Infect Immun, 2003 Jul, 71(7), 4182 - 5 Leptin modulates neutrophil phagocytosis of Klebsiella pneumoniae; Moore SI et al.; Leptin is a pleiotropic hormone-cytokine known to regulate energy homeostasis and immune function . Neutrophils from leptin-deficient mice exhibited impaired phagocytosis of Klebsiella pneumoniae opsonized with serum containing complement and reduced CD11b expression that could be restored with exogenous leptin . These results suggest that leptin is required for normal neutrophil complement-mediated phagocytosis of bacteria. J Am Med Dir Assoc, 2002 Mar-Apr, 3(2), 71 - 2 Rhinoscleroma in an octogenarian woman; Leibovitz A et al.; Rhinoscleroma, a granulomatous infection caused by Klebsiella rhinoscleromatis and usually afflicting young adults, was diagnosed in an 81-year-old woman . Treatment was started with orally administered antibiotics but was soon halted because of side effects . She was treated instead by local spread of a 3% rifampin ointment . After 6 weeks, the lesion cleared up and she improved remarkably . There was no recurrence during 18 months of follow-up . The current report is the second published case of rhinoscleroma in an elderly person and the first report on its cure by local treatment with a 3% rifampin ointment . This favorable experience contributes to the limited knowledge on rhinoscleroma in the elderly. West Indian Med J, 2003 Mar, 52(1), 37 - 40 Hepatic abscesses at the University Hospital of the West Indies . A 24-year autopsy review; Gaskin DA et al.; A retrospective review of all hepatic abscesses identified at autopsy over the 24-year period 1977-2000 at the University Hospital of the West Indies (UHWI), Kingston, Jamaica, was conducted . Post mortem and microbiology records were reviewed . Data collected included age, gender, predisposing factors, organisms isolated, number of abscesses and associated conditions, such as diabetes mellitus . Data for the adults was analyzed separately . Thirty-nine cases of pyogenic abscesses were identified from 7480 post-mortems . Thirty-three occurred in adults of mean age 59.5 years . Sixty per cent of the abscesses were solitary . Biliary tract disease was the predisposing factor in 33% of cases . Six per cent were cryptogenic: 11/33 patients were diabetic and these were significantly older than non-diabetics (p < 0.014) Klebsiella pneumoniae was the most common organism isolated from diabetics (6.9) . Only 2/33 abscesses were diagnosed ante-mortem . The abscesses in children were more frequently multiple and associated with extra-abdominal infection . Gram positive cocci were the commonly isolated organisms in children . There were no cases of amoebic abscess . The prevalence of hepatic abscess was low . Diabetes mellitus was a significant contributing factor . A high index of clinical suspicion is therefore warranted particularly in elderly diabetics. FEBS Lett, 2003 Jun 12, 545(1), 9 - 17 Proton pumping by NADH:ubiquinone oxidoreductase . A redox driven conformational change mechanism? Brandt U, Kerscher S, Drose S, Zwicker K, Zickermann V. The modular evolutionary origin of NADH:ubiquinone oxidoreductase (complex I) provides useful insights into its functional organization . Iron-sulfur cluster N2 and the PSST and 49 kDa subunits were identified as key players in ubiquinone reduction and proton pumping . Structural studies indicate that this 'catalytic core' region of complex I is clearly separated from the membrane . Complex I from Escherichia coli and Klebsiella pneumoniae was shown to pump sodium ions rather than protons . These new insights into structure and function of complex I strongly suggest that proton or sodium pumping in complex I is achieved by conformational energy transfer rather than by a directly linked redox pump. Biosci Biotechnol Biochem, 2003 Apr, 67(4), 933 - 6 Microbial production of 2-deoxyribose 5-phosphate from acetaldehyde and triosephosphate for the synthesis of 2'-deoxyribonucleosides; Ogawa J et al.; 2-Deoxyribose 5-phosphate was produced from acetaldehyde and dihydroxyacetone phosphate via D-glyceraldehyde 3-phosphate by Klebsiella pneumoniae B-4-4 through deoxyriboaldolase- and triosephosphate isomerase-catalyzing reactions . Under the optimum conditions, 98.7 mM 2-deoxyribose 5-phosphate was produced from 200 mM acetaldehyde and 117 mM dihydroxyacetone phosphate in 2 h with a molar yield of 84% . The 2-deoxyriobse 5-phosphate produced was directly transformed to 2'-deoxyribonucleoside by phosphopentomutase- and nucleoside phosphorylase-catalyzing reactions. Int J Immunopathol Pharmacol, 1999 May, 12(2), 103 - 111 Effect of bacterial extracts on the immunologic profile in chronic relapsing brucellosis patients; Boura P et al.; Brucellosis is an intracellular bacterial disease of common incidence in Greece . Existing therapy is inadequate and a considerable proportion of patients become chronically ill and are immunocompromised . Defects of the monocyte-macrophage system and T-lymphocytes have been described in chronic brucellosis and can be restored after immunopotentiation therapy . Bacterial (Klebsiella pneumoniae) extracts exert immunostimulating effects on the monocyte-macrophage system and have already been used successfully in the prevention of common infections of the respiratory track . So we decided to investigate: 1) Leukocyte Migration Index (LMI), 2) Monocyte-macrophage random and directed migration against both nonspecific leukoattractant (casein) and disease specific antigens (Brucella melitensis, Brucella abortus), 3) Monocyte-macrophage phagocytosis index, 4) Delayed-type hypersensitivity (skin tests) against seven antigens, before (TO), during (T2), and after (T3) oral administration of bacterial (Klebsiella pneumoniae) extracts at conventional doses plus antibiotics or not . Our results show that: 1) Concerning the LMI, 4 out of 19 remained anergic at time T3 of the study, 2) Random migration was not affected during treatment, 3) Directed migration increased significantly without reaching control group values, 4) Phagocytosis index increased significantly and reached normal values at T3, 5) Delayed type hypersensitivity reactions (skin tests) increased significantly at the end of the study period . Reaction against Tuberculin and Candida antigens showed the most pronounced increase in skin reactivity . In conclusion, bacterial (Klebsiella pneumoniae) extracts improve peripheral monocyte locomotion and restore phagocytosis index, thus enhancing cellular immunity parameters in immunocompromised chronic brucellosis patients. J Formos Med Assoc, 2003 Mar, 102(3), 189 - 92 Therapeutic effect of interferon-gamma for prevention of severe infection in X-linked chronic granulomatous disease; Ma HR et al.; Chronic granulomatous disease (CGD) is an inherited phagocytic disorder caused by defective oxidative burst activity of neutrophils and monocytes . Patients with CGD may present with recurrent, life-threatening bacterial or fungal infections and often need repeated hospitalization as early as infancy . We report a case of a boy aged 3 years and 1 month with a history of oral thrush, chronic diarrhea, skin abscesses, multiple small joints osteomyelitis, and multiple liver abscesses since 2 months of age . X-linked chronic granulomatous disease was diagnosed by nitroblue tetrazolium test and further confirmed by genetic study using single strand conformation polymorphism (SSCP) analysis and genomic DNA sequencing . Two episodes of aspergillosis with severe aspergillus lung empyema and life-threatening Klebsiella pneumoniae infections were noted in the first 2 years of life despite long-term prophylaxis with trimethoprim-sulfamethoxazole and itraconazole . Recombinant human interferon-gamma (rINF-gamma) 50 microg/m(2) subcutaneous injection 3 times per week was added after his last episode of severe infection . Thereafter, the boy had normal growth and development with no evidence of severe infection during 18 months of follow-up at our outpatient clinic. Klin Monatsbl Augenheilkd, 2003 May, 220(5), 357 - 61 {Long-term follow-up of bilateral endogenous Klebsiella endophthalmitis}; Holak H et al.; BACKGROUND: Endophthalmitis subsequent to Klebsiella sepsis leads to functional blindness in most cases and is very difficult to treat . Every successful therapeutic modality can therefore help in creating an optimal therapeutic plan . CASE REPORT: A 69-year old diabetic patient exhibited bilateral Klebsiella endophthalmitis with sepsis after a pneumonia . Two intravenous antibiotics were used: aminoglycosides (Gentamycin) and cephalosporins (Cefotaxim or Cefuroxim) with local parabulbar injections of Prednisolon . The long-term follow-up of four years provided some overview of morphological aspects of the development of endophthalmitis . Characteristic greyish hypopyon was seen in both eyes, which was more pronounced in the left eye than in the right . The left eye became phthisic . After resorption of the hypopyon in the right eye and prolonged resorption of the subretinal abscess for 9 months a useful visual acuity at 0.2 was achieved . Two years after the endophthalmitis a cataract surgery with implantation of a posterior chamber silicon lens was performed and good visual acuity (0.6) was achieved . After four years, the subretinal abscess left an extremely large, sharp bordered, unpigmented scar up to the sclera . CONCLUSION: An early diagnosis and adequate long-time antibiotic therapy under the co-operative supervision of an ophthalmologist with internist appears to be most important for the therapeutic success in Klebsiella endophthalmitis. Zhonghua Er Bi Yan Hou Ke Za Zhi, 2001 Feb, 36(1), 42 - 3 {Clinicopathologic analysis of rhinoscleroma}; Zhang S et al.; OBJECTIVE: To investigate the clinicopathologic characteristics and reason of tissue injury caused by klebsiella rhinoscleromatis(KR) infection . METHODS: Twenty-four cases of rhinoscleromas in the nasal and pharyngeal region in this hospital from March 1983 to March 1998 were studied retrospectively . Warthin-Starry (W-S) stain was used in all cases and CD43, CD20, CD68, Lysozyme mark were observed in eleven specimens by ABC immunochemical method . Two cases were studied further with transmission electron microscopy . RESULTS: A great number of KR were found in Mikulicz cells and were confirmed further in the phagosomes in cytoplasm of Mikulicz cells, where a small number of endoplasmic reticulums and lysozymes was squeezed to the side of cells . A few of T-lymphocytes was found within the granuloma . CONCLUSION: Cellular immunity function of the patient with rhinoscleroma was depressed and the response to immunization of host which regards to KR was the major cause of tissue injury. FEMS Microbiol Lett, 2003 May 16, 222(1), 93 - 8 Sequence analysis and biochemical characterisation of chromosomal CAV-1 (Aeromonas caviae), the parental cephalosporinase of plasmid-mediated AmpC 'FOX' cluster; Fosse T et al.; Aeromonas caviae CIP 74.32 was resistant to amoxicillin, ticarcillin and cephalothin, and susceptible to cefoxitin, cefotaxime, ceftazidime, aztreonam and imipenem . This strain produced a cephalosporinase (pI 7.2) and an oxacillinase (pI 8.5) . The cephalosporinase gene cav-1 was cloned and sequenced . Unlike A . caviae donor, Escherichia coli pNCE50 transformant producing CAV-1 beta-lactamase was resistant to cefoxitin . The deduced protein sequence CAV-1 contained 382 amino acids, and shared >96% homology with FOX-1 to FOX-5 cephalosporinase . CAV-1 presented only two amino acid substitutions (Thr270Ser and Arg271Ala) with FOX-1 . CAV-1 is the chromosomal putative ancestor of the FOX family, a cluster of class C/group 1 plasmidic cephalosporinases spreading in Klebsiella and E . coli clinical isolates via conjugative plasmids. J Bacteriol, 2003 Jun, 185(11), 3416 - 28 Type IV-like pili formed by the type II secreton: specificity, composition, bundling, polar localization, and surface presentation of peptides; Vignon G et al.; The secreton or type II secretion machinery of gram-negative bacteria includes several type IV pilin-like proteins (the pseudopilins) that are absolutely required for secretion . We previously reported the presence of a bundled pilus composed of the pseudopilin PulG on the surface of agar-grown Escherichia coli K-12 cells expressing the Klebsiella oxytoca pullulanase (Pul) secreton genes at high levels (N . Sauvonnet, G . Vignon, A . P . Pugsley, and P . Gounon, EMBO J . 19:2221-2228, 2000) . We show here that PulG is the only pseudopilin in purified pili and that the phenomenon is not restricted to the Pul secreton reconstituted in E . coli or to PulG . For example, high-level expression of the endogenous E . coli gsp secreton genes caused production of bundled pili composed of the pseudopilin GspG, and the Pul secreton was able to form pili composed of PulG-like proteins from secreton systems of other bacteria . PulG derivatives in which the C terminus was extended by the addition of eight different peptides were also assembled into pili and functioned in secretion . Three of the C-terminal peptides were shown to be exposed along the entire length of the assembled pili . Hence, the C terminus of PulG may represent a permissive site for the insertion of immunogenic epitopes or other peptide sequences . One of these PulG variants, with a six-histidine tag at its C terminus, formed nonpolar, nonbundled pili, suggesting that bundle formation and polar localization are not correlated with the ability of PulG to function in secretion . We propose that the PulG pilus is an artifactual manifestation of a periplasmic "pseudopilus" and that cycles of pseudopilus extension and retraction within the periplasm propel pullulanase through secretin channels in the outer membrane . Abnormally long pili that extend beyond the outer membrane are produced only when pilus length control and retraction are deregulated by overproduction of the major pseudopilus subunit (PulG). Natl Toxicol Program Tech Rep Ser, 1986 Oct, 257, 1 - 222 NTP Toxicology and Carcinogenesis Studies of Diglycidyl Resorcinol Ether (Technical Grade) (CAS No . 