Klebsiella

Pediatr Transplant, 2001 Oct, 5(5), 370 - 3
Pneumatosis intestinalis in an infant undergoing bone marrow transplantation for Wiskott-Aldrich syndrome; Uckan D et al.; A 7-month-old patient with Wiskott-Aldrich syndrome (WAS) developed pneumatosis intestinalis (PI) in the immediate post-transplant period after receiving paternal human leucocyte antigen (HLA) phenotypically matched bone marrow (BM) . PI has been described in patients with congenital or acquired immunodeficiency states and after bone marrow transplantation (BMT) . To our knowledge, the condition has not been described in WAS . The underlying bowel mucosa damage as a result of the history of massive rectal bleeding, the effects of the conditioning regimen, immunosuppression, neutropenia, and infection, may all have contributed to the development of PI . Although the condition resolved by conservative management alone, the patient developed Klebsiella pneumonia sepsis, interstitial pneumonitis, failed to engraft, and died on day +66 following a second infusion of stem cells mobilized from his father's peripheral blood.

Antimicrob Agents Chemother, 2001 Oct, 45(10), 2856 - 61
SHV-1 beta-lactamase is mainly a chromosomally encoded species-specific enzyme in Klebsiella pneumoniae; Chaves J et al.; The nature of the SHV-1 beta-lactamase gene was analyzed in 97 epidemiologically unrelated Klebsiella pneumoniae strains isolated from clinical samples . beta-Lactamase bands that focused at a pI of 7.6 (SHV-1-type) in 74 strains, at a pI of 7.1 (LEN-1-type) in 13 strains, and at a pI of 5.4 (TEM-1-type) in 10 strains were detected by analytical isoelectric focusing (IEF) . Among the 74 SHV-1-producing strains, 40 had, in addition to the pI 7.6 band, an additional band on IEF: 20 had a band with a pI of 7.1 and 20 had a band with a pI of 5.4 . Most of the 74 SHV-1-producing strains (76.7%) carried plasmids . Transfer of beta-lactam resistance by conjugation was possible in only 9.3% of the strains tested . SHV-1 gene-specific PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the chromosomal DNA was positive for 93 of the 97 strains and negative for only 4 of the 10 samples with K . pneumoniae TEM-1 producers . In an attempt to approximate the location of the SHV gene locus by endonuclease restriction analysis, RFLP analysis with Southern blotting of chromosomal DNA with a labeled SHV-1 fragment as a probe was used to study the 97 strains . A trial with EcoRI showed at least one positive hybridization band for 96 strains; two bands were detected for 8 strains . The hybridization was negative for only one TEM-1 beta-lactamase-producing strain . DNA sequence analysis showed no differences in promoter regions or extra stop-triplet sequences; only point mutations determined different allelic variants . The novel SHV-type variants are designated SHV-32 and SHV-33 . As a result of the RFLP and sequencing analyses, it can be postulated that the loci for SHV-1 and LEN-1 genes are arranged in tandem . Our results strongly support the hypothesis that the ancestor of the SHV-1 beta-lactamase originated from the K . pneumoniae chromosome.

Mech Ageing Dev, 2001 Oct, 122(15), 1899 - 913
Evaluation of phagocytosis and arachidonate metabolism by alveolar macrophages and recruited neutrophils from F344xBN rats of different ages; Mancuso P et al.; The incidence of infectious respiratory diseases increases with aging . Resident alveolar macrophages (AMs) and recruited leukocytes (PMNL) mediate cellular defense against bacterial infections in the lung, and phagocytosis and lipid mediator synthesis are important components of their antimicrobial capacity . The objective of this study was to determine if either phagocytic capacity or lipid mediator generation declines with normal aging, in either AMs or PMNL recruited to a site of inflammation . The F344xBN rat hybrid has a lower incidence of pathologies associated with aging, particularly up to 20 months; animals aged 6,12 and 18 months were chosen to evaluate changes associated with normal aging . As previously reported for peripheral blood leukocytes, phagocytosis by recruited PMNL declined with aging: recruited PMNL from 18 months rats showed a significantly decreased capacity to phagocytose live Klebsiella pneumoniae bacteria, compared to PMNL from 6 months rats . Surprisingly, however, the phagocytic capacity of AMs increased with aging: the phagocytic index of AMs from 18 months rats was more than three times that of AMs from 6 months rats . The capacity of AMs and recruited PMNL to release arachidonic acid or synthesize leukotrienes or prostaglandins did not change with aging . This study demonstrates that, although phagocytosis by recruited PMNL declines with aging, other aspects of immune function do not decline, and may actually increase, with normal aging . These results suggest that impaired phagocytosis by recruited PMNL may be an important component of the increased susceptibility to infectious respiratory diseases during normal aging.

East Mediterr Health J, 2000 Mar-May, 6(2-3), 483 - 6
Blood-culture-proven neonatal septicaemia: a review of 36 cases; Misallati A et al.; The cases of 36 newborns seen in the neonatal unit of Al-Fatah Children's Hospital in Benghazi, Libyan Arab Jamahiriya, with blood-culture-proven septicaemia were reviewed to determine clinical profile and outcome . There were 22 males and 14 females . Of these, 12 infants were premature with a gestational age of < 37 weeks and 24 were full term (gestational age > 37 weeks) . At diagnosis, 11 cases were under 4 days of age . The most common symptoms were lethargy and feeding intolerance . Klebsiella was the most common etiological microorganism . Bacterial isolates were resistant to ampicillin and gentamicin but sensitive to cefotaxime . Of the 36 infants, 12 died (fatality rate = 33%).

Infect Immun, 2001 Oct, 69(10), 6434 - 44
Targeting of nasal mucosa-associated antigen-presenting cells in vivo with an outer membrane protein A derived from Klebsiella pneumoniae; Goetsch L et al.; Administration of vaccines by the nasal route has recently proven to be one of the most efficient ways for inducing both mucosal and systemic antibody responses in experimental animals . Our results demonstrate that P40, a well-defined outer membrane protein A from Klebsiella pneumoniae, is indeed a carrier molecule suitable for nasal immunization . Using fragments from the respiratory syncytial virus subgroup A (RSV-A) G protein as antigen models, it has been shown that P40 is able to induce both systemic and mucosal immunity when fused or coupled to a protein or a peptide and administered intranasally (i.n.) to naive or K . pneumoniae-primed mice . Confocal analyses of nasal mucosa-associated lymphoid tissue after i.n . instillation of P40 showed that this molecule is able to cross the nasal epithelium and target CD11c-positive cells likely to be murine dendritic cells or macrophages . More importantly, this targeting of antigen-presenting cells following i.n . immunization with a subunit of the RSV-A molecule in the absence of any mucosal adjuvant results in both upper and lower respiratory tract protection against RSV-A infection.

Infect Immun, 2001 Oct, 69(10), 6364 - 9
Macrophage inflammatory protein 1alpha/CCL3 is required for clearance of an acute Klebsiella pneumoniae pulmonary infection; Lindell DM et al.; The objective of these studies was to determine the role of macrophage inflammatory protein 1alpha/CCL3 in pulmonary host defense during Klebsiella pneumoniae infection . Following intratracheal inoculation, 7-day survival of CCL3(-/-) mice was less than 10%, compared to 60% for CCL3(+/+) mice . Survival of CCR5(-/-) mice was equivalent to that of controls, indicating that the enhanced susceptibility of CCL3(-/-) mice to K . pneumoniae is mediated via another CCL3 receptor, presumably CCR1 . At day 3, CFU burden in the lungs of CCL3(-/-) mice was 800-fold higher than in CCL3(+/+) mice, demonstrating that CCL3 is critical for control of bacterial growth in the lung . Surprisingly, CCL3(-/-) mice had no differences in the recruitment of monocytes/macrophages and even showed enhanced neutrophil recruitment at days 1, 2, and 3 postinfection, compared to CCL3(+/+) mice . Therefore, the defect in clearance was not due to insufficient recruitment of leukocytes . No significant differences in cytokine levels of monocyte chemoattractant protein 1 (MCP-1), interleukin 12, gamma interferon, or tumor necrosis factor alpha in lung lavages were found between CCL3(+/+) and CCL3(-/-) mice . CCL3(-/-) alveolar macrophages were found to have significantly lower phagocytic activity toward K . pneumoniae than CCL3(+/+) alveolar macrophages . These findings demonstrate that CCL3 production is critical for activation of alveolar macrophages to control the pulmonary growth of the gram-negative bacterium K . pneumoniae.

Biophys Chem, 2001 Jul 24, 91(3), 281 - 304
Duplication and extension of the Thorneley and Lowe kinetic model for Klebsiella pneumoniae nitrogenase catalysis using a MATHEMATICA software platform; Wilson PE et al.; The Thorneley and Lowe kinetic model for nitrogenase catalysis was developed in the early to mid 1980s, and has been of value in accounting for many aspects of nitrogenase catalysis . It has also been of value by providing a model for predicting new catalytic behavior . Since its original publication, new results have been obtained and have been successfully incorporated into the model . However, the computer program used for nitrogenase simulations has not been generally available . Using kinetic schemes and assumptions previously outlined by Thorneley and Lowe, we report attempts to duplicate the original T&L kinetic simulation for Klebsiella pneumoniae nitrogenase catalysis using an updated simulation based on the MATHEMATICA programming format, which makes it more user-friendly and more readily available . Comparisons of our simulations with the original T&L simulations are generally in agreement, but in some cases serious discrepancy is observed . Possible reasons for the differences are discussed . In addition to duplicating the original T&L model, we report effects of updating it by including information that has come to light subsequent to its original publication.

Microb Ecol, 1988, 15(2), 165 - 75
Growth and comparative physiology of Klebsiella oxytoca attached to granular activated carbon particles and in liquid media; Davies DG et al.; Experiments were performed to evaluate the comparative growth and physiology of Klebsiella oxytoca grown attached to granular activated carbon particles (GAC) and in liquid medium . Laboratory studies showed that when this organism attached to GAC, the growth rate was enhanced more than 10 times in the presence of glutamate, a substrate that adsorbed to the surface . No differences were observed if the substrate was glucose, which did not adsorb to GAC . Cellular {3H}thymidine uptake was used to estimate DNA biosynthesis . Attached bacteria grown in a minimal nutrient medium containing 20.0 mg/liter glutamate took up 5 times more {3H}thymidine than cells grown in suspension . {3H}uridine was used as a measure of RNA turnover . Attached cells were shown to assimilate 11 times more {3H}uridine than cells in liquid media . Cell size measurements were performed by differential filtration . Cells grown in a minimal medium with 20.0 mg/liter glutamate decreased in size over time, with 62% of the total number passing through a 1.0 micron filter after 9 days incubation . In the same period, 39% of a cell population that was grown on GAC passed through a 1.0 micron filter . These studies indicate that GAC provides an interfacial environment for the enhanced growth of K . oxytoca when glutamate is the substrate.

Nihon Kokyuki Gakkai Zasshi, 2001 Jun, 39(6), 419 - 24
{Severe Klebsiella pneumonia improved by endotoxin adsorption and continuous hemodiafiltration}; Kamata T et al.; A 57-year-old man who had past history of alcoholic hepatitis was admitted to our hospital because of fever, general malaise and dyspnea . Chest X-ray films and computed tomographic scans disclosed lobar pneumonia of the right upper lobe . Laboratory tests showed hypoxemia and renal dysfunction . Artificial ventilation, endotoxin adsorption (PMX) and continuous hemodiafiltration (CHDF) were started on the day after admission because of the appearance of progressive renal failure, hypotension and disturbance of consciousness . A diagnosis of septic multiple organ failure was made on the basis of a blood and sputum culture that revealed a K . pneumoniae infection . After the initiation of intensive therapy, the patient's general condition improved and artificial ventilation, PMX and CHDF were withdrawn . However, MRSA was isolated from sputum specimens and a lung abscess developed . In spite of vancomycin administration, the effect was limited . The patient underwent a right upper lobectomy and was discharged . The prognosis of bacteremic K . pneumoniae pneumonia in alcoholics is reported to be very poor . In our case, it is suggested that PMX and CHDF contributed to the improvement of the clinical course.

Nihon Kokyuki Gakkai Zasshi, 2001 Jun, 39(6), 405 - 9
{A case of pneumonia, septic pulmonary embolisms with cavities, bilateral large lung abscesses, and multiple liver abscesses caused by Klebsiella pneumoniae}; Yoshimoto A et al.; A 64-year-old man suffering from diabetic hyperosmolar non-ketotic coma was admitted for acute lung abscess in the left apical lung field . Sputum culture and blood culture showed a heavy growth of Klebsiella pneumoniae (K . pneumoniae) . He was suffering from sepsis, septic pulmonary embolisms with cavities, bilateral pulmonary consolidations, and multiple liver abscesses . Gradually, the bilateral lung consolidations resolved and areas of consolidation were noted to undergo extensive cavitation bilaterally . Cavitation and abscess formation are frequent complications of K . pneumoniae . Generally, large bilateral lung abscesses caused by K . pneumoniae have a poor prognosis . Cavity nodules are often present in septic pulmonary embolisms . We report a very rare case in a patient with three types of cavities with differing mechanisms . The first was an acute lung abscess, the second, septic pulmonary embolisms with cavities, and the third, large bilateral lung cavities noted in the course of resolving consolidations.

Indian J Chest Dis Allied Sci, 2001 Jul-Sep, 43(3), 157 - 62
Microbial pattern of acute infective exacerbation of chronic obstructive airway disease in a hospital based study; Arora N et al.; Chronic bronchitis is associated with acute exacerbation, most often infective in origin . In order to study the bacteriological profile in such cases a total of 58 patients were enrolled in this study from the chest clinic of our hospital . The male to female ratio was 2 to 1 . Mean age of study group was 47 years . All patients had increased cough and sputum production . Barlett count, gram stain and sputum cultures were done for all patients . IgM and IgG antibodies for M . pneumoniae by ELISA were estimated in all cases . The etiological diagnosis could be established in 72% cases . S . pneumoniae (25.8%), P . aeruginosa (12%), Klebsiella sp (10.3%), B . catarrhalis (3.4%), S . aureus (1.7%) were isolated . Although M . pneumoniae was not cultured it was demonstrated serologically in 20% of cases . H . influenzae was not isolated in any case . The frequency of isolating an etiological agent increased with severity of dysponea.

J Clin Microbiol, 2001 Sep, 39(9), 3193 - 6
Outbreak of infection with extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a Mexican hospital; Silva J et al.; Thirty-one strains of Klebsiella pneumoniae (including 10 duplicates) from 21 septicemic pediatric patients (age, <2 months) were studied during a 4-month period (June to October 1996) in which the fatality rate was 62% (13 of 21) . These isolates identified by the API 20E system yielded the same biotype . Pulsed-field gel electrophoresis experiments revealed the same clone in 31 strains . The isolates were multidrug-resistant but were still susceptible to ciprofloxacin, imipenem, and cefoxitin . A 135-kb plasmid was harbored in all of the isolates . No transconjugants were obtained that were resistant to ampicillin, cefotaxime, tetracycline, or gentamicin . Isoelectric focusing for beta-lactamases was performed on all strains, and three bands with pIs of 5.4, 7.6, and 8.2 were obtained . Of these, the pI 8.2 beta-lactamase had an extended-spectrum beta-lactamase phenotype . PCR amplification of both TEM- and SHV-type genes was obtained . The sequence analysis of the SHV PCR product indicated a mutation corresponding to the SHV-5 beta-lactamase.

J Endotoxin Res, 2001, 7(2), 119 - 24
A monoclonal antibody with specificity for the genus Klebsiella binds to a common epitope located in the core region of Klebsiella lipopolysaccharide; Brade L et al.; A mouse monoclonal antibody (mAb) which has been obtained after immunization of mice with heat-killed Klebsiella pneumoniae strain R20/O1(-) followed by standard plasmacytoma cell fusion protocols was investigated for its ability to identify various species of the genus Klebsiella . Based on the published observation that the antibody binds to an epitope located in the core region of lipopolysaccharide (LPS) of strain R20/O1(-), we tested whether this epitope is shared and exposed by other species of the genus Klebsiella . The antibody was able to bind to LPS of clinical isolates of K . pneumoniae (n = 77), K . oxytoca (n = 50), K . terrigena (n = 49) and K . planticola (n = 50) in 93%, 98%, 96% and 100%, respectively, but did not bind to LPS of other Gram-negative genera (n = 159) as tested by Western blots and dot blots using proteinase K-digested whole cell lysates as antigens . Western blot analyses indicated that the antibody bound only to those LPS molecules which did not carry an O-antigen and that the antibody is thus different from those already published.

FEMS Microbiol Lett, 2001 Aug 21, 202(2), 177 - 80
Glutamine synthetase from Acetobacter diazotrophicus: properties and regulation; Ureta A et al.; Glutamine synthetase from Acetobacter diazotrophicus, an endophyte originally isolated from sugarcane, was studied as a step in the identification of mechanisms underlying the role of A . diazotrophicus as a major supplier of fixed nitrogen to its host plant . The enzyme was purified and partially characterized . It was also shown that the enzyme is regulated by adenylylation in response to the nitrogen source . Interestingly, there is no upregulation of the synthesis of the enzyme under diazotrophic conditions, which is in contrast to the situation in enterics, e.g . Klebsiella pneumoniae.

J Gastroenterol, 2001 Aug, 36(8), 552 - 6
Liver abscess in patients with cirrhosis of the liver: a 12-year experience; Kuo CM et al.; PURPOSE: Liver abscess is rare in patients with cirrhosis of the liver . The aim of this retrospective study was to investigate the incidence, clinical presentation, causal pathogens, and outcome of liver abscess in cirrhotic patients . METHODS: We collected 21 liver abscess specimens (from 14 male patients and 7 female patients; Child A: B: C, 4: 7: 10) from 22,731 admissions of 6450 cirrhotic patients, from 1986 through 1998 . RESULTS: The common clinical symptoms and signs included fever, chills, and abdominal tenderness . The major predisposing factors were biliary tract disease (52%) and diabetes mellitus (48%) . The diagnosis rate with abdominal ultrasonography was 79% . Gram-negative aerobes were the predominant pathogens (Klebsiella pneumoniae, 66.7%; Escherichia coli, 23.8%), and occurred in 80% and 69% of blood and pus cultures, respectively, while 38% of cases showed polymicrobial pathogens . The location of the abscess was predominantly in the right lobe (71.4%), and 47.6% of patients had multiple abscesses . Six patients died (all with Child C cirrhosis) . The overall mortality rate was 28.6% (6/21) . CONCLUSIONS: The incidence of liver abscess in the cirrhotic patients was low, at 0.09% (21/22,731 admissions) . The clinical presentations and pathogens were not different from those in noncirrhotic patients, except that in our cirrhotic patients, there was no significant difference in mortality between those with monomicrobial and those with polymicrobial abscess: nor was there a significant difference in mortality between those with single and those with multiple abscesses . The Child C patients were the high-risk group.

Int J Antimicrob Agents, 2001 Aug, 18(2), 185 - 8
Characterization of extended spectrum beta-lactamase-producing Klebsiella pneumoniae from Beijing, China; Shen D et al.; Fourteen clinical isolates of Klebsiella pneumoniae with extended-spectrum beta-lactamases (ESBLs) were detected by the double disk synergy test and the Etest ESBL strip . Co-resistances included high MICs for aminoglycosides, fluoroquinolones, tetracyclines, and trimethoprim/sulphamethoxazole . Co-resistance was not observed in five of the 14 strains . These isolates were all genetically distinct as determined by the automated ribotyping method . Isoelectric focusing documented the presence of multiple beta-lactamases (one to four per isolate) with pIs ranging from 5.4 to 8.4 . The majority of isolates contained beta-lactamases with pI values of 7.6 and 8.4 consistent with SHV-type ESBLs and an Amp C enzyme, respectively . Emerging ESBL strains in K . pneumoniae compromise the use of agents such as cefotaxime, ceftriaxone, ceftazidime in China; leading to the expansion of quality infection control practices and formulary management programmes to minimize clonal expansion.

Scand J Infect Dis, 2001, 33(7), 512 - 5
Conjugative resistance to tazobactam plus piperacillin among extended-spectrum beta-lactamase-producing nosocomial Klebsiella pneumoniae; Akhan S et al.; We studied the genetic origins of piperacillin-tazobactam resistance among nosocomial Klebsiella pneumoniae strains . A total of 30 nosocomial isolates resistant to piperacillin-tazobactam were obtained from various regions of Turkey . Isoelectric focusing demonstrated at least 2 enzymes common to all strains: I at a pI of 8.0 and the other at 5.4 . Piperacillin-tazobactam resistance was successfully transferred from all of the strains to Escherichia coli . Of the piperacillin-tazobactam-resistant transconjugates, 23 were also resistant to ceftazidime . However, 7 transconjugates were susceptible to ceftazidime but resistant to piperacillin-tazobactam, producing a single enzyme focusing at pI 5.4 . Piperacillin resistance caused by this enzyme was reversed by clavulanate and by increased amounts of tazobactam, which indicates that this enzyme confers resistance due to its high amount . Sequence analysis revealed this enzyme to be TEM-1 . This study demonstrates that transferable hyper-produced TEM-1 causes piperacillin-tazobactam resistance in Klebsiella strains in Turkish hospitals.

J Exp Med, 2001 Aug 20, 194(4), 519 - 27
Requirement of interleukin 17 receptor signaling for lung CXC chemokine and granulocyte colony-stimulating factor expression, neutrophil recruitment, and host defense; Ye P et al.; Bacterial pneumonia is an increasing complication of HIV infection and inversely correlates with the CD4(+) lymphocyte count . Interleukin (IL)-17 is a cytokine produced principally by CD4(+) T cells, which induces granulopoiesis via granulocyte colony-stimulating factor (G-CSF) production and induces CXC chemokines . We hypothesized that IL-17 receptor (IL-17R) signaling is critical for G-CSF and CXC chemokine production and lung host defenses . To test this, we used a model of Klebsiella pneumoniae lung infection in mice genetically deficient in IL-17R or in mice overexpressing a soluble IL-17R . IL-17R-deficient mice were exquisitely sensitive to intranasal K . pneumoniae with 100% mortality after 48 h compared with only 40% mortality in controls . IL-17R knockout (KO) mice displayed a significant delay in neutrophil recruitment into the alveolar space, and had greater dissemination of K . pneumoniae compared with control mice . This defect was associated with a significant reduction in steady-state levels of G-CSF and macrophage inflammatory protein (MIP)-2 mRNA and protein in the lung in response to the K . pneumoniae challenge in IL-17R KO mice . Thus, IL-17R signaling is critical for optimal production of G-CSF and MIP-2 and local control of pulmonary K . pneumoniae infection . These data support impaired IL-17R signaling as a potential mechanism by which deficiency of CD4 lymphocytes predisposes to bacterial pneumonia.

J Bacteriol, 2001 Sep, 183(18), 5248 - 56
Catabolite repression of the citrate fermentation genes in Klebsiella pneumoniae: evidence for involvement of the cyclic AMP receptor protein; Meyer M et al.; Klebsiella pneumoniae is able to grow anaerobically with citrate as a sole carbon and energy source by a fermentative pathway involving the Na(+)-dependent citrate carrier CitS, citrate lyase, and oxaloacetate decarboxylase . The corresponding genes are organized in the divergent citC and citS operons, whose expression is strictly dependent on the citrate-sensing CitA-CitB two-component system . Evidence is provided here that the citrate fermentation genes are subject to catabolite repression, since anaerobic cultivation with a mixture of citrate and glucose or citrate and gluconate resulted in diauxic growth . Glucose, gluconate, and also glycerol decreased the expression of a chromosomal citS-lacZ fusion by 60 to 75%, whereas a direct inhibition of the citrate fermentation enzymes was not observed . The purified cyclic AMP (cAMP) receptor protein (CRP) of K . pneumoniae bound to two sites in the citC-citS intergenic region, which were centered at position -41.5 upstream of the citC and citS transcriptional start sites . Binding was apparently stimulated by the response regulator CitB . These data indicate that catabolite repression of the citrate fermentation genes is exerted by CRP and that in the absence of repressing carbon sources the cAMP-CRP complex serves to enhance the basal, CitB-dependent transcription level.

J Immunol, 2001 Sep 1, 167(5), 2759 - 65
Critical role of lipopolysaccharide-binding protein and CD14 in immune responses against gram-negative bacteria; Le Roy D et al.; LPS-binding protein (LBP) and CD14 potentiate cell activation by LPS, contributing to lethal endotoxemia . We analyzed the contribution of LBP/CD14 in models of bacterial infection . Mice pretreated with mAbs neutralizing CD14 or LBP showed a delay in TNF-alpha production and died of overwhelming infection within 24 h, after a challenge with 250 CFU of virulent Klebsiella pneumoniae . Blockade of TNF-alpha also increased lethality, whereas pretreatment with TNF-alpha protected mice, even in the presence of LBP and CD14 blockade . Anti-LBP or anti-CD14 mAbs did not improve or decrease lethality with a higher inoculum (10(5) K . pneumoniae) and did not affect outcome following injections of low or high inocula of Escherichia coli O111 . These results point to the essential role of LBP/CD14 in innate immunity against virulent bacteria.

Drug Resist Updat, 1999 Feb, 2(1), 38 - 55
Mechanisms of fluoroquinolone resistance; Hooper DC; Mechanisms of bacterial resistance to fluoroquinolones fall into two principal categories, alterations in drug target enzymes and alterations that limit permeation of drug to the target, both resulting from chromosomal mutations . No specific resistance mechanisms of quinolone degradation or modification have been found . The target enzymes, DNA gyrase and topoisomerase IV are most commonly altered in domains near the enzyme active sites and in some cases reduced drug binding affinity has been demonstrated . Drug permeation is altered by mutations that increase expression of endogenous multidrug efflux pumps, alter outer membrane diffusion channels, or both . Recently a new plasmid-mediated resistance of an as yet undefined mechanism was found in clinical isolates of Klebsiella pneumoniae .

Cas Lek Cesk, 2001 Jul 19, 140(14), 427 - 9
{Acute states in gastroenterology: spontaneous bacterial peritonitis and the acute intestinal pseudoobstruction syndrome}; Lukas K; Our article concentrates on two acute states, which develop less dramatically but their after-effects may be very serious: Spontaneous bacterial peritonitis and Ogilvie's syndrome . Spontaneous bacterial peritonitis is a bacterial infection of the ascitic fluid without any intraperitoneal source of infection . Ascites is a condition of the disease but need not be clinically manifested . Spontaneous bacterial peritonitis comes usually during heavy hepatic impairment . Diagnosis can be set according: 1 . Positive cultivation of ascitic fluid, 2 . PMN levels higher than 250/mm3, 3 . No infection, which may require a surgical intervention is apparent . Liver disease, which brings about the spontaneous bacterial peritonitis can be: 1 . Chronic (e.g . alcoholic cirrhosis), 2 . Subacute (e.g . alcoholic hepatitis), 3 . Acute (e.g . fulminant hepatic failure) . Mortality of this form of peritonitis can reach up to 46% . The most frequent etiological factor is alcohol and viral hepatitis, the most frequent agents are E . coli and Klebsiella pneumoniae . The disease is most effectively cured by cefalosporins of the third generation . With inadequate treatment, prognosis may be poor . Intestinal pseudoobstruction syndrome has clinical symptomatology of a serious impairment with ileus without signs of any mechanical intestinal obstruction . Syndrome can be classified according to its development: 1 . Acute form--acute intestinal pseudoobstruction syndrome--Ogilvie's syndrome, 2 . Chronic form--chronic intestinal pseudoobstruction syndrome . Pathogenic mechanism of the syndrome is not known . The disease is related to immobility, administration of some drugs, electrolyte imbalance and concomitant diseases (most frequently malignant tumors) . Clinical symptomatology dominates nausea, vomiting, diffuse abdominal pain, constipation or diarrhoea . For diagnostics the first step should be termination of all medication, which could have causing affects, then taking native abdominal X-ray picture where gaseous intestinal distension can be prominent (coecum distended up to 9-12 cm) . Identification of fluid surfaces is not usual . Endoscopic examination can exclude obstruction in the distal part of gut minimally . The most frequent complication is perforation of coecum . Pharmacological treatment relays on prokinetics . The basic intervention remains decompression by a rectal catheter or an effective coloscopic decompression with subsequent introduction of a cannula . Mortality of the disease fluctuates between 43 and 46%.

