLUSIONS: The study demonstrated that the majority of strains (n=52) harbouring the OXY-2-type beta-lactamase corresponded to two clones . The first clone (n=27) corresponded to ESBL-producing strains . The second clone (n=25) displayed extended-spectrum activity of the chromosomal beta-lactamase. J Immunol, 2004 Oct 15, 173(8), 5148 - 55 CpG oligodeoxynucleotides stimulate protective innate immunity against pulmonary Klebsiella infection; Deng JC et al.; Bacterial pneumonia is a leading cause of mortality in the United States . Innate immune responses, including type-1 cytokine production, are critical to the effective clearance of bacterial pathogens from the lung . Synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotide motifs (CpG ODN), which mimic the effects of bacterial DNA, have been shown to enhance type-1 cytokine responses during infection due to intracellular pathogens, resulting in enhanced microbial clearance . The role of CpG ODN in modulating protective innate immunity against extracellular pathogens is unknown . Using a murine model of Gram-negative pneumonia, we found that CpG ODN administration stimulated protective immunity against Klebsiella pneumoniae . Specifically, intratracheal (i.t.) administration of CpG ODN (30 microg) 48 h before i.t . K . pneumoniae challenge resulted in increased survival, compared with animals pretreated with control ODN or saline . Pretreatment with CpG ODN resulted in enhanced bacterial clearance in lung and blood, and higher numbers of pulmonary neutrophils, NKT cells, gammadelta-T cells, and activated NK1.1+ cells and gammadelta-T lymphocytes during infection . Furthermore, pretreatment with CpG ODN enhanced the production of TNF-alpha, and type-1 cytokines, including IL-12, IFN-gamma, and the IFN-gamma-dependent ELR- CXC chemokines IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma in response to Klebsiella challenge, compared with control mice . These findings indicate that i.t . administration of CpG ODN can stimulate multiple components of innate immunity in the lung, and may form the basis for novel therapies directed at enhancing protective immune responses to severe bacterial infections of the lung. J Wildl Dis, 2004 Jul, 40(3), 588 - 93 Struvite penile urethrolithiasis in a pygmy sperm whale (Kogia breviceps); Harms CA et al.; Massive urolithiasis of the penile urethra was observed in an adult pygmy sperm whale (Kogia breviceps) stranded on Topsail Island, North Carolina, USA . Calculi occupied the urethra from just distal to the sigmoid flexure to the tip of the penis for a length of 43 cm . A urethral diverticulum was present proximal to the calculi . The major portion of the multinodular urolith weighed 208 g and was 16 cm long x 3.7 cm diameter at the widest point . The urolith was composed of 100% struvite (magnesium ammonium phosphate) and on culture yielded Klebsiella oxytoca, a urease-positive bacterium occasionally associated with struvite urolith formation in domestic animals . Reaction to the calculi was characterized histologically by moderate multifocal to coalescing plasmacytic balanitis and penile urethritis . Role of the urethrolithiasis in the whale's stranding is speculative but could have involved pain or metabolic perturbations such as uremia or hyperammonemia. Photochem Photobiol, 2004 Dec, 80(3), 542 - 7 Purification and Initial Characterization of a Putative Blue Light-regulated Phosphodiesterase from Escherichia coli paragraph sign; Rajagopal S et al.; The Escherichia coli protein YcgF contains a photosensory flavin adenine dinucleotide (FAD)-binding BLUF domain covalently linked to an EAL domain, which is predicted to have cyclic-di-guanosine monophosphate (GMP) phosphodiesterase activity . We have cloned, overexpressed and purified this protein, which we refer to as blue light-regulated phosphodiesterase (Blrp) for its putative activity . Blrp undergoes a reversible photocycle after exposure to light in which the spectrum of its photostationary state and kinetics of recovery of the dark state are similar to those of the isolated BLUF domain of the AppA protein . Unlike the AppA BLUF domain, the chromophore environment in the context of full-length Blrp is asymmetric, and the protein does not undergo any detectable global changes on exposure to blue light . When overexpressed in E . coli, Blrp copurifies with certain proteins which suggests that it plays a protective role in response to oxidative stress . Predicted proteins from Klebsiella pneumoniae and from a bacterium in the Sargasso Sea are similar to E . coli Blrp in both their BLUF and EAL domains, which suggests that blue light sensing in these bacteria may follow similar pathways. Surg