101-90-6) In F344/N Rats and B6C3F1 Mice (Gavage Studies); National Toxicology Program ; Diglycidyl resorcinol ether (DGRE), a pale, yellow, translucent, amorphous solid at room temperature, is used as a liquid spray epoxy resin, as a diluent in the production of other epoxy resins used in electrical, tooling, adhesive, and laminating applications, and as a curing agent for polysulfide rubber . Approximately 3,000 workers are exposed to DGRE . The quantity of DGRE produced in the United States is not known . Toxicology and carcinogenesis studies of technical grade diglycidyl resorcinol ether (81% pure) were conducted by administering the chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats at doses of 25 or 50 mg/kg and to groups of 50 male and female B6C3F1 mice at doses of 50 or 100 mg/kg . A supplemental study of similar design in male and female rats (0 or 12 mg/kg) was started approximately 12 months later because of high mortality in the 50 mg/kg dose groups . Doses were administered five times per week for 103 weeks . Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same dosing schedule and served as vehicle controls . Throughout most of the primary study, mean body weights of high dose male and female rats and female mice were lower than those of the corresponding vehicle controls . In the supplemental study, body weights of both sexes of the dosed rats were unaffected by administration of DGRE . Survival of dosed rats of each sex in the primary study was dose related and was shorter (P<0.001) than that of the vehicle controls . No high dose male rats and only 1/50 high dose female rats lived to the end of the study . Bronchopneumonia was the most frequent cause of early death among the rats and may have resulted from the animals' aspiration of corn oil containing diglycidyl resorcinol ether . Survival of the dosed male rats in the supplemental study was reduced (P<0.005) when compared to controls . There was no significant difference in survival between dosed and control female rats in the supplemental study . Survival of dosed and control mice was comparable but poorer in females, with 20/50 (40%) of the controls, 13/50 (26%) of the low dose, and 10/50 (20%) of the high dose groups alive at the end of 2 years . These early deaths were due to suppurative and necrotizing inflammation of the reproductive tract, possibly caused by a Klebsiella sp . infection . The incidences of rats and mice with hyperkeratosis and hyperplasia of the forestomach were compound related . For rats and mice of each sex, incidences of animals with squamous cell papillomas, squamous cell carcinomas, or both occurred with statistically significant positive trends and the incidences observed in other organs in dosed groups relative to the controls . An audit of the experimental data was conducted for the 2-year studies of diglycidyl resorcinol ether . No data discrepancies were found that influenced the final interpretations . Under the conditions of these 2-year gavage studies, technical grade diglycidyl resorcinol ether caused hyperkeratosis and hyperplasia of the forestomach in rats and mice . DGRE was carcinogenic for male and female F344/N rats and for male and female B6C3F1 mice, causing both benign and malignant neoplasms of the forestomach . Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonym: DGRE Int J Food Microbiol, 2003 Jun 25, 83(3), 295 - 305 The effect of abrupt shifts in temperature on the lag phase duration of Escherichia coli and Klebsiella oxytoca; Mellefont LA et al.; The effect of temperature of incubation on lag times of two gram-negative foodborne bacteria was investigated . Bacteria were instantaneously transferred between temperatures within and beyond the normal physiological temperature range (NPTR) . Abrupt temperature shifts induced lag phases, but the degree of the response was dependent on the direction and magnitude of the shift . Temperature downshifts induced larger relative lag times (RLT; the ratio of lag time to generation time), than equivalent upshifts . The hypothesis of Robinson et al . {Int . J . Food Microbiol . 44 (1998) 83} that lag time can be understood in terms of the amount of work to be done to adjust to new environmental conditions and the rate at which that work is done was supported . Deviation of the reported proportionality between lag time and generation time was observed when late-exponential phase cells were subjected to abrupt temperature shifts from beyond the normal physiological range. West Afr J Med, 2002 Jul-Sep, 21(3), 252 - 5 Klebsiella-induced purpura fulminans in a Nigerian child: case report and a review of literature; Olowu WA; Purpura fulminans (PPF) is a very severe but rare acute thrombohaemorrhagic illness of infants and young children . It occurs mainly, in patients with either congenital or acquired deficiencies of proteins C and S and antithrombin III . Features of PPF include disseminated intravascular coagulopathy, symmetrical necrotic purpura and/or ecchymoses and symmetrical peripheral gangrene; digital and/ or limb(s) amputations and end-organ failure(s) may also occur . The case of a 3.5 year-old Nigerian girl, who developed PPF following Klebsiella-rhinoscleromatis septicaemia is reported to illustrate the seriousness of the disease and the need for early diagnosis and management. Biochemistry, 2003 May 20, 42(19), 5917 - 24 Identification of basic amino acid residues important for citrate binding by the periplasmic receptor domain of the sensor kinase CitA; Gerharz T et al.; The sensor kinase CitA and the response regulator CitB of Klebsiella pneumoniae form the paradigm of a subfamily of bacterial two-component regulatory systems that are capable of sensing tri- or dicarboxylates in the environment and then induce transporters for the uptake of these compounds . We recently showed that the separated periplasmic domain of CitA, termed CitAP (encompasses residues 45-176 supplemented with an N-terminal methionine residue and a C-terminal hexahistidine tag), is a highly specific citrate receptor with a K(d) of 5.5 microM at pH 7 . To identify positively charged residues involved in binding the citrate anion, each of the arginine, lysine, and histidine residues in CitAP was exchanged for alanine, and the resulting 17 muteins were analyzed by isothermal titration calorimetry (ITC) . In 12 cases, the K(d) for citrate was identical to that of wild-type CitAP or slightly changed (3.9-17.2 microM) . In one case (R98A), the K(d) was 6-fold decreased (0.8 microM), whereas in four cases (R66A, H69A, R107A, and K109A) the K(d) was 38- to >300-fold increased (0.2 to >1 mM) . The secondary structure of the latter five proteins in their apo-form as deduced from far-UV circular dichroism (CD) spectra did not differ from the apo-form of wild-type CitAP; however, all of them showed an increased thermostability . Citrate increased the melting point (T(m)) of wild-type CitAP and mutein R98A by 6.2 and 9.5 degrees C, respectively, but had no effect on the T(m) of the four proteins with disturbed binding . Three of the residues important for citrate binding (R66, H69, and R107) are highly conserved in the CitA subfamily of sensor kinases, indicating that they might be involved in ligand binding by many of these sensor kinases. Salud Publica Mex, 2003 Mar-Apr, 45(2), 90 - 5 {Neonatal sepsis morbidity and mortality in a tertiary care hospital.}; Rodriguez-Weber MA et al.; OBJECTIVE: To compare the epidemiological, clinical and microbiological profiles between patients with neonatal sepsis who lived or died . MATERIAL AND METHODS: The medical records of patients with neonatal sepsis were retrospectively reviewed at Instituto Nacional de Pediatria (National Pediatric Institute) of Secretaria de Salud (Ministry of Health) in Mexico City, between 1992 and 2000 . Neonatal sepsis cases were classified as surviving or not after 90 days of postnatal follow-up . The survivor and decreased groups were compared using Mann-Whitney's U test for continuous variables, and the chi-squared test or the Fisher's exact test for categorical variables . Significantly associated variables were included in a Cox proportional hazards model . A p-value < 0.05 was considered statistically significant for all analyses . RESULTS: A total of 116 patients with neonatal sepsis were included (65 live and 51 dead) . Multivariate analysis showed that fetal distress, respiratory distress, a delayed capillary fill up, a low platelet count, and a positive hemoculture for Klebsiella pneumoniae were significant risk factors for death . CONCLUSIONS: Epidemiological, clinical, laboratory, and microbiological variables are significant predictors of death in newborns with neonatal sepsis . The English version of this paper is available at: http://www.insp.mx/salud/index.html. Curr Microbiol, 2003 Jun, 46(6), 423 - 31 Altered kinetic properties of tyrosine-183 to cysteine mutation in glutamine synthetase of anabaena variabilis strain SA1 is responsible for excretion of ammonium ion produced by nitrogenase; Healy FG et al.; A L-methionine- D, L-sulfoximine-resistant mutant of the cyanobacterium Anabaena variabilis, strain SA1, excreted the ammonium ion generated from N(2) reduction . In order to determine the biochemical basis for the NH(4)(+)-excretion phenotype, glutamine synthetase (GS) was purified from both the parent strain SA0 and from the mutant . GS from strain SA0 (SA0-GS) had a pH optimum of 7.5, while the pH optimum for GS from strain SA1 (SA1-GS) was 6.8 . SA1-GS required Mn(+2) for optimum activity, while SA0-GS was Mg(+2) dependent . SA0-GS had the following apparent K(m) values at pH 7.5: glutamate, 1.7 m M; NH(4)(+), 0.015 m M; ATP, 0.13 m M . The apparent K(m) for substrates was significantly higher for SA1-GS at its optimum pH (glutamate, 9.2 m M; NH(4)(+), 12.4 m M; ATP, 0.17 m M) . The amino acids alanine, aspartate, cystine, glycine, and serine inhibited SA1-GS less severely than the SA0-GS . The nucleotide sequences of glnA (encoding glutamine synthetase) from strains SA0 and SA1 were identical except for a single nucleotide substitution that resulted in a Y183C mutation in SA1-GS . The kinetic properties of SA1-GS isolated from E . coli or Klebsiella oxytoca glnA mutants carrying the A . variabilis SA1 glnA gene were also similar to SA1-GS isolated from A . variabilis strain SA1 . These results show that the NH(4)(+)-excretion phenotype of A . variabilis strain SA1 is a direct consequence of structural changes in SA1-GS induced by the Y183C mutation, which elevated the K(m) values for NH(4)(+) and glutamate, and thus limited the assimilation of NH(4)(+) generated by N(2) reduction . These properties and the altered divalent cation-mediated stability of A . variabilis SA1-GS demonstrate the importance of Y183 for NH(4)(+) binding and metal ion coordination. Fukuoka Igaku Zasshi, 2003 Feb, 94(2), 31 - 6 Endophthalmitis with Klebsiella pneumoniae liver abscess; Dohmen K et al.; Endogenous endophthalmitis is a rare, but devastating complication of septicemia . The prognosis of maintaining visual acuity in patients with septic endophthalmitis is poor in spite of an early diagnosis and the timely start of conventional therapeutic procedures because the intravitreous drug concentration remains low after the systemic administration of antibiotics due to the blood-ocular barrier . We treated an elderly female patient with endogenous endophthalmitis complicated with disseminated intravascular coagulation associated with a Klebsiella pneumoniae liver abscess . Endophthalmitis developed rapidly and we thus had to perform an enucleation of both eyeballs even though we made an early diagnosis and performed liver abscess drainage as well as the prompt systemic and subconjunctival administration of antibiotics . Our experience in treating this case emphasizes the need to perform the timely intravitreous infusion of antibiotics with a support therapy consisting of the systemic and subconjunctival administration of antibiotics for endogenous endophthalmitis associated with a Klebsiella pneumoniae liver abscess. J Immunol, 2003 May 1, 170(9), 4432 - 6 Cutting edge: roles of Toll-like receptor 4 and IL-23 in IL-17 expression in response to Klebsiella pneumoniae infection; Happel KI et al.; Local production of IL-17 is a significant factor in effective host defense against Gram-negative bacteria . However, the proximal events mediating IL-17 elaboration by T cells remain unclear . In this study, we show in vivo that intact Toll-like receptor 4 signaling in the lung is required for induction of both the p19 transcript of IL-23 and IL-17 protein elaboration in response to Klebsiella pneumoniae . Although IL-17 is widely considered a CD4(+) T cell product, we also demonstrate significant in vitro IL-17 production by CD8(+) T cells after culture in medium from dendritic cells exposed to these bacteria . The dominant portion of this IL-17-inducing activity for both CD4(+) and CD8(+) T cells is IL-23 . These data demonstrate the critical signaling pathway for IL-17 induction in the host response to Gram-negative pulmonary infection and suggest a direct role for IL-23 in CD8(+) T cell IL-17 production. Carbohydr Res, 2003 May 1, 338(10), 1033 - 7 First synthesis of beta-D-Galf-(1-->3)-D-Galp--the repeating unit of the backbone structure of the O-antigenic polysaccharide present in the lipopolysaccharide (LPS) of the genus Klebsiella; Wang H et al.; beta-D-Galactofuranosyl-(1-->3)-D-galactopyranose (1), the repeating unit of the backbone structure of the O-antigenic polysaccharide present in the lipopolysaccharide (LPS) of the genus Klebsiella, has been efficiently synthesized using 