Antimicrob Agents Chemother, 2001 Sep, 45(9), 2480 - 5
SHV-16, a beta-lactamase with a pentapeptide duplication in the omega loop; Arpin C et al.; A clinical isolate of Klebsiella pneumoniae was found to be resistant to ampicillin (MIC of 128 microg/ml), ticarcillin (MIC of 512 microg/ml), and ceftazidime (MIC of 128 microg/ml) and susceptible to all other beta-lactams; a synergistic effect between clavulanate and ceftazidime suggested the presence of an extended-spectrum beta-lactamase (ESBL) . Transconjugants in Escherichia coli were obtained at low levels (10(-7) per donor cell) and exhibited a similar beta-lactam resistance pattern (resistant to ampicillin, ticarcillin, and ceftazidime at 64 microg/ml) . The ESBL, pI 7.6, was encoded by a large plasmid (>100 kb) which did not carry any other resistance determinant . The ESBL-encoding gene was amplified by PCR using bla(SHV)-specific primers and was sequenced . The deduced amino acid sequence of the SHV-16 ESBL showed that it differed from SHV-1 by only a pentapeptide insertion (163DRWET167) corresponding to a tandem duplication in the omega loop . The implication of the 163a-DRWET163b-DRWET sequence in ceftazidime resistance was confirmed by cloning either bla(SHV-1) or bla(SHV-16) in the same vector, subsequently introduced in the same E . coli strain . Under these isogenic conditions, SHV-16 conferred a 32-fold increase in ceftazidime MIC compared to that with SHV-1 . Furthermore, site-directed mutagenesis experiments modifying either E166aA or E166bA revealed that the functional glutamic residue was that located in the first copy of the duplicated sequence . But surprisingly, the second E166b also conferred a low-level resistance to ceftazidime . This work is the first description of a class A enzyme exhibiting an extended substrate specificity due to an insertion instead of a nucleotide substitution(s) in a clinical isolate.

Antimicrob Agents Chemother, 2001 Sep, 45(9), 2407 - 13
Diversity of SHV and TEM beta-lactamases in Klebsiella pneumoniae: gene evolution in Northern Taiwan and two novel beta-lactamases, SHV-25 and SHV-26; Chang FY et al.; A total of 113 blood culture isolates of Klebsiella pneumoniae from 10 hospitals in northern Taiwan were studied for SHV and TEM beta-lactamase production . bla(SHV) was amplified from all isolates by PCR . TEM-type resistance, was found in 32 of the isolates and was of the TEM-1 type in all isolates . SHV-1, -2, -5, -11, and -12 and two novel enzymes were identified . These novel enzymes were designated SHV-25 and SHV-26 and had pIs of 7.5 and 7.6, respectively . Amino acid differences in comparison to the amino acid sequence of bla(SHV-1) were found at positions T18A (ThrACC-->AlaGCC), L35Q (LeuCTA-->GluCAA), and M129V (MetATG-->ValGTG) for SHV-25 and at position A187T (AlaGCC-->ThrACC) for SHV-26 . The results of substrate profiles and MIC determinations showed that the novel enzymes did not hydrolyze extended-spectrum cephalosporins, rendering the isolates susceptible to these agents . Inhibition profiles revealed that the 50% inhibitory concentration for SHV-26 was higher than those for SHV-1 and SHV-25, resulting in an intermediate resistance to amoxicillin-clavulanic acid . Forty-nine ribotypes were identified, suggesting that major clonal spread had not occurred in any of the hospitals . According to the amino acid sequence, SHV beta-lactamases in Taiwan may basically be derived through stepwise mutation from SHV-1 or SHV-11 and further subdivided by four routes . The stepwise mutations initiated from SHV-1 or SHV-11 to SHV-2, SHV-5, and SHV-12 comprise the evolutionary change responsible for extended-spectrum beta-lactamase (ESBL) production in Taiwan . The stepwise mutations that lead to a non-ESBL (SHV-25) and the beta-lactamase (SHV-26) with reduced susceptibility to clavulanic acid are possibly derived from SHV-11 and SHV-1, respectively . The results suggest a stepwise evolution of SHV beta-lactamases in Taiwan.

Biol Signals Recept, 2001 Sep-Oct, 10(5), 294 - 8
The anti-tumour effect of Klebsiella pneumoniae capsular polysaccharides; Kwok TT et al.; K24 capsular polysaccharide (K24-CPS), with a known structure of a repeating unit, was isolated from the capsule of Klebsiella pneumoniae serotype K24 . The polysaccharide was found to suppress the proliferation of Ehrlich ascites tumour (EAT) cells in vitro, but did not alter the cell cycle distribution of cells . K24-CPS treatment reduced the tyrosine phosphorylation of some proteins in EAT cells . Furthermore, the treatment also decreased the expression of c-JUN, but had no effect on the levels of c-FOS and c-MYC . It is speculated that the growth suppression effect of K24-CPS may be related to its effect in down-regulating c-JUN expression .

Anal Biochem, 2001 Aug 15, 295(2), 180 - 5
A novel purification method for histidine-tagged proteins containing a thrombin cleavage site; Hefti MH et al.; A general procedure for the purification of histidine-tagged proteins has been developed using immobilized metal-ion affinity chromatography . This two-step purification method can be used for proteins containing a hexahistidine tag and a thrombin cleavage site, yielding high amounts of purified protein . The advantage of this method is that thrombin is used instead of imidazole in the final purification step . Imidazole can influence NMR experiments, competition studies, or crystallographic trials, and the presence of imidazole often results in protein aggregates . Removal of the His-tag results in a form of the protein of interest in which no additional tags are present, resembling the native form of the protein, with only three additional amino acids at the N-terminal side . Our method is compared with a more conventional method for the purification of the Azotobacter vinelandii NIFL PAS domain, overexpressed in Escherichia coli . It also proves to be successful for three different His-tagged proteins, the Klebsiella pneumoniae NTRC protein, and the A . vinelandii NIFA and NIFL proteins, and therefore it is a general method for the purification of His-tagged proteins .

Vaccine, 2001 Aug 14, 19(31), 4505 - 10
Potential of Klebsiella pneumoniae cytotoxin toxoid as vaccine against klebsiellosis in rabbits and mice; Singh BR et al.; The protective efficacy of toxoids prepared from crude cytotoxin (polymyxin-B extract, PBE) and purified Klebsiella cytotoxins (KCTs) was studied in rabbits and mice . The toxoids of KCT-I and PBE were found to be protective against homologous as well as heterologous Klebsiella challenges, while toxoids of KCT-II and KCT-III afforded protection in mice against homologous Klebsiella infections . KCT-I and PBE toxoids also induced good humoral anti-Klebsiella response in rabbits with ELISA titres ranging from 20480 to 81480 . Immunized female rabbits passed protective anti-Klebsiella immunoglobulins to their offsprings through colostra . Baby rabbits fed on colostrum of immunized rabbits withstood lethal Klebsiella infection up to 1 month of age, but not on the 50th day . Baby rabbits having an anti-Klebsiella IgG titre > or =1280 were fully protected against lethal dose of Klebsiella . The study revealed a significant protective efficacy of KCT-I and PBE toxoids against klebsiellosis in mice and rabbits.

Eur J Pediatr . 2001 Jul;160(7):449.
Endogenous endophthalmitis and necrotising pneumonia caused by Klebsiella pneumoniae in a child with beta-thalassaemia major; Yao TC et al.; A case beta-thalassaemia major complicated with Klebsiella pneumoniae endogenous endophthalmitis is presented to increase physician awareness of the association of the two clinical conditions . Severe morbidity including subretinal abscess and retinal detachment may develop despite aggressive intravenous and intravitreal ceftriaxone therapy, along with vitrectomy and external drainage.

Pharm Res, 2001 Jun, 18(6), 780 - 7
Host factors influencing the preferential localization of sterically stabilized liposomes in Klebsiella pneumoniae-infected rat lung tissue; Schiffelers RM et al.; PURPOSE: To gain insight into the host factors influencing liposome localization at sites of bacterial infection . METHODS: In a unilateral Klebsiella pneumoniae pneumonia rat model, capillary permeability and number of circulating leukocytes was quantified and related to the degree of liposome target localization . RESULTS: Liposome localization was highest in the hemorrhagic zone of infection, a zone characterized by markedly increased capillary permeability and high bacterial numbers . Both liposome localization and capillary permeability correlated positively with severity of infection . Lung instillation of other inflammatory stimuli, such as lipopolysaccharide or 0.1 M HCl inducing increased capillary permeability, also promoted liposome localization . As liposomal target localization in leukopenic rats was similar to that in immunocompetent rats, contribution of circulating leukocytes seems limited . Intrapulmonary distribution of liposomes shows that leukocytes at the target site are involved in liposome uptake after extravasation . CONCLUSIONS: Increased capillary permeability plays a crucial role in liposome localization at the infected site, whereas contribution of leukocytes is limited . These results suggest inflammatory conditions that could benefit from liposomal drug delivery . The involvement of leukocytes in liposome uptake at the target site could be important information in the selection of appropriate drugs.

J Biol Chem, 2001 Oct 5, 276(40), 37415 - 25 Epub 2001 Jul 25.
Metabolism of sucrose and its five linkage-isomeric alpha-D-glucosyl-D-fructoses by Klebsiella pneumoniae . Participation and properties of sucrose-6-phosphate hydrolase and phospho-alpha-glucosidase; Thompson J et al.; Klebsiella pneumoniae is presently unique among bacterial species in its ability to metabolize not only sucrose but also its five linkage-isomeric alpha-d-glucosyl-d-fructoses: trehalulose, turanose, maltulose, leucrose, and palatinose . Growth on the isomeric compounds induced a protein of molecular mass approximately 50 kDa that was not present in sucrose-grown cells and which we have identified as an NAD(+) and metal ion-dependent 6-phospho-alpha-glucosidase (AglB) . The aglB gene has been cloned and sequenced, and AglB (M(r) = 49,256) has been purified from a high expression system using the chromogenic p-nitrophenyl alpha-glucopyranoside 6-phosphate as substrate . Phospho-alpha-glucosidase catalyzed the hydrolysis of a wide variety of 6-phospho-alpha-glucosides including maltose-6'-phosphate, maltitol-6-phosphate, isomaltose-6'-phosphate, and all five 6'-phosphorylated isomers of sucrose (K(m) approximately 1-5 mm) yet did not hydrolyze sucrose-6-phosphate . By contrast, purified sucrose-6-phosphate hydrolase (M(r) approximately 53,000) hydrolyzed only sucrose-6-phosphate (K(m) approximately 80 microm) . Differences in molecular shape and lipophilicity potential between sucrose and its isomers may be important determinants for substrate discrimination by the two phosphoglucosyl hydrolases . Phospho-alpha-glucosidase and sucrose-6-phosphate hydrolase exhibit no significant homology, and by sequence-based alignment, the two enzymes are assigned to Families 4 and 32, respectively, of the glycosyl hydrolase superfamily . The phospho-alpha-glucosidase gene (aglB) lies adjacent to a second gene (aglA), which encodes an EII(CB) component of the phosphoenolpyruvate-dependent sugar:phosphotransferase system . We suggest that the products of the two genes facilitate the phosphorylative translocation and subsequent hydrolysis of the five alpha-d-glucosyl-d-fructoses by K . pneumoniae.

J Biol Chem, 2001 Sep 28, 276(39), 36514 - 9 Epub 2001 Jul 24.
Characterization and mechanism of action of a reactivating factor for adenosylcobalamin-dependent glycerol dehydratase; Kajiura H et al.; Adenosylcobalamin-dependent glycerol dehydratase undergoes mechanism-based inactivation by its physiological substrate glycerol . We identified two genes (gdrAB) of Klebsiella pneumoniae for a glycerol dehydratase-reactivating factor (Tobimatsu, T., Kajiura, H., Yunoki, M., Azuma, M., and Toraya, T . (1999) J . Bacteriol . 181, 4110-4113) . Recombinant GdrA and GdrB proteins formed a tight complex of (GdrA)(2)(GdrB)(2), which is a putative reactivating factor . The purified factor reactivated the glycerol-inactivated and O(2)-inactivated glycerol dehydratases as well as activated the enzyme-cyanocobalamin complex in vitro in the presence of ATP, Mg(2+), and adenosylcobalamin . The factor mediated the exchange of the enzyme-bound, adenine-lacking cobalamins for free, adenine-containing cobalamins in the presence of ATP and Mg(2+) through intermediate formation of apoenzyme . The factor showed extremely low ATP-hydrolyzing activity and formed a tight complex with apoenzyme in the presence of ADP . Incubation of the enzyme-cyanocobalamin complex with the reactivating factor in the presence of ADP brought about release of the enzyme-bound cobalamin . The resulting tight inactive complex of apoenzyme with the factor dissociated upon incubation with ATP, forming functional apoenzyme and a low affinity form of factor . Thus, it was established that the reactivation of the inactivated holoenzymes takes place in two steps: ADP-dependent cobalamin release and ATP-dependent dissociation of the apoenzyme-factor complex . We propose that the glycerol dehydratase-reactivating factor is a molecular chaperone that participates in reactivation of the inactivated enzymes.

J Hosp Infect, 2001 Aug, 48(4), 281 - 8
Neonatal intensive care unit outbreak caused by a strain of Klebsiella oxytoca resistant to aztreonam due to overproduction of chromosomal beta-lactamase; Jeong SH et al.; Klebsiella oxytoca strains resistant to both aztreonam and ceftriaxone were isolated from six neonates in a neonatal intensive care unit and water reservoirs of two humidifiers attached to the neonatal incubators . These isolates were assumed to be of the same clone because they were characterized by the same antimicrobial susceptibility and pulsed field gel electrophoresis patterns . It was established that the drug resistance was attributed to overproduction of chromosomally encoded Kl beta-lactamase . It was determined that an isolate (K . oxytoca H1) contained a high enzyme concentration (27microg/100microg of protein in enzyme extracts), at least 27 times higher than the control K . oxytoca N1 . It was also demonstrated that isolates had a point mutation in the - 35 concensus region of the promotor gene of bla(OXY-2)leading to enzyme overproduction . Outbreaks caused by K1 hyperproducers have not previously been described .

Antimicrob Agents Chemother, 2001 Aug, 45(8), 2368 - 71
Identification of a plasmid encoding SHV-12, TEM-1, and a variant of IMP-2 metallo-beta-lactamase, IMP-8, from a clinical isolate of Klebsiella pneumoniae; Yan JJ et al.; A multidrug-resistant plasmid encoding TEM-1, SHV-12, and a variant of IMP-2 metallo-beta-lactamase, designated IMP-8, was identified from a clinical isolate of Klebsiella pneumoniae . There are four nucleotide differences between bla(IMP-2) and bla(IMP-8), resulting in two amino acid differences . bla(IMP-8) was also found to be carried by an integron-borne gene cassette similar to the bla(IMP-2) cassette.

Enferm Infecc Microbiol Clin, 2001 Jun-Jul, 19(6), 261 - 6
{Genital infection and infertility}; Rodriguez R et al.; BACKGROUND: Genital infection is the most important cause of infertility worldwide, affecting not only Fallopian tubes but all anatomic urogenital sites, both male and female . METHODS: We present a randomized,prospective and normalized study about sexually transmitted diseases(STD) . We include 487 patients, 376 of whom were infertile and the remaining 111 were not and act as control group . RESULTS: 47.3% of infertile patients showed at least one infection: 10.7% had Chlamydia trachomatis infection, whereas only 0.3% had gonococal infection . We found none syphilis . 12.9% of the patients showed yeast belonging to genus Candida, 5% bacterial vaginosis, 3.8% Escherichia coli and 0.3% Klebsiella pneumoniae . The percentage of isolation of Ureaplasma urealyticum and Mycoplasma hominis were 23.5% and 4.8%, respectively.We detected antibodies against Hepatitis B (any serological marker) in 7.8% of the cases . Chlamydial infection and the presence of U . urealyticum were related with infertility (X 12=6,070, p<0.005 and X12=8,782, p<0.005,respectively) . CONCLUSIONS: We think these results conclude that is necessary to perform routine tests to screen for C . trachomatis, N . gonorrhoeae, and for infections caused by micoplasma as well, among infertile patients.These patients must be considered of being at risk of acquiring STD, since the percentage of these diseases is higher than the percentage we found among the control group . It is important to insist on prevention and early diagnosis and treatment as a main goal to decrease the number of tubal occlusion and infertile couples.

J Leukoc Biol, 2001 Jul, 70(1), 135 - 41
Surfactant protein D-coated Klebsiella pneumoniae stimulates cytokine production in mononuclear phagocytes; Keisari Y et al.; Encapsulated Klebsiella pneumoniae strains K21a, K10, and K50, all of which contain dimannose sequences in their capsular polysaccharides that are recognized by the mannose receptor of macrophages, stimulated interleukin secretion and cytokine mRNA expression by human monocyte-derived macrophages . By contrast, the corresponding unencapsulated phase variants and the K2 strain, which lack the dimannose sequence, did not . Coating of unencapsulated phase variants of Klebsiella strains with surfactant protein (SP)-D resulted in marked stimulation of cytokine mRNA accumulation . The induction of cytokine mRNA via the mannose receptor occurred only in monocyte-derived macrophages, whereas that caused by SP-D-coated Klebsiella strains occurred in both macrophages and peripheral-blood monocytes.The results suggested that innate immunity against pulmonary pathogens might be mediated by SP-D, which acts as an opsonin to enhance the interaction of macrophages with unencapsulated phase variants originating from the upper respiratory tract, and by macrophage mannose receptors, which recognize encapsulated variants expressing capsular dimannose residues.

Appl Environ Microbiol, 2001 Jul, 67(7), 3325 - 7
Incidence of Klebsiella species in surface waters and their expression of virulence factors; Podschun R et al.; To investigate the occurrence of different Klebsiella spp . in aquatic environments, a total of 208 samples of natural surface waters was examined . From half (53%) of these samples, 123 Klebsiella strains were isolated, the most common species being Klebsiella pneumoniae . A comparison of these isolates to a group of 207 clinical K . pneumoniae isolates demonstrated that water isolates of K . pneumoniae, unlike those of K . oxytoca and K . planticola, are as capable as clinical isolates of expressing putative virulence factors such as serum resistance and capsular polysaccharides, pili, and siderophores.

Pediatr Infect Dis J, 2001 Jun, 20(6), 557 - 60
Late onset infection in very low birth weight infants in Malaysian Level 3 neonatal nurseries . Malaysian Very Low Birth Weight Study Group; Ho JJ; BACKGROUND: The purpose of this study was to examine the rate and mortality from late onset infection occurring in very low birth weight infants admitted to Malaysian nurseries . METHODS: Data on all infants 1500 g or below admitted to the 20 participating Level 3 nurseries were analyzed for late onset infection (clinical infection and positive blood or cerebrospinal fluid culture occurring after 48 h of life) . RESULTS: The overall survival of the 962 study infants was 69% . The rate of late onset infection was 19.3% . The mortality in those with late onset infection was 30.8% . The most common infecting organism was Klebsiella pneumoniae, accounting for 38.3% of infections and 46.9% of deaths in infants with infection, followed by coagulase-negative staphylocci, 17.6 and 12.2%, respectively . On logistic regression analysis risk factors for late onset gram-negative compared with gram-positive infection were endotracheal intubation at birth and blood transfusion . Hypoglycemia was associated with gram-positive infection . CONCLUSION: The late onset infection rate in Malaysian very low birth weight infants does not differ from that reported from developed countries, but the mortality is higher . This could be because of an excess of gram-negative infections.

Arch Bronconeumol, 2001 May, 37(5), 265 - 6
{Recurrent pneumonia and empyema due to Klebsiella pneumoniae as a complication of laparoscopic cholecystectomy}; Villena Garrido V et al.; We report a patient with pneumonia and empyema secondary to an abdominal abscess due to Klebsiella pneumoniae from stones released into the abdominal cavity during laparoscopic cholecystectomy.

Int J Syst Evol Microbiol, 2001 May, 51(Pt 3), 915 - 24
Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping; Brisse S et al.; The infra-specific phylogenetic diversity and genetic structure of both Klebsiella pneumoniae and Klebsiella oxytoca was investigated using a combination of randomly amplified polymorphic DNA (RAPD) analysis, sequencing of gyrA and parC genes, and automated ribotyping . After RAPD analysis with four independent primers of 120 clinical isolates collected from 22 European hospitals in 13 countries, K . pneumoniae isolates fell into three clusters and K . oxytoca isolates fell into two clusters, while Klebsiella planticola isolates formed a sixth cluster . Each cluster was geographically widespread . K . pneumoniae cluster I (KpI) accounted for 80% of the isolates of this species and included reference strains of the three subspecies K . pneumoniae subsp . pneumoniae, K . pneumoniae subsp . ozaenae and K . pneumoniae subsp . rhinoscleromatis . Clusters KpII and KpIII were equally represented, as were the two K . oxytoca clusters . Individualization of each cluster was fully confirmed by phylogenetic analysis of gyrA and parC gene sequences . In addition, sequence data supported the evolutionary separation of K . pneumoniae from a phylogenetic group including K . oxytoca, Klebsiella terrigena, K . planticola and Klebsiella ornithinolytica . Automated ribotyping using Mlu I appeared suitable for identification of each Klebsiella cluster . The adonitol fermentation test was found to be useful for cluster identification in K . pneumoniae, since it was negative in all strains of clusters KpIII and in some KpII strains, but always positive in cluster KpI . The usefulness of gyrA and parC sequence data for population genetics and cluster identification in bacteria was demonstrated, even for the phylogenetic positioning of quinolone-resistant isolates.

Alcohol Clin Exp Res, 2001 Jun, 25(6), 872 - 81
Effect of alcohol consumption on host release of interleukin-17 during pulmonary infection with Klebsiella pneumoniae; Shellito JE et al.; BACKGROUND: A link between alcohol abuse and bacterial pneumonia has been recognized for centuries, but mechanisms to explain this relationship are unclarified . Interleukin-17 (IL-17) is a lymphocyte-derived cytokine that is part of the inflammatory cytokine cascade . Previous studies from our laboratory indicated that IL-17 is released in lung tissue in a murine model of bacterial pneumonia caused by Klebsiella pneumoniae . The effects of alcohol consumption on pulmonary release of IL-17 are unknown . METHODS: Mice were maintained on 20% ethanol in drinking water or on a control diet without alcohol . After 2 weeks, alcohol and control mice were challenged with intratracheal K . pneumoniae . Mice were followed for survival after bacterial challenge, neutrophil recruitment was assayed as myeloperoxidase, and IL-17 was measured in lung lavage fluid by enzyme-linked immunosorbent assay . In additional experiments, splenocytes from control mice were incubated with ethanol in vitro, and release of IL-17 was measured in culture supernatants . Finally, control and alcohol mice received intrapulmonary gene transfer of E-1-deleted adenovirus containing the murine IL-17 gene . These mice were then challenged with K . pneumoniae and followed for survival and neutrophil recruitment . RESULTS: In these studies, we demonstrate that a 2-week history of ethanol consumption in mice suppresses release of IL-17 into lung tissue, decreases neutrophil recruitment, and increases mortality from experimental K . pneumonia . In vitro experiments confirm a direct suppressive effect of ethanol on the release of IL-17 from splenocytes . In vivo administration of the IL-17 gene in an adenoviral vector to alcohol-consuming mice results in release of IL-17 into lavage fluid and normalizes neutrophil recruitment and mortality after bacterial challenge . CONCLUSIONS: The results of these experiments strongly implicate IL-17 as an important pathway for the immunosuppression associated with alcohol abuse and support gene therapeutic approaches to augment immune function in the alcoholic host or to treat infections associated with alcoholism.

Laryngoscope, 2001 Jun, 111(6), 1020 - 6
An interpretation of the structural changes responsible for the chronicity of rhinoscleroma; Canalis RF et al.; OBJECTIVE: To identify the structural changes of the proliferative phase of rhinoscleroma which could be responsible for the chronicity of the disease . STUDY DESIGN: Observational research . METHODS: Samples of friable tissue taken from the nasal mucosa of nine untreated patients were processed for light and ultrastructural microscopy . RESULTS: The majority of changes contributing to the chronicity of the disease occurred in the subepithelium and followed three closely related but distinct events . In the first (infiltrative), subepithelial invasion by the Klebsiella was followed by its active multiplication and proliferation of capillaries . In the second (neutrophilic), large numbers of neutrophils were delivered into this space . Neutrophils actively phagocytized the Klebsiella but appeared to die at an accelerated rate without completing digestion of the microorganisms . In the third event (histiocytic), histiocytes entered the subepithelium and engaged in unrestrained phagocytosis of decaying neutrophils, Klebsiella, and debris . During this process, the histiocytes' phagosomes underwent massive dilation, thus becoming Mikulicz cells . Mikulicz cells were unable to consistently destroy the Klebsiella and eventually ruptured, releasing them into the interstitium . Evidence was found that an autophagic process might contribute to phagosome distention and to the rupture of the vacuolar membranes and cell wall . CONCLUSIONS: Several critical changes responsible for the chronicity of rhinoscleroma occur during the proliferative phase of the disease . The majority of these take place in the subepithelium and include: 1) factors leading to the transformation of histiocytes into Mikulicz cells, 2) the inability of these cells to consistently destroy the Klebsiella, 3) their rupture releasing viable Klebsiella, and 4) the intrinsic resistance of the pathogen.

J Pharmacol Exp Ther, 2001 Jul, 298(1), 369 - 75
In vivo synergistic interaction of liposome-coencapsulated gentamicin and ceftazidime; Schiffelers RM et al.; Antimicrobial agents may interact synergistically . But to ensure synergy in vivo, the drugs should both be present at the site of infection at sufficiently high concentrations for an adequate period of time . Coencapsulation of the drugs in a drug carrier may ensure parallel tissue distributions . Since liposomes localize preferentially at sites of infection, this mode of drug delivery could, in addition, increase drug concentrations at the focus of infection . The therapeutic efficacy of gentamicin and ceftazidime coencapsulated into liposomes was examined by monitoring survival in a rat model of an acute unilateral pneumonia caused by antibiotic-susceptible and antibiotic-resistant Klebsiella pneumoniae strains . It is shown that administration of gentamicin in combination with ceftazidime in the free form either as single dose or as 5-day treatment resulted in an additive effect on rat survival in both models . In contrast, targeted delivery of liposome-coencapsulated gentamicin and ceftazidime resulted in a synergistic interaction of the antibiotics in both models . Consequently, liposome coencapsulation of gentamicin and ceftazidime allowed both a shorter course of treatment at lower cumulative doses compared with administration of the antibiotics in the free form to obtain complete survival of rats . Liposomal coencapsulation of synergistic antibiotics may open new perspectives in the treatment of severe infections.

Antimicrob Agents Chemother, 2001 Jul, 45(7), 2141 - 3
Molecular characterization of a novel plasmid-encoded cefotaximase (CTX-M-12) found in clinical Klebsiella pneumoniae isolates from Kenya; Kariuki S et al.; Nine Klebsiella pneumoniae isolates, six from blood and three from cerebrospinal fluid of newborn babies at Kenyatta National Hospital, Nairobi, Kenya, were analyzed for the mechanism of cephalosporin resistance . By using pulsed-field gel electrophoresis of XbaI-digested chromosomal DNA, all the nine isolates were found to be clonal . PCR and direct sequencing revealed a novel extended-spectrum beta-lactamase, which we designated CTX-M-12 . It has a more potent hydrolytic activity against cefotaxime than against ceftazidime and a pI of 9.0 and is encoded on a large self-transferable ca . 160-kbp plasmid.

Plasmid, 2001 May, 45(3), 209 - 21
Molecular characterization of the klebicin B plasmid of Klebsiella pneumoniae; Riley MA et al.; The nucleotide sequence of a bacteriocin-encoding plasmid isolated from Klebsiella pneumoniae (pKlebB-K17/80) has been determined . The encoded klebicin B protein is similar in sequence to the DNase pyocins and colicins, suggesting that klebicin B functions as a nonspecific endonuclease . The klebicin gene cluster, as well as the plasmid backbone, is a chimera, with regions similar to those of pore-former colicins, nuclease pyocins and colicins as well as noncolicinogenic plasmids . Similarities between pKlebB plasmid maintenance functions and those of the colicin E1 plasmid suggest that pKlebB is a member of the ColE1 plasmid replication family .