Neurol, 2004 Oct, 62(4), 362 - 5; discussion 365 Massive intracerebral air embolism associated with meningitis and lumbar spondylitis: case report; Kuo TH et al.; BACKGROUND: Massive intracerebral air embolism is a rare pathologic state and never in association with meningitis and lumbar spondylitis . To the best of our knowledge, our presented case is the first of a massive intracerebral air embolism associated with meningitis and lumbar spondylitis of Klebsiella pneumonia . CASE DESCRIPTION: A 55-year-old man presented with a high fever and low back pain . Blood culture showed Klebsiella pneumonia . Lumbar computed tomography (CT) revealed discitis at L1-2 and L2-3 levels and paraspinal abscess in which air was found . Despite management with antibiotics, patient's consciousness deteriorated, and brain CT revealed diffuse intravenous air embolism and severe brain swelling . Cerebrospinal fluid (CSF) examination demonstrated bacterial meningitis, and the CSF culture showed Klebsiella pneumonia . Later, septic shock occurred and patient expired . CONCLUSION: Intracerebral air embolism can occur in the Klebsiella pneumonia meningitis that resulted from lumbar spondylitis and sepsis. Nepal Med Coll J, 2004 Jun, 6(1), 67 - 8 A blackening lady--case report; Biswas R et al.; A 40 year old lady presented with the classical clinical features of Addison's disease which on further investigations with an ultrasound abdomen showed a right suprarenal mass . This was subjected to a fine needle aspiration which revealed pus which on culture grew Klebsiella pneumoniae . Patient responded well to steroids and antibiotics . To the best of our knowledge this is the first report of Klebsiella pneumoniae in association with Addison's disease. Antimicrob Agents Chemother, 2004 Oct, 48(10), 3720 - 8 Epidemiology and clinical features of bloodstream infections caused by AmpC-type-beta-lactamase-producing Klebsiella pneumoniae; Pai H et al.; Cases of bacteremia caused by AmpC-type-beta-lactamase-producing Klebsiella pneumoniae isolates were retrospectively studied to determine the epidemiologic features and clinical outcomes of bloodstream infections . Among 389 blood isolates recovered from 1998 to 2002, 65 isolates (16.7%) were found to be extended-spectrum beta-lactamase (ESBL) or AmpC beta-lactamase producers . The beta-lactamases from 61 of the 65 isolates were characterized; 28 of 61 isolates produced AmpC-type enzymes (14 isolates each produced DHA-1 and CMY-1-like enzymes), 32 isolates produced TEM or SHV-related ESBLs, and 1 isolate produced a CTX-M-14-like enzyme . To compare the clinical features and outcomes of bloodstream infections caused by AmpC producers with those caused by TEM- or SHV-related ESBL producers, 27 patients infected with isolates producing AmpC-type enzymes (AmpC group) and 25 patients infected with isolates producing TEM- or SHV-related enzymes (ESBL group) were analyzed . There was no significant difference between the AmpC and the ESBL groups in terms of risk factors . When the initial response was assessed at 72 h after antimicrobial therapy, the treatment failure rate for the AmpC group was 51.9% (14 of 27 patients) and the 7- and 30-day mortality rates were 14.8 and 29.6%, respectively, which were similar to those for the ESBL group . When the mortality rate for the patients who received extended-spectrum cephalosporins as definitive treatment was assessed, all four patients in the DHA-1 group and one of three patients in the CMY-1-like group died . In summary, the prevalence of AmpC enzyme-producing K . pneumoniae was high at the Seoul National University Hospital, and the clinical features and outcomes for the patients infected with AmpC-producing organisms were similar to those for the patients infected with TEM- or SHV-related ESBL producers. Curr Microbiol, 2004 Oct, 49(4), 267 - 73 The electron transfer flavoprotein fixABCX gene products from Azospirillum brasilense show a NifA-dependent promoter regulation; Sperotto RA et al.; The complete nucleotide sequence of the A . brasilense fixA, fixB, fixC, and fixX genes is reported here . Sequence similarities between the protein sequences deduced from fixABCX genes and many electron transfer flavoproteins (ETFs) have been noted . Comparison of the amino acid sequences of both subunits of ETF with the A . brasilense fixA and fixB gene products exhibits an identity of 30% . The amino acid sequence of the other two genes, fixC and fixX, revealed similarity with the membrane-bound electron transfer flavoprotein ubiquinone oxidoreductase (ETF-QO) . Using site-directed mutagenesis, mutations were introduced in the fixA promoter element of the A . brasilense fixABCX operon and chimeric p