1,2:5,6-di-O-isopropylidine-alpha-D-galactofuranose (3) as the glycosyl acceptor and 2,3,5,6-tetra-O-benzoyl-beta-D-galactofuranosyl trichloroacetimidate (6) as the glycosyl donor with TMSOTf as catalyst by the well-known Schmidt glycosylation method . The preparation of 3 was improved by increasing the ratio of DMF to acetone and employing a solid-supported catalyst. J Bacteriol, 2003 May, 185(9), 2920 - 6 Nitrogen regulation of the codBA (cytosine deaminase) operon from Escherichia coli by the nitrogen assimilation control protein, NAC; Muse WB et al.; Transcription of the cytosine deaminase (codBA) operon of Escherichia coli is regulated by nitrogen, with about three times more codBA expression in cells grown in nitrogen-limiting medium than in nitrogen-excess medium . Beta-galactosidase expression from codBp-lacZ operon fusions showed that the nitrogen assimilation control protein NAC was necessary for this regulation . In vitro transcription from the codBA promoter with purified RNA polymerase was stimulated by the addition of purified NAC, confirming that no other factors are required . Gel mobility shifts and DNase I footprints showed that NAC binds to a site centered at position -59 relative to the start site of transcription and that mutants that cannot bind NAC there cannot activate transcription . When a longer promoter region (positions -120 to +67) was used, a double footprint was seen with a second 26-bp footprint separated from the first by a hypersensitive site . When a shorter fragment was used (positions -83 to +67), only the primary footprint was seen . Nevertheless, both the shorter and longer fragments showed NAC-mediated regulation in vivo . Cytosine deaminase expression in Klebsiella pneumoniae was also regulated by nitrogen in a NAC-dependent manner . K . pneumoniae differs from E . coli in having two cytosine deaminase genes, an intervening open reading frame between the codB and codA orthologs, and a different response to hypoxanthine which increased cod expression in K . pneumoniae but decreased it in E . coli. J Appl Physiol, 2003 Aug, 95(2), 491 - 6 Epub 2003 Apr 11. Active hexose correlated compound enhances resistance to Klebsiella pneumoniae infection in mice in the hindlimb-unloading model of spaceflight conditions; Aviles H et al.; Previous studies have demonstrated that resistance to infection is decreased in Swiss Webster female mice maintained in the hindlimb-unloading model (Aviles H, Belay T, Fountain K, Vance M, and Sonnenfeld G . J Appl Physiol 95: 73-80, 2003; Belay T, Aviles H, Vance M, Fountain K, and Sonnenfeld G . J Allergy Clin Immunol 110: 262-268, 2002) . This is a model of some of the aspects of spaceflight conditions, including lack of load bearing on hindlimbs and a fluid shift to the head . Active hexose correlated compound (AHCC), extracted from Basidiomycete mushrooms, has been shown to induce enhancement of immune responses, including enhanced natural killer activity . In the present study, AHCC was orally administered to mice to determine whether the treatment could decrease immunosuppression and mortality of mice maintained in the hindlimb-unloaded model and infected with Klebsiella pneumoniae . The results of the present study showed that administration of AHCC by gavage for 1 wk (1 g/kg body wt) before suspension and throughout the 10-day suspension period yielded significant beneficial effects for the hindlimb-unloaded group, including 1) . decreased mortality, 2) . increased time to death, and 3) . increased ability to clear bacteria . The results suggest that AHCC can decrease the deleterious effects of the hindlimb-unloading model on immunity and resistance to infection. Conn Med, 2003 Mar, 67(3), 149 - 52 Utilization of extended-spectrum beta-lactamase (ESBL) detection systems in microbiology laboratories: survey of Connecticut hospitals from 1998-2002; Dandekar PK et al.; There has been recent concern raised over the detection, prevalence, and clinical implications of infection with Escherichia coli and Klebsiella spp . which produce extended-spectrum beta-lactamases (ESBLs) . These enzymes hydrolyze beta-lactams including the cephalosporins (i.e., ceftazidime, ceftriaxone, cefotaxime), thus frequently leading to treatment failures . Standard in vitro testing may report these isolates as susceptible when in fact they are resistant in vivo . As a result of this phenomenon, additional testing for suspected isolates is recommended nationally . We surveyed 28 Connecticut hospitals from 1998-2002 to determine if these institutions utilized screening and confirmation methods for suspected isolates . The number of hospitals which have implemented ESBL detection systems doubled from 11 to 22 over the study period . Currently, 15 of the 22 laboratories conduct both screening and confirmatory testing . This expanded testing will be of great assistance to clinicians in optimizing the clinical care of patients with Gramnegative infections. J Biol Chem, 2003 Jun 20, 278(25), 22717 - 25 Epub 2003 Apr 08. Structural rationalization for the lack of stereospecificity in coenzyme B12-dependent diol dehydratase; Shibata N et al.; Adenosylcobalamin-dependent diol dehydratase of Klebsiella oxytoca is apparently not stereospecific and catalyzes the conversion of both (R)- and (S)-1,2-propanediol to propionaldehyde . To explain this unusual property of the enzyme, we analyzed the crystal structures of diol dehydratase in complexes with cyanocobalamin and (R)- or (S)-1,2-propanediol . (R)- and (S)-isomers are bound in a symmetrical manner, although the hydrogen-bonding interactions between the substrate and the active-site residues are the same . From the position of the adenosyl radical in the modeled "distal" conformation, it is reasonable for the radical to abstract the pro-R and pro-S hydrogens from (R)- and (S)-isomers, respectively . The hydroxyl groups in the substrate radicals would migrates from C(2) to C(1) by a suprafacial shift, resulting in the stereochemical inversion at C(1) . This causes 60 degrees clockwise and 70 degrees counterclockwise rotations of the C(1)-C(2) bond of the (R)- and (S)-isomers, respectively, if viewed from K+ . A modeling study of 1,1-gem-diol intermediates indicated that new radical center C(2) becomes close to the methyl group of 5'-deoxyadenosine . Thus, the hydrogen back-abstraction (recombination) from 5'-deoxyadenosine by the product radical is structurally feasible . It was also predictable that the substitution of the migrating hydroxyl group by a hydrogen atom from 5'-deoxyadenosine takes place with the inversion of the configuration at C(2) of the substrate . Stereospecific dehydration of the 1,1-gem-diol intermediates can also be rationalized by assuming that Asp-alpha335 and Glu-alpha170 function as base catalysts in the dehydration of the (R)- and (S)-isomers, respectively . The structure-based mechanism and stereochemical courses of the reaction are proposed. Infect Control Hosp Epidemiol, 2003 Mar, 24(3), 172 - 9 Use of an alcohol-based hand rub and quality improvement interventions to improve hand hygiene in a Russian neonatal intensive care unit; Brown SM et al.; BACKGROUND: Hand hygiene (HH) is critical to infection control, but compliance is low . Alcohol-based antiseptics may improve HH . HH practices in Russia are not well described, and facilities are often inadequate . SETTING: Four 6-bed units in a neonatal intensive care unit in St . Petersburg, Russia . METHODS: Prospective surveillance of HH compliance, nosocomial colonization, and antibiotic administration was performed from January until June 2000 . In February 2000, alcohol-based hand rub was provided for routine HH use . Eight weeks later, a quality improvement intervention was implemented, consisting of review of interim data, identification of opinion leaders, posting of colonization incidence rates, and regular feedback . Means of compliance, colonization, and antibiotic use were compared for periods before and after each intervention . RESULTS: A total of 1,027 events requiring HH were observed . Compliance was 44.2% before the first intervention, 42.3% between interventions, and 48% after the second intervention . Use of alcohol rose from 15.2% of HH indications to 25.2% between interventions and 41.5% after the second intervention . The incidence of nosocomial colonization (per 1,000 patient-days) with Klebsiella pneumoniae was initially 21.5, decreased to 4.7, and then was 3.2 in the final period . Rates of antibiotic and device use also decreased . CONCLUSIONS: HH may have increased slightly, but the largest effect was a switch from soap and water to alcohol which may have been associated with decreased cross-transmission of Klebsiella, although this may have been confounded by lower device use . Alcohol-based antiseptic may be an improvement over current practices, but further research is required. J Antimicrob Chemother, 2003 Apr, 51(4), 957 - 62 Epub 2003 Feb 25. Clinical features and in vitro antimicrobial susceptibilities of community-acquired Klebsiella pneumoniae meningitis in Taiwan; Lee PY et al.; Twenty-seven adult patients were identified as having community-acquired Klebsiella pneumoniae meningitis . The K . pneumoniae isolates, collected from cerebrospinal fluid samples, were tested for in vitro antimicrobial susceptibilities . The prognostic factors of these 27 patients were also analysed . All of the third- and fourth-generation cephalosporins tested, as well as monobactam, carbapenem and ciprofloxacin, had good activities against the isolated K . pneumoniae strains . None of the clinical isolates was detected as being an extended-spectrum beta-lactamase-producing pathogen . Among the third- and fourth-generation cephalosporins, ceftizoxime, cefepime, ceftriaxone and cefotaxime had superior activities, with MIC90s about four- to eight-fold lower than those of ceftazidime and moxalactam . Mortality rates of patients classified by different antimicrobial agents were as follows: ceftazidime 38% (8/21) and cefepime 16.7% (1/6) . The presence of septic shock and the initial level of consciousness at the start of appropriate antimicrobial therapy were the major determinants of survival and neurological outcomes in these 27 patients . Early diagnosis and choice of appropriate antibiotics according to antimicrobial susceptibilities may improve therapeutic outcomes. Am J Respir Cell Mol Biol, 2003 Apr, 28(4), 443 - 50 Hyperoxia impairs antibacterial function of macrophages through effects on actin; O'Reilly PJ et al.; Oxidative stress may impair alveolar macrophage function in patients with inflammatory lung diseases or those exposed to high concentrations of oxygen . We investigated putative mechanisms of injury to macrophages by oxidative stress, using RAW 264.7 cells exposed to 95% oxygen for 48 h . Hyperoxia-exposed macrophages were less able to phagocytose and kill Klebsiella pneumoniae than normoxic controls, despite increased production of nitric oxide, a free radical important in pathogen killing . Exposure of macrophages to hyperoxia had marked effects on the actin cytoskeleton, including increased actin polymerization, loss of cortical actin, formation of stress fibers, de novo synthesis of actin, and actin oxidation . Hyperoxia induced changes in cell morphology, with increased cell size and pseudopod formation . Exposure of macrophages to jasplakinolide, an agent that increases actin polymerization, also impaired their ability to phagocytose Klebsiella . Alveolar macrophages isolated from mice exposed to 100% oxygen for 84 h also demonstrated impaired phagocytic function, as well as similar effects on the actin cytoskeleton and cell morphology to macrophages exposed to hyperoxia in vitro . We conclude that oxidative stress in vitro and in vivo impairs macrophage antibacterial function through effects on actin. Eur J Biochem, 2003 Apr, 270(7), 1555 - 66 Oxygen control of nif gene expression in Klebsiella pneumoniae depends on NifL reduction at the cytoplasmic membrane by electrons derived from the reduced quinone pool; Grabbe R et al.; In Klebsiella pneumoniae, the flavoprotein, NifL regulates NifA mediated transcriptional activation of the N2-fixation (nif) genes in response to molecular O2 and ammonium . We investigated the influence of membrane-bound oxidoreductases on nif-regulation by biochemical analysis of purified NifL and by monitoring NifA-mediated expression of nifH'-'lacZ reporter fusions in different mutant backgrounds . NifL-bound FAD-cofactor was reduced by NADH only in the presence of a redox-mediator or inside-out vesicles derived from anaerobically grown K . pneumoniae cells, indicating that in vivo NifL is reduced by electrons derived from membrane-bound oxidoreductases of the anaerobic respiratory chain . This mechanism is further supported by three lines of evidence: First, K . pneumoniae strains carrying null mutations of fdnG or nuoCD showed significantly reduced nif-induction under derepressing conditions, indicating that NifL inhibition of NifA was not relieved in the absence of formate dehydrogenase-N or NADH:ubiquinone oxidoreductase . The same effect was observed in a heterologous Escherichia coli system carrying a ndh null allele (coding for NADH dehydrogenaseII) . Second, studying nif-induction in K . pneumoniae revealed that during anaerobic growth in glycerol, under nitrogen-limitation, the presence of the terminal electron acceptor nitrate resulted in a significant decrease of nif-induction . The final line of evidence is that reduced quinone derivatives, dimethylnaphthoquinol and menadiol, are able to transfer electrons to the FAD-moiety of purified NifL . On the basis of these data, we postulate that under anaerobic and nitrogen-limited conditions, NifL inhibition of NifA activity is relieved by reduction of the FAD-cofactor by electrons derived from the reduced quinone pool, generated by anaerobic