Fetal Diagn Ther, 2001 Jul-Aug, 16(4), 245 - 7
Sepsis in second trimester of pregnancy due to an infected myoma . A case report and a review of the literature; Grune B et al.; Considering the high incidence of uterine myomata in women in reproductive age, myomata are only found in 2% of all pregnancies . Although they frequently lead to complications in pregnancy, cases of pyomyomata during pregnancy are rarely reported . A 44-year-old gravida 1 in her 26th week of gestation was admitted to the hospital for septic temperatures of unknown cause . A 12-cm leiomyoma with solid structures of heterogenic sonographic pattern and cystic spaces had been documented on a prior first trimester sonogram . The myoma now appeared with the same size but an increased echogenicity of the liquid parts . Ultrasound guided aspiration of the fluid within the myoma showed an infection with Klebsiella pneumoniae . A cesarian section with myo- mectomy confirmed the diagnosis of a pyomyoma .

Enzyme Microb Technol, 2001 Jun 7, 28(9-10), 735 - 743
Purification and properties of a novel raw starch degrading-cyclodextrin glycosyltransferase from Klebsiella pneumoniae AS- 22; Gawande BN et al.; A novel raw starch degrading alpha-cyclodextrin glycosyltransferase (CGTase; E.C . 2.4.1.19), produced by Klebsiella pneumoniae AS-22, was purified to homogeneity by ultrafiltration, affinity and gel filtration chromatography . The specific cyclization activity of the pure enzyme preparation was 523 U/mg of protein . No hydrolysis activity was detected when soluble starch was used as the substrate . The molecular weight of the pure protein was estimated to be 75 kDa with SDS-PAGE and gel filtration . The isoelectric point of the pure enzyme was 7.3 . The enzyme was most active in the pH range 5.5-9.0 whereas it was most stable in the pH range 6-9 . The CGTase was most active in the temperature range 35-50 degrees C . This CGTase is inherently temperature labile and rapidly loses activity above 30 degrees C . However, presence of soluble starch and calcium chloride improved the temperature stability of the enzyme up to 40 degrees C . In presence of 30% (v/v) glycerol, this enzyme was almost 100% stable at 30 degrees C for a month . The K(m) and k(cat) values for the pure enzyme were 1.35 mg ml(-1) and 249 &mgr;M mg(-1) min(-1), respectively, with soluble starch as the substrate . The enzyme predominantly produced alpha-cyclodextrin without addition of any complexing agents . The conditions employed for maximum alpha-cyclodextrin production were 100 g l(-1) gelatinized soluble starch or 125 g l(-1) raw wheat starch at an enzyme concentration of 10 U g(-1) of starch . The alpha:beta:gamma-cyclodextrins were produced in the ratios of 81:12:7 and 89:9:2 from gelatinized soluble starch and raw wheat starch, respectively.

Int J STD AIDS, 2001 Jul, 12(7), 423 - 7
Donovanosis: an update; O'Farrell N; Donovanosis has been ignored for many years until recently . The condition still has a limited geographical distribution . A significant epidemic of donovanosis has been identified in KwaZulu/Natal, South Africa where it may be a risk factor for acquiring HIV in men . After a gap of more than 30 years, the organism was cultured by researchers in Durban, South Africa and Darwin, Australia . Polymerase chain reaction (PCR) techniques for donovanosis were developed soon after, most recently using a colorimetric detection system . Similarities between the causative organism, Calymmatobacterium granulomatis and Klebsiella spp . were confirmed . A proposal that the organism be reclassified under the genus Klebsiella has been put forward . Azithromycin has been confirmed as the drug of choice but is yet to be accepted universally because of cost issues . Treatment in patients with significant HIV induced immune deficiency may need to be prolonged . A donovanosis eradication programme is underway amongst the aboriginal community in Australia . Elsewhere, management through current syndromic guidelines for genital ulcers are yet to be validated in areas where donovanosis is endemic . PCR testing should enable further recognition of donovanosis and lead to more concerted efforts in disease control and possible eradication.

J Formos Med Assoc, 2001 Feb, 100(2), 142 - 4
Pneumocephalus secondary to septic thrombosis of the superior sagittal sinus: report of a case; Luo CB et al.; Pneumocephalus secondary to septic superior sagittal sinus thrombosis (SSSST) is extremely rare . We report computed tomography (CT) findings in a 63-year-old man with SSSST caused by the gas-forming organism Klebsiella pneumoniae . The patient presented with fever, chills, general weakness, and spontaneous progressive swelling of the right frontoparietal scalp . CT revealed a gas-containing abscess over the right frontoparietal subgaleal region and in the superior sagittal sinus . Surgical drainage of the subgaleal abscess was performed and blood and pus cultures grew Klebsiella pneumoniae . The patient died of sepsis on the 6th day of hospitalization.

J Antimicrob Chemother, 2001 Jun, 47(6), 755 - 61
Outer membrane protein change combined with co-existing TEM-1 and SHV-1 beta-lactamases lead to false identification of ESBL-producing Klebsiella pneumoniae; Wu TL et al.; Nine isolates of Klebsiella pneumoniae, obtained from one colonized and eight bacteraemic patients on a paediatric ward, were shown to be identical by PFGE, indicating an outbreak . Screening for extended-spectrum beta-lactamase (ESBL) production using the double-disc synergy test, Etest for ESBLs and agar diffusion tests indicated ESBL production . The isolates showed reduced susceptibility to cefotaxime but not to other third-generation cephalosporins . Molecular studies revealed production of TEM-1 and SHV-1 but no ESBLs were identified . Deficiency in expression of an outer membrane protein (OmpK35) was also observed . These observations led us to postulate that the extremely low level of OmpK35 expression and the co-existence of TEM-1 and SHV-1 resulted in an increased MIC of cefotaxime and the false designation of the isolates as ESBL producers . All the infected infants were treated with either third-generation cephalosporins alone or multiple antibiotics including a third-generation cephalosporin, and recovered and were discharged without sequelae.

Am J Kidney Dis, 2001 Jun, 37(6), 1253 - 9
Impact of co-amoxiclav on polymorphonuclear granulocytes from chronic hemodialysis patients; Cuffini AM et al.; Phagocyte-dependent host defenses are frequently impaired in maintenance hemodialysis patients who show an increased susceptibility to infections . In these individuals, the course of infections can be more aggressive than in normal hosts, and the antibiotic of choice should have a high antimicrobial effect without impairing host defenses . Hence, in uremic patients, the antibiotic enhancement of phagocyte functions may be of potential clinical importance in the outcome of bacterial infections . Because we demonstrated previously that co-amoxiclav had beneficial properties that result in enhancement of the microbicidal functions of human polymorphonuclear cells (PMNs) from healthy subjects, we investigated the influence of this combination on the activities of PMNs from chronic hemodialysis patients against Klebsiella pneumoniae, a human pathogen that can pose severe problems in patients whose immunity is impaired . PMNs from chronic dialysis patients showed a diminished in vitro phagocytic efficiency with a reduced phagocytosis and bactericidal activity towards intracellular K . pneumoniae compared with that seen in PMNs from healthy subjects . When co-amoxiclav was added to PMNs from chronic hemodialysis patients, it was able to restore the depressed primary functions of PMNs, resulting in a significant high increase in both phagocytosis or killing activity . A similar pattern was detected with PMNs collected from hemodialysis patients treated with co-amoxiclav . The results of the present study provide evidence that co-amoxiclav is able to induce stimulation of depressed phagocytic response of PMNs from patients on chronic hemodialysis, restoring their primary functions both in vitro and in vivo.

J Clin Microbiol, 2001 Jun, 39(6), 2206 - 12
Outcome of cephalosporin treatment for serious infections due to apparently susceptible organisms producing extended-spectrum beta-lactamases: implications for the clinical microbiology laboratory; Paterson DL et al.; Although extended-spectrum beta-lactamases (ESBLs) hydrolyze cephalosporin antibiotics, some ESBL-producing organisms are not resistant to all cephalosporins when tested in vitro . Some authors have suggested that screening klebsiellae or Escherichia coli for ESBL production is not clinically necessary, and when most recently surveyed the majority of American clinical microbiology laboratories did not make efforts to detect ESBLs . We performed a prospective, multinational study of Klebsiella pneumoniae bacteremia and identified 10 patients who were treated for ESBL-producing K . pneumoniae bacteremia with cephalosporins and whose infecting organisms were not resistant in vitro to the utilized cephalosporin . In addition, we reviewed 26 similar cases of severe infections which had previously been reported . Of these 36 patients, 4 had to be excluded from analysis . Of the remaining 32 patients, 100% (4 of 4) patients experienced clinical failure when MICs of the cephalosporin used for treatment were in the intermediate range and 54% (15 of 28) experienced failure when MICs of the cephalosporin used for treatment were in the susceptible range . Thus, it is clinically important to detect ESBL production by klebsiellae or E . coli even when cephalosporin MICs are in the susceptible range (<or = 8 microg/ml) and to report ESBL-producing organisms as resistant to aztreonam and all cephalosporins (with the exception of cephamycins).

Acta Crystallogr D Biol Crystallogr, 2001 Jun, 57(Pt 6), 857 - 9 Epub 2001 May 25.
Crystallization and preliminary X-ray studies of meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae IAM1063; Otagiri M et al.; Meso-2,3-butanediol dehydrogenase (meso-BDH) has been crystallized and preliminary X-ray crystallographic characterization of meso-BDH crystals has been performed . Single crystals of meso-BDH were prepared in two forms by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant . Form I crystals belong to space group C2, with unit-cell parameters a = 215.5, b = 79.4, c = 134.8 A, beta = 98.22 degrees, and form II crystals belong to space group P2(1), with unit-cell parameters a = 69.16, b = 109.78, c = 127.28 A, beta = 102.29 degrees . The crystals diffracted to 2.0 and 1.7 A resolutions, respectively, using synchrotron radiation.

Crit Care Med, 2001 Mar, 29(3), 609 - 17
Cardiac functional and structural alterations induced by endotoxin in rats: importance of platelet-activating factor; Iwase M et al.; OBJECTIVE: In this study, we evaluated the time course of the alterations in left ventricular (LV) dimensions, LV wall thickness, and LV systolic function in rats with endotoxemia by using echocardiography as well as myocardial histopathologic assessments . Our second goal was to examine whether pretreatment with a platelet-activating factor (PAF) antagonist would ameliorate the lipopolysaccharide (LPS)-induced cardiovascular collapse during the early phase . DESIGN: A prospective, controlled, in vivo animal laboratory study . SETTING: Research laboratory at a university . SUBJECTS: Male, Wistar rats (8-9 wks old; n = 83) . INTERVENTIONS: In pentobarbital-anesthetized rats, the right carotid artery was cannulated to measure the arterial blood pressure and to sample blood . The right jugular vein also was catheterized for the administration of drugs . LPS (2 mg/kg) derived from Klebsiella pneumoniae or physiologic saline was administered in the presence or absence of pretreatment with TCV-309, a specific potent PAF antagonist . Echocardiographic studies were performed with an 8- to 13-MHz transducer . MEASUREMENTS AND MAIN RESULTS: LPS administration immediately induced progressive hypotension . The maximal hypotensive response was observed at 10 mins after LPS infusion with mean arterial pressure decreasing from 119 +/- 2 to 56 +/- 3 mm Hg (p < .001) . LV end-diastolic internal dimensions decreased from 6.4 +/- 0.1 to 3.1 +/- 0.1 mm (p < .001) at 30 mins after LPS and remained significantly reduced compared with control rats . LV end-systolic dimensions also decreased dramatically from 3.5 +/- 0.2 to 0.5 +/- 0.1 mm (p < .001) at 30 mins after LPS and remained significantly reduced throughout the experiment . LV fractional shortening increased from 45 +/- 1% to 84 +/- 2% (p < .001) at 30 mins after LPS and remained elevated compared with control rats . LV wall thickness increased strikingly from 15 mins until 2 hrs after LPS infusion . Pathologic studies demonstrated marked congestion of capillaries and mild edema in the LV myocardium . The hematocrit increased after the administration of LPS . LPS markedly increased sympathetic tone as demonstrated by the elevation of plasma concentrations of epinephrine and norepinephrine . There was no elevation of concentrations of nitrite and nitrate . Pretreatment with TCV-309, a specific potent PAF antagonist, reduced LPS-induced hypotension and attenuated LV functional and structural changes . TCV-309 administration reduced the LPS-induced adrenergic activation and hemoconcentration . CONCLUSIONS: The hypotension that occurred during the initial phase of LPS-induced shock was accompanied by LV functional and structural alterations . The marked increase in LV wall thickness can be ascribed to the congestion of capillaries and edema in the LV myocardium . Pretreatment with a PAF antagonist reduced LPS-induced alterations . PAF may play a pivotal role during the initial phase of LPS-induced cardiovascular responses.

Biochemistry, 2001 May 29, 40(21), 6379 - 87
Mechanistic studies of two dioxygenases in the methionine salvage pathway of Klebsiella pneumoniae; Dai Y et al.; Two dioxygenases (ARD and ARD') were cloned from Klebsiella pneumoniae that catalyze different oxidative decomposition reactions of an advanced aci-reductone intermediate, CH(3)SCH(2)CH(2)COCH(OH)=CH(OH) (I), in the methionine salvage pathway . The two enzymes are remarkable in that they have the same polypeptide sequence but bind different metal ions (Ni(2+) and Fe(2+), respectively) . ARD converts I to CH(3)SCH(2)CH(2)COOH, CO, and HCOOH . ARD' converts I to CH(3)SCH(2)CH(2)COCOOH and HCOOH . Kinetic analyses suggest that both ARD and ARD' have ordered sequential mechanisms . A model substrate (II), a dethio analogue of I, binds to the enzyme first as evidenced by its lambda(max) red shift upon binding . The dianion formation from II causes the same lambda(max) red shift, suggesting that II bind to the enzyme as a dianion . The electron-rich II dianion likely reacts with O(2) to form a peroxide anion intermediate . Previous (18)O(2) and (14)C tracer experiments established that ARD incorporates (18)O(2) into C(1) and C(3) of II and C(2) is released as CO . ARD' incorporates (18)O(2) into C(1) and C(2) of II . The product distribution seems to necessitate the formation of a five-membered cyclic peroxide intermediate for ARD and a four-membered cyclic peroxide intermediate for ARD' . A model chemical reaction demonstrates the chemical and kinetic competency of the proposed five-membered cyclic peroxide intermediate . The breakdown of the four-membered and five-membered cyclic peroxide intermediates gives the ARD' and ARD products, respectively . The nature of the metal ion appears to dictate the attack site of the peroxide anion and, consequently, the different cyclic peroxide intermediates and the different oxidative cleavages of II . A cyclopropyl substrate analogue inactivates both enzymes after multiple turnovers, providing evidence that a radical mechanism may be involved in the formation of the peroxide anion intermediate.

Chang Gung Med J, 2001 Feb, 24(2), 103 - 13
Outbreaks of nosocomial bloodstream infections associated with multiresistant Klebsiella pneumoniae in a pediatric intensive care unit; Su LH et al.; BACKGROUND: Between June and October 1997, and during April 1998, a cluster of nosocomial bloodstream infections (BSIs) associated with Klebsiella pneumoniae was observed in 8 premature neonates from 1 pediatric intensive care unit (TPICU) in a 4000-bed medical center in northern Taiwan . An investigation was conducted to identify the possible reservoirs and mode of transmission . METHODS: Epidemiologic surveillance and infection control interventions were executed . The environment was checked by submitting several swab samples for microbiological studies . The antibiograms and results from 2 molecular typing methods (pulsed-field gel electrophoresis and infrequent-restriction site polymerase chain reaction) of all bacteremic and environmental isolates of K . pneumoniae were compared . RESULTS: Totally 39 K . pneumoniae isolates, including 9 from bacteremia, 26 from the environment, and 4 controls, were analyzed . One major pattern was found in 21 isolates, which included 8 bacteremic isolates with identical antibiograms, a single isolate from rectal swab screening, 2 of 8 isolates from hand cultures of medical staff, and 10 of 17 isolates from swabs of sinks in the TPICU . All 21 isolates illustrated identical antibiograms, while the other 18 isolates shared 4 antibiograms and 15 unique patterns . CONCLUSIONS: The nosocomial BSIs appeared to be an outbreak induced by 1 multiresistant K . pneumoniae strain . The sinks may have acted as reservoirs for this outbreak strain . During washing, splattered water droplets containing the bacterial particles may have contaminated the hands of medical personnel and were then further transmitted to patients.

Vaccine, 2001 May 14, 19(25-26), 3499 - 508
Unusual persistence in healthy volunteers and ill patients of hyperimmune immunoglobulin directed against multiple Pseudomonas O-chain and Klebsiella serotypes after intravenous infusion; McClain JB et al.; Persistence of intravenous (i.v.) hyperimmune immunoglobulin (100 mg/kg) directed against clinically predominant serotypes of Pseudomonas and Klebsiella in ill, febrile patients was compared to healthy volunteers to determine if ill patients have a decreased Ig half-life resulting in an increased immunoglobulin requirement . Type-specific antibodies were measured by ELISA for 83 days in eight healthy volunteers and for 35 days in eight ill patients with surgical complications or hematologic malignancy . Mean values and fold rises of antibody concentrations for the two groups were above preinfusion values at 35 days . The antibody fold rises in patients and in healthy volunteers were similar . Type-specific antibody levels in some patients increased after illness coincident with elevation of total immunoglobulins . We conclude that the duration of potentially therapeutic levels of infused type-specific hyperimmune immunoglobulin may persist for a longer period of time than what has been measured for total immunoglobulin . While the mechanism of this persistence remains to be characterized, the possibility of type-specific antibody synthesis induced by immunoglobulin administration must be considered.

Folia Microbiol (Praha), 2000, 45(5), 387 - 90
The in vitro effect of imipenem and ofloxacin on enzymic activity of Klebsiella strains; Hostacka A; Fifty seven Klebsiella strains, viz . K . pneumoniae (28), K . planticola (19), K . oxytoca (6), K . ornithinolytica (3) and K . terrigena (1) possessed lipolytic and urealytic activity . The effect of imipenem and ofloxacin at subinhibitory concentrations (sub-MICs) on these enzymic activities of 4 strains was studied . At all the concentrations tested (mainly at 1/4 of the MICs) imipenem enhanced lipase activity manifested by cleavage of the substrate Tween 20 . The effect of ofloxacin was strain- and concentration-dependent but in most cases lipolytic activity was also increased . The antibiotics practically did not affect the urease activity of the strains.

Bull Soc Pathol Exot, 2001 Mar, 94(1), 32 - 5
{Contribution of heparin therapy in the medical treatment of Buruli ulcer apropos of 1 case}; Kanga JM et al.; Medical treatment of Buruli ulcer is mostly disappointing even if Mycobacterium ulcerans is susceptible to many antibacterial drugs . The inefficiency in vivo of the drugs may be due to the tissue vascularisation disorders caused by the toxin that Mycobacterium ulcerans produces . This toxin causes an endarteritis followed by a thrombosis of the dermal vessels responsible for an ischemia which prevents the antibacterial drugs from reaching the infected area . Removal or prevention of that thrombosis should allow the drugs to be more effective . To verify this assumption, we used a combined therapy with two gold standard medicines in an oedematous form of Buruli ulcer on the face which could not be surgically treated: heparin for its activity on thrombosis and rifampin for its bactericidal activity on Mycobacterium ulcerans . Rifampin was administered at 300 mg dose per day . Based on the management of envenomisation cases due to viper bites, we used standard heparin at 500 Ul dose per kg repeatedly administered by an electrical syringe releasing 1 cc per hour in the tubulure of isotonic glucose infusion . The results were encouraging . The dreadful oedema of the face started to reduce on the 15th day and disappeared on the 30th day . A small area at the temple dried . But we were obliged to stop the use of standard heparin due to an occurrence of Klebsiella oxytoca septicaemia from permanent vein route . Rifampin was still administered at the same dose . The face oedema reappeared quickly, followed by a full closure of the eyelids and an ulceration at the right temple level . The standard heparin was therefore substituted by low weight molecular heparin, enoxaparin, administered at 40 mg twice per day by subcutaneous route . 45 days later the oedema reduced and ulceration did not develop . After 90 days of treatment, usual signs of mycobacterial infection progression disappeared . We stopped therefore the use of enoxaparin but continued with rifampin until healing of the ulceration which occurred after 12 months of treatment . We observed no recurrence over a period of 16 months after complete healing . We can conclude that heparin combined with antimycobacterial drugs--which are active in vitro on Mycobacterium ulcerans--could provide an effective medical treatment for Buruli ulcer.

Isr Med Assoc J, 2000 Dec, 2(12), 908 - 11
Epidemiology of ceftazidime-resistant Klebsiella pneumoniae in a large university hospital in Tel Aviv; Noy A et al.; BACKGROUND: An increase in multiple drug-resistant Klebsiella pneumoniae due to extended spectrum beta-lactamase production has recently been reported from many centers around the world . There is no information in the literature regarding this problem in Israel . A high prevalence of ceftazidime-resistant K . pneumoniae was noted in our Intensive Care Unit in the first few months of 1995 . OBJECTIVE: To describe the epidemiology of ceftazidime-resistant K . pneumoniae in our medical center, as representing the situation in tertiary care hospitals in Israel . METHODS: We vigorously restricted the use of ceftazidime in the ICU and enforced barrier precautions . The susceptibility rate of K . pneumoniae was surveyed in the ICU and throughout the hospital before and after the intervention in the ICU . RESULTS: Following the intervention, the susceptibility rate of K . pneumoniae increased from 11% (3/28) to 47% (14/30) (P < 0.01) among ICU isolates, from 55% (154/280) to 62% (175/281) (P = 0.08) among total hospital isolates, and from 61% (50/82) to 74% (84/113) (P < 0.05) among total hospital blood isolates, although no additional control measures were employed outside the ICU . CONCLUSIONS: The epidemiology of ceftazidime-resistant K . pneumoniae in our medical center is similar to that reported from other centers around the world . Early awareness to the emergence of this resistance, identification of the source of the epidemic, and prompt action at the putative source site may reduce the rate of acquisition and spread of such resistance inside and outside of the source unit.

Infection, 2001 Mar-Apr, 29(2), 81 - 6
Klebsiella brain abscess in adults; Liliang PC et al.; BACKGROUND: We analyzed the clinical manifestations and therapeutic outcomes of Klebsiella brain abscesses . PATIENTS AND METHODS: The clinical data of 15 patients with Klebsiella brain abscesses, retrospectively collected over a 14-year period, were studied . RESULTS: The 15 patients included 13 cases of Klebsiella pneumoniae infection and two cases of Klebsiella oxytoca . All but one case were community-acquired infections . Locations of all of these abscesses were supratentorial . 12 cases involved a single abscess, and three involved multiple abscesses . Gas formation was also found in two cases (13%) . Common predisposing factors included metastatic spread, chronic otitis media and neurosurgical procedures . Among these 15 patients, 11 were treated surgically and four received antibiotics alone . In total, 11 patients survived and four died, with an overall mortality rate of 26.7% . CONCLUSION: The clinical presentations and therapeutic outcomes varied according to the different Klebsiella species . While debilitating diseases were common in K . pneumoniae infections, they were not common in K . oxytoca infection . And while metastatic septic abscess is a well-known, devastating complication of K . pneumoniae septicemia, usually seen as a brain abscess with a gas-forming appearance, all of these K . oxytoca infections had both otogenic infections and more favorable outcomes.

J Med Microbiol, 2001 May, 50(5), 456 - 61
Induction of interleukin-10 and down-regulation of cytokine production by Klebsiella pneumoniae capsule in mice with pulmonary infection; Yoshida K et al.; The role of the capsule of Klebsiella pneumoniae in inducing cytokine production was investigated by comparing the responses of mice with experimentally induced pneumonia caused by capsulate (strain DT-S) or non-capsulate (mutant strain DT-X) K . pneumoniae . Anaesthetised ICR mice were inoculated intranasally . Whereas all DT-S-infected mice died within 3 days, no deaths were observed in DT-X-infected mice by 14 days after infection . During the early stage of infection, interferon-gamma (IFN-gamma) levels in bronchoalveolar lavage fluid (BALF) of DT-X-infected mice were significantly higher than those in DT-S-infected mice . In contrast, in the late stage of infection, serum levels of granulocyte macrophage-colony stimulating factor (GM-CSF) and IFN-gamma in DT-S-infected mice were significantly higher than those in DT-X-infected mice . Levels of interleukin10 (IL-10) in BALF and serum of DT-S-infected mice were significantly and persistently higher than those of DT-X-infected mice . The IL-10/TNF-alpha (tumour necrosis factor-a) ratios in BALF and serum indicated that higher levels of IL-10 production were induced in mice infected with strain DT-S than in those infected with strain DT-X . The results suggest that the capsule of K . pneumoniae may induce IL-10 production at the site of infection and, thereafter, these high IL-10 levels may serve to down-regulate the expression of pro-inflammatory cytokines.

J Biol Chem, 2001 Jul 13, 276(28), 26276 - 84 Epub 2001 May 02.
Evaluation of the substrate specificity of human mast cell tryptase beta I and demonstration of its importance in bacterial infections of the lung; Huang C et al.; Human pulmonary mast cells (MCs) express tryptases alpha and beta I, and both granule serine proteases are exocytosed during inflammatory events . Recombinant forms of these tryptases were generated for the first time to evaluate their substrate specificities at the biochemical level and then to address their physiologic roles in pulmonary inflammation . Analysis of a tryptase-specific, phage display peptide library revealed that tryptase beta I prefers to cleave peptides with 1 or more Pro residues flanked by 2 positively charged residues . Although recombinant tryptase beta I was unable to activate cultured cells that express different types of protease-activated receptors, the numbers of neutrophils increased >100-fold when enzymatically active tryptase beta I was instilled into the lungs of mice . In contrast, the numbers of lymphocytes and eosinophils in the airspaces did not change significantly . More important, the tryptase beta I-treated mice exhibited normal airway responsiveness . Neutrophils did not extravasate into the lungs of tryptase alpha-treated mice . Thus, this is the first study to demonstrate that the two nearly identical human MC tryptases are functionally distinct in vivo . When MC-deficient W/W(v) mice were given enzymatically active tryptase beta I or its inactive zymogen before pulmonary infection with Klebsiella pneumoniae, tryptase beta I-treated W/W(v) mice had fewer viable bacteria in their lungs relative to zymogen-treated W/W(v) mice . Because neutrophils are required to combat bacterial infections, human tryptase beta I plays a critical role in the antibacterial host defenses of the lung by recruiting neutrophils in a manner that does not alter airway reactivity.

Epidemiol Mikrobiol Imunol, 2001 Apr, 50(2), 92 - 6
{Klebsiella species from the viewpoint of nosocomial infections and virulence factors}; Hostacka A; The author reviewed data on Klebsiella species mainly in connection with their participation in nosocomial infections, with their antibiotic resistance and production of ESBL . Potential virulence factors of Klebsiella are also evaluated.

J Bacteriol, 2001 May, 183(10), 3076 - 82
Role of Escherichia coli nitrogen regulatory genes in the nitrogen response of the Azotobacter vinelandii NifL-NifA complex; Reyes-Ramirez F et al.; The redox-sensing flavoprotein NifL inhibits the activity of the nitrogen fixation (nif)-specific transcriptional activator NifA in Azotobacter vinelandii in response to molecular oxygen and fixed nitrogen . Although the mechanism whereby the A . vinelandii NifL-NifA system responds to fixed nitrogen in vivo is unknown, the glnK gene, which encodes a PII-like signal transduction protein, has been implicated in nitrogen control . However, the precise function of A . vinelandii glnK in this response is difficult to establish because of the essential nature of this gene . We have shown previously that A . vinelandii NifL is able to respond to fixed nitrogen to control NifA activity when expressed in Escherichia coli . In this study, we investigated the role of the E . coli PII-like signal transduction proteins in nitrogen control of the A . vinelandii NifL-NifA regulatory system in vivo . In contrast to recent findings with Klebsiella pneumoniae NifL, our results indicate that neither the E . coli PII nor GlnK protein is required to relieve inhibition by A . vinelandii NifL under nitrogen-limiting conditions . Moreover, disruption of both the E . coli glnB and ntrC genes resulted in a complete loss of nitrogen regulation of NifA activity by NifL . We observe that glnB ntrC and glnB glnK ntrC mutant strains accumulate high levels of intracellular 2-oxoglutarate under conditions of nitrogen excess . These findings are in accord with our recent in vitro observations (R . Little, F . Reyes-Ramirez, Y . Zhang, W . Van Heeswijk, and R . Dixon, EMBO J . 19:6041-6050, 2000) and suggest a model in which nitrogen control of the A . vinelandii NifL-NifA system is achieved through the response to the level of 2-oxoglutarate and an interaction with PII-like proteins under conditions of nitrogen excess.