fixA-lacZ reporter gene fusions were constructed . The activation of the fixA promoter of A . brasilense is dependent upon the presence of the NifA protein being approximately 7 times less active than the A . brasilense nifH promoter . These results indicate that NifA from Klebsiella pneumoniae activates the fix promoter of A . brasilense and provide further evidence in support of the regulatory model of NifA activation in A . brasilense . Although no specific function has been assigned to the fixABCX gene products they are apparently required for symbiotic nitrogen fixation . An electron-transferring capacity in the nitrogen fixation pathway has been suggested for the fix gene products based on sequence homologies to the ETFs and ETF-QO proteins and by the absence of orthologous electron transfer proteins NifJ and NifF in A . brasilense. Scand J Immunol, 2004 Oct, 60(4), 351 - 5 The improved survival of experimental animals fed with fish oil is suppressed by a leukotriene inhibitor; Thors VS et al.; Fish oil is believed to alter the immune response and improve survival after infections in experimental animals . This effect may be due to altered production of the leukotrienes (LT) . We, therefore, performed a study in order to evaluate whether the effect of fish oil on the immune response of experimental animals is mediated through altered production of the LT . Female NMRI mice in four groups were fed with fish oil, fish oil with 5-lipoxygenase (5-LO) inhibitor (Zileuton, Abbott Laboratories, Chicago, IL, USA), corn oil or corn oil with 5-LO inhibitor . After 6 weeks, the mice were infected with Klebsiella pneumoniae and the survival was monitored . The experiment was performed twice . Analysis was performed mainly on data pooled from both experiments . The survival of the groups fed with fish oil was increased, compared to that of all the other groups and when compared to the groups fed with fish oil with 5-LO inhibitor (log-rank test) the difference was significant (P = 0.007) . It has been postulated that the effect of fish oil on the immune system is mediated through altered production of LT . In our study, blocking of the production of the LT eliminated the beneficial effects of fish oil . Our results are in concord with the hypothesis that the effect of fish oil is, at least partly, mediated through altered production of LT. Chang Gung Med J, 2004 May, 27(5), 390 - 3 Nasal septal abscess as a complication of laser inferior turbinectomy; Lo SH et al.; Postoperative infections are infrequent following laser inferior turbinate surgery . We report a 52-year-old man with a Klebsiella pneumoniae nasal septal abscess as a complication of potassium-titanium-phosphate 532-nm laser turbinectomy . To our knowledge, this is the first report of such a potentially serious complication resulting from minor ambulatory intranasal surgery . The clinical presentation, pathogenesis, and management of nasal septal abscesses are discussed. J Infect Chemother, 2004 Aug, 10(4), 212 - 5 Changes of antimicrobial resistance and extended-spectrum beta-lactamase production in Klebsiella spp . strains; Timko J; Changes of antimicrobial resistance and extended-spectrum beta-lactamase (ESBL) production of Klebsiella spp . strains isolated at the Central Military Hospital, Ruzomberok, Slovakia, with a special focus on the Anesthesiology--Resuscitation and Intensive Medicine Department during the years 1998-2002, were analyzed . Of 3920 gram-negative strains isolated from clinical materials during this period, Klebsiella spp . represented 8% . The incidence of ESBL-producing Klebsiella spp . isolates increased from 29% in 1998 to 69% in 2002 . Of 17 antibiotics tested, meropenem was found to be the most effective drug (100%) . The overall efficacy of cefotaxime was 31%, that of gentamicin 23%, and that of ciprofloxacin 54% . Analyzed Klebsiella isolates were characterized also by a high degree of multiresistance (53%) . The high incidence of reduced antibiotic susceptibility among Klebsiella spp . strains isolated at the intensive-care department suggests that more effective strategies are necessary to control the selection and spread of resistant organisms in this hospital. J Immunol, 2004 Sep 15, 173(6), 4075 - 83 STAT4 is a critical mediator of early innate immune responses against pulmonary Klebsiella infection; Deng JC et al.; Bacterial pneumonia is a leading cause of morbidity and mortality in the U.S . An effective innate immune response is critical for the clearance of bacteria from the lungs . IL-12, a key T1 cytokine in innate immunity, signals through STAT4 . Thus, understanding how STAT4 mediates pulmonary immune responses against bacterial pathogens will have