respiration, that favours membrane association of NifL . We further hypothesize that the quinol/quinone ratio is important for providing the signal to NifL. J Bacteriol, 2003 Apr, 185(7), 2267 - 76 Mutations which uncouple transport and phosphorylation in the D-mannitol phosphotransferase system of Escherichia coli K-12 and Klebsiella pneumoniae 1033-5P14; Otte S et al.; Mutants of Escherichia coli K-12 were isolated which lack the normal phosphotransferase system-dependent catabolic pathway for D-mannitol (Mtl) . In some mutants the pts genes for the general proteins enzyme I and histidine protein of the phosphoenolpyruvate-dependent carbohydrate phosphotransferase systems were deleted . Other mutants expressed truncated mannitol-specific enzymes II (II(Mtl)) which lacked the IIA(Mtl) or IIBA(Mtl) domain(s), and the mtlA genes originated either from E . coli K-12 or from Klebsiella pneumoniae 1033-5P14 . The dalD gene from Klebsiella oxytoca M5a1 was cloned on single-copy plasmids and transformed into the strains described above . This gene encodes an NAD-dependent D-arabinitol dehydrogenase (DalD) which converts D-arabinitol into D-xylulose and also converts D-mannitol into D-fructose . The different strains were used to isolate mutations which allow efficient transport of mannitol through the nonphosphorylated II(Mtl) complexes by selecting for growth on this polyhydric alcohol . More than 40 different mutants were analyzed to determine their ability to grow on mannitol, as well as their ability to bind and transport free mannitol and, after restoration of the missing domain(s), their ability to phosphorylate mannitol . Four mutations were identified (E218A, E218V, H256P, and H256Y); all of these mutations are located in the highly conserved loop 5 of the IIC membrane-bound transporter, and two are located in its GIHE motif . These mutations were found to affect the various functions in different ways . Interestingly, in the presence of all II(Mtl) variants, whether they were in the truncated form or in the complete form, in the phosphorylated form or in the nonphosphorylated form, and in the wild-type form or in the mutated form, growth occurred on the low-affinity analogue D-arabinitol with good efficiency, while only the uncoupled mutated forms transported mannitol at a high rate. Clin Imaging, 2003 Mar-Apr, 27(2), 129 - 31 Clinically occult isolated right iliac mycotic aneurysm with duodenal involvement in a diabetic elderly man: multislice CT diagnosis; Ko SF et al.; A diabetic old man presented with vague abdominal discomfort and intermittent tarry stools for 2 days and gastric ulcers with bleeding was diagnosed after endoscopy . Multislice computed tomography (MSCT) clearly depicted an isolated right iliac mycotic aneurysm with retroperitoneal extension and duodenal involvement . Timely operation and effective antibiotic treatment resulted in complete recovery . To our knowledge, this is the first report of an isolated mycotic iliac artery aneurysm (IAA) complicated with an aneurysmo-duodenal fistula induced by Klebsiella pneumoniae . Int J Antimicrob Agents, 2003 Mar, 21(3), 285 - 8 Extended-spectrum beta-lactamase types in Klebsiella pneumoniae and Escherichia coli in two Greek hospitals; Tzelepi E et al.; Seventy-nine Klebsiella pneumoniae and 124 Escherichia coli clinical strains, isolated consecutively during August-October 2001 in two Greek hospitals, were examined for production of extended-spectrum beta-lactamases (ESBLs) . Seventy-one (35%) isolates (46 K . pneumoniae and 25 E . coli) were ESBL-positive by phenotypic methods . Isoelectric focusing of beta-lactamases and PCR assays for bla genes showed that SHV-5-type ESBLs were the most frequent (45 isolates, 22%) followed by CTX-M (24 isolates, 12%) and IBC (three isolates, 1.5%) . The latter two ESBL types may have been established recently in this setting. FEMS Immunol Med Microbiol, 2003 Mar 20, 35(2), 93 - 8 K-antigen-specific, but not O-antigen-specific natural human serum antibodies promote phagocytosis of Klebsiella pneumoniae; Lepper PM et al.; Infections due to Klebsiella pneumoniae and other Klebsiella spp . are a leading cause of hospital-associated morbidity, especially in the intensive care setting . In this study, the hypothesis that normal human sera contain sufficient concentrations of K-antigen-specific antibodies to promote phagocytic killing of encapsulated, highly virulent Klebsiella organisms was tested . K2-antigen-specific IgG and IgM antibodies were detected in each of 10 normal sera, and such antibodies were functionally active in a phagocytic killing assay . Phagocytosis depended critically on sufficient numbers of neutrophils and was impaired by the presence of soluble Klebsiella capsular polysaccharide (CPS) . Thus, insufficient numbers of neutrophils and circulation of soluble CPS but not lack of K-specific antibodies may be detrimental in Klebsiella sepsis . The efficacy of hyperimmune sera might be based not on enhancement of phagocytosis but on the neutralization of these detrimental effects of circulating CPS and LPS. Microbiology, 2003 Feb, 149(Pt 2), 451 - 7 Identification of the essential histidine residue for high-affinity binding of AlbA protein to albicidin antibiotics; Weng LX et al.; The albA gene from Klebsiella oxytoca encodes a protein that binds albicidin phytotoxins and antibiotics with high affinity . Previously, it has been shown that shifting pH from 6 to 4 reduces binding activity of AlbA by about 30%, indicating that histidine residues might be involved in substrate binding . In this study, molecular analysis of the albA coding region revealed sequence discrepancies with the albA sequence reported previously, which were probably due to sequencing errors . The albA gene was subsequently cloned from K . oxytoca ATCC 13182(T) to establish the revised sequence . Biochemical and molecular approaches were used to determine the functional role of four histidine residues (His(78), His(125), His(141) and His(189)) in the corrected sequence for AlbA . Treatment of AlbA with diethyl pyrocarbonate (DEPC), a histidine-specific alkylating reagent, reduced binding activity by about 95 % . DEPC treatment increased absorbance at 240-244 nm by an amount indicating conversion to N-carbethoxyhistidine of a single histidine residue per AlbA molecule . Pretreatment with albicidin protected AlbA against modification by DEPC, with a 1 : 1 molar ratio of albicidin to the protected histidine residues . Based on protein secondary structure and amino acid surface probability indices, it is predicted that His(125) might be the residue required for albicidin binding . Mutation of His(125) to either alanine or leucine resulted in about 32 % loss of binding activity, and deletion of His(125) totally abolished binding activity . Mutation of His(125) to arginine and tyrosine had no effect . These results indicate that His(125) plays a key role either in an electrostatic interaction between AlbA and albicidin or in the conformational dynamics of the albicidin-binding site. J Clin Microbiol, 2003 Mar, 41(3), 1161 - 6 Patterns of resistance associated with integrons, the extended-spectrum beta-lactamase SHV-5 gene, and a multidrug efflux pump of Klebsiella pneumoniae causing a nosocomial outbreak; Gruteke P et al.; Multiresistant Klebsiella pneumoniae caused a nosocomial outbreak . Resistance patterns of the presumed outbreak isolates varied among and within patients . In order to control the outbreak, screening for extended-spectrum beta-lactamase (ESBL)-producing K . pneumoniae was commenced . A number of susceptible K . pneumoniae strains were stored to serve as controls in genetic strain typing . Typing by pulsed-field gel electrophoresis proved the clonality of the strains in the recognized outbreak patients . Typing of the control strains by pulsed-field gel electrophoresis showed that at least one patient had been missed by the ESBL screening procedure . Further genetic typing confirmed the presence of the SHV-5 ESBL gene in all but one of the outbreak strains . Variable presence of integrons that carried the aminoglycoside resistance genes aadB and aadA2 was found . A gyrA mutation in codon 83 was present in all outbreak strains tested, despite considerable differences in ciprofloxacin MICs . The MICs of ciprofloxacin and the chemically unrelated drug cefoxitin were correlated (r = 0.86, P < 0.01) and were compatible with the overexpression of an efflux pump in a subset of the outbreak strains . We conclude that outbreak strains that express an ESBL gene only at a low level may pass unnoticed in a screening procedure, when the laboratory is unaware of variable ESBL expression . In this particular outbreak, screening for strains for which ciprofloxacin MICs were > or =0.25 micro g/ml would in retrospect have been the most sensitive method for detection of the K . pneumoniae outbreak strain. J Antimicrob Chemother, 2003 Mar, 51(3), 711 - 4 Plasmid-mediated, carbapenem-hydrolysing beta-lactamase, KPC-2, in Klebsiella pneumoniae isolates; Smith Moland E et al.; Four isolates of Klebsiella pneumoniae obtained from patients at a Maryland medical centre exhibited reduced susceptibility to carbapenems and were found to produce the novel, class A, plasmid-mediated, carbapenem-hydrolysing enzyme, KPC-2 . This enzyme has 99% identity with the plasmid-mediated, carbapenem-hydrolysing enzyme KPC-1, reported previously in a North Carolina K . pneumoniae isolate . The KPC-2-producing isolates were either susceptible or intermediate to imipenem and meropenem, unlike the KPC-1-producing isolate, which was resistant to these agents . Detection of KPC-2 may be a problem for clinical laboratories because in this study it was associated with positive extended-spectrum beta-lactamase (ESBL) confirmation tests (clavulanate-potentiated activities of ceftriaxone, ceftazidime, cefepime and aztreonam) . Therefore, a failure to recognize the significance of reduced carbapenem susceptibility in the isolates that remained susceptible to imipenem or meropenem could have resulted in the isolates being incorrectly identified as ESBL producers. J Antimicrob Chemother, 2003 Mar, 51(3), 605 - 12 Variable susceptibility to piperacillin/tazobactam amongst Klebsiella spp . with extended-spectrum beta-lactamases; Babini GS et al.; MICs of piperacillin/tazobactam are conventionally determined by varying the concentration of piperacillin in the presence of a fixed 4 mg/L tazobactam . When tested in this way, the MIC distribution for Klebsiella isolates with extended-spectrum beta-lactamases (ESBLs) is strongly bimodal, such that many producers are inhibited at 16 + 4 mg/L whilst others require MICs of > or =512 + 4 mg/L . When, however, piperacillin/tazobactam was tested as a fixed 8:1 ratio, the MIC distribution became unimodal . If clavulanate 4 mg/L was combined with piperacillin, a unimodal MIC distribution was seen for ESBL-producing Klebsiella spp . but a bimodal distribution arose if the clavulanate concentration was reduced to 0.25 mg/L . These data for alternative combinations suggested that the bimodal MIC distribution seen for piperacillin + tazobactam 4 mg/L was a titration effect, not a reflection of some ESBLs being resistant to tazobactam . Even within single strains, as defined by serotype and DNA fingerprints, there was considerable variation in susceptibility to piperacillin + tazobactam 4 mg/L, with some representatives highly susceptible and others highly resistant . Some of the more resistant representatives produced more of their ESBL, or had a greater number of beta-lactamase types, but these associations were not universal . Elevated resistance to piperacillin + tazobactam was not associated with porin change in any ESBL producer examined, but has been found by others. Biophys J, 2003 Mar, 84(3), 1651 - 9 The citrate carrier CitS probed by single-molecule fluorescence spectroscopy; Kastner CN et al.; A prominent region of the Na(+)-dependent citrate carrier (CitS) from Klebsiella pneumoniae is the highly conserved loop X-XI, which contains a putative citrate binding site . To monitor potential conformational changes within this region by single-molecule fluorescence spectroscopy, the target cysteines C398 and C414 of the single-Cys mutants (CitS-sC398, CitS-sC414) were selectively labeled with the thiol-reactive fluorophores AlexaFluor 546/568 C(5) maleimide (AF(546), AF(568)) . While both single-cysteine mutants were catalytically active citrate carriers, labeling with the fluorophore was only tolerated at C398 . Upon citrate addition to the functional protein fluorophore conjugate CitS-sC398-AF(546), complete fluorescence quenching of the majority of molecules was observed, indicating a citrate-induced conformational change of the fluorophore-containing domain of CitS . This quenching was specific for the physiological substrate citrate and therefore most likely reflecting a conformational change in the citrate transport mechanism . Single-molecule studies with dual-labeled CitS-sC398-AF(546/568) and dual-color detection provided strong evidence for a homodimeric association of CitS. J Liposome Res, 2002 Feb-May, 12(1-2), 121 - 7 Liposome-enabled synergistic interaction of antimicrobial agents; Schiffelers RM et al.; Antimicrobial agents may interact synergistically when both drugs are present at the infected site for an adequate period of time at sufficient concentrations . Generally speaking, the agents in the combination show different tissue distributions and pharmacokinetics . By co-encapsulation of the drugs in a drug carrier, like liposomes, parallel tissue distributions of both drugs may be ensured and drug concentrations at the site of infection may be increased . In this presentation therapeutic efficacy of liposome-co-encapsulated gentamicin (GN) and ceftazidime (CZ) will be shown in a GN-CZ-susceptible and GN-CZ-resistant