Carbohydr Res, 2001 Mar 22, 331(2), 149 - 61
Phosphorylation and metabolism of sucrose and its five linkage-isomeric alpha-D-glucosyl-D-fructoses by Klebsiella pneumoniae; Thompson J et al.; Not only sucrose but the five isomeric alpha-D-glucosyl-D-fructoses trehalulose, turanose, maltulose, leucrose, and palatinose are utilized by Klebsiella pneumoniae as energy sources for growth, thereby undergoing phosphorylation by a phosphoenolpyruvate-dependent phosphotransferase system uniformly at 0-6 of the glucosyl moiety . Similarly, maltose, isomaltose, and maltitol, when exposed to these conditions, are phosphorylated regiospecifically at O-6 of their non-reducing glucose portion . The structures of these novel compounds have been established unequivocally by enzymatic analysis, acid hydrolysis, FAB negative-ion spectrometry, and 1H and 13C NMR spectroscopy . In cells of K . pneumoniae, hydrolysis of sucrose 6-phosphate is catalyzed by sucrose 6-phosphate hydrolase from Family 32 of the glycosylhydrolase superfamily . The five 6'-O-phosphorylated alpha-D-glucosyl-fructoses are hydrolyzed by an inducible (approximately 49-50 Kda) phospho-alpha-glucosidase from Family 4 of the glycosylhydrolase superfamily.

Infect Control Hosp Epidemiol, 2001 Mar, 22(3), 148 - 51
Nosocomial colonization of premature babies with Klebsiella oxytoca: probable role of enteral feeding procedure in transmission and control of the outbreak with the use of gloves; Berthelot P et al.; OBJECTIVE: To investigate the persistence of colonization of premature babies by Klebsiella oxytoca, with special emphasis on the mode of transmission of the bacterium and evaluation of Standard Precautions to stop the epidemic . DESIGN: Retrospective analysis of cases and prospective study of systematic bacteriological samples (stools and throat) from babies, healthcare workers (HCWs), and environment, with genotyping of strains by arbitrarily primed polymerase chain reaction . SETTING: A premature baby unit (PBU) and a neonatal intensive care unit in the university hospital of Saint-Etienne, France . RESULTS: An outbreak of K oxytoca was suspected in two pediatric wards after the occurrence of a fatal bacteremia in a newborn hospitalized in the PBU and the colonization of other babies 2 months later . Retrospective analysis showed that 24 babies' digestive tract had been colonized . No environmental reservoir was recovered in the units nor in enteral feeding . No K oxytoca was isolated from HCW samples . Genotyping confirmed the presence of epidemic strains, although independent clones were responsible for infections or colonizations in each of the two units . The chronology and the site of babies' colonization (isolation of K oxytoca in stools before throat) were determined during a prospective study and suggested that enteral feeding procedures could be the source of contamination . Therefore, use of gloves during this practice by HCWs was recommended and, after readjustment of Standard Precautions, stopped the outbreak . CONCLUSION: To prevent cross-contamination among high-risk babies, careful attention must be paid to Standard Precautions . Bacteriological surveillance of the digestive tract of neonates could help to check compliance with these guidelines

Antimicrob Agents Chemother, 2001 May, 45(5), 1487 - 92
Improved efficacy of ciprofloxacin administered in polyethylene glycol-coated liposomes for treatment of Klebsiella pneumoniae pneumonia in rats; Bakker-Woudenberg IA et al.; Animal and clinical data show that high ratios of the area under the concentration-time curve and the peak concentration in blood to the MIC of fluoroquinolones for a given pathogen are associated with a favorable outcome . The present study investigated whether improvement of the therapeutic potential of ciprofloxacin could be achieved by encapsulation in polyethylene glycol (PEG)-coated long-circulating sustained-release liposomes . In a rat model of unilateral Klebsiella pneumoniae pneumonia (MIC = 0.1 microg/ml), antibiotic was administered at 12- or 24-h intervals at twofold-increasing doses . A treatment period of 3 days was started 24 h after inoculation of the left lung, when the bacterial count had increased 1,000-fold and some rats had positive blood cultures . The infection was fatal within 5 days in untreated rats . Administration of ciprofloxacin in the liposomal form resulted in delayed ciprofloxacin clearance and increased and prolonged ciprofloxacin concentrations in blood and tissues . The ED(50) (dosage that results in 50% survival) of liposomal ciprofloxacin was 3.3 mg/kg of body weight/day given once daily, and that of free ciprofloxacin was 18.9 mg/kg/day once daily or 5.1 mg/kg/day twice daily . The ED(90) of liposomal ciprofloxacin was 15.0 mg/kg/day once daily compared with 36.0 mg/kg/day twice daily for free ciprofloxacin; 90% survival could not be achieved with free ciprofloxacin given once daily . In summary, the therapeutic efficacy of liposomal ciprofloxacin was superior to that of ciprofloxacin in the free form . PEG-coated liposomal ciprofloxacin was well tolerated in relatively high doses, permitting once daily administration with relatively low ciprofloxacin clearance and without compromising therapeutic efficacy.

Microbios, 2001, 104(408), 115 - 24
Antibiotic susceptibility, serum response and surface properties of Klebsiella species; Hostacka A et al.; Altogether 130 clinical isolates of five Klebsiella species (K . pneumoniae, K . planticola, K . oxytoca, K . ornithinolytica and K . terrigena) were characterized, for their susceptibility to five antibiotics, for susceptibility to serum bactericidal activity and for their hydrophobic properties . All strains exhibited ampicillin resistance . Ampicillin/sulbactam, gentamicin and ofloxacin showed effectiveness in 63.1, 67.7 and 71.5% of the Klebsiella isolates . K . planticola manifested the highest level of resistance to these antibiotics . The majority of Klebsiella strains (93.9%) were susceptible to cefuroxime . Sixty-four strains (49.2%) were serum resistant and intermediate serum sensitivity was shown by 57 strains (43.8%) . A high percentage of serum resistant strains (65%) was found in K . planticola . Moderately hydrophobic properties determined by adherence of bacteria to xylene were demonstrated in 25 strains (19.2%).

Emerg Infect Dis, 2001 Mar-Apr, 7(2), 333 - 6
Controversies about extended-spectrum and AmpC beta-lactamases; Thomson KS; Many clinical laboratories have problems detecting extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases . Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions . Failure to detect these enzymes has contributed to their uncontrolled spread and sometimes to therapeutic failures . Although National Committee for Clinical Laboratory Standards recommendations exist for detecting ESBL- producing isolates of Escherichia coli and Klebsiella spp., no recommendations exist for detecting ESBLs in other organisms or for detecting plasmid-mediated AmpC beta-lactamases in any organisms . Clinical laboratories need to have adequate funding, equipment, and expertise to provide a rapid and clinically relevant antibiotic testing service in centers where these resistance mechanisms are encountered.

Pediatr Hematol Oncol, 2001 Apr-May, 18(3), 229 - 32
Klebsiella pneumoniae meningitis in thalassemia major patients; Li CK et al.; Two thalassemia major patients received regular blood transfusion and desferrioxamine chelation, and 1 patient had a splenectomy at 9 years of age . Both patients developed Klebsiella pneumoniae meningitis at age of 27 and 15 years . They died within a short time despite appropriate antibiotic treatment . Klebsiella meningitis may be more common in transfusion-dependent thalassemia patients.

J Miss State Med Assoc, 2001 Mar, 42(3), 67 - 70
Pyogenic liver abscesses due to Klebsiella pneumoniae in a diabetic patient; Ayinala SR et al.; Pyogenic liver abscess due to Klebsiella pneumoniae is a rare clinical entity . It has emerged as an important infection complication in diabetics and its incidence in diabetics without intraabdominal or biliary tract infections is increasing . We present herein a case of multiple pyogenic liver abscesses due to K . pneumoniae in a diabetic patient and discuss clinical course, treatment and possible reasons for association between K . pneumoniae liver abscess and diabetes.

Plant Mol Biol, 2001 Jan, 45(2), 169 - 79
Functional characterisation of urease accessory protein G (ureG) from potato; Witte CP et al.; The activation of the nickel metalloenzyme urease is a complex process . In bacteria, several urease accessory proteins are essential for incorporation of nickel into the active centre of urease . Comparatively little is known about the activation process and the proteins involved in plants . We cloned five different cDNAs encoding isoforms of urease accessory protein G (ureG) in potato . The 5'-coding region of these cDNAs is highly polymorphic within Solanum tuberosum ssp . tuberosum, containing mainly a simple sequence repeat encoding histidine and aspartate . Mapping on an ultrahigh-density map of the potato genome and Southern blot analysis showed that the isoforms arise from allelic differences of a single-copy gene which was located on chromosome 2 . Expression analysis at the mRNA and protein levels indicated the presence of ureG in almost all tissues examined, consistent with the ubiquitous expression of urease . An attempt to correlate urease activity with ureG expression levels in different tissues was made . Allelic copies of ureG were expressed in a tissue-specific manner . UreG from potato and the Klebsiella aerogenes urease operon defective in bacterial ureG were co-expressed in Escherichia coli . The plant gene complements the K . aerogenes ureG mutation, demonstrating that it encodes a urease accessory protein and indicating a structural conservation between the plant and the bacterial urease activation complexes.

Clin Infect Dis, 2001 Apr 15, 32(8), 1162 - 71 Epub 2001 Mar 26.
Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae: risk factors for infection and impact of resistance on outcomes; Lautenbach E et al.; The prevalence of antibiotic resistance among extended-spectrum beta-lactamase (ESBL)--producing Escherichia coli and Klebsiella pneumoniae has increased markedly in recent years . Thirty-three patients with infection due to ESBL-producing E . coli or K . pneumoniae (case patients) were compared with 66 matched controls . Total prior antibiotic use was the only independent risk factor for ESBL-producing E . coli or K . pneumoniae infection (odds ratio, 1.10; 95% confidence interval, 1.03--1.18; P=.006) . Case patients were treated with an effective antibiotic a median of 72 hours after infection was suspected, compared with a median of 11.5 hours after infection was suspected for controls (P<.001) . ESBL-producing E . coli or K . pneumoniae infection was associated with a significantly longer duration of hospital stay and greater hospital charges (P=.01 and P<.001, respectively) . Finally, many ESBL-producing E . coli and K . pneumoniae isolates were closely related . ESBL-producing E . coli and K . pneumoniae infections have a significant impact on several important clinical outcomes, and efforts to control outbreaks of infection with ESBL-producing E . coli and K . pneumoniae should emphasize judicious use of all antibiotics as well as barrier precautions to reduce spread.

Appl Environ Microbiol, 2001 Apr, 67(4), 1911 - 21
Genomic interspecies microarray hybridization: rapid discovery of three thousand genes in the maize endophyte, Klebsiella pneumoniae 342, by microarray hybridization with Escherichia coli K-12 open reading frames; Dong Y et al.; In an effort to efficiently discover genes in the diazotrophic endophyte of maize, Klebsiella pneumoniae 342, DNA from strain 342 was hybridized to a microarray containing 96% (n = 4,098) of the annotated open reading frames from Escherichia coli K-12 . Using a criterion of 55% identity or greater, 3,000 (70%) of the E . coli K-12 open reading frames were also found to be present in strain 342 . Approximately 24% (n = 1,030) of the E . coli K-12 open reading frames are absent in strain 342 . For 1.6% (n = 68) of the open reading frames, the signal was too low to make a determination regarding the presence or absence of the gene . Genes with high identity between the two organisms are those involved in energy metabolism, amino acid metabolism, fatty acid metabolism, cofactor synthesis, cell division, DNA replication, transcription, translation, transport, and regulatory proteins . Functions that were less highly conserved included carbon compound metabolism, membrane proteins, structural proteins, putative transport proteins, cell processes such as adaptation and protection, and central intermediary metabolism . Open reading frames of E . coli K-12 with little or no identity in strain 342 included putative regulatory proteins, putative chaperones, surface structure proteins, mobility proteins, putative enzymes, hypothetical proteins, and proteins of unknown function, as well as genes presumed to have been acquired by lateral transfer from sources such as phage, plasmids, or transposons . The results were in agreement with the physiological properties of the two strains . Whole genome comparisons by genomic interspecies microarray hybridization are shown to rapidly identify thousands of genes in a previously uncharacterized bacterial genome provided that the genome of a close relative has been fully sequenced . This approach will become increasingly more useful as more full genome sequences become available.

Int J Pharm, 2001 Feb 19, 214(1-2), 103 - 5
Therapeutic efficacy of liposomal gentamicin in clinically relevant rat models; Schiffelers RM et al.; Sterically stabilized liposomes are able to localize selectively at sites of infection, potentially permitting targeted drug delivery . Up to now, the majority of studies investigating therapeutic efficacy of liposomes have been conducted in animals with an intact host defense infected with high antibiotic-susceptible bacteria . In the present study, the therapeutic efficacy of gentamicin encapsulated in sterically stabilized liposomes, alone or in combination with the free drug was studied in rats with intact host defense as well as leukopenic rats . Rats were inoculated with a high gentamicin-susceptible or low-gentamicin susceptible Klebsiella pneumoniae in the left lung, resulting in an acute unilateral pneumonia . Survival rates demonstrate the valuable therapeutic properties of the liposome-encapsulated drug in these clinically relevant animal models.

Singapore Med J, 2000 Oct, 41(10), 489 - 92
Pyogenic liver abscess--a tropical centre's experience in management with review of current literature; Liew KV et al.; AIM OF STUDY: To perform a retrospective study, with the help of literature review, of the management of patients with pyogenic liver abscess in a general hospital . METHOD: A retrospective study of 73 consecutive patients treated atTanTock Seng Hospital between January 1994 and December 1997 was conducted to determine the demographic, clinical, laboratory, radiological and microbiological characteristics of these patients, as well as the management strategies employed . RESULTS: Liver abscess was more common in males, occurring more frequently in the right hepatic lobe . Most patients presented with non-specific clinical and biochemical features . A raised alkaline phosphatase level was the most common biochemical abnormality found in about two-thirds of patients . Ultrasonography was not as sensitive as computed tomographic scans in detecting abscesses . Klebsiella pneumoniae was the most common etiological agent detected in cultures of blood and abscess aspirates . All patients were treated with intravenous antibiotics . Twenty-two (30%) needed percutaneous catheter drainage and five (7%) required surgical management.There was no hospital mortality in our series . Prolonged hospitalisation was associated with advanced age, degree of loculation within the abscess, concomitant diabetes mellitus and Klebsiella septicaemia . CONCLUSION: Pyogenic liver abscesses require a high index of suspicion for early diagnosis . When appropriate therapy in the form of antibiotics in combination with percutaenous drainage or surgery is administered, mortality is very low . However, significant morbidity is still a problem, particularly in the elderly, diabetic patient.

Indian J Med Res, 2001 Jan, 113, 1 - 4
Rise in free intracellular calcium in HeLa cells infected with aggregative Klebsiella pneumoniae strains isolated from cases of diarrhoea; Pal A et al.; BACKGROUND & OBJECTIVES: Klebsiella pneumoniae strains occasionally cause diarrhoea in humans . This study was done to determine the involvement of calcium in the pathogenesis of aggregative K . pneumoniae strains . METHODS: A total of nine strains of K . pneumoniae were tested for adherence assay in HeLa cell line . A representative strain CO-1215 was used for {Ca2+}i study using Fura-2 fluorescence . RESULTS: Infection of cultured HeLa cells with aggregative K . pneumoniae strain resulted in five-fold elevation of intracellular level of free calcium ({Ca2+}i) with maximum Ca2+ influx at 3 h after bacterial infection . Chelation of extracellular Ca2+ with {ethylenebis(oxyethylenenitrile)} tetraacetic acid and suspension of cells in Ca2+ free buffer suggested that the rise of Ca2+ in aggregative K . pneumoniae infected HeLa cells was due to influx of Ca2+ from extracellular medium . INTERPRETATION & CONCLUSIONS: This study showed aggregative adherence in HeLa cells and this adherence leads to influx of extracellular Ca2+ . The unrestricted passage of calcium ions across cell membranes could cause phosphorylation of proteins involved in ion transport across the membrane, which could result in secretory diarrhoea . Further work is in progress to study the enterotoxicity of these strains in an in vitro rabbit intestinal model.

J Bacteriol, 2001 Apr, 183(8), 2709 - 14
Growth inhibition caused by overexpression of the structural gene for glutamate dehydrogenase (gdhA) from Klebsiella aerogenes; Janes BK et al.; Two linked mutations affecting glutamate dehydrogenase (GDH) formation (gdh-1 and rev-2) had been isolated at a locus near the trp cluster in Klebsiella aerogenes . The properties of these two mutations were consistent with those of a locus containing either a regulatory gene or a structural gene . The gdhA gene from K . aerogenes was cloned and sequenced, and an insertion mutation was generated and shown to be linked to trp . A region of gdhA from a strain bearing gdh-1 was sequenced and shown to have a single-base-pair change, confirming that the locus defined by gdh-1 is the structural gene for GDH . Mutants with the same phenotype as rev-2 were isolated, and their sequences showed that the mutations were located in the promoter region of the gdhA gene . The linkage of gdhA to trp in K . aerogenes was explained by postulating an inversion of the genetic map relative to other enteric bacteria . Strains that bore high-copy-number clones of gdhA displayed an auxotrophy that was interpreted as a limitation for alpha-ketoglutarate and consequently for succinyl-coenzyme A (CoA) . Three lines of evidence supported this interpretation: high-copy-number clones of the enzymatically inactive gdhA1 allele showed no auxotrophy, repression of GDH expression by the nitrogen assimilation control protein (NAC) relieved the auxotrophy, and addition of compounds that could increase the alpha-ketoglutarate supply or reduce the succinyl-CoA requirement relieved the auxotrophy.

J Wildl Dis, 2001 Jan, 37(1), 201 - 3
Ehrlichiosis in a moose calf in Norway; Jenkins A et al.; A case of granulocytic ehrlichiosis in a moose calf (Alces alces) in Norway is described . The animal was heavily infested with ticks (Ixodes ricinus), and died from a Klebsiella pneumoniae septicemia . Examination of blood smears from the calf revealed cytoplasmic inclusions (morulae) typical of infection with Ehrlichia phagocytophila in the granulocytes . Ehrlichia sp . was detected by polymerase chain reaction (PCR) in blood from the calf, and in the ticks . Sequence determination identified it as E . phagocytophila . This is the first report of ehrlichiosis in moose.

Microb Drug Resist, 2000 Winter, 6(4), 297 - 303
Detection of multiresistant ceftazidime-susceptible Klebsiella pneumoniae isolates lacking TEM-26 after class restriction of cephalosporins; Urban C et al.; A multitude of extended spectrum beta-lactamases (ESBLs) have evolved in response to the use of late generation cephalosporins . In those hospitals where Klebsiella pneumoniae and other bacteria possessing these enzymes flourish, many interventions have been applied to reduce this trend . We instituted a policy of class restriction of cephalosporins in our hospital in 1996 that led to a 44% reduction in ceftazidime-resistant K . pneumoniae hospital-wide and an 87% decrease in the surgical intensive care unit . Another interesting outcome of this strategy was the identification of multiresistant K . pneumoniae, which was now susceptible to ceftazidime . Characterization of these novel isolates demonstrated that the TEM-26 enzyme, which was responsible for ceftazidime resistance in our earlier described outbreak, was lacking in most of the isolates examined . Among the remaining ceftazidime-resistant K . pneumoniae, TEM-26 was also absent, and new enzymes that hydrolyze ceftazidime were detected . Loss of ceftazidime-hydrolyzing beta-lactamases was observed after in vitro passage of ceftazidime-resistant K . pneumoniae on antibiotic-free media . These findings suggest that class restriction of cephalosporins may increase susceptibility among extended-spectrum beta-lactamase-producing pathogens.

J Antimicrob Chemother, 2001 Apr, 47(4), 463 - 5
SHV-27, a novel cefotaxime-hydrolysing beta-lactamase, identified in Klebsiella pneumoniae isolates from a Brazilian hospital; Corkill JE et al.; From a collection of cefotaxime-resistant Klebsiella pneumoniae isolated from neonatal blood culture specimens in a maternity hospital in Aracaju, Brazil, two isolates (strains KPBRZ-842 and -843, indistinguishable by pulsed-field gel electrophoresis) were found to produce beta-lactamases with isoelectric points (pI) of 5.4 and 8.2, respectively . Using a gel overlay method, cefotaxime hydrolysis was shown to be associated with the pI 8.2 protein . Nucleotide sequencing of the gene encoding the pI 8.2 beta-lactamase revealed a bla(SHV-ESBL)-type gene differing from the gene encoding SHV-1 by three silent point mutations, and a fourth that resulted in an amino acid substitution, aspartate for glycine, at position 156 . This novel SHV-type extended-spectrum beta-lactamase is designated SHV-27.

Nat Immunol, 2000 Dec, 1(6), 502 - 9
OmpA targets dendritic cells, induces their maturation and delivers antigen into the MHC class I presentation pathway; Jeannin P et al.; We analyzed the interaction between a bacterial cell wall protein and dendritic cells (DCs) . Outer membrane protein A from Klebsiella pneumoniae (kpOmpA) specifically bound to professional antigen presenting cells and was endocytosed by immature DCs via a receptor-dependent mechanism . kpOmpA signaled through Toll-like receptor 2, induced DCs to produce interleukin 12 and induced maturation of DCs . Whole antigen that was coupled to kpOmpA and injected into mice was taken up by DCs and delivered to the conventional cytosolic MHC class I presentation pathway . kpOmpA also primed antigen-specific CD8+ CTLs in the absence of CD4+ T cell help or adjuvant and elicited therapeutic immunity to antigen-expressing tumors . Thus, OmpA belongs to a class of proteins that are able to elicit CTL responses to exogenous antigen.

Transplantation, 2001 Feb 27, 71(4), 575 - 7
Improved phagocyte response by co-amoxiclav in renal transplant recipients; Cuffini AM et al.; BACKGROUND: Infectious diseases are a major source of morbidity and mortality for immunosuppressed transplant recipients and the antimicrobial chemotherapy can be often less effective in these individuals, because the contribution of underlying host defenses is absent . METHODS: The influence of co-amoxiclav on the functions of polymorphonuclear granulocytes (PMNs) from renal transplant recipients were investigated . RESULTS: PMNs from renal transplant recipients showed a diminished phagocytic activity with reduced phagocytosis and bactericidal activity against intracellular Klebsiella pneumoniae, compared to that seen with PMNs from healthy subjects . Co-amoxiclav significantly elicited the functions of PMNs from uremic patients, resulting in an increased percentage of ingested klebsiellae and in a higher bactericidal effect (98-99%), compared with the drug-free control system . When PMNs were collected from renal transplant recipients treated with co-amoxiclav a significant high increase in both phagocytosis and killing activity were detected, showing the co-amoxiclav capability of "restoring" even in vivo the depressed primary functions of PMNs . CONCLUSIONS: The interesting beneficial properties of co-amoxiclav, which result in restoring the phagocyte-dependent response in renal transplant patients both in vitro and in vivo, may make this drug more suitable for the treatment of infections in patients with defects of phagocyte functions.

Antimicrob Agents Chemother, 2001 Apr, 45(4), 1287 - 91
In vitro and in vivo activities of amikacin, cefepime, amikacin plus cefepime, and imipenem against an SHV-5 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strain; Szabo D et al.; The in vitro and in vivo effectiveness of amikacin, cefepime, and imipenem was studied using a high inoculum of an extended-spectrum beta-lactamase-producing Klebsiella pneumoniae strain . An in vitro susceptibility test at the standard inoculum predicted the in vivo outcome of amikacin or imipenem while it did not do so for cefepime due to the inoculum effect.

J Immunol, 2001 Apr 1, 166(7), 4612 - 9
Cancer vaccine design: a novel bacterial adjuvant for peptide-specific CTL induction; Miconnet I et al.; The recent identification of tumor Ags as potential vaccines has prompted the search for efficient adjuvants and delivery systems, especially in the case of peptide-based vaccination protocols . Here, we investigated the adjuvant potential of the recombinant 40-kDa outer membrane protein of Klebsiella pneumoniae (P40) for specific CTL induction . We studied the CTL response induced in HLA-A*0201/K(b) transgenic mice immunized with peptides derived from two melanoma-associated differentiation Ags, the HLA-A*0201-restricted decapeptide Melan-A(26--35) substituted at position 2 and the K(b)-restricted tyrosinase-related protein 2(181--188) T cell epitope . We found that both peptides are able to generate a specific CTL response when mixed with the protein in the absence of conventional adjuvant . This CTL response is a function of the amount of P40 used for immunization . Moreover, the CTL response generated against the tyrosinase-related protein 2(181-188) peptide in presence of P40 is associated with tumor protection in two different experimental models and is independent of the presence of CD4(+) T lymphocytes . Thus, the recombinant bacterial protein P40 functions as a potent immunological adjuvant for specific CTL induction.

Infect Immun, 2001 Apr, 69(4), 2011 - 6
Leukotriene B4 augments neutrophil phagocytosis of Klebsiella pneumoniae; Mancuso P et al.; Neutrophils play a critical role in the clearance of bacteria from the lung and other organs by their capacity for phagocytosis and killing . Previously, we identified an important role for the leukotrienes in rat alveolar macrophage phagocytosis of Klebsiella pneumoniae . In this report, we explored the possibility that the leukotrienes play an important role in phagocytosis by neutrophils as well . Inhibition of endogenous leukotriene synthesis by 5-lipoxygenase knockout in mice or by pharmacologic means in human peripheral blood neutrophils attenuated phagocytosis of opsonized K . pneumoniae . Reduced phagocytosis was also observed in human neutrophils pretreated with a leukotriene B4 receptor but not a cysteinyl-leukotriene receptor antagonist . While leukotriene B4 reconstituted defective phagocytosis in leukotriene-deficient neutrophils and enhanced phagocytosis in neutrophils capable of leukotriene synthesis, leukotriene C4, leukotriene D4, 5-hydroperoxyeicosatetraenoic acid, and 5-oxo-eicosatetraenoic acid were ineffective . To determine the opsonin dependence of the leukotriene B4 augmentation of phagocytosis, we assessed the ability of leukotriene B4 to modulate neutrophil phagocytosis and the adherence of sheep erythrocytes opsonized with immunoglobulin G or the complement fragment C3bi . While leukotriene B4 augmented both Fc receptor- and complement receptor-mediated phagocytosis, increased adherence to leukotriene B4-treated neutrophils was limited to complement opsonized targets . In conclusion, we have identified a novel role for leukotriene B4 in the augmentation of neutrophil phagocytosis mediated by either the Fc or complement receptor.

Eur J Biochem, 2001 Mar, 268(6), 1722 - 9
The structure of the core region of the lipopolysaccharide from Klebsiella pneumoniae O3 . 3-deoxy-alpha-D-manno-octulosonic acid (alpha-Kdo) residue in the outer part of the core, a common structural element of Klebsiella pneumoniae O1, O2, O3, O4, O5, O8, and O12 lipopolysaccharides; Vinogradov E et al.; The structure of lipid A-core region of the lipopolysaccharide (LPS) from Klebsiella pneumoniae serotype O3 was determined using NMR, MS and chemical analysis of the oligosaccharides, obtained by mild acid hydrolysis, alkaline deacylation, and deamination of the LPS: {carbohydrate structure see text} where P is H or alpha-Hep; J is H or beta-GalA; R is H or P (in the deacylated oligosaccharides).Screening of the LPS from K . pneumoniae O1, O2, O4, O5, O8, and O12 using deamination showed that they also contain alpha-Hep-(1-->4)-alpha-Kdo-(2-->6)-GlcN and alpha-Kdo-(2-->6)-GlcN fragments.

Postepy Hig Med Dosw, 2000, 54(6), 727 - 47
{Fimbriae as a pathologic factor of bacteria and a carrier in conjugate vaccines}; Staniszewska M et al.; Fimbriae play important role as pathogenic factors in many bacteria by their adhesive properties . Adhesin is located at the tip of fimbriae but also in other parts of fimbriae . Recent findings on structure of fimbriae genes and their expression for the biosynthesis and formulation of complete fimbriae have been described . Special attention was focused on the participation of fimbriae in the mechanism of pathogenesis and their specificity towards tissue receptors . Most recent studies have been performed on E . coli and Klebsiella and those data predominate in this work . Fimbriae can be used for the construction of vaccine as a proteinous carrier for haptenic carbohydrate epitopes . In conjugates fimbriae express distinct immunogenic, adjuvant and protective properties.