important implications for the development of rational immunotherapy targeted at augmenting innate immunity . We intratracheally administered Klebsiella pneumoniae to wild-type BALB/c and STAT4 knockout (STAT4-/-) mice . Compared with wild-type controls, STAT4-/- mice had decreased survival following intratracheal Klebsiella administration, which was associated with a higher lung and blood bacterial burden . STAT4-/- animals also displayed impaired pulmonary IFN-gamma production and decreased levels of proinflammatory cytokines, including the ELR- CXC chemokines IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma . Although total lung leukocyte populations were similar between STAT4-/- and wild-type animals following infection, alveolar macrophages isolated from infected STAT4-/- mice had decreased production of proinflammatory cytokines, including IFN-gamma, compared with infected wild-type mice . The intrapulmonary overexpression of IFN-gamma concomitant with the systemic administration of IFN-gamma partially reversed the immune deficits observed in STAT4-/- mice, resulting in improved bacterial clearance from the blood . Collectively, these studies demonstrate that STAT4 is required for the generation of an effective innate host defense against bacterial pathogens of the lung . J Mater Sci Mater Med, 1999, 10(10/11), 649 - 52 Investigation into the mechanism of bacterial adhesion to hydrogel-coated surfaces; Kunz R et al.; As a model for hydrogel-coated biomaterials, self-assembled monolayers of polyoxyethylene (POE) derivatives on sheets of polymeric biomaterials were prepared . The POE derivatives consisted of hydrophilic chains with different lengths and a long-chain alkyl group that served as an anchor function . The coatings obtained were analyzed with XPS and contact angle measurements showing hydrophilic chains of different lengths extending away from the surface . Bacterial adhesion was measured with a clinically relevant Klebsiella pneumoniae type strain and measurements reproduced 12 times . Bacterial adhesion decreased markedly with increasing hydrophilic chain length . Based upon these findings a new model for bacterial adhesion to hydrogel-coated surfaces is suggested: steric repulsion effects that increase with increasing chain length of grafted hydrophilic chains play an important role in bacterial adhesion to hydrogel-coated surfaces . Appl Environ Microbiol, 2004 Sep, 70(9), 5441 - 6 Comparison of atomic force microscopy interaction forces between bacteria and silicon nitride substrata for three commonly used immobilization methods; Vadillo-Rodriguez V et al.; Atomic force microscopy (AFM) has emerged as a powerful technique for mapping the surface morphology of biological specimens, including bacterial cells . Besides creating topographic images, AFM enables us to probe both physicochemical and mechanical properties of bacterial cell surfaces on a nanometer scale . For AFM, bacterial cells need to be firmly anchored to a substratum surface in order to withstand the friction forces from the silicon nitride tip . Different strategies for the immobilization of bacteria have been described in the literature . This paper compares AFM interaction forces obtained between Klebsiella terrigena and silicon nitride for three commonly used immobilization methods, i.e., mechanical trapping of bacteria in membrane filters, physical adsorption of negatively charged bacteria to a positively charged surface, and glutaraldehyde fixation of bacteria to the tip of the microscope . We have shown that different sample preparation techniques give rise to dissimilar interaction forces . Indeed, the physical adsorption of bacterial cells on modified substrata may promote structural rearrangements in bacterial cell surface structures, while glutaraldehyde treatment was shown to induce physicochemical and mechanical changes on bacterial cell surface properties . In general, mechanical trapping of single bacterial cells in filters appears to be the most reliable method for immobilization. Bacteriol Virusol Parazitol Epidemiol, 2003 Apr-Sep, 48(2-3), 130 - 4 {The sensitivity of some strains of Klebsiella spp . collected in a neonatal department}; Craciunescu M et al.; In order to analyse the evolution of the sensitivity to antibiotics of same strains with nosocomial potential such as Klebsiella isolated in a hospital, we took into study a number of 976 samples, collected in a new-born care department of the "Dr . D . Popescu" hospital Timisoara . The study took place between November-December 2002 . The collected samples were pharyngeal swabs, nasal swabs, gastric aspirates, ocular and umbilical secretions, vernix, urines, faeces and blood . From all these samples 803 strains with nosocomial potential were isolated, 84 strains being Klebsiella spp . Most of the isolated strains presented multiple phenotypes of resistance to antibiotics. Bacteriol Virusol Parazitol Epidemiol, 2003 Apr-Sep, 48(2-3), 118 - 22 {Bacterial strains isolated in high nosocomial risk departments}; Muntean D et al.; We collected 312 samples from hospitalized patients in two hospitals in Timisoara between September-December 2003 . We isolated 83 strains with nosocomial potential . Identification of the germs was performed using the automatic API system, and the susceptibility tests were performed using disc-diffusion and the agar dilution test . By analyzing the extended antibiograms we categorized the germs considering their phenotypes of resistance and we remarked a high percentage of E . coli, Klebsiella pneumoniae pneumoniae and S . aureus with multiple resistance to antibiotics. J Microbiol Immunol Infect, 2004 Aug, 37(4), 208 - 15 Evidence for arylamine N-acetyltransferase activity in Klebsiella pneumoniae; Hui CS et al.; Arylamine N-acetyltransferase (NAT) enzymes have been found in laboratory animals, humans, microorganisms (fungi, bacteria and parasites), and in plants . But the characteristics of NAT from Klebsiella pneumoniae are not clear . NAT activities with p-aminobenzoic acid (PABA) and 2-aminofluorene (AF) as substrates were examined in the cytosol of K . pneumoniae . NAT activity (N-acetylation of substrates) was determined using an acetyl coenzyme A recycling assay and high performance liquid chromatography for determining the amounts of acetylated or non-acetylated PABA or AF . NAT activities from a number of K . pneumoniae isolates were found to be 0.72 +/- 0.08 nmol/min/mg protein for AF, and 0.49 +/- 0.04 nmol/min/mg protein for PABA . The kinetic parameters of apparent Michaelis constant (Km) and maximum velocity (Vmax) obtained were 2.92 +/- 0.48 mM and 7.89 +/- 0.82 nmol/min/mg protein, respectively, for AF and 2.42 +/- 0.28 mM and 9.87 +/- 0.64 nmol/min/mg protein, respectively, for PABA . The optimal pH value for the NAT activity was 7.0 for AF and PABA . The optimal temperature for NAT activity was 37 degrees C for both substrates . The NAT activity was inhibited by 50% with 0.25 mM iodoacetamide, and by more than 90% at 1.0 mM . Among a series of divalent cations and salts, Cu2+ and Zn2+ were the most potent inhibitors of NAT activity. Jpn J Infect Dis, 2004 Aug, 57(4), 150 - 5 Protective role of liposome incorporated lipopolysaccharide antigen of Klebsiella pneumoniae in a rat model of lobar pneumonia; Chhibber S et al.; In a lobar pneumonia model of Klebsiella pneumoniae, the immunoprotective role of free lipoploysaccharide (LPS) and liposome-incorporated LPS was studied . An alteration in the biological activity of the LPS molecule, in terms of its pyrogenicity and lethal toxicity, was observed on incorporation in the liposome . Compared at equal doses, liposome-incorporated LPS was found to be non-pyrogenic and 10 times less toxic than free LPS . Liposome-incorporated LPS was more effective in providing protection against K . pneumoniae induced lobar pneumonia in rats . The immunological mechanism underlying protection revealed involvement of both nonspecific and specific immune response . Alveolar macrophage activation was observed after 4 and 14 days of treatment with the free and liposome-entrapped forms of LPS, respectively . Specific immunity in terms of plaque-forming cells was seen with both forms of LPS . Delayed type hypersensitivity reaction was observed only with liposome-incorporated LPS . It is concluded that a non-toxic and immunogenic form of K . pneumoniae LPS can be obtained by incorporation of the native preparation into liposomes. J Dairy Sci, 2004 Aug, 87(8), 2420 - 32 Characterization of the bovine innate immune response to intramammary infection with Klebsiella pneumoniae; Bannerman DD et al.; Gram-negative bacteria are responsible for almost one-half of the clinical cases of mastitis that occur annually . Of those gram-negative bacteria that induce mastitis, Klebsiella pneumoniae remains one of the most prevalent . Detection of infectious pathogens and the induction of a proinflammatory response are critical components of host innate immunity . The objective of the current study was to characterize several elements of the bovine innate immune response to intramammary infection with Klebsiella pneumoniae . The inflammatory cytokine response and changes in the levels of soluble CD14 (sCD14) and lipopolysaccharide (LPS)-binding protein (LBP), 2 proteins that contribute to host recognition of gram-negative bacteria, were studied . The contralateral quarters