Klebsiella pneumoniae-pneumonia in rats. Zhejiang Da Xue Xue Bao Yi Xue Ban, 2002 Aug, 31(6), 457 - 460 {Encoding genes and genotypes of gamma-lactamases produced by a multiple resistant Klebsiella pneumoniae}; Yu YS et al.; OBJECTIVE: To determine the sequence of gene for encoding beta-lactamase produced by Klebsiella pneumoniae E3 isolated from Jiaxing Area in Zhejiang Province . METHODS The Klebsiella pneumoniae strain E3 was identified as an ESBLs-producing bacterium by inhibitor-potentiated broth dilution test . The gene encoding gamma-lactamase of the strain was amplified by PCR . The purified PCR product was cloned and sequenced by Sanger's dideoxy chain termination composition method . RESULTS The Klebsiella pneumoniae strain E3 produced both TEM and SHV gamma lactamases . The SHV encoding gene had 812 nucleotide residues responsible for encoding SHV-11 gamma-lactamase and the TEM encoding gene had 973 nucleotide residues responsible for encoding TEM-1 gamma-lactamase . CONCLUSION The Klebsiella pneumoniae strain E3 isolated from a patient in Jiaxing Area in Zhejiang Province is able to produce both TEM-1 and SHV-11 gamma-lactamases. Hua Xi Yi Ke Da Xue Xue Bao, 2001 Jun, 32(2), 157 - 62 {K . pneumoniea endotoxin induced mice beta-defensin-4 mRNA expression and its signaling transduction}; Cai S et al.; OBJECTIVE: To investigate the in vivo effects of Klebsiella pneumoniae endotoxin(LPS) on beta-defensin expression and the relevant signaling transduction pathway . METHODS: A LPS tolerant mouse C3H/HeJ with a point mutation at Toll-like receptor-4 (TLR4) gene and its wild type strain C3H/HeN were used in this study . C3H/HeJ and C3H/HeN were injected with 4 mg/kg of LPS intraperitoneally . The tracheas, lungs and kidneys of the C3H/HeJ and C3H/HeN were collected respectively at different LPS-treated time points, and the total RNA of each sample was extracted . The expression of mice beta-defensin-3 and/or beta-defensin-4 mRNA in these tissues was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) . The sequence of cDNA amplified from the lung of C3H/HeN treated by LPS for 24 h was analyzed . By using western blot, p-I kappa B alpha (phosphorylated I kappa B alpha) and I kappa B alpha of in the lungs of C3H/HeJ and C3H/HeN were detected at different time points after treatment with LPS or without LPS . RESULTS: 1 . beta-defensin-4 mRNA was detected in the lungs of C3H/HeN after 24 h treatment with LPS . In contrast, no signal was determined in C3H/HeJ mice with LPS treatment and the C3H/HeN mice without LPS treatment . 2 . Compared with the control, increas of the p-I kappa B alpha was observed in the lungs of C3H/HeN at 4 h after treatment with LPS, while both the p-I kappa B alpha and I kappa B alpha contents showed a tendency to go down at 8 h after treatment and dramatically decreased at 24 h . But there were no changes in the of p-I kappa B alpha and I kappa B alpha content the lungs of C3H/HeJ under the same conditions . CONCLUSION: K . pneumoniea endotoxin could induce the expression of beta-defensin-4 mRNA in the lung of C3H/HeN, and TLR4-mediated NF-kappa B activation signaling pathway may be responsible for this event. Infect Immun, 2003 Mar, 71(3), 1306 - 15 Intrapulmonary expression of macrophage inflammatory protein 1alpha (CCL3) induces neutrophil and NK cell accumulation and stimulates innate immunity in murine bacterial pneumonia; Zeng X et al.; Macrophage inflammatory protein 1alpha (MIP-1alpha) (CCL3) is an important mediator of leukocyte recruitment and activation in a variety of inflammatory states, including infection . A recombinant human type 5 adenovirus containing the murine MIP-1alpha cDNA (AdMIP-1alpha) was constructed to determine the effect of transient intrapulmonary expression of MIP-1alpha on leukocyte recruitment, activation, and bacterial clearance in a murine model of Klebsiella pneumoniae pneumonia . The intratracheal administration of AdMIP-1alpha resulted in both time- and dose-dependent expression of MIP-1alpha mRNA and protein within the lung . Importantly, the intrapulmonary overexpression of MIP-1alpha resulted in a maximal 35- and 100-fold reduction in lung and blood bacterial burden, respectively, in animals cochallenged with K . pneumoniae, which was associated with a significant increase in neutrophil and activated NK cell accumulation . Furthermore, the transgenic expression of MIP-1alpha during bacterial pneumonia resulted in enhanced expression of gamma interferon mRNA, compared to that observed in Klebsiella-challenged animals pretreated with control vector . These findings indicate an important role for MIP-1alpha in the recruitment and activation of selected leukocyte populations in vivo and identify this cytokine as a potential immunoadjuvant to be employed in the setting of localized bacterial infection. Clin Infect Dis, 2003 Mar 1, 36(5), 575 - 9 Epub 2003 Feb 07. Klebsiella infection in patients with thalassemia; Chung BH et al.; Klebsiella infection has previously been reported in a few patients with transfusion-dependent thalassemia . The incidence and clinical spectrum of this infection in our cohort of patients were reviewed retrospectively . Among 160 patients observed for 12 years, there were 15 episodes of Klebsiella infection that occurred in 12 patients (7.5%), resulting in an incidence of 0.78 infections per 100 patient-years . The clinical spectrum included sinusitis (4 cases), intracranial infection (5 cases), septicemia (4 cases), and abscesses of the liver, lung, kidney, and parotid gland (1 case each) . Three patients had recurrent infections involving different sites, 2 (16%) died of fulminant septicemia, and 3 (25%) had significant permanent neurological deficits . The antibiotic susceptibility pattern for the isolates was similar to the pattern for isolates recovered in the community . With regard to predisposing factors, iron overload and liver function derangement were found to be significant on univariate analysis (P=.046 and P=.049, respectively) but insignificant on multivariate analysis . Klebsiella infection was a serious and frequently encountered complication in our patients with transfusion-dependent thalassemia, resulting in high mortality and morbidity rates. Enferm Infecc Microbiol Clin, 2003 Feb, 21(2), 72 - 6 {Risk factors associated with ceftazidime-resistant Klebsiella pneumoniae infection}; Bermejo J et al.; INTRODUCTION: Risk factors associated with ceftazidime-resistant Klebsiella pneumoniae (CAZ-R Kp) infection may vary among hospitals and in the same hospital at different time points . Knowledge of these factors is required to establish suitable infection control programs . METHODS: A case-control study was conducted to assess risk factors for CAZ-R Kp infection . Thirty-two cases were compared with 28 controls admitted to a 200-bed general hospital during 1999 and 2000 . RESULTS: In the univariate analysis Kp CAZ-R isolates were significantly associated with nosocomial acquisition (OR 5 17.40), prior antibiotic use (OR 5 14.94), particularly ciprofloxacin use (OR 5 5), and hospitalization stay of more than 6 days (OR 5 6.72) . Significantly associated variables in the logistic regression analysis included nosocomial acquisition (OR 5 9.29), prior antibiotic use (OR 5 6.21), and particularly, ciprofloxacin use (OR 5 10.84) . CONCLUSIONS: Efforts toward more rational overall antibiotic use and particularly ciprofloxacin use, combined with infection control measures are necessary to decrease the prevalence of CAZ-R Kp in our hospital. FEMS Microbiol Lett, 2003 Jan 21, 218(1), 121 - 6 Identification and characterization of KvgAS, a two-component system in Klebsiella pneumoniae CG43; Lai YC et al.; A two-component system encoding gene cluster kvgAS that is present only in virulent Klebsiella pneumoniae CG43 was isolated and its sequence determined . RT-PCR and Southern analysis demonstrated that kvgAS is organized as an operon . No apparent effect of a kvgS deletion on bacterial virulence was observed in a mouse peritonitis model . In the presence of paraquat or 2,2-dipyridyl, the activity of kvgAS promoter in the kvgS mutant was found to be reduced to half of the level in the wild-type strain . The data suggest that the KvgAS system is autoregulated and plays a role in countering free radical stresses and sensing iron-limiting conditions. Zhonghua Yi Xue Za Zhi (Taipei), 2002 Nov, 65(11), 534 - 9 Risk factors for endogenous endophthalmitis secondary to Klebsiella pneumoniae liver abscess; Sheu SJ et al.; BACKGROUND: To identify the risk factors for endogenous endophthalmitis secondary to Klebsiella pneumoniae (KP) liver abscess . METHODS: Retrospective chart review of 200 patients with KP liver abscess from 1990 to 2000 was performed . Data variables included age, sex, past history, systemic condition, initial and final visual acuity, slit lamp biomicroscopy, intraocular pressure, fundus, course and treatment . RESULTS: Extrahepatic metastasis developed in 44 patients, including 18 cases (23 eyes) of endophthalmitis . Seventeen eyes had final vision less than counting fingers . Diabetes had significant association with the development of extrahepatic metastasis (p = 0.045) and the poor visual outcome of endophthalmitis (p = 0.022), whereas, neither the initial vision nor vitrectomy was significantly related to the outcome of endophthalmitis . CONCLUSIONS: The visual prognosis of endogenous endophthalmitis secondary to KP liver abscess is generally poor, and diabetes might be a significant risk factor for the poor visual outcome . Even when the initial vision is as bad as light perception only, some eyes can be saved . Both internists and ophthalmologists should be alert enough to detect and treat these patients early. Indian J Pathol Microbiol, 2000 Jan, 43(1), 35 - 40 Immunoglobulin-M estimation and C-reactive protein detection in neonatal septicemia; Krishna BV et al.; Immunoglobulin-M estimation, and C-Reactive Protein test and blood culture were performed on 57 neonates, clinically suspected to have septicemia . IgM level of > or = 20 mg/dl was found in 58.62% cases and CRP test was positive in 68.98% of culture proven sepsis . Blood culture was positive in 29 cases, with the predominant organism being Klebsiella . Of the 2 tests, CRP had a higher sensitivity (68.97%) while IgM estimation had a higher specificity (82.14%) . When the 2 tests were considered together the sensitivity and specificity further increased . Death rate among neonates with IgM levels < 20 mg/dl was 5 times higher than those with elevated IgM levels. Eur J Biochem, 2003 Feb, 270(4), 635 - 45 Relation between domain evolution, specificity, and taxonomy of the alpha-amylase family members containing a C-terminal starch-binding domain; Janecek S et al.; The alpha-amylase family (glycoside hydrolase family 13; GH 13) contains enzymes with approximately 30 specificities . Six types of enzyme from the family can possess a C-terminal starch-binding domain (SBD): alpha-amylase, maltotetraohydrolase, maltopentaohydrolase, maltogenic alpha-amylase, acarviose transferase, and cyclodextrin glucanotransferase (CGTase) . Such enzymes are multidomain proteins and those that contain an SBD consist of four or five domains, the former enzymes being mainly hydrolases and the latter mainly transglycosidases . The individual domains are labelled A {the catalytic (beta/alpha)8-barrel}, B, C, D and E (SBD), but D is lacking from the four-domain enzymes . Evolutionary trees were constructed for domains A, B, C and E and compared with the 'complete-sequence tree' . The trees for domains A and B and the complete-sequence tree were very similar and contain two main groups of enzymes, an amylase group and a CGTase group . The tree for domain C changed substantially, the separation between the amylase and CGTase groups being shortened, and a new border line being suggested to include the Klebsiella and Nostoc CGTases (both four-domain proteins) with the four-domain amylases . In the 'SBD tree' the border between hydrolases (mainly alpha-amylases) and transglycosidases (principally CGTases) was not readily defined, because maltogenic alpha-amylase, acarviose transferase, and the archaeal CGTase clustered together at a distance from the main CGTase cluster . Moreover the four-domain CGTases were rooted in the amylase group, reflecting sequence relationships for the SBD . It appears that with respect to the SBD, evolution in GH 13 shows a transition in the segment of the proteins C-terminal to the catalytic (beta/alpha)8-barrel(domain A). J Clin Microbiol, 2003 Feb, 41(2), 798 - 802 Use of multienzyme multiplex PCR amplified fragment length polymorphism typing in analysis of outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit; van der Zee A et al.; We developed and optimized a new modified amplified fragment length polymorphism (AFLP) typing method to obtain a multibanding fingerprint that can be separated by agarose gel electrophoresis . Both to maximize the discriminatory power and to facilitate the computer-assisted analysis, bacterial DNA was digested with four different restriction enzymes . After ligation of adaptors to the DNA fragments, PCR testing of various single primers was performed . Two single primers that gave optimal results with regard to band resolution and discriminatory power were selected and combined . The computer-assisted analysis of fingerprint patterns was performed with Pearson's product-moment correlation values of densitometric curves, without assigning bands to peaks . Thus, the analysis is not subject to human interpretation errors . With this method, we investigated two outbreaks of multiresistant Klebsiella pneumoniae in an intensive care unit and various sporadic isolates of K . pneumoniae and Klebsiella oxytoca . Cluster analysis of isolates analyzed in different experiments and on different gels showed that fingerprint patterns