Semin Respir Infect, 2000 Dec, 15(4), 299 - 307
Extended-spectrum beta-lactamases; Patterson JE; Resistance to broad-spectrum cephalosporins among Klebsiella pneumoniae has increased significantly, particularly in the intensive care unit setting, in the past decade.The problem has been noted not only in the United States, but around the world . A major mechanism responsible for this is the emergence of extended-spectrum beta-lactamases (ESBLs).These plasmid-mediated beta-lactamases confer resistance to broad-spectrum beta-lactam antibiotics, including third- and fourth-generation cephalosporins, aztreonam, and extended-spectrum penicillins . Other resistances, such as aminoglycoside and trimethoprim-sulfamethoxazole resistance, are often cotransferred on the same plasmid . Fluoroquinolone resistance is also frequently associated, resulting in an organism resistant to most broad-spectrum options . The carbapenems are currently considered the drug of choice for these pathogens . Prevention and control measures are important because of the multiresistant nature of these pathogens . Such traditional infection control measures as contact precautions are recommended . In addition, because this type of antimicrobial resistance appears to be particularly influenced by antibiotic use, antibiotic control measures may also be a very important intervention in controlling the spread of ESBLs.

J Autoimmun, 2001 Feb, 16(1), 29 - 36
Production of experimental autoimmune sialadenitis in mice immunized with homologous salivary gland extract and Klebsiella O3 lipopolysaccharide; Mu MM et al.; An experimental murine model for autoimmune sialadenitis was produced by repeated immunization of homologous salivary gland extract together with Klebsiella O3 lipopolysaccharides as an immunological adjuvant . The cell infiltration was observed in the salivary glands of mice immunized more than twice . Inflammatory cells consisting mainly of CD4+ T cells and CD8+ T cells accumulated at the perivascular regions . There was hyperplasia and enlargement of ductal epithelial cells in the secretory acinar units in salivary glands of repeatedly immunized mice . The repeated immunization developed delayed-type hypersensitivity and autoantibody production to the homologous salivary gland extract . The immunohistochemical analysis showed positive staining on the cuboidal cells in the intercalated ducts, and the columnar pseudostratified cells in the striated ducts . Organ-specific antigens with molecular weights ranging from 20 to 90 kDa were recognized by the sera from immunized mice . Therefore, it was suggested that the sialadenitis was produced by the autoimmune mechanism and might be a new experimental model for characterization of the pathogenesis of autoimmune sialadenitis.

J Radiat Res (Tokyo), 2000 Sep, 41(3), 259 - 77
Ionizing radiation and bacterial challenge alter splenic cytokine gene expression; Chang CM et al.; Irradiation increases susceptibility to bacterial infection . Exogenous proinflammatory cytokines can alter the response of mice to gamma radiation, but the role of endogenous inflammatory cytokines after bacterial infection in irradiated animals is not known . Gene expression of hematopoietic (GM-CSF) and proinflammatory (IL-1 beta, IL-6 and TNF-alpha) cytokines were examined in spleens of B6D2F1/J female mice after irradiation alone (1.0- and 7.0-Gy), and after irradiation followed by Klebsiella pneumoniae s.c . challenge 4 days postirradiation by using the reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot hybridization . At 4, 8, and 24 h after bacterial challenge in 7.0-Gy-irradiated mice, GM-CSF mRNA increased (p < 0.05) . TNF-alpha mRNA in irradiated mice were slightly decreased, whereas after bacterial challenge, TNF-alpha mRNA elevated at 30 h in 7.0-Gy-irradiated mice; at 4, and 8 h in 1.0-Gy-irradiated mice, and at 1 h in sham-irradiated mice (p < 0.05) . IL-6 mRNA displayed a biphasic response in 7.0-Gy-irradiated mice, and, after bacterial challenge, in both irradiated mice (1.0- and 7.0-Gy) and sham-irradiated mice . IL-1 beta mRNA remained at or below normal for 8 h and increased at 24 h after bacterial challenge on day 4 in 7.0-Gy-irradiated mice . These results indicate that sublethal gamma radiation alters the patterns of the hematopoietic and proinflammatory cytokine responses to bacterial challenge in vivo . Consequently, treatment protocols may need to take into account changes in cytokine gene responses to resolve infection after irradiation.

Braz Dent J, 2000, 11(2), 161 - 5
A rare case of lip ulcer infected by Klebsiella pneumoniae: case report; Pinheiro AL et al.; The aetiology of mouth ulcers is diverse and may include several types of trauma, systemic disease and infection . The size, depth, outline, base, aspect of the floor, pain, time of evolution and resolution of oral ulcers are discussed . Both past and present medical history, biopsy, blood tests and microbiological tests are also considered in order to assure precise identification . This paper reports a case of a lip ulcer caused by Klebsiella pneumoniae.

New Microbiol, 2001 Jan, 24(1), 35 - 45
Molecular analysis of Klebsiella pneumoniae strains isolated in pediatric wards by ribotyping, pulsed field gel electrophoresis and antimicrobial susceptibilities; Sechi LA et al.; The purpose of this study was to investigate the usefulness of different molecular typing techniques in the surveillance and control of the spread of extended-spectrum-beta-lactamase-(ESBL) producing Klebsiella pneumoniae in the pediatric department of the "Agostino Gemelli" hospital of the Catholic University in Rome, over a period of nine months . The strains were characterized by ribotyping using HindIII as restriction enzyme and pulsed field gel electrophoresis (PFGE) using XbaI as endonuclease . Sixty six K . pneumoniae clinical strains were isolated during this period, the first 32 were isolated in the summer of 1998 . Among these first isolates, ribotyping generated 26 different patterns whereas PFGE produced 16 patterns . The remaining 34 strains were isolated during January and April 1999 and all of them were ESBL producers . Ribotyping clustered the strains into 6 patterns whereas PFGE generated only 3 patterns . PCR revealed the presence in 10 isolates of both bla(TEM) and bla(SHV) genes and 24 strains carried only the bla(SHV) gene . In our experience ribotyping revealed a higher power of differentiation with respect to PFGE and was of great help in the surveillance of the infection.

Indian J Med Res, 2000 Oct, 112, 133 - 4
Enteroaggregative Klebsiella pneumoniae in association with childhood diarrhoea; Niyogi SK et al.; A total of 19 strains of Klebsiella pneumoniae isolated as sole pathogen from children with diarrhoea were used to study their virulence mechanism using different assays . Eith strains of K . pneumoniae exhibited aggregative adherence that was distinct from the stacked brick enteroaggregative pattern shown by Escherichia coli . The study suggests the presence of a new virulence mechanism in the pathogenesis of Klebsiella-associated diarrhoea.

J Immunol, 2001 Mar 1, 166(5), 3362 - 8
Impaired pulmonary host defense in mice lacking expression of the CXC chemokine lungkine; Chen SC et al.; Lungkine (CXCL15) is a novel CXC chemokine that is highly expressed in the adult mouse lung . To determine the biologic function of Lungkine, we generated Lungkine null mice by targeted gene disruption . These mice did not differ from wild-type mice in their hematocrits or in the relative number of cells in leukocyte populations of peripheral blood or other tissues, including lung and bone marrow . However, Lungkine null mice were more susceptible to Klebsiella pneumonia infection, with a decreased survival and increased lung bacterial burden compared with infected wild-type mice . Histologic analysis of the lung and assessment of leukocytes in the bronchioalveolar lavage revealed that neutrophil numbers were normal in the lung parenchyma, but reduced in the airspace . The production of other neutrophil chemoattractants in the Lungkine null mice did not differ from that in wild-type mice, and neutrophil migration into other tissues was normal . Taken together, these findings demonstrate that Lungkine is an important mediator of neutrophil migration from the lung parenchyma into the airspace.

Int J Infect Dis, 2000, 4(3), 123 - 8
Epidemiology and molecular characterization of nosocomially transmitted multidrug-resistant Klebsiella pneumoniae; Parasakthi N et al.; OBJECTIVES: To describe the epidemiology, antimicrobial susceptibility, genomic profiles, and control of a nosocomial outbreak of multidrug-resistant Klebsiella pneumoniae (MRKP) that occurred in the pediatric oncology unit of the University of Malaya Medical Centre in Kuala Lumpur . MATERIALS AND METHODS: A prospective epidemiologic and microbiologic study was conducted of MRKP isolated from the blood and wound of a boy with necrotizing fasciitis after a 7-day course of ceftazidime and amikacin . In the following 2 weeks, phenotypically similar MRKP were isolated from the blood cultures of four other patients and rectal swabs of another three patients and two liquid soap samples located in the same ward . RESULTS: Antimicrobial profiles demonstrated that all the isolates were resistant to ceftazidime, sensitive to imipenem and ciprofloxacin, and confirmed to be extended-spectrum beta-lactamase producers . Plasmids of varying molecular weights were present in all isolates . In eight of these isolates, which included four from blood, there were common large molecular weight plasmids ranging from 80 kb to 100 kb . Pulsed-field gel electrophoresis analysis using XbaI demonstrated six different DNA profiles, A to F . Profile A was shared by two blood culture isolates and were related by 91% . Profile B was found in one rectal swab isolate and one isolate from liquid soap and were related by 94% . Profile C was shared by one blood isolate and one liquid soap isolate and showed 100% relatedness . Profiles D, E, and F each were demonstrated by one blood isolate and two rectal swab isolates, respectively . These showed only 65% relatedness . CONCLUSIONS: The MRKP strains in this outbreak were not clonal in origin . The decline of the outbreak after 4 weeks was attributed to the reemphasis of standard infection control procedures and the implementation of a program that addressed sites of environmental contamination.

Acta Crystallogr D Biol Crystallogr, 2001 Feb, 57(Pt 2), 260 - 2
Crystallization of full-length CysB of Klebsiella aerogenes, a LysR-type transcriptional regulator; Verschueren KH et al.; CysB is a positive regulator of transcription of genes involved in cysteine biosynthesis in Gram-negative bacteria and belongs to the large family of LysR-type transcriptional regulators . The full-length protein from Klebsiella aerogenes has been crystallized from solutions containing PEG 8000 in the presence and in the absence of the inducer N-acetylserine by the method of vapour diffusion in hanging drops . For the complexed protein different crystal forms appear in the same drops.

Eur J Biochem, 2001 Feb, 268(3), 809 - 18
Transition state complexes of the Klebsiella pneumoniae nitrogenase proteins . Spectroscopic properties of aluminium fluoride-stabilized and beryllium fluoride-stabilized MgADP complexes reveal conformational differences of the Fe protein; Miller RW et al.; Stable inactive 2 : 1 complexes of the Klebsiella pneumoniae nitrogenase components (Kp2/Kp1) were prepared with ADP or the fluorescent ADP analogue, 2'(3')-O-{N-methylanthraniloyl} ADP and AlF(4)(-) or BeF(3)(-) ions . By analogy with published crystallographic data {Schindelin et al . (1997) Nature 387, 370-376)}, we suggest that the metal fluoride ions replaced phosphate at the two ATP-binding sites of the iron protein, Kp2 . The beryllium (BeF(x)) and aluminium (AlF(4)(-)) containing complexes are proposed to correspond to the ATP-bound state and the hydrolytic transition states, respectively, by analogy with the equivalent complexes of myosin {Fisher et al . (1995) Biochemistry 34, 8960-8972} . (31)P NMR spectroscopy showed that during the initial stages of complex formation, MgADP bound to the complexed Kp2 in a manner similar to that reported for isolated Kp2 . This process was followed by a second step that caused broadening of the (31)P NMR signals and, in the case of the AlF4- complex, slow hydrolysis of some of the excess ADP to AMP and inorganic phosphate . The purified BeFx complex contained 3.8 +/- 0.1 MgADP per mol Kp1 . With the AlF(4)(-) complex, MgAMP and adenosine (from MgAMP hydrolysis) replaced part of the bound MgADP although four AlF(4)(-) ions were retained, demonstrating that full occupancy by MgADP is not required for the stability of the complex . The fluorescence emission maximum of 2'(3')-O-{N-methylanthraniloyl} ADP was blue-shifted by 6-8 nm in both metal fluoride complexes and polarization was 6-9 times that of the free analogue . The fluorescence yield of bound 2'(3')-O-{N-methylanthraniloyl} ADP was enhanced by 40% in the AlF(4)(-) complex relative to the solvent but no increase in fluorescence was observed in the BeFx complex . Resonance energy transfer from conserved tyrosine residues located in proximity to the Kp2 nucleotide-binding pocket was marked in the AlF(4)(-) complex but minimal in the BeFx fluoride complex, illustrating a clear conformational difference in the Fe protein of the two complexes . Our data indicate that complex formation during the nitrogenase catalytic cycle is a multistep process involving at least four conformational states of Kp2: similar to the free Fe protein; as initially complexed with detectable (31)P NMR; as detected in mature complexes with no detectable (31)P NMR; in the AlF(4)(-) complex in which an altered tyrosine interaction permits resonance energy transfer with 2'(3')-O-{N-methylanthraniloyl} ADP.

Clin Microbiol Infect, 2000 May, 6(5), 233 - 8
Twenty-three years of Klebsiella phage typing: a review of phage typing of 12 clusters of nosocomial infections, and a comparison of phage typing with K serotyping; Sechter I et al.; OBJECTIVE: To review phage typing of 12 clusters of nosocomial Klebsiella infections which occurred between 1974 and 1997, and to compare phage typing and K serotyping . Materials and methods A total of 489 clinical and laboratory Klebsiella isolates were phage typed using 110 different phage preparations and K typed by counter current immunoelectrophoresis against 77 K antisera . RESULTS: A total of 152 phage types (PT) and 82 K types were found . Thirty-six phage types and 14 K types were represented only by the reference type strains . Of the remaining 68 K types, 60 could be subdivided into from two to 10 phage types . Ten out of 12 clusters of nosocomial Klebsiella infections could be verified as outbreaks by phage typing, whereas two clusters were found to be accumulations of sporadic cases . K typing performed retrospectively confirmed these results . In addition, for a subset of 104 epidemiologically unrelated isolates, O typing and pulsed field gel electrophoresis typing data were available . Based on these results the discriminative power of phage typing was found to be comparable with that of K typing, but phage types were less stable and reproducible . CONDITIONS: In an outbreak situation, phage typing was found to be very useful, although it seems less suitable for long-term surveillance purposes.

FEMS Microbiol Lett, 2001 Jan 15, 194(2), 221 - 7
The mtl genes and the mannitol-1-phosphate dehydrogenase from Klebsiella pneumoniae KAY2026; Otte S et al.; The mtl operon of Klebsiella pneumoniae KAY2026 (formerly Aerobacter aerogenes 1033-5P14) was shown to contain as the promoter-proximal gene mtlA, encoding a D-mannitol-specific enzyme II transporter (IICBA(Mtl)) . This gene is followed by mtlD, coding for a mannitol-1-phosphate dehydrogenase (MtlD, 382 amino acid residues), and mtlR (MtlR, 195 amino acid residues) coding for a putative repressor, gene mtlR overlaps the termination codon of mtlD . The DNA and protein sequences are highly similar to the corresponding genes (81% identical bp) and proteins (79-85% identical amino acids) of Escherichia coli K-12 . A truncated form of MtlD lacking the 162 C-terminal amino acid residues still shows 10% dehydrogenase activity which may explain the controversy in the literature concerning the properties of mannitol-phosphate and other medium-length dehydrogenases.

J Hosp Infect, 2001 Jan, 47(1), 53 - 9
An outbreak of hospital-acquired Klebsiella pneumoniae bacteraemia, including strains producing extended-spectrum beta-lactamase; Pena C et al.; This study describes the clinical outcome of an outbreak of extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KP) bacteraemia . Ninety-two episodes of hospital-acquired K . pneumoniae bacteraemia were studied, 49 ESBL-KP and 43 non-ESBL-KP, from May 1993 to June 1995 . Of these, 44 (90%) episodes of ESBL-KP vs . 20 (46%) episodes of non-ESBL-KP occurred in intensive care unit (ICU) patients . The incidence of K . pneumoniae bacteraemia (mainly due to ESBL-KP) increased in the ICU during the outbreak . A significant association was found between intravascular catheter-related bacteraemia and isolation of ESBL-KP {27 (56%) in the ESBL-KP group vs . 13 (30%) in the non-ESBL-KP group;P= 0.01} . The worst prognostic features were identified as age > 65 years (P= 0.02), septic shock (P< 0.001) and secondary bacteraemia (P= 0.04) . High rates of resistance to beta-lactam/beta-lactamase inhibitors observed in our ESBL-KP isolates, as well as variable activity of aminoglycosides, restricts the empirical use of these antibiotics . Carbapenems should be the treatment of choice since they are uniformly active against these strains . Our study shows that ESBL-KP bacteraemia occurring in an epidemic ICU setting is mainly catheter-related . We did not find ESBL strains to be associated with a significantly poor outcome .

J Bacteriol, 2001 Mar, 183(5), 1610 - 20
Effect of P(II) and its homolog GlnK on reversible ADP-ribosylation of dinitrogenase reductase by heterologous expression of the Rhodospirillum rubrum dinitrogenase reductase ADP-ribosyl transferase-dinitrogenase reductase-activating glycohydrolase regulatory system in Klebsiella pneumoniae; Zhang Y et al.; Reversible ADP-ribosylation of dinitrogenase reductase, catalyzed by the dinitrogenase reductase ADP-ribosyl transferase-dinitrogenase reductase-activating glycohydrolase (DRAT-DRAG) regulatory system, has been characterized in Rhodospirillum rubrum and other nitrogen-fixing bacteria . To investigate the mechanisms for the regulation of DRAT and DRAG activities, we studied the heterologous expression of R . rubrum draTG in Klebsiella pneumoniae glnB and glnK mutants . In K . pneumoniae wild type, the regulation of both DRAT and DRAG activity appears to be comparable to that seen in R . rubrum . However, the regulation of both DRAT and DRAG activities is altered in a glnB background . Some DRAT escapes regulation and becomes active under N-limiting conditions . The regulation of DRAG activity is also altered in a glnB mutant, with DRAG being inactivated more slowly in response to NH4+ treatment than is seen in wild type, resulting in a high residual nitrogenase activity . In a glnK background, the regulation of DRAT activity is similar to that seen in wild type . However, the regulation of DRAG activity is completely abolished in the glnK mutant; DRAG remains active even after NH4+ addition, so there is no loss of nitrogenase activity . The results with this heterologous expression system have implications for DRAT-DRAG regulation in R . rubrum.

Antimicrob Agents Chemother, 2001 Feb, 45(2), 464 - 70
Therapeutic efficacy of liposome-encapsulated gentamicin in rat Klebsiella pneumoniae pneumonia in relation to impaired host defense and low bacterial susceptibility to gentamicin; Schiffelers RM et al.; Long-circulating liposomes (LCL) may be used as targeted antimicrobial drug carriers as they localize at sites of infection . As a result, LCL-encapsulated gentamicin (LE-GEN) has demonstrated superior antibacterial activity over the free drug in a single-dose study of immunocompetent rats with Klebsiella pneumoniae pneumonia . In the present study, the therapeutic efficacy of LE-GEN was evaluated by monitoring rat survival and bacterial counts in blood and lung tissue in clinically relevant models, addressing the issue of impaired host defense and low bacterial antibiotic susceptibility . The results show that in immunocompetent rats infected with the high-GEN-susceptibility K . pneumoniae strain, a single dose of LE-GEN is clearly superior to an equivalent dose of free GEN . Yet complete survival can also be obtained with multiple doses of free GEN . In leukopenic rats infected with the high-GEN-susceptible K . pneumoniae strain, free GEN at the maximum tolerated dose (MTD) was needed to obtain survival . However, with the addition of a single dose of LE-GEN to free-GEN treatment, complete survival can be obtained using a sevenfold-lower cumulative amount of GEN than with free-GEN treatment alone . In leukopenic rats infected with low-GEN-susceptible K . pneumoniae cells, free GEN at the MTD did not result in survival . The use of LE-GEN is needed for therapeutic success . Increasing LE-GEN bilayer fluidity resulted in an increased GEN release from the liposomes and hence improved rat survival, thus showing the importance of the liposome lipid composition for therapeutic efficacy . These results warrant further clinical studies of liposomal formulations of aminoglycosides in immunocompromised patients with severe infections.

J Bacteriol, 2001 Feb, 183(4), 1385 - 93
Fnr Is required for NifL-dependent oxygen control of nif gene expression in Klebsiella pneumoniae; Grabbe R et al.; In Klebsiella pneumoniae, NifA-dependent transcription of nitrogen fixation (nif) genes is inhibited by NifL in response to molecular oxygen and combined nitrogen . We recently showed that K . pneumoniae NifL is a flavoprotein, which apparently senses oxygen through a redox-sensitive, conformational change . We have now studied the oxygen regulation of NifL activity in Escherichia coli and K . pneumoniae strains by monitoring its inhibition of NifA-mediated expression of K . pneumoniae phi (nifH'-'lacZ) fusions in different genetic backgrounds . Strains of both organisms carrying fnr null mutations failed to release NifL inhibition of NifA transcriptional activity under oxygen limitation: nif induction was similar to the induction under aerobic conditions . When the transcriptional regulator Fnr was synthesized from a plasmid, it was able to complement, i.e., to relieve NifL inhibition in the fnr mutant backgrounds . Hence, Fnr appears to be involved, directly or indirectly, in NifL-dependent oxygen regulation of nif gene expression in K . pneumoniae . The data indicate that in the absence of Fnr, NifL apparently does not receive the signal for anaerobiosis . We therefore hypothesize that in the absence of oxygen, Fnr, as the primary oxygen sensor, activates transcription of a gene or genes whose product or products function to relieve NifL inhibition by reducing the flavin adenine dinucleotide cofactor under oxygen-limiting conditions.

J Bacteriol, 2001 Feb, 183(4), 1312 - 9
Disulfide bond formation in secreton component PulK provides a possible explanation for the role of DsbA in pullulanase secretion; Pugsley AP et al.; When expressed in Escherichia coli, the 15 Klebsiella oxytoca pul genes that encode the so-called Pul secreton or type II secretion machinery promote pullulanase secretion and the assembly of one of the secreton components, PulG, into pili . Besides these pul genes, efficient pullulanase secretion also requires the host dsbA gene, encoding a periplasmic disulfide oxidoreductase, independently of disulfide bond formation in pullulanase itself . Two secreton components, the secretin pilot protein PulS and the minor pseudopilin PulK, were each shown to posses an intramolecular disulfide bond whose formation was catalyzed by DsbA . PulS was apparently destabilized by the absence of its disulfide bond, whereas PulK stability was not dramatically affected either by a dsbA mutation or by the removal of one of its cysteines . The pullulanase secretion defect in a dsbA mutant was rectified by overproduction of PulK, indicating reduced disulfide bond formation in PulK as the major cause of the secretion defect under the conditions tested (in which PulS is probably present in considerable excess of requirements) . PulG pilus formation was independent of DsbA, probably because PulK is not needed for piliation.

Rheumatology (Oxford), 2001 Jan, 40(1), 15 - 23
Correlation between the immune responses to collagens type I, III, IV and V and Klebsiella pneumoniae in patients with Crohn's disease and ankylosing spondylitis; Tiwana H et al.; BACKGROUND: Increased levels of collagen types I, III and V are found in strictures of patients with Crohn's disease (CD) compared with normal gut tissue . Type IV collagen is present in the basement membranes, basal lamina, retina and cornea . Elevated levels of antibody to Klebsiella pneumoniae are found in both active CD and active ankylosing spondylitis (AS) patients compared with healthy controls . METHODS: Reactivities for immunoglobulin class-specific antibodies (IgM, IgG and IgA) against collagen types I, III, IV, V and whole K . pneumoniae were measured by ELISA in nine patients with early CD and 10 with late CD from King's College Hospital and 12 late CD patients and 36 HLA-B27-positive AS patients from Middlesex Hospital and was compared with values for 26 healthy controls from the Blood Transfusion Service in London . RESULTS: Levels of class-specific IgM, IgG and IgA antibodies to collagen types I, III, IV, V and K . pneumoniae were significantly elevated in early and late CD patients compared with healthy controls (P<0.001) . Levels of IgM, IgG antibody to the four collagen types and K . pneumoniae were also significantly elevated (P<0.001) in AS patients compared with healthy controls . In addition, the level of IgA antibody to K . pneumoniae was elevated in AS patients (P<0.001) . Furthermore, a positive correlation between antibody levels to collagen types I, III, IV and K . pneumoniae was demonstrated in both early and late CD patients and in those with AS, whilst a positive correlation to type V was found in early CD . CONCLUSION: The role of K . pneumoniae and anti-collagen antibodies in the aetiopathogenesis of CD and AS requires further study.

J Biol Chem, 2000 Apr 14, 275(15), 10731 - 7
In vivo and in vitro kinetics of metal transfer by the Klebsiella aerogenes urease nickel metallochaperone, UreE; Colpas GJ et al.; The urease accessory protein encoded by ureE from Klebsiella aerogenes is proposed to deliver Ni(II) to the urease apoprotein during enzyme activation . Native UreE possesses a histidine-rich region at its carboxyl terminus that binds several equivalents of Ni(2+); however, a truncated form of this protein (H144*UreE) binds only 2 Ni(2+) per dimer and is functionally active (Brayman, T . G., and Hausinger, R . P . (1996) J . Bacteriol . 178, 5410-5416) . The urease activation kinetics were studied in vivo by monitoring the development of urease activity upon adding Ni(2+) to spectinomycin-treated Escherichia coli cells that expressed the complete K . aerogenes urease gene cluster with altered forms of ureE . Site-specific alterations of H144*UreE decrease the rate of in vivo urease activation, with the most dramatic changes observed for the H96A, H110A, D111A, and H112A substitutions . Notably, urease activity in cells producing H96A/H144*UreE was lower than cells containing a ureE deletion . Prior studies had shown that H110A and H112A variants each bound a single Ni(2+) per dimer with elevated K(d) values compared with control H144*UreE, whereas the H96A and D111A variants bound 2 Ni(2+) per dimer with unperturbed K(d) values (Colpas, G . J., Brayman, T . G., Ming, L.-J., and Hausinger, R . P . (1999) Biochemistry 38, 4078-4088) . To understand why cells containing the latter two proteins showed reduced rates of urease activation, we characterized their metal binding/dissociation kinetics and compared the results to those obtained for H144*UreE . The truncated protein was shown to sequentially bind two Ni(2+) with k(1) approximately 18 and k(2) approximately 100 M(-1) s(-1), and with dissociation rates k(-1) approximately 3 x 10(-3) and k(-2) approximately 10(-4) s(-1) . Similar apparent rates of binding and dissociation were noted for the two mutant proteins, suggesting that altered H144*UreE interactions with Ni(2+) do not account for the changes in cellular urease activation . These conclusions are further supported by in vitro experiments demonstrating that addition of H144*UreE to urease apoprotein activation mixtures inhibited the rate and extent of urease formation . Our results highlight the importance of other urease accessory proteins in assisting UreE-dependent urease maturation.

APMIS, 2000 Mar, 108(3), 237 - 40
Comparison of screening methods for detection of extended-spectrum beta-lactamases and their prevalence among Escherichia coli and Klebsiella species in Hong Kong; Ho PL et al.; Three tests, the disk diffusion test, the double-disc synergy test and the inhibitor-potentiated disc diffusion test, were compared for their abilities to detect production of extended-spectrum beta-lactamases (ESBL) in 702 Escherichia coli and 472 Klebsiella spp . strains from four hospitals . Eleven percent E . coli and 13% Klebsiella spp . were found to produce ESBL . As an indicator of ESBL activity, the sensitivities of the five extended-spectrum beta-lactams were as follows: cefotaxime (100%), cefpodoxime (99.3%), ceftriaxone (98.6%), aztreonam (93%) and ceftazidime (57.7%) when interpreted using the National Committee for Clinical Laboratory Standards criteria . Their positive predictive values ranged from 67.8-83.8% . Both the inhibitor-potentiated disc diffusion test and the double-disc synergy test (at three inter-disc widths of 20, 25 and 30 mm) were capable of identifying all the ESBL-producers . However, at a single inter-disc width of 30 mm, the double-disc synergy test has limited sensitivity (83.8%) . As a second test for confirming ESBL activity in strains with reduced susceptibility to beta-lactams, the inhibitor-potentiated disc diffusion test is therefore a simple and reliable option.