of 7 late-lactating Holstein cows were challenged with either saline or K . pneumoniae, and milk and blood samples were collected . Initial increases in the chemoattractants C5a and IL-8, as well as TNF-alpha, were evident in infected quarters within 16 h of challenge and were temporally coincident with increases in milk somatic cells . Augmented levels of TNF-alpha and IL-8 were observed in infected quarters until >48 h postchallenge, respectively . Elevated levels of IL-12, IFN-gamma, and the antiinflammatory cytokine, IL-10, which were first detected between 12 and 20 h postinfection, persisted in infected quarters throughout the study (>96 h) . Initial increases in milk LBP and sCD14 were detected 16 and 20 h, respectively, after challenge . Together, these data demonstrate that intramammary infection with K . pneumoniae elicits a host response characterized by the induction of proinflammatory cytokines and elevation of accessory molecules involved in LPS recognition. Antimicrob Agents Chemother, 2004 Sep, 48(9), 3477 - 82 Increased serum resistance in Klebsiella pneumoniae strains producing extended-spectrum beta-lactamases; Sahly H et al.; The aim of this study was to determine whether there is an association between serum resistance, O serotypes, and the production of extended-spectrum beta-lactamases (ESBLs) in Klebsiella pneumoniae . Ninety ESBL-producing and 178 non-ESBL-producing K . pneumoniae isolates gathered in five European countries were O serotyped and tested for sensitivity to the serum's bactericidal effect . The frequency of serum-resistant isolates was higher among ESBL-producing strains (30%; 27/90 isolates) than among non-ESBL-producing strains (17.9%; 32/178 isolates) (P = 0.037; odds ratio {OR} = 1.96; 95% confidence interval {95% CI} = 1.08 to 3.53) . Although O1 was the most common O serotype in both Klebsiella groups, its frequency among ESBL-producing strains was significantly higher (59%; 53/90 isolates) than among non-ESBL producers (36%; 64/178 isolates) (P = 0.0006; OR = 2.5; 95% CI = 1.52 to 4.29) . Furthermore, the prevalence of the O1 serotype was higher among serum-resistant strains of both ESBL-producing (74%; 20/27isolates) and non-ESBL producers (75%; 24/32 isolates) than among serum-sensitive ESBL producers (52.4%; 33/63 isolates) and non-ESBL producers (27.4%; 40/146 isolates) . Serum resistance among ESBL-producing strains (36%; 17/47 isolates) versus non-ESBL-producing strains (16%; 27/166 isolates) was also significantly higher after the exclusion of clonal strains (P = 0.0056; OR = 2.9; 95% CI = 1.41 to 6.01) . Sixteen ESBL types were detected, among which the frequency of serum resistance was significantly lower among the SHV-producing strains (9/48 isolates) than among the TEM producers (16/35 isolates) (P = 0.016; OR = 3.65; CI = 1.3 to 9.7) . Curing ESBL-coding plasmids did not influence the serum resistance of the bacteria; all six plasmid-cured derivatives maintained serum resistance . The present findings suggest that ESBL-producing strains have a greater pathogenic potential than non-ESBL-producing strains, but the linkage between O serotypes, serum resistance, and ESBL production remains unclear at this stage. Zhonghua Nei Ke Za Zhi, 2004 Jul, 43(7), 487 - 90 {Plasmid-mediated cephalosporinase among extended-spectrum-beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae}; Wang QT et al.; OBJECTIVE: To investigate the prevalence and genotype of plasmid-mediated cephalosporinase (AmpC) beta-lactamase in extended-spectrum-beta-lactamase-producing (ESBL) Escherichia coli and Klebsiella pneumoniae . METHODS: 24 strains of cefoxitin-resistant ESBL-producing E . coli and 8 strains of K . pneumoniae were collected from January to December 2001 at Beijing Chaoyang Hospital . Analytical isoelectric focusing electrophoresis was used to measure the pI of the beta-lactamase . Conjugation experiment was used to study the transfer of cefoxitin resistance . The homology of the isolates was determined by pulsed field gel electrophoresis (PFGE) . Plasmid-mediated AmpC enzyme genes were amplified and sequenced by using multiplex PCR . RESULTS: The prevalence of ESBL-producing E . coli and K . pneumoniae were 16.8% (49/292) and 16.5% (35/212), respectively . The prevalence of AmpC enzyme among ESBL-producing E . coli and K . pneumoniae isolates were 2.0% (1/49) and 17.1% (6/35), respectively . These 7 isolates produced DHA-1 AmpC enzyme . One strain of K . pneumoniae could transfer cefoxition resistance to the recipient . Among 7strains, 5 strains produced CTX-M-3 and 2 produced SHV-12 ESBL . These 7 strains also produced TEM-1 broad-spectrum enzymes . These strains harbored 2 - 5 plasmids and one of them