clustered correctly according to subspecies or to the outbreaks . Multienzyme multiplex PCR AFLP revealed that the first outbreak was caused by two different types of strains . Outbreak two was caused by yet another strain of K . pneumoniae . In conclusion, the typing method used here is easy to perform and highly reproducible, and due to generation of complex banding patterns, it has a higher discriminatory power . Furthermore, the multienzyme multiplex PCR fingerprints are easy to analyze, and a reliable database can be stored in the computer to facilitate comparison of future isolates of Klebsiella spp . The method can be performed in every clinical microbiology laboratory. J Clin Microbiol, 2003 Feb, 41(2), 772 - 7 Occurrence of extended-spectrum and AmpC beta-lactamases in bloodstream isolates of Klebsiella pneumoniae: isolates harbor plasmid-mediated FOX-5 and ACT-1 AmpC beta-lactamases; Coudron PE et al.; We tested 190 Klebsiella pneumoniae bloodstream isolates recovered from 189 patients in 30 U.S . hospitals in 23 states to determine the occurrence of extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase producers . Based on growth inhibition by clavulanic acid by disk and MIC test methods, 18 (9.5%) of the isolates produced ESBLs . Although the disk diffusion method with standard breakpoints identified 28 cefoxitin-nonsusceptible isolates, only 5 (18%) of these were confirmed as AmpC producers . Of two AmpC confirmatory tests, the three-dimensional extract test was easier to perform than was the double-disk approximation test using a novel inhibitor, Syn2190 . Three of the five AmpC producers carried the bla(FOX-5) gene, while the other two isolates harbored the bla(ACT-1) gene . All AmpC genes were transferable . In vitro susceptibility testing with standard inocula showed that all five AmpC-producing strains were susceptible to cefepime, imipenem, and ertapenem but that with a high inoculum, more of these strains were susceptible to the carbapenems than to cefepime . All but 1 of 14 screen-positive AmpC nonproducers (and ESBL nonproducers) were susceptible to ceftriaxone and cefepime at the standard inoculum as were 6 of 6 isolates that were randomly selected and tested with a high inoculum . These results indicate that (i) . a significant number of K . pneumoniae bloodstream isolates harbor ESBL or AmpC beta-lactamases, (ii) . confirmatory tests are necessary to identify true AmpC producers, and (iii) . in vitro, carbapenems are active against AmpC-producing strains of K . pneumoniae. Diagn Microbiol Infect Dis, 2003 Jan, 45(1), 29 - 34 Comparison of BDPhoenix and VITEK2 automated antimicrobial susceptibility test systems for extended-spectrum beta-lactamase detection in Escherichia coli and Klebsiella species clinical isolates; Sturenburg E et al.; The present study compares the ability to detect extended-spectrum beta-lactamases (ESBL) among a collection of 34 ESBL producing clinical isolates belonging to Escherichia coli and Klebsiella species with two new rapid susceptibility and identification instruments-VITEK2 (bioMerieux, Marcy l'Etoile, France) vs . BDPhoenix (BD Biosciences, Sparks, MD) . ESBL content in these isolates was previously characterized on the basis of PCR amplification and sequencing results which were used as the reference method in our evaluation . BDPhoenix correctly determined the ESBL outcome for all strains tested (100% detection rate), whereas VITEK2 was not able to detect the ESBL status in 5 isolates (85% detection rate) . Detailed analysis revealed that the discrepancies were mainly observed with 'difficult-to-detect' strains . Misidentification was either due to low oximino cephalosporin MIC in these strains or was associated with pronounced 'cefotaximase' or 'ceftazidimase' phenotypes . Klebsiella oxytoca chromosomal beta-lactamase (K1) is phenotypically quite similar to ESBL enzymes . In order to evaluate whether the K1 and ESBL enzymes could be discriminated, we expanded our analysis by 8 clinical K . oxytoca strains with K1 phenotypes . VITEK2 gave excellent identification of these strains whereas 7 out of 8 were falsely labeled ESBL-positive by the BDPhoenix system. Eur J Immunol, 2003 Feb, 33(2), 326 - 33 Outer membrane protein A renders dendritic cells and macrophages responsive to CCL21 and triggers dendritic cell migration to secondary lymphoid organs; Jeannin P et al.; Outer membrane protein A (OmpA) is a class of bacterial cell wall protein that is immunogenic without adjuvant . As specific immune responses are initiated in the lymph nodes (LN, we analyzed the effect of the OmpA from Klebsiella pneumoniae (KpOmpA) onchemokine/ chemokine receptor expression by APC and on cell migration to the LN . Upon contact with KpOmpA, human immature DC and macrophages acquire CCR7 expression and responsiveness to CCL21 . In parallel, CCR1 and CCR5 expression is down-regulated and CXCL8, CCL2, CCL3 and CCL5 production is up-regulated . Mice injected subcutaneously with KpOmpA present a transient inflammatory reaction at the site of injection accompanied by an enlargement of the draining LN with a higher proportion of DC and macrophages . Lastly, when exposed to KpOmpA prior injection, DC but not macrophages migrate to the draining LN . In conclusion, KpOmpA confers a migratory phenotype to DC and triggers their migration to the regional LN . This property contributes to explain how innate cells initiate adaptive immune response upon recognition of conserved bacterial components and also why OmpA is immunogenic in the absence of adjuvant. Antimicrob Agents Chemother, 2003 Feb, 47(2), 755 - 8 Emergence in Klebsiella pneumoniae of a chromosome-encoded SHV beta-lactamase that compromises the efficacy of imipenem; Poirel L et al.; A Klebsiella pneumoniae isolate was identified that had reduced susceptibility to several expanded-spectrum cephalosporins and imipenem . That isolate produced a chromosome-encoded SHV-type beta-lactamase, SHV-38, that had an alanine to valine substitution in position Ambler 146 compared to beta-lactamase SHV-1 . The kinetic parameters for purified beta-lactamases SHV-38 and SHV-1 showed that the hydrolytic spectrum of SHV-38 included only ceftazidime and imipenem . This report is the first example of an SHV-type beta-lactamase capable of hydrolyzing imipenem. Antimicrob Agents Chemother, 2003 Feb, 47(2), 559 - 62 Prevalence of plasmid-mediated quinolone resistance; Jacoby GA et al.; Quinolone resistance encoded by the qnr gene and mediated by plasmid pMG252 was discovered in a clinical strain of Klebsiella pneumoniae that was isolated in 1994 at the University of Alabama at Birmingham Medical Center . The gene codes for a protein that protects DNA gyrase from quinolone inhibition and that belongs to the pentapeptide repeat family of proteins . The prevalence of the gene has been investigated by using PCR with qnr-specific primers with a sample of more than 350 gram-negative strains that originated in 18 countries and 24 states in the United States and that included many strains with plasmid-mediated AmpC or extended spectrum beta-lactamase enzymes . qnr was found in isolates from the University of Alabama at Birmingham only during 6 months in 1994, despite the persistence of the gene for FOX-5 beta-lactamase, which is linked to qnr on pMG252 . Isolates from other locations were negative for qnr . The prevalence of mcbG in the same sample was also examined . mcbG encodes another member of the pentapeptide repeat family and is involved in immunity to microcin B17, which, like quinolones, targets DNA gyrase . A single clinical isolate contained mcbG on a transmissible R plasmid . This plasmid and one carrying the complete microcin B17 operon slightly decreased sparfloxacin susceptibility but had a much less protective effect than pMG252 . Plasmid-mediated quinolone resistance was thus rare in the sample examined. Se Pu, 2000 Jul, 18(4), 328 - 31 {Research of indole-3-acetic acid biosynthetic pathway of Klebsiella oxytoca SG-11 by HPLC and GC-MS}; Lu ZX et al.; The plant growth promoting bacteria are closely associated to plant . The bacteria are used to adhering to plant rhizoplane, promoting plant growth by fixing nitrogen from atmosphere, secreting stimulating substances or producing antagonistic to plant pathogens . It was indicated that the biological nitrogen fixation played an important role in plant growth promoting function . In fact, it was verified recently by overall research that IAA does it . Therefore research of IAA production and biosynthetic pathway of plant growth-promoting bacteria is much more important . The various ways of IAA production indicated the strong or weak promoting function of bacterium to plants in general . The purpose of this paper is to determine whether IAA exists in cultured medium of Klebsiella axytoca SG-11 and biosynthetic pathway of IAA, in order to opimize cultural conditions for IAA production . Klebsiella axytoca SG-11 is a plant growth promoting bacterium, isolated from rice rhizoplane, which can fix nitrogen . The supernatant of SG-11 cultured medium determined by HPLC showed that 47.4 mg/L of IAA existed in LB medium and 1.2 mg/L of IAA, in basal medium . IAA in metabolite was identified by GC/MS as well . The intermediate determination of tryptamine, indole-3-acetamide, tryptophol and indole-3-acetonitrile indirectly indicated that IAA was biosynthesized in a pathway of indole-3-pyruvic acid . Meanwhile, tryptophol in metabolite of SG-11 was verified by GC/MS . The direct intermediates of indole-3-pyruvic acid and indole-3-acetaldehyde in the pathway can not be determined, because both are unstable under normal condition . As reversible conversion existed between indole-3 pyruvic aldehyde and tryptophol, the presence of tryptophol also proved the pathway of indole-3-pyruvic acid in the synthesis of IAA by Klebsiella axytoca SG-11 . The results laid basis for further research of plant growth-promoting function of the bacterium. Lin Chuang Er Bi Yan Hou Ke Za Zhi, 2000 Apr, 14(4), 173 - 4 {HSP70 expression of middle ear mucosa in acute suppurative otitis media}; Bi AF et al.; OBJECTIVE: To study HSP70 expression of middle ear mucosa in acute suppurative otitis media . METHOD: An animal model of acute middle ear infection was established by inoculating Klebsiella Pneumoniae into middle ear of guinea pigs . Animals were sacrificed 1, 3, 5 and 7 days after inoculation . The expression of Hsp70 epitope related proteins in middle ear mucosa were analysed by modified western blot test . RESULT: Very light 70 kD bands were recognized by anti Hsp70 monoclonal antibody in the unstressed group . However, middle ear mucosa of the stressed group not only showed relatively strong 70 kD bands, but also expressed more intensely 31 kD and 17 kD bands . The infected mucosa expressed one 70 kD band at 1st day, the strongest band at 3rd day, falling at 5th day; 31 kD was recognized two bands at 3rd day, the strongest bands at 5th day, falling at 7th day; two bands of 17 kD were detected in all days, with the strongest bands at 7th day . CONCLUSION: The results suggested that middle ear mucosa expressed Hsp70 and Hsp70 epitope related proteins like 31 kD and 17 kD in acute otitis media. J Inorg Biochem, 2003 Jan 1, 93(1-2), 18 - 32 Effects of substrates (methyl isocyanide, C2H2) and inhibitor (CO) on resting-state wild-type and NifV(-)Klebsiella pneumoniae MoFe proteins; McLean PA et al.; We report the use of electron nuclear double resonance (ENDOR) spectroscopy to examine how the metal sites in the FeMo-cofactor cluster of the resting nitrogenase MoFe protein respond to addition of the substrates acetylene and methyl isocyanide and the inhibitor carbon monoxide . 1H, 57Fe and 95Mo ENDOR measurements were performed on the wild-type and the NifV(-)proteins from Klebsiella pneumoniae . Among the molecules tested, only the addition of acetylene to either protein induced widespread changes in the 57Fe ENDOR spectra . Acetylene also induced increases in intensity from unresolved protons in the proton ENDOR spectra . Thus we conclude that acetylene may bind to the resting-state MoFe protein to perturb the FeMo-cofactor environment . On the other hand, the present results show that methyl isocyanide and carbon monoxide do not substantially alter the FeMo cofactor's geometric and electronic structures . We interpret this as lack of interaction between those two molecules and the FeMo cofactor in the resting state MoFe protein . Thus, although it is generally accepted that substrates or inhibitors bind to the FeMo-cofactor only under turnover condition, this work provides evidence that at least one substrate can perturb the active site of nitrogenase under non-catalytic conditions. Ann Fr Anesth Reanim, 2002 Dec, 21(10), 807 - 11 {Discitis after spinal anesthesia for transurethral resection of the prostate}; Malinovsky JM et al.; We described a case of discitis and meningitis following spinal anaesthesia for transurethral resection of the prostate . The patient received antibiotics for a month before surgery, because of Klebsiella prostatitis . Spinal anaesthesia was performed in L3-L4 interspace by using 22G Quincke needle . Bacteriaemia occurred during the first postoperative hours . Ten days after spinal anaesthesia, patient suffered from lumbar pain, exacerbated by vertebral percussion, and motor weakness within lower limb, which was marked on right side . MRI examination showed L3-L4 discitis with psoas abcess in regard, and epiduritis marked around L3 right spinal root . CSF examination confirmed meningitis but no bacteria was found . Antibiotics were administered over a 6 weeks period, and then patient discharged from hospital without neurological sequellae . Infectious discitis related to disk puncture during spinal anaesthesia and postoperative bacteriaemia was likely in our patient. J Bacteriol, 2003 Feb, 185(3), 788 - 800 RmpA2, an activator of capsule biosynthesis in Klebsiella pneumoniae CG43, regulates K2 cps gene expression at the transcriptional level; Lai YC et al.; The rmpA2 gene, which encodes an activator for capsular polysaccharide (CPS) synthesis, was isolated from a 200-kb virulence plasmid of Klebsiella pneumoniae CG43 . Based on the sequence homology with LuxR at the carboxyl-terminal DNA-binding motif, we hypothesized that RmpA2 exerts its effect by activating the expression of cps genes that are responsible for CPS biosynthesis . Two luxAB transcriptional fusions, each containing a putative promoter region of the K . pneumoniae K2 cps genes, were constructed and were found to be activated in the presence of multicopy rmpA2 . The activation is likely due to direct binding of RmpA2 to the cps gene promoter through its C-terminal DNA binding motif . Moreover, the loss of colony mucoidy in a K . pneumoniae strain deficient in RcsB, a regulator for cps gene expression, could be recovered by complementing the strain with a multicopy plasmid carrying rmpA2 . The CPS production in Lon protease-deficient K . pneumoniae significantly increased, and the effect was accompanied by an increase of RmpA2 stability . The expression of the rmpA2 gene was negatively autoregulated and could be activated when the organism was grown in M9 minimal medium . An IS3 element located upstream of the rmpA2 was required for the full activation of the rmpA2 promoter . In summary, our results suggest that the enhancement of K2 CPS synthesis in K . pneumoniae CG43 by RmpA2 can be attributed to its transcriptional activation of K2 cps genes, and the expression level of rmpA2 is autoregulated and under the control of Lon protease. Chemosphere, 2003 Mar, 50(8), 1055 - 61 Biotransformation of cyanide to methane and ammonia by Klebsiella oxytoca; Kao CM et al.; Klebsiella oxytoca, isolated from cyanide-containing industrial wastewater, was shown to be able to biodegrade cyanide to non-toxic endproducts using cyanide as the sole nitrogen source . In this study, ammonia was one of the detected endproduct of cyanide biodegradation by the concentrated resting cells of K . oxytoca . Moreover, cyanide has been shown to be biotransformed to methane through the actions of concentrated resting cells . Biodegradation of cyanide by cell-free extracts was not observed, which might be due to the inactivation of nitrogenase (an oxygen-labial enzyme) caused by the oxygen exposure after cell disruption . Results show that the cyanide consumption by resting cells of K . oxytoca was induced when the pretreatment of these cells with cyanide was conducted . However, the cyanide-degrading capability of resting cells pretreated with ammonia was inhibited . The inhibition of cyanide degradation by resting cells of K . oxytoca was affected by the ammonia concentration . This might result from the suppression of nitrogenase activity of K . oxytoca by ammonia since nitrogenase was suggested to be the sole cyanide-degrading enzyme during the cyanide degradation process . Results from this study also show that the processes of cyanide biodegradation and ammonia production by resting cells occurred simultaneously . This suggests that the utilization of cyanide as nitrogen source by K . oxytoca might proceed using ammonia as an assimilatory substrate . Vaccine, 2003 Jan 17, 21(5-6), 458 - 67 B- and T-cell responses to the mycobacterium surface antigen PstS-1 in the respiratory tract and adjacent tissues . Role of adjuvants and routes of immunization; Rodriguez A et al.; Induction of mucosal immunity in the respiratory tract is crucial for protection against respiratory infections . Here, we have investigated the effects of the routes of immunization as well as of three different adjuvants on the induction of mucosal immune responses . Mice were immunized using intranasal (i.n.) or intraperitoneal (i.p.) routes with the mycobacterium PstS-1 antigen . Cholera toxin (CT), detoxified pertussis toxin (detPT) and RU 41.740 from Klebsiella pneumoniae were compared as mucosal adjuvants . Our data showed that i.n . route of immunization induced the most favorable stimulation of mucosal antigen-specific IgA responses supported by mixed Th cells producing IL-4, IL-5, IFN-gamma . In contrast, i.p . immunizations elicited only enhancement of systemic responses, predominantly of the Th2 type . Furthermore, the use of CT as mucosal adjuvant resulted in the stimulation of a mixed Th cell response whereas detPT evoked mainly Th2 type of responses . Likewise CT, the RU 41.740 adjuvant elicited a mixed Th cell response, albeit supported by much lower numbers of CD4(+) T-cells . Thus, i.n . route of immunization favors the induction of mucosal and systemic immune responses, while the Th cell development at mucosal inductive site is influenced by the adjuvant used for immunizations. Zh Mikrobiol Epidemiol Immunobiol, 2002 Sep-Oct, (5), 48 - 53 {Specific biological features of opportunistic bacteria determining dysbacteriosis in the composition of the large intestine normal microflora}; Levanova LA et al.; Dysbiotic manifestations in the gastrointestinal tract are widely spread . They are characterized by a prolonged, persistent course with the tendency to transition to the chronic form and poorly respond to corrective treatment . The occurrence of high concentrations is 70% for Klebsiella oxytoca, while for K . pneumoniae--within the limit of 30% . The most topical problem is colonization of the intestinal mucosa in children aged up to 1 year by Klebsiella, the seeded bacteria retaining their capacity for growth for up to 2-3 years despite the use of probiotics . As shown in this study, the occurrence of Klebsiella and the level of the antilysozyme and "antiinterferon" activity of these bacteria, their resistance to antimicrobial preparations increase, correlating with the level of dysbiotic disturbances. Pharm Res, 2002 Dec, 19(12), 1801 - 7 Locally delivered polyclonal antibodies potentiate intravenous antibiotic efficacy against gram-negative infections; Barekzi NA et al.; PURPOSE: Comparison of the anti-microbial efficacy of locally delivered antibodies in tandem with conventional systemic administration of ceftazidime antibiotic therapy in two lethal gram-negative animal infection models . METHODS: Previously published lethal E . coli-induced closed peritonitis and Klebsiella-induced burn wound infections were generated in outbred female CF-1 mice cohorts . Pooled human polyclonal antibodies were injected locally into sites of infection in these mice simultaneously with intravenous infusions of the broad-spectrum antibiotic, ceftazidime . Mouse survival was compared in sham control cohorts vs . both ceftazidime-alone or antibody-alone systemically infused cohorts as well as local antibody-systemic ceftazidime combination therapy cohorts . Microbial burdens in blood and tissue samples (by agar plating), as well as interleukin-6 cytokine levels (using ELISA) correlated with sepsis, were monitored in sacrificed animals as a function of antimicrobial treatment regimen . RESULTS: Local delivery of human polyclonal antibodies to infection sites was shown to produce synergistic therapeutic efficacy in combination with systemic antibiotic administration in these lethal wound infection models in mice . Enhanced benefits of the unique combination therapy included host survival, bacterial burden both locally and systemically, and IL-6 levels in host serum . CONCLUSIONS: Commercial pooled human antibodies contain a broad spectrum of antimicrobial activity against gram-negative pathogens . Prevention of systemization of infection correlates with host survival in these models . Local control of infection using doses of local, high-titer polyclonal antibodies can enhance traditional approaches to curb systemic spread of infection using intravenous antibiotics . Antibodies provide antimicrobial efficacy independent of known pathogen resistance mechanisms. Structure (Camb), 2003 Jan, 11(1), 109 - 19 Structure of glycerol dehydratase reactivase: a new type of molecular chaperone; Liao DI et al.; The function of glycerol dehydratase (GDH) reactivase is to remove damaged coenzyme B(12) from GDH that has suffered mechanism-based inactivation . The structure of GDH reactivase from Klebsiella pneumoniae was determined at 2.4 A resolution by the single isomorphous replacement with anomalous signal (SIR/AS) method . Each tetramer contains two elongated 63 kDa alpha subunits and two globular 14 kDa beta subunits . The alpha subunit contains structural features resembling both GroEL and Hsp70 groups of chaperones, and it appears chaperone like in its interactions with ATP . The fold of the beta subunit resembles that of the beta subunit of glycerol dehydratase, except that it lacks some coenzyme B(12) binding elements . A hypothesis for the reactivation mechanism of reactivase is proposed based on these structural features. J Synchrotron Radiat, 2003 Jan 1, 10(Pt 1), 71 - 5 Epub 2002 Dec 24. XAFS studies of nitrogenase: the MoFe and VFe proteins and the use of crystallographic coordinates in three-dimensional EXAFS data analysis; Strange RW et al.; This paper reports a three-dimensional EXAFS refinement of the Mo coordination sphere of the FeMoco cluster of the dithionite-reduced MoFe protein from Klebsiella pneumoniae nitrogenase (Kp1) using the 1.6 A-resolution crystallographic coordinates . At this resolution, the positions of the heavy (Fe and S) atoms of the cluster are well determined and there is excellent agreement between the crystallographic and EXAFS models . However, the lighter homocitrate and histidine ligands are poorly determined in the crystal structure, and it is shown that the application of EXAFS-derived distance restraints during the early stages of crystallographic refinement provides a means of substantially improving (by approximately 0.1 A) the final crystallographic model . The consistency of the EXAFS analysis with the crystallographic information in this case justifies applications of EXAFS to cases where protein crystal structures are absent . Thus, the VFe protein of V-nitrogenase has been shown by EXAFS to possess a V-atom site catalytically similar to the well characterized MoFe-nitrogenases, with V replacing Mo. J Bacteriol, 2003 Jan, 185(2), 688 - 92 Isolation of a negative control mutant of the nitrogen assimilation control protein, NAC, in Klebsiella aerogenes; Janes BK et al.; A negative control mutant of the nitrogen assimilation control protein, NAC, has been isolated . Mutants with the leucine at position 111 changed to a nonhydrophobic residue activate transcription from hut and ure promoters, but fail to repress gdhA expression . This failure does not result from failure to bind to either of the two sites required for gdhA repression, but the binding at those sites is altered in the mutant . It appears that the NAC negative control mutants fail to form the complex structures (probably tetramers) formed by wild-type NAC at the gdhA promoter. Zh Mikrobiol Epidemiol Immunobiol, 2002 Nov-Dec, (6), 13 - 8 {Effect of metal oxides on the growth, hemolytic and serologic properties of Klebsiella pneumoniae}; Aleksakhina NN et al.; Silicon, dysprosium, germanium, yttrium, iron, cobalt, samarium, lutecium oxides, as well as the mixture of 8 metal oxides, at a concentration of 20 g/l were found to produce a stimulating or inhibiting effect on the growth of K . pneumoniae strains 204 and K-9 . Silicon, dysprosium, germanium and yttrium oxides were shown to stimulate the growth of K . pneumoniae strain 204 . Iron, cobalt, samarium and lutecium oxides, as well as the mixtures of all oxides under study, inhibited the growth of this strain . Silicon, samarium and lutecium oxides produced no effect on the growth of K . pneumoniae strain K-9; at the same time germanium and yttrium oxides stimulated the growth of these bacteria, while dysprosium, iron, cobalt oxides, as well as the mixture of all oxides, inhibited their growth . The presence of metal oxides did not change the serological activity of the cultures of both strains growing old, i.e . by 24 hours of their growth . The addition of silicon, germanium and iron oxides to the culture medium increased the hemolytic activity of K . pneumoniae strain K-9 seven to ninefold in comparison with the control grown in a synthetic nutrient medium without metal oxides . The comparison of these two strains (K-9 and 204) revealed that K . pneumoniae strain K-9 possessed greater hemolytic activity. J Biol Chem, 2003 Mar 7, 278(10), 7863 - 74 Epub 2002 Dec 28. Kinetic and catalytic properties of dimeric KpnI DNA methyltransferase; Bheemanaik S et al.; KpnI DNA-(N(6)-adenine)-methyltransferase (KpnI MTase) is a member of a restriction-modification (R-M) system in Klebsiella pneumoniae and recognizes the sequence 5'-GGTACC-3' . It modifies the recognition sequence by transferring the methyl group from S-adenosyl-l-methionine (AdoMet) to the N(6) position of adenine residue . KpnI MTase occurs as a dimer in solution as shown by gel filtration and chemical cross-linking analysis . The nonlinear dependence of methylation activity on enzyme concentration indicates that the functionally active form of the enzyme is also a dimer . Product inhibition studies with KpnI MTase showed that S-adenosyl-l-homocysteine is a competitive inhibitor with respect to AdoMet and noncompetitive inhibitor with respect to DNA . The methylated DNA showed noncompetitive inhibition with respect to both DNA and AdoMet . A reduction in the rate of methylation was observed at high concentrations of duplex DNA . The kinetic analysis where AdoMet binds first followed by DNA, supports an ordered bi bi mechanism . After methyl transfer, methylated DNA dissociates followed by S-adenosyl-l-homocysteine . Isotope-partitioning analysis showed that KpnI MTase-AdoMet complex is catalytically