Diagn Microbiol Infect Dis, 2000 Dec, 38(4), 255 - 7
Cefpodoxime screening of Escherichia coli and Klebsiella spp . by Vitek for detection of organisms producing extended-spectrum beta-lactamases; Gibb AP et al.; We used a Vitek custom card to detect cefpodoxime MIC of--2 mg/L as a screen for extended-spectrum beta-lactamase (ESBL) production in E . coli and Klebsiella spp . Of 2873 organisms tested, 60 were screen positive, but only 3 were confirmed to be ESBL producers . Cefpodoxime is believed to be a sensitive screen for ESBL production, but a more specific test is desirable.

J Bacteriol, 2001 Jan, 183(2), 637 - 43
Urea utilization in the phototrophic bacterium Rhodobacter capsulatus is regulated by the transcriptional activator NtrC; Masepohl B et al.; The phototrophic nonsulfur purple bacterium Rhodobacter capsulatus can use urea as a sole source of nitrogen . Three transposon Tn5-induced mutations (Xan-9, Xan-10, and Xan-19), which led to a Ure(-) phenotype, were mapped to the ureF and ureC genes, whereas two other Tn5 insertions (Xan-20 and Xan-22) were located within the ntrC and ntrB genes, respectively . As in Klebsiella aerogenes and other bacteria, the genes encoding urease (ureABC) and the genes required for assembly of the nickel metallocenter (ureD and ureEFG) are clustered in R . capsulatus (ureDABC-orf136-ureEFG) . No homologues of Orf136 were found in the databases, and mutational analysis demonstrated that orf136 is not essential for urease activity or growth on urea . Analysis of a ureDA-lacZ fusion showed that maximum expression of the ure genes occurred under nitrogen-limiting conditions (e.g., serine or urea as the sole nitrogen source), but ure gene expression was not substrate (urea) inducible . Expression of the ure genes was strictly dependent on NtrC, whereas sigma(54) was not essential for urease activity . Expression of the ure genes was lower (by a factor of 3.5) in the presence of ammonium than under nitrogen-limiting conditions, but significant transcription was also observed in the presence of ammonium, approximately 10-fold higher than in an ntrC mutant background . Thus, ure gene expression in the presence of ammonium also requires NtrC . Footprint analyses demonstrated binding of NtrC to tandem binding sites upstream of the ureD promoter . Phosphorylation of NtrC increased DNA binding by at least eightfold . Although urea is effectively used as a nitrogen source in an NtrC-dependent manner, nitrogenase activity was not repressed by urea.

Biosci Biotechnol Biochem, 2000 Oct, 64(10), 2040 - 6
Regulation by organic acids of polysaccharide-mediated microbe-plant interactions; Nakata K et al.; A polysaccharide flocculant of Klebsiella pneumoniae H12 has been suggested to mediate microbe-plant interactions with the aid of Ca2+ {K . Nakata et al., Biosci . Biotechnol . Biochem., 64, 459-465, 2000} . Here, two-way regulation of polysaccharide-mediated interactions between K . pneumoniae and Raphanus sativus was studied using organic acids . Namely, 10 mM equivalents of organic acids promoted production of the polysaccharide by the bacterium, but inhibited flocculation of bacterial cells by the polysaccharide . These phenomena were counterbalanced by equi-molar equivalents of Ca2+, suggesting competition for Ca2+ between the carboxylic residues of the polysaccharide and those of the aliphatic acids . By electron microscopy observations, bacterial cell aggregates were sparsely distributed over the main roots and root hairs, had various sizes, and seemed to tightly adhere to root tissues . Their shapes seemed to be distorted and abundant in cavities . In brief, these microscopical observations may be explained by a two-way regulation system of bacterial adhesion to a plant by organic acids.

Chemotherapy, 2001 Jan-Feb, 47(1), 29 - 38
A 13-year study of antimicrobial susceptibility of common gram-negative bacteria isolated from the bloodstream in a teaching hospital; Ling TK et al.; The choice of antimicrobial therapy for the treatment of bacteremia is often empirical and based on the knowledge of susceptibility profiles of the most common bacteria . We analyzed blood culture isolates from a teaching hospital for 13 years prospectively . This study examined the susceptibility profiles of 6,616 gram-negative bacteria . Escherichia coli ranked among the commonest bacteria, representing 43.6% (2,890) of all gram-negative isolates . Klebsiella sp . ranked second, 17.7% (1,171) and 16.7% were resistant to ceftazidime in 1997 . There was a trend towards an increase in resistance to fluoroquinolones in the common gram-negative bacteria . Imipenem was the most active agent against gram-negative bacteria . The results of the susceptibility of gram-negative bacteria causing bacteremia provide valuable information for implementing the appropriate chemotherapy for bacteremia .

Antimicrob Agents Chemother, 2001 Jan, 45(1), 309 - 11
SHV-14, a novel beta-lactamase variant in Klebsiella pneumoniae isolates from Nijmegen, The Netherlands; Yuan M et al.; Four ceftazidime-resistant isolates of a Klebsiella pneumoniae strain were collected from intensive care unit patients in Nijmegen, The Netherlands . These isolates had TEM-29 and SHV-14 beta-lactamases . SHV-14 is a novel variant, with two substitutions compared with the sequence of SHV-1: Ile8Phe and Arg43Ser . Its gene also had a silent C-->T mutation at nucleotide 481 . The SHV-14 enzyme had slightly higher V(max) rates than SHV-1 for oxyimino-aminothiazolyl cephalosporins, but this activity was insufficient for the enzyme to count as an extended-spectrum beta-lactamase.

Antimicrob Agents Chemother, 2001 Jan, 45(1), 88 - 95
Complexity and diversity of Klebsiella pneumoniae strains with extended-spectrum beta-lactamases isolated in 1994 and 1996 at a teaching hospital in Durban, South Africa; Essack SY et al.; beta-Lactamase production was investigated in cultures of 25 Klebsiella pneumoniae isolates isolated at a hospital in Durban, South Africa, in 1994 and 1996 . Twenty of these isolates gave ceftazidime MIC/ceftazidime plus clavulanate MIC ratios of >/=8, implying production of extended-spectrum beta-lactamases (ESBLs), and DNA sequencing identified an ESBL gene (bla(TEM-53)) in a further two isolates . Pulsed-field gel electrophoresis (PFGE) defined 4 distinct strains among the 12 isolates collected in 1994 and 9 distinct strains among the 13 isolates collected in 1996 . In three cases, multiple isolates from single patients varied in their PFGE profiles and antibiograms, implying mixed colonization or infection . Isoelectric focusing and DNA hybridization found both TEM and SHV enzymes and their genes in all 25 isolates . Many isolates had multiple identical or different beta-lactamase gene variants, with at least 84 bla(SHV) and bla(TEM) gene copies among the 25 organisms . Sequencing identified the genes for the SHV-1, -2, and -5 enzymes and for four new SHV types (SHV-19, -20, -21, and -22) . These new SHV variants had novel mutations remote from sites known to affect catalytic activity . Sequencing also found the genes for TEM-1, TEM-53, and one novel type, TEM-63 . All the isolates had multiple and diverse plasmids . These complex and diverse patterns of ESBL production and strain epidemiology are far removed from the concept of an ESBL outbreak and suggest a situation in which ESBL production has become endemic and in which evolution is generating a wide range of enzyme combinations . This complexity and diversity complicates patient management and the design of antibiotic use policies.

Infect Immun, 2001 Jan, 69(1), 24 - 33
Surfactant protein D enhances phagocytosis and killing of unencapsulated phase variants of Klebsiella pneumoniae; Ofek I et al.; Pulmonary surfactant protein D (SP-D) is a collagenous C-type lectin (collectin) that is secreted into the alveoli and distal airways of the lung . We have studied the interactions of SP-D and alveolar macrophages with Klebsiella pneumoniae, a common cause of nosocomial pneumonia . SP-D does not agglutinate encapsulated K . pneumoniae but selectively agglutinates spontaneous, unencapsulated phase variants, such as Klebsiella strain K50-3OF, through interactions with their lipopolysaccharides (LPS) . These effects are calcium dependent and inhibited with maltose but not lactose, consistent with involvement of the SP-D carbohydrate recognition domain . Precoating of K50-3OF with SP-D enhances the phagocytosis and killing of these organisms by rat alveolar macrophages in cell culture and stimulates the production of nitric oxide by the NR-8383 rat alveolar macrophage cell line . SP-D similarly enhances the NO response to K50-3OF LPS adsorbed to Latex beads under conditions where soluble LPS or SP-D, or soluble complexes of SP-D and LPS, do not stimulate NO production . Our studies demonstrate that interactions of SP-D with exposed arrays of Klebsiella LPS on a particulate surface can enhance the host defense activities of alveolar macrophages and suggest that activation of macrophages by SP-D requires binding to microorganisms or other particulate ligands . Because unencapsulated phase variants are likely to be responsible for the initial stages of tissue invasion and infection, we speculate that SP-D-mediated agglutination and/or opsonization of K . pneumoniae is an important defense mechanism for this respiratory pathogen in otherwise healthy individuals.

Eur J Clin Microbiol Infect Dis, 2000 Oct, 19(10), 765 - 72
Extended-spectrum TEM- and SHV-type beta-lactamase-producing Klebsiella pneumoniae strains causing outbreaks in intensive care units in Italy; Pagani L et al.; The aim of the present study was to investigate the production of extended-spectrum beta-lactamases (ESbetaLs) and the epidemiological correlations in a total of 107 Klebsiella pneumoniae strains resistant to third- and fourth-generation cephalosporins . The strains were collected from patients in four intensive care units (3 neonatal and 1 general) in three hospitals in Italy between March 1996 and July 1997 . All strains were found to produce ESbetaLs . Phenotypic (antibiotyping and ESbetaL patterns) and genotypic (plasmid profile and pulsed-field gel electrophoresis) analyses showed that a single strain had been responsible for each outbreak in each of the four intensive care units . Isoelectric focusing, activity on substrates and gene sequencing showed that the strains produced SHV-5, SHV-2a, SHV-12 and TEM-52 beta-lactamases . This is not only the first time that ESbetaL-producing Klebsiella pneumoniae strains have been reported as causing epidemics in Italian hospitals, it is also, to the best of our knowledge, the first time that an outbreak caused by a TEM-52 ESbetaL-producing Klebsiella pneumoniae strain has been reported . The data presented here illustrate the complexity of determining the epidemiological pattern of ESbetaL producers in large hospitals that do not have an ESbetaL-monitoring program.

Biochim Biophys Acta, 2000 Dec 15, 1524(2-3), 247 - 52
Synthesis of {(14)C}pyrroloquinoline quinone (PQQ) in E . coli using genes for PQQ synthesis from K . pneumoniae; Stites TE et al.; Radiochemical forms of pyrroloquinoline quinone (PQQ) are of utility in studies to determine the metabolic role and fate of PQQ in biological systems . Accordingly, we have synthesized {(14)C}PQQ using a tyrosine auxotrophic strain of Escherichia coli (AT2471) . A construct containing the six genes required for PQQ synthesis from Klebsiella pneumoniae was used to transform the auxotrophic strain of E . coli . E . coli were then grown in minimal M9 medium containing 3.7x10(9) Bq/mmol {(14)C}tyrosine . At confluence, the medium was collected and applied to a DEAE A-25 anionic exchange column; {(14)C}PQQ was eluted using a KCl gradient (0-2 M in 0.1 M potassium phosphate buffer, pH 7.0) . Radioactivity co-eluting as PQQ was next pooled, acidified and passed through a C-18 column; {(14)C}PQQ was eluted with a phosphate buffer (0.1 M, pH 7.0) . Reverse phase HPLC (C-18) using either the ion-pairing agent trifluoroacetic acid (0 . 1%) and an acetonitrile gradient or phosphoric acid and a methanol gradient were used to isolate {(14)C}PQQ . Fractions were collected and analyzed by liquid scintillation counting . (14)C-labelled compounds isolated from the medium eluted corresponding to the elution of various tyrosine-derived products or PQQ . The radioactive compound corresponding to PQQ was also reacted with acetone to form 5-acetonyl-PQQ, which co-eluted with a 5-acetonyl-PQQ standard, as a validation of {(14)C}PQQ synthesis . The specific activity of synthesized {(14)C}PQQ was 3.7x10(9) Bq/mmol {(14)C}PQQ, equal to that of {U-(14)C}tyrosine initially added to the medium.

Int Microbiol, 1998 Mar, 1(1), 53 - 7
Efficient phagocytosis of Klebsiella pneumoniae strains that poorly bind to human polymorphonuclear leukocytes; Pruzzo C et al.; The phagocytosis process of unencapsulated MIAT-negative strains that, although binding very poorly to human polymorphonuclear leukocytes (PMN) at 4 degrees C, are efficiently killed by these cells at 37 degrees C, was studied . At 37 degrees C the number of bacteria bound to the PMN external surface was similar to that observed at 4 degrees C (about 100 bacteria/100 PMN after 60 min); on the contrary the number of internalized bacteria was much higher (from 500 bacteria/100 PMN after 60 min) . Interactions between phagocytosis-sensitive Klebsiella pneumoniae strains (PSK) and PMN were then compared with those of two isogenic Escherichia coli strains with and without type 1 fimbriae . Whereas PSK strain binding to blocked PMN was very slow and became significant only after 5-6 h, that of phagocytosis-sensitive fimbriated E . coli was rapid and efficient . Phagocytosis-resistant, non fimbriated E . coli strain bound with an efficiency that, within the first 60 min, was not very different from that of the PSK strains . However, longer incubations led to increases in PSK binding, whereas unfimbriated E . coli remained constant . PSK and fimbriated E . coli strains were efficiently internalized and killed, whereas the unfimbriated E . coli strain was not . It is suggested that PMN can phagocytize unopsonized bacteria through two different mechanisms . By one mechanism, observed with the fimbriated E . coli strain, PMN bind many more bacteria than those they can internalize . By the other, observed with PSK strains, PMN bind only the bacteria they can immediately internalize.

Am J Respir Cell Mol Biol, 2000 Dec, 23(6), 727 - 33
Modulation of alveolar macrophage phagocytosis by leukotrienes is Fc receptor-mediated and protein kinase C-dependent; Mancuso P et al.; We have previously established an important role for leukotrienes (LTs) in augmenting rat alveolar macrophage (AM) phagocytosis of Klebsiella pneumoniae opsonized with complete immune serum (IS), which contains the two well-known opsonins, immunoglobulin (Ig) G and complement (C) . In this report, the specific opsonin requirements for LT modulation of AM phagocytosis and the dependence of this response on protein kinase (PK) C activity were investigated . Phagocytosis of K . pneumoniae opsonized with IS, non-immune serum, or heat-inactivated immune serum and of inert targets (IgG-opsonized fluorescent microspheres or C-opsonized sheep red blood cells) was examined . Inhibition of endogenous LT synthesis or action attenuated, whereas the addition of exogenous LTs augmented, phagocytosis only of targets opsonized with IgG . LTs had no effect on phagocytosis of C-opsonized or unopsonized targets . LTs did not affect adherence of IgG-opsonized targets, implying instead an enhancement of internalization . Macrophage internalization of phagocytic targets has previously been shown to require PKC activity . Pretreatment of AMs with the PKC inhibitors staurosporine or calphostin C, or with phorbol 12-myristate 13-acetate to deplete PKC, completely inhibited the ability of LTB(4) and largely inhibited the ability of LTC(4) to augment phagocytosis of IgG-opsonized microspheres . These results demonstrate that LT enhancement is confined to Fc receptor (FcR)-mediated phagocytosis . Moreover, PKC activation represents an important mechanism by which LTs promote FcR-mediated phagocytosis.

Braz J Infect Dis, 1997 Aug, 1(4), 196 - 203
Antimicrobial Susceptibility of Klebsiella pneumoniae Producing Extended-Spectrum beta-lactamase (ESBL) Isolated in Hospitals in Brazil; Gales AC et al.; The prevalence of klebsiella pneumoniae producing extended-spectrum beta-lactamase (ESBL) has been increasing all over the world . Infections caused by ESBL producing isolates are difficult to detect with current susceptibility tests, and are difficult to treat . ESBLs confer resistance to all currently available beta-lactam, except carbapenems . In addition, ESBL production is usually associated with resistance to other classes of antimicrobial agents such as aminoglycosides and quinolones . The objective of this study was to evaluate the in vitro susceptibility patterns of ESBL producing K pneumoniae isolated in Brazil . Seventy-two strains were tested using E test against 30 antimicrobial agents, including carbapenems, second and third generation cephalosporins, aminoglycosides, quinolones, and some new compounds . The most active compounds (i.e . 100% susceptibility) were meropenem (MIC90, 0.125microg/mL), imipenem (MIC90, 0.25microg/mL), and cefotetan (MIC90, 2microg/mL) . Ciprofloxacin (MIC90, 1microg/mL, 94% susceptibility) and cefepime (MIC90, 6microg/mL, 92% susceptibility), were also very active against our collection of ESBL producing K pneumoniae . None of the six aminoglycosides showed good activity against these strains (16% to 41% susceptibility) and only 39% of the isolates were susceptible to piperacillin/tazobactam . The results of our study indicated that the carbapenems are the most active compounds against ESBL producing L pneumoniae in Brazil, and ciprofloxacin remains very active against these strains . Cefotetan and cefepime were also very active against ESBL producing K.pneumoniaein Brazil; however, further studies are necessary to evaluate the role of these cephalosporins in the treatment of infections due to ESBL producing strains.

Braz J Infect Dis, 1997 Jun, 1(3), 138 - 141
Antimicrobial Susceptibility of Klebsiella pneumoniae Producing Extended-Spectrum beta-lactamase (ESBL) Isolated in Hospitals in Brazil; Tajiki MH et al.; Twelve patients with leptospirosis were enrolled in a study to evaluate whether a correlation exists between disease severity and plasma levels of TNF-alpha and IL-10 as determined by enzyme-linked immunoassay . Six patients had less severe disease by analysis of liver, kidney and lung involvement, and 6 patients had severe disease, 2 of whom died . The results of the study confirm an association between high levels of TNF-alpha and poor outcome in leptospirosis . Levels of IL-10 were in a similar range in the 10 surviving and the 2 non-surviving patients . The ratio of IL-10/TNF-a correlated with disease severity in that a high ratio was associated with less severe disease and survival . The possibility is raised that treatment approaches to change this cytokine profile may be more effective in this type of inflammatory condition than it has been in bacterial sepsis.

J Antimicrob Chemother, 2000 Dec, 46(6), 895 - 900
Emergence of imipenem resistance in Klebsiella pneumoniae owing to combination of plasmid-mediated CMY-4 and permeability alteration; Cao VT et al.; Klebsiella pneumoniae BM2974 isolated from an abdominal abcess was resistant to high concentrations of all available beta-lactams, including recently developed third-generation cephalosporins and carbapenems . Isoelectric focusing of beta-lactamases and amplification, cloning and sequencing of the corresponding genes, together with conjugation and transformation experiments, indicated that, in addition to the chromosomally encoded beta-lactamase, the strain produced three plasmid-mediated beta-lactamases with pIs of 5.4, 8.2 and 9.0, which corresponded to TEM-1, SHV-5 and AmpC-type CMY-4, respectively . Strain BM2974 also lacked a major outer membrane protein of c . 40 kDa which was present in the spontaneous imipenem-susceptible revertant BM2974-1 . We suggest that imipenem resistance in strain BM2974 is attributable to production of CMY-4 beta-lactamase combined with permeability alteration.

Clin Infect Dis, 2000 Dec, 31(6), 1368 - 72 Epub 2000 Nov 10.
Characterization of an outbreak due to extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a pediatric intensive care unit transplant population; Rebuck JA et al.; Limited information exists regarding Klebsiella pneumoniae's production of an extended-spectrum beta-lactamase (KP-ESBL) in pediatric patients, particularly solid-organ transplant recipients . This study characterized the microbiological, epidemiological, and clinical features of a KP-ESBL outbreak in children receiving a liver transplant, an intestinal transplant, or both . All children found to have microbiologically confirmed K . pneumoniae during a 21-month period were reviewed . ESBL production was defined by double-disk diffusion, and 6 distinct pulsed-field gel electrophoresis patterns were identified . Fifty-six percent of the transplant patients we studied developed KP-ESBL, representing 87% of all microbiologically confirmed cases at our institution . As compared with 16 control transplant patients who were negative for KP-ESBL, the 20 transplant patients who acquired KP-ESBL were younger (aged < or = 5 years; 80.0% vs . 43.8%, P = .038) and experienced placement of > or = 3 central venous catheters before recovery of the first K . pneumoniae isolate (73.7% vs . 18.8%, P = .002) . This study suggests that children who receive liver or intestinal transplants are at high risk for KP-ESBL acquisition.

Chang Gung Med J, 2000 Sep, 23(9), 566 - 71
Bilateral endogenous Klebsiella pneumoniae endophthalmitis associated with meningitis-useful vision regained after treatment: case report; Liao SB et al.; Endogenous endophthalmitis is a rare but devastating complication of bacteremia . Klebsiella pneumoniae is reported to be the leading organism of endogenous endophthalmitis in Taiwan, and the prognosis of endogenous Klebsiella pneumoniae endophthalmitis is extremely poor . A 46-year-old male patient was hospitalized because of fever, chills, and consciousness disturbance for 1 day . Meningitis was the impression by clinical presentation and findings of cerebrospinal fluid analysis . Parenteral ceftriaxone (4 g/day) and gentamicin (180 mg/day) were given, and his consciousness gradually cleared . On hospital day 6, he complained of blurred vision in both eyes and floaters in the left eye for 1 day . After ophthalmic examination, bilateral endogenous endophthalmitis was diagnosed . After aggressive treatment with intravitreal antibiotics and trans pars plana vitrectomy, the visual outcome of both eyes was better than those of other reported cases.

J Biol Chem, 2001 Mar 2, 276(9), 6582 - 90 Epub 2000 Nov 17.
Evidence for the selective population of FeMo cofactor sites in MoFe protein and its molecular recognition by the Fe protein in transition state complex analogues of nitrogenase; Grossmann JG et al.; We have collected synchrotron x-ray solution scattering data for the MoFe protein of Klebsiella pneumoniae nitrogenase and show that the molecular conformation of the protein that contains only one molybdenum per alpha(2)beta(2) tetramer is different from that of the protein that has full occupancy i.e . two molybdenums per molecule . This structural finding is consistent with the existence of MoFe protein molecules that contain only one FeMo cofactor site occupied and provides a rationale for the 50% loss of the specific activity of such preparations . A stable inactive transition state complex has been shown to form in the presence of MgADP and AlF(4)(-) . Gel filtration chromatography data show that the MoFe protein lacking a full complement of the cofactor forms initially a 1:1 complex before forming a low affinity 1:2 complex . A similar behavior is found for the MoFe protein with both cofactors occupied, but the high affinity 1:2 complex is formed at a lower ratio of Fe protein/MoFe protein . The 1:1 complex, MoFe protein-Fe protein x (ADP x AlF(4)(-))(2), formed with MoFe protein that lacks one of the cofactors, is stable . X-ray scattering studies of this complex have enabled us to obtain its low resolution structure at approximately 20-A resolution, which confirms the gel filtration finding that only one molecule of the Fe protein binds the MoFe protein . By comparison with the low resolution structure of purified MoFe protein that contains only one molybdenum per tetramer, we deduce that the Fe protein interacts with the FeMo cofactor-binding alpha-subunit of the MoFe protein . This observation demonstrates that the conformation of the alpha-subunit or the alpha beta subunit pair that lacks the FeMo cofactor is altered and that the change is recognized by the Fe protein . The structure of the 1:1 complex reveals a similar change in the conformation of the Fe protein as has been observed in the low resolution scattering mask and the high resolution crystallographic study of the 1:2 complex where both cofactors are occupied and with the Fe protein bound to both subunits . This extensive conformational change observed for the Fe protein in the complexes is, however, not observed when MgATP or MgADP binds to the isolated Fe protein . Thus, the large scale conformational change of the Fe protein is associated with the complex formation of the two proteins.

Infect Immun, 2000 Dec, 68(12), 6744 - 9
Capsule impedes adhesion to and invasion of epithelial cells by Klebsiella pneumoniae; Sahly H et al.; The adhesion of K21a, K26, K36, and K50 capsulated Klebsiella strains to ileocecal (HCT-8) and bladder (T24) epithelial cell lines was significantly lower than that of their corresponding spontaneous noncapsulated variants K21a/3, K26/1, K36/3, and K50/3, respectively . Internalization of the bacteria by both epithelial cell lines was also significantly reduced . Similarly, a capsule-switched derivative, K2(K36), that exhibited a morphologically larger K36 capsule and formed more capsular material invaded the ileocecal epithelial cell line poorly compared to the corresponding K2 parent strain . None of the capsulated strains exhibited significant mannose-sensitive type 1 fimbriae, whereas two of the noncapsulated variants K21a/3 and K50/3 exhibited potent mannose-sensitive hemagglutinating activity . Although hemagglutinating activity that could be attributed to mannose-resistant Klebsiella type 3 fimbriae was weak in all strains, in several cases the encapsulated parent strains exhibited lower titers than their corresponding noncapsulated variants . Although the level of adhesion to the ileocecal cells is not different from adhesion to bladder cells, bacterial internalization by bladder cells was significantly lower than internalization by ileocecal cells, suggesting that bladder cells lack components required for the internalization of Klebsiella.

Ginekol Pol, 2000 Sep, 71(9), 975 - 8
{Central nervous system infections in neonates caused by multiresistant Klebsiella pneumoniae}; Wojsyk-Banaszak I et al.; OBJECTIVE: To evaluate the incidence, clinical course and outcome of central nervous system (CNS) infections caused by multiresistant Klebsiella pneumoniae (MRKP) in critically ill neonates . METHODS: Retrospective study of neonates treated in Neonatal Department of Poznan University of Medical Science during a three years period from 1st January 1997 to 31st December 1999 . Isolates were identified as Klebsiella pneumoniae with the ID32Gn test (bioMerieux), and antibiotic susceptibility was determined with ATBG test (bioMerieux) and with disc-diffusion technique . RESULTS: In this period there were 27 cases of CNS infections in our Department 17--65.4% of which (16 meningitis, one ventriculitis and one brain abscess), were caused by Klebsiella pneumoniae . 81.25% occurred in prematures (< 31 weeks of gestational age and < 1500 grams) . In 10 (58.8%) cases CNSI developed in the course of sepsis . 9 children died . In all the isolates ESBL expression in vitro was detected . Only carbapenems and fluoroquinolones were active in all the cases . Ventriculitis was treated successfully with imipenem administered intraventrically and brain abscess surgically . CONCLUSIONS: CNS infections in neonates caused by MRKP are quite frequent, severe and possibly life threatening . Eradication of these strains from hospital environment by introducing adequate sanitary regime and reasonable antibiotic policies is the only method of conquering the infections.

Laryngoscope, 2000 Nov, 110(11), 1857 - 60
Intranasal surfactant aerosol therapy for otitis media with effusion; Venkatayan N et al.; OBJECTIVE: To determine the effect of surfactant alone and with other medications delivered intranasally as a metered dose inhaler (MDI) aerosol on the resolution of experimentally induced otitis media with effusion (OME) . BACKGROUND: Eustachian tube dysfunction is a primary factor in the pathogenesis of OME . Intranasal surfactant via MDI has been shown in this laboratory to reduce passive opening pressure of the eustachian tube in normal gerbils and mice . STUDY DESIGN: OME was developed in 35 gerbils by transtympanic injection of 10 microg lipopolysaccharide from Klebsiella pneumoniae . Pretreatment otomicroscopy and tympanometry were performed to exclude pre-existing middle ear disease, and postinfection evaluations were performed on alternate days for a period of 30 days . Five animals received no treatment (control group); four were treated with propellant only (placebo); seven received surfactant alone; eight received surfactant and betamethasone; and six received surfactant with phenylephrine . All medications were sprayed intranasally as an aerosolized MDI and administered daily from postinfection day 2 onward . RESULTS: OME resolved after 16.0 +/- 0.44 days (mean + SD) in controls . There was no difference seen in the placebo or the surfactant with phenylephrine groups . Treatment with surfactant yielded resolution in 10.57 +/- 0.37 days; this was reduced to 8.57 +/- 0.37 days with surfactant plus betamethasone . These differences are statistically significant . There was no recurrence of OME in any group . CONCLUSION: This study demonstrates that using an aerosolized MDI surfactant with and without betamethasone decreases the duration of OME in this in vivo gerbil model.