were 33 - 36 kb . PFGE showed these strains came from a variety of clones . CONCLUSIONS: In this hospital, 7 strains of the ESBL-positive E . coli and K . pneumoniae produced both DHA-1AmpC enzyme and CTX-M-3/SHV-12 ESBL . These 7 strains were from different clones. Lik Sprava, 2001 Jan-Feb, (1), 97 - 102 {Pathomorphological aspects and outcome of influenza virus and Klebsiella pneumonia in the experiment}; Anisimova IuN; Mice infected with K . pneumoniae against the background of prior introduction of influenza A/Hong-Kong/68 virus developed focal bronchial pneumonia with leukocytic exudate that started resolving at day 10 of the experiment . At about this time and later there appeared in the lungs spots of apparent proliferative processes characteristic of productive inflammation . These were manifested by diffuse and focal infiltration of the interstice with plasmablasts, plasmacytes, histiocytes, lymphocytes, by proliferation of the vascular endothelium epithelium, of the bronchi, bronchioles, alveolar cells that were seen to be desquamative and developing obliterating macrophagal alveolitis, focal proliferation of fibroblasts . Abnormalities in immunoregulatory mechanisms were induced by immunodepressive action of the influenza virus, early chronization of viral-bacterial pneumonia, with pneumosclerosis developing in its wake. J Korean Med Sci, 2004 Aug, 19(4), 608 - 10 Klebsiella pneumoniae Septic Arthritis in a Cirrhotic Patient with Hepatocellular Carcinoma; Park CH et al.; Despite septic arthritis is increasingly being reported in elderly patients with diabetes or alcoholism, reported cases of spontaneous bacterial arthritis in cirrhotic patients are extremely rare . We present the first reported case of K . pneumoniae septic arthritis and spontaneous bacterial peritonitis in a cirrhotic patient with hepatocellular carcinoma . K . pneumoniae, one of the most common causative organisms of spontaneous bacterial peritonitis in cirrhotic patients, was isolated from both the blood and the joint fluid, which suggests that the route of infection was hematogenous . After the treatment with cefotaxime and closed tube drainage, the condition of the patient was improved, and subsequently, the joint fluid became sterile and the blood cultures were proved negative . Therefore, this case provides further evidence for the mode of infection being bacteremia in cirrhotic patients and suggests that the enteric bacteremia in cirrhotics may cause infection in different organ systems . Copyright The Korean Academy of Medical Sciences Clin Microbiol Infect, 2004 Aug, 10(8), 760 - 2 Results of two worldwide surveys into physician awareness and perceptions of extended-spectrum beta-lactamases; Goossens H et al.; An omnibus survey of microbiologists (n = 400) and a survey of participants (n = 49) in the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) programme were conducted to determine the awareness and prevalence of extended-spectrum beta-lactamases (ESBLs), and the regularity and method of screening . Of the omnibus survey participants, 69% screened regularly for ESBLs, compared with 83% of MYSTIC participants . In both surveys, ESBLs were more common in Klebsiella pneumoniae (73% and 79%, respectively) and Escherichia coli (63% and 81%, respectively) than in other bacteria . The surveys demonstrated that awareness of, and testing for, ESBLs is inconsistent. Acta Crystallogr D Biol Crystallogr, 1997 Mar, 53(Pt 2), 227 - 8 Crystallization and preliminary X-ray studies of nitrogenase component 1 (the MoFe protein) from Klebsiella pneumoniae; Roe SM; Two crystal forms of component 1 (the MoFe protein) of nitrogenase from Klebsiella pneumoniae have been isolated and characterized . The triclinic form has cell dimensions a = 76.0, b = 109.6, c = 144.6 A, alpha = 80.3, beta = 74.9 and gamma = 69.6 degrees, diffracts to around 3.0 A and has two molecules in the asymmetric unit . The monoclinic form belongs to space group P2(1) with a = 76.6, b = 127.8, c = 109.1 A and beta = 104.6 degrees (frozen at 100 K), diffracts to 1.5 A and has one molecule in the asymmetric unit . At this resolution the outstanding questions concerning the structure and the operation of the enzyme, in particular the linkage between the Fe(4)S(4) units in the P clusters, the true geometry of the apparently trigonal Fe atoms in the FeMoco and the reduction site itself, should be answerable. J Clin Microbiol, 2004 Aug, 42(8), 3388 - 98 Molecular serotyping of Klebsiella species isolates by restriction of the amplified capsular antigen gene cluster; Brisse S et al.; The objective of the present work was to develop a molecular method that would enable determination of the capsular