active. Acta Crystallogr D Biol Crystallogr, 2003 Jan, 59(Pt 1), 150 - 1 Epub 2002 Dec 19. Expression, crystallization and preliminary X-ray analysis of isomaltulose synthase (PalI) from Klebsiella sp . LX3; Li N et al.; Isomaltulose synthase (PalI) catalyzes the hydrolysis of the alpha-1,2 bond between the glucose and fructose moieties of sucrose and the formation of alpha-1,6 and alpha-1,1 bonds between the two components to produce isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofranose) and trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofranose), respectively . The PalI protein has been overexpressed, purified and crystallized at 295 K using the hanging-drop vapour-diffusion method . The crystals diffract to 2.2 A resolution using synchrotron radiation and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 59.239, b = 94.153, c = 111.294 A. Antimicrob Agents Chemother, 2003 Jan, 47(1), 342 - 9 In34, a complex In5 family class 1 integron containing orf513 and dfrA10; Partridge SR et al.; A complex class 1 integron, In34, found in a conjugative plasmid from a multidrug-resistant Klebsiella pneumoniae strain isolated in 1997 at a hospital in Sydney, Australia, was shown to have a backbone related to that of In2, which belongs to the In5 family . In In34, the aadB gene cassette replaces the aadA1a cassette in In2, and two additional resistance genes, dfrA10 and aphA1, that are not part of a gene cassette are present . The aphA1 gene is in a Tn4352-like transposon that is located in the tniA gene . The dfrA10 gene lies adjacent to a 2,154-bp DNA segment, known as the common region, that contains an open reading frame predicting a product of 513 amino acids (Orf513) . Orf513 is 66 and 55% identical to the products of two further open reading frames that, like the common region, are found adjacent to antibiotic resistance genes . A 27-bp conserved sequence was found at one end of each type of common region . The loss of dfrA10 due to homologous recombination between flanking direct repeats and incorporation of the excised circle by homologous recombination were demonstrated . Part of In34 is identical to the sequenced portion of In7, which is from a multidrug-resistant Escherichia coli strain that had been isolated 19 years earlier in the same hospital . In34 and In7 are in plasmids that contain the same six resistance genes conferring resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, neomycin, tobramycin, trimethoprim, and sulfonamides, but the plasmid backbones appear to be unrelated, suggesting that translocation of a multiple-drug-resistance-determining region as well as horizontal transfer may have occurred. Zhonghua Jie He He Hu Xi Za Zhi, 2002 Oct, 25(10), 607 - 9 {Drug resistance and genotyping of Klebsiella pneumoniae in lower respiratory tract infection}; Shi J et al.; OBJECTIVE: To understand the current drug resistance of Klebsiella pneumoniae in lower respiratory tract infection and the prevalence trend of extended spectrum beta-lactamase (ESBLs)-producing Klebsiella pneumoniae . METHODS: Drug resistance in 86 strains of Klebsiella pneumoniae and ESBLs-producing strains were genotyped by randomly amplified DNA polymorphisms (RAPD) . RESULTS: ESBLs-producing strains accounted for 20.9% of the 86 strains, and 83.3% of the ESBLs-producing strains were from respiratory ICU . The drug resistance rate of ESBLs positive bacteria was much higher than that of ESBLs negative bacteria . ESBLs positive bacteria were resistant to both the quinolones and a minoglylosides . Eighteen ESBLs-producing strains were divided into 12 types . 3 strains had the same profile (genotype A), and 5 strains had an identical profile (genotype B), but other strains had different profiles . CONCLUSIONS: Chromosomal DNA analysis proved that certain types of ESBLs-producing Klebsiella pneumoniae were epidemic in respiratory ICU . RAPD was an economic, simple but reliable method in epidemic study. Chin Med J (Engl), 2002 Oct, 115(10), 1479 - 82 Epidemiological and antibiotic resistant study on extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in Zhejiang Province; Yu Y et al.; OBJECTIVE: To investigate the epidemiological status of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E . coli) and Klebsiella pneumoniae (K . pneumoniae) and the drug resistance profiles of such organisms . METHODS: A total of 282 clinical isolates of E . coli and 180 of K . pneumoniae were collected from different districts of Zhejiang Province . Inhibitor potentiated broth dilution tests were performed for detecting extended-spectrum beta-lactamases . Etests were performed to detect the drug resistance of these strains against nine commonly used antibiotics . RESULTS: The prevalence of extended-spectrum beta-lactamases in E . coli and K . pneumoniae was 34.0% and 38.3%, respectively . The average prevalence of extended-spectrum beta-lactamases in E . coli and K . pneumoniae was 35.7% . The resistance prevalence of extended spectrum beta-lactamase producing strains to ceftazidime and cefotaxime was 40% and 26% respectively, so were those to cefepime, cefoxitin, piperacillin-tazobactam, cefoperazone-sulbactam, amikacin and ciprofloxacin . All these strains were sensitive to imipenem . CONCLUSION: The results in this study showed that the prevalence of extended-spectrum beta-lactamases was high, while extended-spectrum beta-lactamase producing strains were resistant to most antimicrobial agents except imipenem. J Korean Med Sci, 2002 Dec, 17(6), 778 - 83 Glutamine on the luminal microbial environment after massive small bowel resection; Choi SH et al.; To evaluate the oral glutamine (GLN) on the luminal microbes and bacterial translocation (BT) in short bowel, 45 Wistar rats were utilized in three groups; A (control), and B and C (short bowel, 85% of small bowel resected) . The group A was fed with elemental diet (EmD), B with EmD+2% glycine, and C with EmD+2% GLN . The groups B and C were isocaloric and isonitrogenous . Wet weight, DNA, protein, and histomorphometry of the mucosa and parallel microbial culture from cecal contents, caval blood, and tissue blocks of the liver, spleen, and mesenteric lymph nodes were performed on the 5th, 10th, and 15th day . Mucosal growth was higher in group C than B . Colony forming units (CFU) from cecal contents increased more in group B than in C . BTs in A, B, and C were 7/15, 8/15, and 2/15, respectively . Total CFUs in blood and tissues were 5.8 X 10(4)/g, 5.5 X 10(6)/g, and 1.8 X 10(4)/g, respectively . As for BT, the most frequent organism was Klebsiella in A (79.3%), but E . coli in B and C (94.2% and 55.6%) . GLN seems to suppress luminal microbes, and reduces BT in short bowel due to enforced barrier function and proliferation of the mucosa. Am J Trop Med Hyg, 2002 Nov, 67(5), 524 - 7 Hepatic hydatid cysts with superinfection in a non-endemic area in Taiwan; Chen YC et al.; Hepatic hydatid cysts are extremely rare in Taiwan . We report a case of complicated, multivesicular, hydatid cysts in the liver and a secondary infection with Klebsiella pneumoniae . The cysts were detected in an early stage by ultrasonography and computed tomography and treated successfully with radical resection. Kaohsiung J Med Sci, 2002 Aug, 18(8), 386 - 92 Space infection of the head and neck; Wang LF et al.; Deep neck infection may be lethal, especially when life-threatening complications occur . We conducted a retrospective analysis of 184 patients with deep neck infection who were treated at Kaohsiung Medical University Hospital during the past 6 years . Factors such as age, sex, hospitalization days, clinical presentations, involved spaces, imaging studies, microbiology, and treatment method were analyzed . There were 122 men and 62 women with a mean age of 41.7 years . The average hospitalization was 8.4 days . The involved spaces, determined by physical examination and radiologic findings, were the peritonsillar space (59 patients), parapharyngeal space (77 patients), submandibular space (55 patients), and retropharyngeal space (20 patients) . Of the 49 patients for whom the origin of infection was identified, 29 were infected via the upper respiratory tract and 13 had infection of odontogenic origin . The most common isolated organism was Klebsiella pneumoniae . One hundred and thirty-seven patients (74.5%) underwent surgery, including repeated needle aspiration (87 patients) and surgical drainage (50 patients) . The remaining 47 patients recovered uneventfully with antibiotic therapy alone . Eighteen patients developed life-threatening complications, such as descending mediastinitis, sepsis, airway obstruction, and jugular vein thrombosis . Two patients died of septic shock . The combination of accurate diagnosis, effective antibiotic therapy, airway maintenance, and intensive surgical debridement for those who fail to respond to conservative treatment will lead to a good prognosis. Sex Transm Infect, 2002 Dec, 78(6), 452 - 7 Donovanosis; O'Farrell N; Donovanosis, a chronic cause of genital ulceration, has recently been the subject of renewed interest after a long period of relative obscurity . The causative organism, Calymmatobacterium granulomatis, has been cultured for the first time in many years and a polymerase chain reaction diagnostic using a colorimetric detection system has been developed . Phylogenetic analysis confirms close similarities with the genus Klebsiella and a proposal made that C granulomatis be reclassified as Klebsiella granulomatis comb nov . Azithromycin has emerged as the drug of choice and should be used if the diagnosis is confirmed or suspected . In donovanosis endemic areas, syndromic management protocols for genital ulceration may need to be adapted locally . A significant donovanosis epidemic was reported in Durban from 1988-97 but the current status of this epidemic is unclear . The donovanosis elimination programme among Aboriginals in Australia appears successful and is a model that could be adopted in other donovanosis endemic areas . Overall, the incidence of donovanosis seems to be decreasing . Increased attention would undoubtedly be paid to donovanosis if policy makers recognised more readily the importance of genital ulcers in fuelling the HIV epidemic. J Clin Microbiol, 2002 Dec, 40(12), 4666 - 9 Molecular epidemiology of extended-spectrum beta-lactamase-producing, fluoroquinolone-resistant isolates of Klebsiella pneumoniae in Taiwan; Yu WL et al.; Strains of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KP) have emerged worldwide . Concomitant ciprofloxacin resistance with ESBL production in K . pneumoniae isolates would severely restrict treatment options . Among 39 (18.5%) of 211 ESBL-KP isolates resistant to ciprofloxacin (MIC, >/=4 micro g/ml), 37 (95%) were high level resistant (MIC, >/=16 micro g/ml) . These isolates were also cross resistant to the newer fluoroquinolones, including levofloxacin, gatifloxacin, gemifloxacin, and garenoxacin (BMS 284756) . Sitafloxacin was most active against these ciprofloxacin-resistant ESBL-KP isolates with MICs for 67% of the isolates being </=2 micro g/ml . The molecular epidemiology of these multiresistant isolates was investigated by automated ribotyping and pulsed-field gel electrophoresis (PFGE) . Ribotyping identified 18 different strains among the 39 ciprofloxacin-resistant ESBL-KP isolates . The majority (67%) of these isolates were contained in six ribogroups which were further confirmed by PFGE . The distribution of the six major strains of ciprofloxacin-resistant ESBL-KP within Taiwan included one (ribogroup 255.3-PFGE type E) with a nationwide distribution and several institution-specific strains . Interhospital cooperation appears necessary, with strict infection control practices coupled with restriction of fluoroquinolone and extended-spectrum beta-lactam use to control both the major epidemic strain and the more diverse strains observed within individual institutions. Intensive Care Med, 2002 Dec, 28(12), 1718 - 23 Epub 2002 Oct 17. Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae bloodstream infection: risk factors and clinical outcome; Du B et al.; OBJECTIVES: To study the risk factor for nosocomial bacteremia caused by Escherichia coli or Klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) and the influence on patient outcome . DESIGN: Retrospective, single-center study of consecutive bacteremic patients . SETTINGS: A university-affiliated teaching hospital . PATIENTS: A total of 85 patients with nosocomial bacteremia due to E . coli or K . pneumoniae were enrolled . INTERVENTION: None . MEASUREMENTS AND MAIN RESULTS: The demographic characteristics and clinical information including treatment were recorded upon review of patients' records . The primary end point was hospital mortality . Twenty-seven percent of isolates produced ESBLs . Previous treatment with 3rd-generation cephalosporins was the only independent risk factor for bacteremia due to ESBL-producing pathogens {odds ratio (OR) 4.146, P=0.008} . Antibiotic treatment was considered appropriate in 71 cases (83%), and failed in 23 patients (27%) . Twenty-one patients (25%) died in the hospital . Antibiotic treatment failure was the only independent risk factor for hospital mortality (OR 15.376, P=0.001) . Inappropriate antibiotic treatment might lead to significantly higher mortality rate (7/14 vs 14/71, P=0.016) . Patients treated with imipenem were more likely to survive while those receiving cephalosporin treatment tended to have a poorer outcome (1/19 vs 14/40, P=0.023) . CONCLUSIONS: More judicious use of cephalosporins, especially 3rd-generation cephalosporins, may decrease ESBL-producing E . coli or K . pneumoniae bacteremia, and also improve patient outcome. J Bacteriol, 2002 Dec, 184(24), 6966 - 75 Repression of glutamate dehydrogenase formation in Klebsiella aerogenes requires two binding sites for the nitrogen assi