Zhonghua Yi Xue Za Zhi (Taipei), 2000 Oct, 63(10), 721 - 8
Klebsiella pneumoniae renal abscess
Chang SW, Yen DH, Fung CP, Liu CY, Chen KK, Tiu CM, Wang LM, Lee CH.
BACKGROUND: The clinical characteristics of renal abscess caused by Klebsiella pneumoniae have not been previously reported in case-series studies . The purpose of this study is to demonstrate the clinical characteristics of K pneumoniae renal abscess for early diagnosis and to identify the clinical risk factors associating with poor prognosis . METHODS: We retrospectively reviewed the medical records of 24 patients with K pneumoniae renal abscess, from April, 1982 through February, 1998 . The clinical presentations, including the demographic characteristics, predisposing disorders, initial signs and symptoms, laboratory test results, diagnostic radiology studies, therapeutic modalities and risk factors associated with mortality were studied . RESULTS: The mean age was 58.7 years and the male to female ratio was 10:14 . The most common predisposing factors were diabetes mellitus (58%), urolithiasis (25%) and immunosuppression (17%) . Fever, chills and flank pain were the most common symptoms and signs, whereas pyuria, elevation of leukocyte count, glucose, blood urea nitrogen and creatinine were the common laboratory features . The distinct complications of K pneumoniae renal abscess were bacteremia in 13 (54%), emphysematous pyelonephritis in five (21%), and metastatic septic infection in three (12.5%) . The cure rate was 52% (11/21) in patients treated with a combination of antibiotics and percutaneous drainage; however, six (35%) patients who survived required another surgical procedure for complete recovery . The overall mortality rate was 25% . The clinical factors of elderly age (> 65 years) at presentation, lethargy, elevation of serum blood urea nitrogen and pulmonary complications were associated with poor prognoses . CONCLUSIONS: Focusing on the early diagnosis of K pneumoniae renal abscess and recognition of the prognostic factors for a poor prognosis, we highlight the specific clinical characteristics that include elderly age, lethargy, impairment of renal function, metastatic septic lesions and pulmonary complications . All patients with K pneumoniae renal abscesses should receive empiric antibiotics and percutaneous drainage or aspiration, and surgical intervention as necessary for patients with intractable disease.

J Med Microbiol, 2000 Nov, 49(11), 1003 - 10
Role of bacterial capsule in local and systemic inflammatory responses of mice during pulmonary infection with Klebsiella pneumoniae; Yoshida K et al.; The role of bacterial capsule in inflammatory responses in experimentally induced pneumonia caused by Klebsiella pneumoniae was evaluated by comparing the host immunological responses in mice infected with capsulate strain DT-S and non-capsulate mutant strain DT-X . Anaesthetised ICR mice were infected intranasally with inocula of strain DT-S or DT-X . Mice infected with strain DT-X survived significantly longer than those inoculated with strain DT-S . Viable bacterial counts in lungs and blood increased rapidly in mice infected with strain DT-S, in contrast to the gradual decrease in their density in lungs and intermittent bacteraemia in mice infected with strain DT-X . The number of broncho-alveolar lavage (BAL) cells in mice infected with strain DT-X at 24 h after inoculation was significantly higher than in those infected with strain DT-S . In the early stages of infection, the levels of tumour necrosis factor-a and interleukin-6 in BAL fluid of mice infected with strain DT-X were significantly higher than those of mice infected with strain DT-S . In contrast, in the late stage of infection, the levels of these cytokines in serum of mice infected with strain DT-S were significantly higher than in mice infected with strain DT-X . These results suggest that K . pneumoniae capsule may suppress the host immunological responses,thus allowing the bacteria to grow, causing pneumonia, septicaemia and death.

Nucleic Acids Res, 2000 Nov 15, 28(22), 4419 - 27
Single amino acid substitution mutants of Klebsiella pneumoniae sigma(54) defective in transcription; Pitt M et al.; Transcription initiation by the sigma(54) RNA polymerase requires specialised activators and their associated nucleoside triphosphate hydrolysis . To explore the roles of sigma(54) in initiation we used random mutagenesis of rpoN and an in vivo activity screen to isolate functionally altered sigma(54) proteins . Five defective mutants, each with a different single amino acid substitution, were obtained . Three failed in transcription after forming a closed complex . One such mutant mapped to regulatory Region I of sigma(54), the other two to Region III . The Region I mutant allowed transcription independently of activator and showed reduced activator-dependent sigma(54) isomerisation . The two Region III mutants displayed altered behaviour in a sigma(54) isomerisation assay and one failed to stably bind early melted DNA as the holoenzyme; they may contribute to a communication pathway linking changes in sigma to open complex formation . Two further Region III mutants showed gross defects in overall DNA binding . For one, sufficient residual DNA binding activity remained to allow us to demonstrate that other activities were largely unaffected . Changes in DNA binding preferences and core polymerase-dependent properties were evident amongst the mutants.

Lancet . 2000 Jul 22;356(9226):310.
Disinfectant contaminated with Klebsiella oxytoca as a source of sepsis in babies; Reiss I et al.; We report an outbreak of sepsis from contaminated disinfectant in a neonatal and paediatric intensive-care unit . 28 infants were infected with Klebsiella oxytoca and basic measures to control the outbreak failed . The resistance of K . oxytoca against the disinfectant was probably mediated by capsule formation, visible as mucoid colonies.

J Clin Microbiol, 2000 Nov, 38(11), 4228 - 32
Detection of extended-spectrum beta-lactamases in klebsiellae with the Oxoid combination disk method; Carter MW et al.; The Oxoid combination disk method for detecting extended-spectrum beta-lactamases (ESBLs) depends on comparing the inhibition zones of cefpodoxime (10-microg) and cefpodoxime-plus-clavulanate (10- plus 1-microg) disks . The presence of clavulanate enlarged the zones for all of 180 ESBL-producing klebsiellae by >/=5 mm, whereas zones for cefpodoxime-susceptible isolates and cefpodoxime-resistant isolates with AmpC and K1 beta-lactamases were enlarged by </=1 mm . Good discrimination was achieved with either the NCCLS or the British disk method.

Biochemistry, 2000 Oct 31, 39(43), 13233 - 40
Identification of the active site of phosphoribosyl-dephospho-coenzyme A transferase and relationship of the enzyme to an ancient class of nucleotidyltransferases; Hoenke S et al.; Malonate decarboxylase from Klebsiella pneumoniae contains an acyl carrier protein (MdcC) to which a 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group is attached via phosphodiester linkage to serine 25 . We have shown in the preceding paper in this issue that the formation of this phosphodiester bond is catalyzed by a phosphoribosyl-dephospho-coenzyme A transferase MdcG with the substrate 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA that is synthesized from ATP and dephospho-coenzyme A by the triphosphoribosyl transferase MdcB . The reaction catalyzed by MdcG is related to nucleotidyltransfer reactions, and the enzyme indeed catalyzes unphysiological nucleotidyltransfer, e.g., adenylyltransfer from ATP to apo acyl carrier protein (ACP) . These unspecific side reactions are favored at high Mg(2+) concentrations . A sequence motif including D134 and D136 of MdcG is a signature of all nucleotidyltransferases . It is known from the well-characterized mammalian DNA polymerase beta that this motif is at the active site of the enzyme . Site-directed mutagenesis of D134 and/or D136 of MdcG to alanine abolished the transfer of the prosthetic group to apo ACP, but the binding of triphosphoribosyl-dephospho-CoA to MdcG was not affected . Evidence is presented that similar to MdcG, MadK encoded by the malonate decarboxylase operon of Malonomonas rubra and CitX from the operon encoding citrate lyase in Escherichia coli are phosphoribosyl-dephospho-CoA transferases catalyzing the attachment of the phosphoribosyl-dephospho-CoA prosthetic group to their specific apo ACPs.

Biochemistry, 2000 Oct 31, 39(43), 13223 - 32
Biosynthesis of triphosphoribosyl-dephospho-coenzyme A, the precursor of the prosthetic group of malonate decarboxylase; Hoenke S et al.; Malonate decarboxylase from Klebsiella pneumoniae consists of four subunits MdcA, D, E, and C and catalyzes the cleavage of malonate to acetate and CO(2) . The smallest subunit MdcC is an acyl carrier protein to which acetyl and malonyl thioester residues are bound via a 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group and turn over during the catalytic mechanism . We report here on the biosynthesis of holo acyl carrier protein from the unmodified apoprotein . The prosthetic group biosynthesis starts with the MdcB-catalyzed condensation of dephospho-CoA with ATP to 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA . In this reaction, a new alpha (1' ' --> 2') glycosidic bond between the two ribosyl moieties is formed, and thereby, the adenine moiety of ATP is displaced . MdcB therefore is an ATP:dephospho-CoA 5'-triphosphoribosyl transferase . The second protein involved in holo ACP synthesis is MdcG . This enzyme forms a strong complex with the 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA prosthetic group precursor . This complex, called MdcG(i), is readily separated from free MdcG by native polyacrylamide gel electrophoresis . Upon incubation of MdcG(i) with apo acyl carrier protein, holo acyl carrier protein is synthesized by forming the phosphodiester bond between the 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group and serine 25 of the protein . MdcG corresponds to a 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA:apo ACP 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA transferase . In absence of the prosthetic group precursor, MdcG catalyzes at a low rate the adenylylation of apo acyl carrier protein using ATP as substrate . The adenylyl ACP thus formed is an unphysiological side product and is not involved in the biosynthesis of holo ACP . The 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA precursor of the prosthetic group has been purified and its identity confirmed by mass spectrometry and enzymatic analysis.

Br J Biomed Sci, 2000, 57(3), 185 - 91
Clinical and community strains of Klebsiella pneumoniae: multiple and increasing rates of antibiotic resistance in Abha, Saudi Arabia; Bilal NE et al.; Klebsiella pneumoniae is the most commonly isolated bacterial species in a maternity hospital in Saudi Arabia . Here, 380 strains isolated in 1997 and 480 strains in 1999 were studied for their resistance antibiograms, using the standardised disc diffusion test . Of 16 antibiotics tested, four in 1997 and six in 1999 were ineffective against > 50% of the respective isolates, and resistance rates to 11 antibiotics increased over the two-year period (P = 0.05-< 0.0001) . With resistance rates of < 20%, imipenem, ciprofloxacin, gentamicin and cefotaxime were more effective in 1997; only imipenem and ciprofloxacin remained as effective in 1999 . In addition, 105 community strains were tested and > 50% were resistant to four antibiotics . Resistance rates to most antibiotics were lower than those of clinical strains (P = 0.0285-< 0.0001) . Imipenem resistance was detected among both clinical and community isolates . Multiresistance was 64.5% in 1997 and 79.2% in 1999 (P < 0.0001), and 83.8% in community strains in 1999 . Using the double-disc synergy test, extended-spectrum beta-lactamase (ESBL) was detected in 27.5% of ceftazidime-resistant clinical strains isolated in 1999 . Among the clinical strains, seven (65%) and 11 (67.9%) resistance antibiograms occurred frequently in 1997 and 1999, respectively . Such frequency was not observed among community isolates . These findings confirm the alarmingly high rates of multiresistance and the emergence of ESBL-producing strains, highlighting the urgent need to restrict over-the-counter availability of antibiotics, and increase awareness in the local medical community.

J Hosp Infect, 2000 Oct, 46(2), 110 - 7
Molecular investigation of two clusters of hospital-acquired bacteraemia caused by multi-resistant Klebsiella pneumoniae using pulsed-field gel electrophoresis and in frequent restriction site PCR.Infection Control Group; Su LH et al.; Two molecular typing methods, DNA macrorestriction analysis with XbaI resolved by pulsed-field gel electrophoresis (PFGE) and infrequent restriction site PCR (IRS-PCR) assay with adapters designed for XbaI and HhaI restriction sites, were used to investigate two clusters of hospital-acquired bacteraemia associated with multi-resistant Klebsiella pneumoniae which occurred in a paediatric intensive care unit (PICU) . A total of 56 K . pneumoniae isolates were analysed . These included 10 bacteraemic isolates from eight patients, 26 isolates obtained during an epidemiological survey, and 20 epidemiologically non-related isolates incorporated as controls . One major pattern was demonstrated in 22 of the 56 isolates analysed . These included nine of the 10 bacteraemic isolates, a single rectal isolate, two hand culture isolates and 10 sink isolates . All of these 22 isolates illustrated identical antibiograms, whilst the other 34 isolates shared six antibiograms and 31 unique patterns by either PFGE or IRS-PCR assay . The two clusters of bacteraemia appeared to be outbreaks induced by the same strain of K . pneumoniae which may have utilized sinks as reservoirs and been transmitted through the hands of medical personnel to patients . IRS-PCR demonstrates concordant results with PFGE analysis in studying the genetic relationships among K . pneumoniae isolates, and serves as an excellent epidemiological tool for this bacterium .

Pediatr Res, 2000 Nov, 48(5), 685 - 90
Structural and functional features of modified heat-stable toxins produced by enteropathogenic Klebsiella cells; Albano F et al.; Heat-stable enterotoxins (STs) are 18- or 19-amino acid peptides (STa or ST1) produced by enteropathogenic bacteria with small differences in their amino acid sequence and a highly conserved carboxy terminus . All STs contain a core of three disulfide bridges whose integrity is believed to be necessary for full biologic activity . We previously reported that strains of Klebsiella pneumoniae transformed by the plasmid pSLM004 produce a modified toxin not recognized by MAb raised against genuine Escherichia coli ST . Investigation of the chemical structure of the modified toxins revealed that three new toxins were present . These were purified to homogeneity by a series of sequential chromatography on reverse-phase columns using guanylate cyclase to monitor the enterotoxic activity during purification procedures . The sequence of the modified toxins was obtained by a combination of Edman degradation and mass spectrometry, showing that they are proteolytically processed forms of E . coli ST1b . In particular, toxin A-2 lacks the cysteine at position 18 and then is not able to form the disulfide bridge cysteine-10-cysteine-18 . All three toxins showed the ability to stimulate guanylate cyclase and to elicit chloride secretion in Caco-2 cell monolayers mounted in Ussing chambers . Toxin A-1 and toxin B demonstrated greatly reduced immunoreactivity whereas toxin A-2 was not recognized at all in the ELISA . It is likely that the three modified toxins were generated by Klebsiella specific proteolytic processing of the original pretoxin . These results have important implications for the diagnosis and prevention of heat-stable toxin-induced diarrhea.

Epidemiol Mikrobiol Imunol, 1999 Feb, 48(1), 28 - 30
{Effect of ofloxacin, pefloxacin, and tobramycin on the hydrophobicity of Klebsiella pneumoniae}; Hostacka A; The effect of ofloxacin, pefloxacin and tobramycin at subinhibitory concentrations (1/4, 1/8 and 1/16 of the MICs) on surface hydrophobicity of the Klebsiella pneumoniae strain was studied . The antibiotics tested decreased cell surface hydrophobicity in a dose-dependent manner . The most significant reduction of surface hydrophobicity was found after treatment with antibiotics at 1/4 of their MICs . Surface hydrophobicity of K . pneumoniae after exposure to these concentrations was decreased to 44% (ofloxacin), 50.7% (pefloxacin) and 56.1% (tobramycin) compared with controls.

J Theor Biol, 2000 Nov 7, 207(1), 1 - 14
Mathematical treatment of transport data of bacterial transport systems to estimate limitation in diffusion through the outer membrane; Tralau C et al.; Bacterial transport systems are traditionally treated as enzymes exhibiting a saturable binding site giving rise to an apparent K(m)of transport, whereas the maximal rate of transport is regarded equivalent to the V(max)of enzymatic reactions . Thus, the Michaelis-Menten theory is usually applied in the analysis of transport data and K(m)and V(max)are derived from the treatment of data obtained from the rate of transport at varying substrate concentrations . Such an analysis tacitly assumes that the substrate recognition site of the transport system is freely accessible to substrate . However, this is not always the case . In systems endowed with high affinity in the micro M range or those recognizing large substrates or those exhibiting high V(max), the diffusion through the outer membrane may become rate determining, particularly at low external substrate concentrations . In such a situation the dependence of the overall rate of transport (from the medium into the cytoplasm) on the substrate concentration in the medium will no longer follow Michaelis-Menten kinetics . By analysing the deviation of transport data from the corresponding ideal Michaelis-Menten plot we developed a method that allows us to determine diffusion limitation through the outer membrane . The method allows us to find the correct K(m)of the transport system functioning at the inner membrane even under conditions of strong diffusion limitation through the outer membrane . The model was tested and validified with the Escherichia coli binding protein-dependent ABC transporter for maltose . The corresponding systems for sn -glycerol-3-phospate of Escherichia coli and the alpha -cyclodextrin transport of Klebsiella oxitoca were used as test systems .

Nihon Kokyuki Gakkai Zasshi, 2000 Jul, 38(7), 557 - 60
{A case of Klebsiella pneumoniae infection causing a buccal abscess complicated with multiple lung abscesses}; Kosugi E et al.; A 51 year-old man fitted with a dental prosthesis was hospitalized with buccal swelling, fever and chest pain . Laboratory data showed marked inflammatory changes, and chest radiography and CT scanning revealed small nodular shadows within the lung . A diagnosis of multiple lung abscesses secondary to a buccal abscess possibly caused by the prosthesis was made from needle aspiration biopsies of the lung nodules and of a buccal lesion . Klebsiella pneumoniae was isolated from these lesions and from a blood culture . The patient was successfully treated with antibiotics and by surgical drainage of the buccal abscess . It is important to note that the patient was immunodeficient at the time as a result of diabetes and alcohol intoxication.

Biochim Biophys Acta, 2000 Sep 29, 1468(1-2), 253 - 61
Localization of sterically stabilized liposomes in experimental rat Klebsiella pneumoniae pneumonia: dependence on circulation kinetics and presence of poly(ethylene)glycol coating; Schiffelers RM et al.; Preferential localization of liposomes at sites of infection or inflammation has been demonstrated in a variety of experimental models . Most studies report enhanced localization at the target site of poly(ethyelene) glycol (PEG)-coated liposomes as compared to conventional non-coated liposomes . It is generally accepted that the prolonged circulation time of PEG-coated liposomes increases target site exposure, which results in increased target localization . A quantitative relationship between circulation kinetics and localization at the pathological site has not been defined as yet . Besides, an effect of the PEG coating itself has been suggested, as theoretically the PEG coating may facilitate liposome extravasation . In the present study, in a rat model of an acute unilateral Klebsiella pneumoniae pneumonia, circulation kinetics of PEG-coated liposomes were manipulated by incorporation of different amounts of phosphatidylserine (PS) and variation of lipid dose, additionally allowing evaluation of the saturability of the localization process . In addition, this paper addresses the effect of the PEG coating, by comparing the circulation kinetics and target localization of long-circulating 'PEG-free' and PEG-coated liposomes . It is shown that the degree of liposome localization at the target site is positively linearly related to the area under the blood concentration time curve (AUC) of the liposome formulations, irrespective of PEG coating . This finding is discussed in relation to the equation of Kedem and Katchalsky, which describes protein influx into sites of infection or inflammation.

Biochemistry, 2000 Oct 10, 39(40), 12435 - 40
UreE stimulation of GTP-dependent urease activation in the UreD-UreF-UreG-urease apoprotein complex; Soriano A et al.; The activation of metal-containing enzymes often requires the participation of accessory proteins whose roles are poorly understood . In the case of Klebsiella aerogenes urease, a nickel-containing enzyme, metallocenter assembly requires UreD, UreF, and UreG acting as a protein chaperone complex and UreE serving as a nickel metallochaperone . Urease apoprotein within the UreD-UreF-UreG-urease apoprotein complex is activated to wild-type enzyme activity levels under physiologically relevant conditions (100 microM bicarbonate and 20 microM Ni2+) in a process that requires GTP and UreE . The GTP concentration needed for optimal activation is greatly reduced in the presence of UreE compared to that required in its absence . The amount of UreE provided is critical, with maximal activation observed at a concentration equal to that of Ni2+ . On the basis of its ability to facilitate urease activation in the presence of chelators, UreE is proposed to play an active role in transferring Ni2+ to urease apoprotein . Studies involving site-directed variants of UreE provide evidence that His96 has a direct role in metal transfer . The results presented here parallel those obtained from previous in vivo studies, demonstrating the relevance of this in vitro system to the cellular metallocenter assembly process.

Curr Microbiol, 2000 Nov, 41(5), 357 - 62
Internal glutamine and glutamate pools in Klebsiella pneumoniae grown under different conditions of nitrogen availability; Schmitz RA; Internal pool sizes of glutamine and glutamate in Klebsiella pneumoniae grown under nitrogen limitation or nitrogen sufficiency were measured to study the signal transduction of external nitrogen limitation . K . pneumoniae cells were grown in an anaerobic, ammonium-limited chemostat culture . At a growth rate of 0.217 h(-1), the steady state ammonium concentration in the culture was 55 microm, correlating with repression of the nitrogen fixation (nif) genes . At growth rates below 0.138 h(-1), the ammonium concentration in the culture dropped below 0.5 microm and the nif genes became derepressed . During the transition from nitrogen sufficiency to nitrogen limitation, the internal glutamine pool in K . pneumoniae decreased by a factor of approximately 6 . The glutamate pool, however, remained stable . Similarly, in anaerobic batch cultures with different limiting nitrogen sources, the glutamine pool generally decreased by a factor of 7 to 9 when nif gene derepression was achieved . All the limiting nitrogen sources used resulted in decreased growth rates compared with growth under nitrogen excess, suggesting an inverse relationship between glutamine pool size and doubling time . These studies indicate that K . pneumoniae perceives external nitrogen limitation as internal glutamine limitation.

Electrophoresis, 2000 Aug, 21(14), 2932 - 9
Using electrophoresis to observe the interaction of nitrogenase with ions; Zou L et al.; The two protein components of nitrogenase from Klebsiella pneumoniae were shown to interact with metal ions and ADP, altering their electrophoretic mobility in polyacrylamide gel electrophoresis . Both Mg+2 and Mn+2 caused reduced mobility of Fe protein relative to other proteins . The effect was about 50% complete at concentrations around 0.2 mM . Other ions including Fe+2, Ni+2 and Co+2 had no observable effect at levels up to 1 _mM . Both Cd+2 and Zn+2 appeared to interact with the protein; Cd+2 at 0.5 mM dramatically destabilized the protein . The effects of more than a dozen different mutations of the Fe protein on Mg+2 interaction were examined . All mutated proteins appeared to interact with Mg+2 similarly to wild-type . Using relative mobility differences of charge-changed mutants it was estimated that two to three Mg+2 interact with each Fe protein monomer . The MoFe protein also showed interaction with metal ions but the alteration of mobility was much smaller than for the Fe protein because it is larger and less acidic, so that it runs much more slowly than the Fe protein in standard gels . The interaction of ADP with Fe protein was examined in the presence of Mg+2 . Increasing ADP partially reversed the mobility decrease observed on Mg+2 binding, and produced a more diffuse protein band indicative of a reaction zone of interconverting conformers . No alteration of MoFe protein mobility was observed with ADP added during electrophoresis.

J Assoc Physicians India, 1999 Mar, 47(3), 338 - 9
Splenic abscess as a complication of acute pancreatitis; Duggal RK et al.; Though there is close anatomical proximity between pancreas and the spleen, the involvement of latter is extremely uncommon in acute inflammation of the pancreas . In this report, we present a case of splenic abscess as a complication of acute pancreatitis . The aspirate from the abscess yielded the organism, Klebsiella . We were able to successfully treat this case with percutaneous drainage along with parenteral antibiotics, and we did not have a resort to splenectomy.

Eur J Clin Invest, 2000 Sep, 30(9), 818 - 22
Apolipoprotein E-deficient mice have an impaired immune response to Klebsiella pneumoniae; de Bont N et al.; BACKGROUND: All lipoproteins are able to bind to bacterial lipopolysaccharide (LPS), thereby neutralizing its deleterious effects . However, we demonstrated, recently, that in the absence of apolipoprotein E (apoE), eight-fold increased very-low-density lipoprotein levels were not sufficient to protect apoE-deficient (apoE-/-) mice against LPS . During a live Gram-negative infection, mechanisms other than LPS-neutralization may play a role in the pathogenesis of the disease . In the present study we further examined the role of apoE in Gram-negative sepsis . METHODS: Survival, bacterial outgrowth in liver, spleen, kidneys and blood, and tumour necrosis factor-alpha (TNF-alpha) production were measured in apoE-/- mice and control C57BL/6J mice, after an intravenous infection with Klebsiella pneumoniae . RESULTS: Mice that lack apoE showed higher mortality in response to K . pneumoniae than control mice (90% vs . 23% respectively after 2 weeks) . ApoE-/- mice had 10-100 times more outgrowth of the bacteria in their organs than controls . Furthermore, circulating TNF-alpha concentrations 90 min after a challenge, were almost twice as high in the apoE-/- mice compared to controls (13.0 +/- 2.9 ng mL-1 vs . 7.6 +/- 3.8 ng mL-1) . When apoE-/- and control mice were rendered neutropenic, the discrepancy in survival and outgrowth of K . pneumoniae disappeared . CONCLUSIONS: The apoE-/- mice were more susceptible than control C57BL/6 mice to a K . pneumoniae infection . The absence of apoE may render these mice more susceptible, since this protein is of importance in the detoxification of lipopolysaccharide of Gram-negative bacteria . On the other hand, the phagocytic capacity of granulocytes seems to be decreased in apoE-/- mice, resulting in increased outgrowth and mortality.

Immunopharmacology, 2000 Sep, 49(3), 325 - 33
Prevention of collagen-induced arthritis in DBA/1 mice by oral administration of AZ-9, a bacterial polysaccharide from Klebsiella oxytoca; Sugihara R et al.; Collagen-induced arthritis (CIA) is an excellent model of rheumatoid arthritis (RA) in humans that is induced in DBA/1 mice immunized with bovine type II collagen (CII) . Here, we report that the induction of CIA was effectively suppressed by oral administration of AZ-9, a purified polysaccharide with the average molecular weight of approximately 200 kDa that was produced by a soil bacterium, Klebsiella oxytoca . When AZ-9 was administered at 125-250 mg/kg/day orally for 9 consecutive days after immunization with CII followed by its administration every 3 days, resulted in a marked reduction of the incidence and the severity of CIA . The serum level of anti-CII IgG2a and the production of IFN-gamma and IL-12 in the draining lymph node (LN) cells were significantly lower in AZ-9-administered mice than the untreated control . These findings suggest that orally administered AZ-9 suppressed CIA through attenuating a Th1-type response to CII . AZ-9 could be fragmented into smaller molecules (3-4 kDa) without losing its suppressive activity.

Arch Microbiol, 2000 Jul-Aug, 174(1-2), 81 - 8
Specificities of reactivating factors for adenosylcobalamin-dependent diol dehydratase and glycerol dehydratase; Tobimatsu T et al.; Adenosylcobalamin-dependent glycerol and diol dehydratases undergo inactivation by the physiological substrate glycerol during catalysis . In the permeabilized cells of Klebsiella pneumoniae, Klebsiella oxytoca, and recombinant Escherichia coli, glycerol-inactivated glycerol dehydratase and diol dehydratase are reactivated by their respective reactivating factors in the presence of ATP, Mg2+, and adenosylcobalamin . Both of the reactivating factors consist of two subunits . To examine the specificities of the reactivating factors, their genes or their hybrid genes were co-expressed with dehydratase genes in E . coli cells in various combinations . The reactivating factor of K . oxytoca for diol dehydratase efficiently cross-reactivated the inactivated glycerol dehydratase, whereas the reactivating factor of K . pneumoniae for glycerol dehydratase hardly cross-reactivated the inactivated diol dehydratase . Both of the two hybrid reactivating factors rapidly reactivated the inactivated glycerol dehydratase . In contrast, the hybrid reactivating factor containing the large subunit of the glycerol dehydratase reactivating factor hardly reactivated the inactivated diol dehydratase . These results indicate that the glycerol dehydratase reactivating factor is much more specific for the dehydratase partner than the diol dehydratase reactivating factor and that a large subunit of the reactivating factors principally determines the specificity for a dehydratase.