serotypes of Klebsiella isolates without the use of antiserum . PCR amplification of the capsular antigen gene cluster (cps) was followed by digestion with the restriction enzyme HincII (cps PCR-restriction fragment length polymorphism {RFLP} analysis) . The profiles (C patterns) obtained for 224 strains representing the 77 known K serotypes showed 3 to 13 fragments ranging in size from 0.2 to 4.4 kb . A total of 97 distinct C patterns were obtained; 100% of 61 pairs of samples tested twice showed reproducible C patterns . The C patterns were K-type specific; i.e., the C pattern(s) of any K serotype was distinct from the C patterns of all other K serotypes, with the only exceptions being serotypes K22 and K37, which are known to cross-react . For 12 of 17 K types for which at least two strains were included, C-pattern variations were found among strains with the same K serotype . Therefore, cps PCR-RFLP analysis has a higher discriminatory power than classical K serotyping . C-pattern identity was observed among strains with a given K type that were collected many years apart and from distinct sources, indicating C-pattern stability . Only 4.5% of the strains were nontypeable, because of unsuccessful PCR amplification (whereas 8 to 23% are nontypeable by classical K serotyping) . Three of four noncapsulated strains analyzed showed recognizable C patterns . The K serotypes of 18 (82%) of 22 recent Klebsiella pneumoniae clinical isolates could be deduced from their C patterns . In conclusion, cps PCR-RFLP analysis allows determination of the K serotype, while it is easier to perform and more discriminatory than classical serotyping. Biotechnol Prog, 2004 Jul-Aug, 20(4), 1069 - 75 Biokinetic evaluation and modeling of continuous thiocyanate biodegradation by Klebsiella sp; Ahn JH et al.; Biokinetics for autotrophic degradation of thiocyanate using batch culture of Klebsiella sp . were evaluated both analytically and numerically . A sequential approach with an analytical method followed by a numerical approximation was used to evaluate and to ensure the accuracy of the parameter estimation . The nonlinear least-squares method with a 95% confidence interval was employed . The growth conditions were maintained at pH 7 and 38 degrees C for all experiments . With an automated incubation and turbidity reader, a total of 16 different initial thiocyanate concentrations, ranging from 10 to 300 mg L(-1), were used to develop a kinetic expression of specific growth rate as a function of substrate concentration . The biodegradation of thiocyanate with Klebsiella sp . followed a substrate inhibition pattern . Three identical automated bioreactors with working volumes of 1.5 L, equipped with sterilizable sampling ports, were also used for the numerical approximation of the biokinetic parameters in batch mode . A fourth order Runge-Kutta method was used to approximate the substrate inhibition kinetics of the Klebsiella sp . utilizing thiocyanate . Although the kinetic coefficients estimated by analytical and numerical methods were not statistically different at a 0.05 alpha level, model responses of numerical approximation generated a better prediction of changes in thiocyanate and cell mass concentrations . The hypothetical maximum growth rate, micro m, half saturation coefficient, Ks, microbial yield coefficient, Y, cell mass decay rate coefficient, kd, and substrate inhibition coefficient, Ksi, were evaluated as being 0.62 +/- 0.05 d(-1), 85 +/- 8 mg SCN- L(-1), 0.076 +/- 0.011 mg cell mass (mg SCN)(-1), 0.03 +/- 0.002 d(-1), and 131 +/- 22 mg SCN- L(-1), respectively . The calculated maximal substrate concentration, Sm, and apparent maximum specific growth rate, micro'm, were 105.5 +/- 8.7 mg SCN- L(-1) and 0.24 +/- 0.01 d(-1), respectively . Using these estimated parameters, the theoretical performance of the continuous operation was also illustrated, which depicts the residual thiocyanate and Klebsiella sp . concentrations in the non-steady and steady states at different hydraulic retention times (HRTs) . Assuming the influent concentration of 250 mg SCN- L(-1), the expected treatment efficiency ranged from 94.9% to 69.4% between 20 and 5 days HRT, respectively . Klebsiella sp . was expected to be washed out at 4.8 days HRT, thus resulting in no treatment of thiocyanate. Eur J Biochem, 2004 Aug, 271(16), 3379 - 88 GlnK effects complex formation between NifA and NifL in Klebsiella pneumoniae; Stips J et al.; In Klebsiella pneumoniae, the nif specific transcriptional activator NifA is inhibited by NifL in response to molecular oxygen and ammonium . Here, we demonstrate complex formation between NifL and NifA (approximately 1 : 1 ratio), when synthesized in the