Arch Microbiol, 2000 Jul-Aug, 174(1-2), 67 - 73
The Na+-dependent citrate carrier of Klebsiella pneumoniae: high-level expression and site-directed mutagenesis of asparagine-185 and glutamate-194; Kastner CN et al.; The Na+-dependent citrate carrier of Klebsiella pneumoniae (CitS) is a member of the 2-hydroxycarboxylate transporter family . Within the highly conserved helix Vb region, Asn-185 of CitS was mutated to Val and Glu-194 was mutated to Gln . The wild-type and mutant proteins were synthesised in Escherichia coli DH5alpha or C43(DE3) as biotinylated or His-tagged CitS-fusions, respectively . The synthesis and purification procedure yielded 6.5 mg pure CitS per litre culture . The fusion proteins were characterised with E . coli cell suspensions or after reconstitution of the purified enzymes into proteoliposomes . The E194Q mutation had almost no effect on the kinetics of Na+ or citrate transport . In contrast, aberrant citrate transport kinetics were found for the N185V mutant . The apparent K(m) value for the citrate species H-citrate(2-) was increased about nine-fold, whereas the apparent Vmax value and the effect of Na+ on the transport kinetics were comparable to the wild-type . Asn-185 of CitS appears therefore to participate in the binding of H-citrate(2-).

Diagn Microbiol Infect Dis, 2000 Aug, 37(4), 279 - 84
Klebsiella pneumoniae isolates causing liver abscess in Taiwan; Chang SC et al.; Klebsiella pneumoniae has been the leading cause of pyogenic liver abscess in Taiwan during the period from 1985 to 1999, which is different from other countries . The present study investigated the in vitro antimicrobial susceptibilities of 51 K . pneumoniae isolates collected from blood cultures of patients with liver abscess in Taiwan during the period from 1993-1997, and typed by pulsed-field gel electrophoresis (PFGE) . All 51 isolates were resistant to ampicillin, but susceptible to other antimicrobial agents . The minimum inhibitory concentrations (MICs) were less than 1 microg/ml for the third- and fourth-generation cephalosporins, monobactam, carbapenems, and ciprofloxacin . In comparison, 62 isolates of K . pneumoniae from community-acquired bacteremic patients without liver abscess had similar antimicrobial susceptibilities, while 142 isolates from patients with hospital-acquired bacteremia without liver abscess were much less susceptible to all of the tested antimicrobial agents . PFGE molecular epidemiologic analysis found 20 out of 51 isolates belonged to eight clusters of genetically related strains, with two or three isolates in each clusters . The other 31 isolates were genetically distinct strains . This study demonstrated that K . pneumoniae isolates which cause liver abscess in Taiwan remained susceptible to a wide range of antimicrobial agents and that they were not genetically related.

Chemotherapy, 2000 Sep-Oct, 46(5), 371 - 8
Safety and efficacy of cefpirome in comparison with ceftazidime in Chinese patients with sepsis due to bacterial infections; Fang CT et al.; BACKGROUND: The safety and efficacy of cefpirome (CPM), a fourth-generation cephalosporin, has not yet been studied in an Asian population . METHODS: Sixty-nine adult Chinese patients with sepsis due to bacterial infections were enrolled in the study and randomly assigned to receive CPM (2 g i.v . every 12 h) or ceftazidime (2 g i.v . every 8 h) . RESULTS: Both groups were comparable in terms of demographic, clinical and microbiological features . The causative pathogen was Escherichia coli or Klebsiella pneumoniae in the majority of microbiologically documented cases . The rates of clinical response (94 vs . 94%) and bacteriological response (65 vs . 68%) were similar in both groups . Similar adverse events, including rashes, gastrointestinal upset and asymptomatic elevation of hepatic enzymes, occurred in a minority of patients in both groups . All events were self-limited after discontinuation of the drugs . CONCLUSION: CPM is as safe and effective as ceftazidime in the treatment of sepsis due to bacterial infections in Chinese patients .

Pediatr Neurol, 2000 Jul, 23(1), 69 - 72
Non-Hodgkin's lymphoma of the uterus and CNS; Marjanovic B et al.; We report a case of a non-Hodgkin's lymphoma of the uterus and central nervous system in an 8-year-old female . The neurologic signs included blurred vision, neck stiffness, and walking difficulties but no abdominal problems . She deteriorated further, and repeated lumbar punctures revealed the presence of malignant cells in the cerebrospinal fluid . A repeated ultrasound scan of the abdomen demonstrated a markedly enlarged uterus . Biopsy revealed B-cell non-Hodgkin's lymphoma . Treatment according to the Berlin-Frankfurt-Munster protocol was initiated, but she developed hyperventilation syndrome and required mechanical ventilation . Her condition improved after 1 week but then deteriorated again, and despite additional chemotherapy she developed myelosuppression and septicemia with multiresistant Klebsiella pneumoniae and eventually died 13 months after her first admission to the hospital . No clinical or laboratory signs of relapse were evident at the time of death.

Cas Lek Cesk, 2000 May 10, 139(9), 277 - 9
{The chemiluminescence test in the laboratory evaluation of effects of immunostimulatory therapy}; Macurova H et al.; BACKGROUND: Number of patients treated by general practitioners with various immunomodulatory drugs has recently increased . Effects of such medication on the immune system were not usually monitored . The aim of our work was to evaluate effect of selected immunomodulatory drugs on the phagocytic and metabolic activities of the phagocytes . METHODS AND RESULTS: 51 patients (18 males and 33 females) of the average age 36 years with repeating respiratory, mycotic and herpetic infections were investigated . Immunomodulatory treatment included: Decaris (Lavamizolum), Isoprinosine (Methisoprinolum), Imudon (Lysatum bacteriale mixtum), Biostim (Klebsiella pneumoniae), and Immodin (Leukocyti dialysati lyophylysatum) . Before and after treatment all patients underwent basic immunological examination IgG, IgA, IgM, C3, C4 complement components, PEG, CD3, CD4, CD8 and CD19) . Phagocytotic activity was estimated by means of FAGO MSHP test with HEMA particles and by chemiluminiscence test . Chemiluminiscence was measured using ML 3000 Microtiter Plate Luminometer (Dynex), 26 healthy individuals of the corresponding age were the controls . Results were statistically evaluated by Student's t-test . Significant increase of the cellular metabolic activity was found in Decaris and Immodin treated patients (P < 0.001) . CONCLUSIONS: Chemiluminiscence test, which evaluates the metabolic activity of phagocytes, can be used for the accurate laboratory monitoring of the effects of some immunomodulatory drugs on the natural immunity of patients.

J Biol Chem, 2000 Dec 8, 275(49), 38452 - 6
Two residues in the T-loop of GlnK determine NifL-dependent nitrogen control of nif gene expression; Arcondeguy T et al.; X-ray crystallographic analysis of the Escherichia coli P(II) protein paralogues GlnB and GlnK has shown that they share a superimposable structural core but can differ in conformation of the T-loop, a region of the protein (residues 37-54) that has been shown to be important for interaction with other proteins . In Klebsiella pneumoniae GlnK has been shown to have a clearly defined function in regulating NifL-mediated inhibition of NifA activity in response to the nitrogen status, and GlnB, when expressed from the chromosome, does not substitute for GlnK . Because the T-loops of K . pneumoniae and E . coli GlnB and GlnK differ at just three residues, 43, 52, and 54, we have used a previously constructed heterologous system, in which K . pneumoniae nifLA is expressed in E . coli, to investigate the importance of GlnK residues 43, 52, and 54 for regulation of the NifLA interaction . By site-directed mutagenesis of glnB we have shown that residue 54 is the single most important amino acid in the T-loop in the context of the regulation of NifA activity . Furthermore, a combination of just two changes, in residues 54 and 43, allows GlnB to function as GlnK and completely relieve NifL inhibition of NifA activity.

Antimicrob Agents Chemother, 2000 Sep, 44(9), 2382 - 8
Characterization of the extended-spectrum beta-lactamase reference strain, Klebsiella pneumoniae K6 (ATCC 700603), which produces the novel enzyme SHV-18; Rasheed JK et al.; Klebsiella pneumoniae K6 (ATCC 700603), a clinical isolate, is resistant to ceftazidime and other oxyimino-beta-lactams . A consistent reduction in the MICs of oxyimino-beta-lactams by at least 3 twofold dilutions in the presence of clavulanic acid confirmed the utility of K . pneumoniae K6 as a quality control strain for extended-spectrum beta-lactamase (ESBL) detection . Isoelectric-focusing analysis of crude lysates of K6 demonstrated a single beta-lactamase with a pI of 7.8 and a substrate profile showing preferential hydrolysis of cefotaxime compared to ceftazidime . PCR analysis of total bacterial DNA from K6 identified the presence of a bla(SHV) gene . K6 contained two large plasmids with molecular sizes of approximately 160 and 80 kb . Hybridization of plasmid DNA with a bla(SHV)-specific probe indicated that a bla(SHV) gene was encoded on the 80-kb plasmid, which was shown to transfer resistance to ceftazidime in conjugal mating experiments with Escherichia coli HB101 . DNA sequencing of this bla(SHV)-related gene revealed that it differs from bla(SHV-1) at nine nucleotides, five of which resulted in amino acid substitutions: Ile to Phe at position 8, Arg to Ser at position 43, Gly to Ala at position 238, and Glu to Lys at position 240 . In addition to the production of this novel ESBL, designated SHV-18, analysis of the outer membrane proteins of K6 revealed the loss of the OmpK35 and OmpK37 porins.

J Immunol, 2000 Sep 1, 165(5), 2643 - 50
Gamma delta-T cells are critical for survival and early proinflammatory cytokine gene expression during murine Klebsiella pneumonia; Moore TA et al.; Although cells of the innate inflammatory response, such as macrophages and neutrophils, have been extensively studied in the arena of Gram-negative bacterial pneumonia, a role for T cells remains unknown . To study the role of specific T cell populations in bacterial pneumonia, mice deleted of their TCR beta- and/or delta-chain were intratracheally inoculated with Klebsiella pneumoniae . Gamma delta T cell knockout mice displayed increased mortality at both early and late time points . In contrast, mice specifically lacking only alpha beta-T cells were no more susceptible than wild-type mice . Pulmonary bacterial clearance in gamma delta-T cell knockout mice was unimpaired . Interestingly, these mice displayed increased peripheral blood dissemination . Rapid up-regulation of IFN-gamma and TNF-alpha gene expression, critical during bacterial infections, was markedly impaired in lung and liver tissue from gamma delta-T cell-deficient mice 24 h postinfection . The increased peripheral blood bacterial dissemination correlated with impaired hepatic bacterial clearance following pulmonary infection and increased hepatic injury as measured by plasma aspartate aminotransferase activity . Combined, these data suggest that mice lacking gamma delta-T cells have an impaired ability to resolve disseminated bacterial infections subsequent to the initial pulmonary infection . These data indicate that gamma delta-T cells comprise a critical component of the acute inflammatory response toward extracellular Gram-negative bacterial infections and are vital for the early production of the proinflammatory cytokines IFN-gamma and TNF-alpha.

J Biol Chem, 2000 Nov 3, 275(44), 34287 - 92
Phg1p is a nine-transmembrane protein superfamily member involved in dictyostelium adhesion and phagocytosis; Cornillon S et al.; To identify the molecular mechanisms involved in phagocytosis, we generated random insertion mutants of Dictyostelium discoideum and selected two mutants defective for phagocytosis . Both represented insertions in the same gene, named PHG1 . This gene encodes a polytopic membrane protein with an N-terminal lumenal domain and nine potential transmembrane segments . Homologous genes can be identified in many species; however, their function is yet to be elucidated . Disruption of PHG1 caused a selective defect in phagocytosis of latex beads and Escherichia coli, but not Klebsiella aerogenes bacteria . This defect in phagocytosis was caused by a decrease in the adhesion of mutant cells to phagocytosed particles . These results indicate that the Phg1 protein is involved in the adhesion of Dictyostelium to various substrates, a crucial event of phagocytosis and demonstrate the usefulness of a genetic approach to dissect the molecular events involved in the phagocytic process.

J Drug Target, 1999, 6(6), 415 - 26
Bioavailability of a small unilamellar low-clearance liposomal amikacin formulation after extravascular administration; Fielding RM et al.; Amikacin in small, low-clearance liposomes (MiKasome) has prolonged plasma and tissue residence and in vivo activity against extracellular infections, including Klebsiella pneumonia and Pseudomonas endocarditis . Small liposomes may cross endothelial barriers, and enter the systemic circulation after extravascular administration . We compared the systemic bioavailability (F) of low-clearance liposomal amikacin in rats following intravenous (i.v.), intraperitoneal (i.p.), intramuscular (i.m.) and subcutaneous (s.c.) injection (20 mg/kg) and intratracheal (i.t.) instillation (10 mg/kg) . Drug-containing liposomes were extensively absorbed after i.p . (F = 87-146%) and i.t . (F = 64%) administration, with maximum amikacin plasma concentrations of 171 micrograms/ml at 9 h and 80 micrograms/ml at 18 h, respectively . Absorption was slower and less extensive following s.c . (plasma Tmax: 20.3 micrograms/ml at 48 h) and i.m . (plasma Tmax: 49.6 micrograms/ml at 19 h) injection, but a significant fraction (12-27%) of the liposomes was absorbed . The plasma AUCs of liposomal amikacin exceeded the AUC of conventional i.v . amikacin by at least 25-fold for all routes . Amikacin AUCs in regional lymph nodes exceeded plasma AUCs by 4-fold after s.c . and i.m . injection of liposomal amikacin . AUCs in tissues surrounding the injection sites were 20- and 191-fold higher than plasma AUCs after i.m . and s.c . injection, respectively . Thus, small low-clearance liposomes produced sustained levels of liposome-encapsulated amikacin in plasma, local tissues and lymph nodes after extravascular administration, suggesting applications in perioperative prophylaxis, pneumonias and intralesional therapy as well as sustained systemic delivery of encapsulated drugs.

J Antimicrob Chemother, 2000 Aug, 46(2), 273 - 7
Relationship between outer membrane alterations and susceptibility to antimicrobial agents in isogenic strains of Klebsiella pneumoniae; Hernandez-Alles S et al.; The activities of beta-lactams, chloramphenicol, tetracycline, fluoroquinolones and aminoglycosides against Klebsiella pneumoniae C3 (O1:K66, producing porins OmpK35 and OmpK36) and a set of isogenic mutants derived from it lacking the O antigen of lipopolysaccharide (LPS), capsular K antigen, or one or both porins were determined . MICs remained within one dilution step in mutants deficient in antigen O, in capsule or in one of the two porins . No increases in the MICs of aminoglycosides, fluoroquinolones, tetracycline and chloramphenicol were observed for strains deficient in the two porins, but the MICs of ampicillin, cephalothin, cefoxitin, cefotaxime and ceftazidime for this type of mutant increased four- to >256-fold . The highest MICs of beta-lactams were obtained in a porin-deficient mutant expressing increased beta-lactamase activity . It is concluded that isolated outer membrane alterations in K . pneumoniae are not decisive factors in increasing resistance to antimicrobial agents, but porin loss co-operates with beta-actamase production to increase resistance to beta-lactams.

J Antimicrob Chemother, 2000 Aug, 46(2), 241 - 7
Influence of a new fluoroquinolone, AF3013 (the active metabolite of prulifloxacin), on macrophage functions against Klebsiella pneumoniae: an in vitro comparison with pefloxacin; Tullio V et al.; The efficacy of an antibiotic in the treatment of bacterial infections depends upon the interaction of bacterium, drug and phagocytes . In this study we have investigated the influence of AF3013, a new fluoroquinolone, on the activities of mouse peritoneal macrophages against Klebsiella pneumoniae, in comparison with the influence of pefloxacin . Bacterial susceptibility to phagocytosis and intracellular killing were determined after klebsiellae and macrophages had been incubated simultaneously with inhibitory concentrations of both AF3013 and pefloxacin and following pre-exposure of the microorganisms and the macrophages individually to the same concentrations of each drug . Under the experimental conditions used, both AF3013 and pefloxacin potentiated the phagocytic and microbicidal activities of the macrophages, although different mechanisms may be involved.

J Antimicrob Chemother, 2000 Aug, 46(2), 215 - 21
Cross-resistance patterns among clinical isolates of Klebsiella pneumoniae with decreased susceptibility to cefuroxime; Schumacher H et al.; The frequency of decreased susceptibility to cefuroxime and quinolones and the correlation between these drug resistance traits was investigated in clinical isolates of Klebsiella pneumoniae from two Danish counties . Eighty-three randomly selected clinical isolates of K . pneumoniae with decreased susceptibility to cefuroxime were examined for cross-resistance patterns and the production of beta-lactamases . The frequency of resistance to cefuroxime and ciprofloxacin has increased from <5% in 1990 to 15% and 7% in 1998, respectively . Two of the 83 isolates were multiply resistant and seemed to produce extended-spectrum beta-lactamases . However, cross-resistance to ciprofloxacin and other classes of drug in 68% of the remaining isolates indicates that other resistance mechanisms, such as penetration barriers, had probably been selected in these Danish isolates . The susceptibility to ciprofloxacin decreased successively with decreasing susceptibility to cefuroxime for K . pneumoniae . This did not occur in cefuroxime-resistant Escherichia coli.

Rev Latinoam Microbiol, 1998 Jul-Dec, 40(3-4), 136 - 40
Functional Bradyrhizobium japonicum NifA expression under a hybrid nptII-nifH promoter in E . coli and Acetobacter diazotrophicus SRT4; Menendez C et al.; A hybrid promoter consisting of the in tandem fusion of the Tn5 nptII and the Klebsiella pneumoniae nifH promoters was constructed to study the functionality of the nif genes transcriptional activator NifA from Bradyrhizobium japonicum in two different host bacteria . beta-galactosidase experiments in E . coli revealed that the hybrid nptII-nifH promoter can behave as a constitutive or a NifA-inducible promoter depending on the aeration conditions . Expression of the B . japonicum NifA from the hybrid nptII-nifH promoter (plasmid pBPF204) induced "in trans" lacZ transcription from the Azotobacter chroococcum nifH promoter in E . coli and A . diazotrophicus cells grown at low pO2 . Similarly, the plasmid pBPF204 increased nitrogenase activity in A . diazotrophicus cells grown under microaerobic conditions . Based on these results, we suggest that the B . japonicum NifA could function as an efficient O2-sensitive transcriptional activator of nif genes in genetically distant diazotrophic bacteria.

Am J Perinatol, 2000, 17(1), 35 - 9
Septicemia due to multiresistant Klebsiella pneumoniae in a neonatal unit: a case-control study; Roilides E et al.; An unusually high incidence of septicemia due to multiresistant Klebsiella pneumoniae occurred in the Aristotle University Neonatology Department . Forty neonates suffered from 42 episodes of septicemia . Mortality was 43% ranging from 32% in neonates with birth weight (BW) - 1,500 g to 55% with < 1,500 g . No differences were found between 17 neonates who died and 23 survivors . All isolates were resistant to aminoglycosides, third-generation cephalosporins, and aztreonam, but susceptible to imipenem and ciprofloxacin . The neonates with septicemia due to K . pneumoniae were matched 1:1 with neonates without septicemia (31 pairs) or with neonates with septicemia due to other organisms (8 pairs) according to BW and time of admission . Factors associated with septicemia were mechanical ventilation (p = 0.004) and ongoing parenteral nutrition (p = 0.027) . In a multivariate model, nutrition exhibited no independent association after adjusting for ventilation . No differences were detected between the patients with Klebsiella septicemia and those with septicemia due to other organisms . Enhanced Infection Control measures and a temporary change of antibiotic policy reduced this serious complication . Three small outbreaks of multiresistant K . pneumoniae previously reported in neonates are reviewed.

Am J Ophthalmol, 2000 Jun, 129(6), 778 - 85
Bacterial subretinal abscess: a case report and review of the literature; Harris EW et al.; PURPOSE: To report a case of Klebsiella subretinal abscess with a successful visual outcome with treatment and to review the literature pertaining to focal intraocular infection in bacterial endophthalmitis . METHODS: Clinical data including medical history, findings on physical examination, blood cultures, and an abdominal computed tomographic scan were collected in a 32-year-old man with Klebsiella sepsis, liver abscesses, and a focal subretinal abscess . Ocular data including visual acuity, fundus photographs, fluorescein angiography, and ultrasound were evaluated, as were results of culture and histopathologic studies . RESULTS: Despite immediate intervention, including vitreous tap and intravitreal antibiotics, the eye deteriorated, with enlargement of the abscess . A pars plana vitrectomy was performed in which the subretinal abscess material was removed after an extensive retinectomy of the involved area in association with an endophotocoagulative barrier and intravitreal amikacin without gas or oil tamponade . Culture confirmed Klebsiella subretinal infection . A retinal detachment occurred 1 month postoperatively and was successfully repaired . Visual acuity was 20/30 and has remained stable for 14 months . CONCLUSION: Klebsiella endophthalmitis with subretinal abscess formation is a rare but devastating ocular condition . In the present case, prompt intervention with extensive retinectomy, complete abscess excision, and intravitreal antibiotic therapy resulted in unprecedented visual recovery.

Infect Control Hosp Epidemiol, 2000 Jul, 21(7), 455 - 8
Association of antibiotic utilization measures and control of multiple-drug resistance in Klebsiella pneumoniae; Patterson JE et al.; OBJECTIVE: To study the association of antibiotic-utilization measures and control of multidrug-resistant (MDR) Klebsiella pneumoniae after emergence in two hospitals in our medical center . DESIGN AND SETTING: Rates of MDR K . pneumoniae at two hospitals were compared before and after acute interventions, including emphasis on Contact Precautions and education in antibiotic utilization . Antipseudomonal beta-lactam antibiotic use was measured before and after the interventions at both hospitals . Pulsed-field gel electrophoresis of whole cell DNA was used as a marker of strain identity . RESULTS: Clonal strain dissemination was the major mechanism of emergence at hospital A; emergence was polyclonal at hospital B . Antibiotic-utilization interventions at both institutions included physician education regarding the association of ceftazidime use and MDR K . pneumoniae . At hospital A, ceftazidime use decreased from 4,301 g in the preintervention period, to 1,248 g in the postintervention period . Piperacillin-tazobactam use increased from 12,455 g to 17,464 g . Ceftazidime resistance in K . pneumoniae decreased from 110 (22%) of 503 isolates to 61 (15%) of 407 isolates (P<.05); piperacillin-tazobactam resistance decreased from 181 (36%) of 503 to 77 (19%) of 407 isolates (P<.05) . At hospital B, ceftazidime use decreased from 6,533 g in the preintervention period to 4,792 g in the postintervention period . Piperacillin-tazobactam use increased from 58,691 g to 67,027 g . Ceftazidime resistance in K . pneumoniae decreased from 42 (10%) of 415 isolates to 19 (5%) of 383 isolates (P<.05) . Piperacillin-tazobactam resistance decreased from 91 (22%) of 415 isolates to 54 (14%) of 383 isolates (P<.05) . Follow-up data showed continued decrease in piperacillin-tazobactam resistance despite increased use at both hospitals . CONCLUSIONS: Antibiotic-use measures may be particularly important for control of MDR K . pneumoniae, whether emergence is clonal or polyclonal.

Biochemistry, 2000 Aug 8, 39(31), 9438 - 50
Biosynthesis of the prosthetic group of citrate lyase; Schneider K et al.; Citrate lyase (EC 4.1.3.6) catalyzes the cleavage of citrate to acetate and oxaloacetate and is composed of three subunits (alpha, beta, and gamma) . The gamma-subunit serves as an acyl carrier protein (ACP) and contains the prosthetic group 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA, which is attached via a phosphodiester linkage to serine-14 in the enzyme from Klebsiella pneumoniae . In this work, we demonstrate by genetic and biochemical studies with citrate lyase of Escherichia coli and K . pneumoniae that the conversion of apo-ACP into holo-ACP is dependent on the two proteins, CitX (20 kDa) and CitG (33 kDa) . In the absence of CitX, only apo-ACP was synthesized in vivo, whereas in the absence of CitG, an adenylylated ACP was produced, with the AMP residue attached to serine-14 . The adenylyltransferase activity of CitX could be verified in vitro with purified CitX and apo-ACP plus ATP as substrates . Besides ATP, CTP, GTP, and UTP also served as nucleotidyl donors in vitro, showing that CitX functions as a nucleotidyltransferase . The conversion of apo-ACP into holo-ACP was achieved in vitro by incubation of apo-ACP with CitX, CitG, ATP, and dephospho-CoA . ATP could not be substituted with GTP, CTP, UTP, ADP, or AMP . In the absence of CitG or dephospho-CoA, AMP-ACP was formed . Remarkably, it was not possible to further convert AMP-ACP to holo-ACP by subsequent incubation with CitG and dephospho-CoA . This demonstrates that AMP-ACP is not an intermediate during the conversion of apo- into holo-ACP, but results from a side activity of CitX that becomes effective in the absence of its natural substrate . Our results indicate that holo-ACP formation proceeds as follows . First, a prosthetic group precursor {presumably 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA} is formed from ATP and dephospho-CoA in a reaction catalyzed by CitG . Second, holo-ACP is formed from apo-ACP and the prosthetic group precursor in a reaction catalyzed by CitX.

Hepatogastroenterology, 2000 May-Jun, 47(33), 626 - 30
Conversion factors in laparoscopic cholecystectomy for acute cholecystitis; Teixeira JP et al.; BACKGROUND/AIMS: Although laparoscopic cholecystectomy already occupies an important role in acute cholecystitis, a high rate of conversion continues to be referred to in the series published . One of the objectives of this study is to assess the preoperative factors that might lead to conversion . METHODOLOGY: 100 laparoscopic cholecystectomies for acute cholecystitis were studied of which 24 were converted . Several parameters are taken into consideration: age, sex, ASA, echographic data, bacteriological examination of the bile, time elapsing between diagnosis and surgery, morbidity and mortality . RESULTS: The analysis made in relation to the preoperative, predetermining factors with regards to the converted cases provided the following elements in unvaried analysis: Preoperative existence of: "plastron" gallbladder (P < 0.002), temperature above 38 degrees C (P < 0.04), leucocytosis (P < 0.02)), time elapsing between diagnosis and surgery (P < 0.005), presence in the echography of pericholecystic liquid (P < 0.0005) and edema (P < 0.001); Klebsiella in the gallbladder bile (P < 0.005) . Age (P = 0.136), sex (P = 0.992), associated diseases (P = 0.961), and gallbladder pathology (P = 0.282) did not present statistical differences that would prevent valid conclusions . In multi-varied analysis it was only possible to consider as independent factors the leucocytosis and the time between diagnosis and surgery . CONCLUSIONS: Laparoscopic cholecystectomy is a safe and efficient technique in the treatment of acute cholecystitis, which should constitute the first choice of treatment for this disease . However, it should be carried out within the first 4 days following surgical diagnosis.

Pediatr Infect Dis J, 2000 Jul, 19(7), 625 - 30
Mycobacterium tuberculosis infection in pediatric liver transplant recipients; Verma A et al.; OBJECTIVES: To study the incidence, clinical presentation, management, complications and outcome of tuberculosis in pediatric liver transplant recipients . METHODS: A retrospective review of the medical records of children who underwent liver transplantation between 1991 and 1998 . RESULTS: Mycobacterium tuberculosis infection occurred in 6 of 254 (2.4%) children undergoing liver transplantation between 1991 and 1998 . Cough, pyrexia and poor appetite were common presentations; one-half had normal chest radiographs . The median time to confirmation of diagnosis was 8 months (range, 1 to 17 months) . Tests contributing to diagnosis included: Ziehl-Neelsen (ZN) stain (2 patients), M . tuberculosis polymerase chain reaction (1 patient), Mantoux test (1 patient) and histopathology (4 patients) . Family health screening was productive for 4 patients . Duration of treatment varied from 9 to 18 months . Isoniazid-induced hepatitis was observed in 2 patients but resolved with dose reduction . Two patients died while receiving treatment, one of Klebsiella spp . septicemia and the other of pulmonary hemorrhage . CONCLUSIONS: Tuberculosis after liver transplantation has a significant morbidity and mortality . Pretransplantation a personal and family history of tuberculosis must be sought, and screening of patients and their families should be considered . Standard regimens incorporating isoniazid and rifampin are effective, but regular monitoring of liver function is essential to detect drug-induced hepatotoxicity.

Lijec Vjesn, 2000 Jan-Feb, 122(1-2), 8 - 13
{Effect of broad spectrum beta-lactamases on in vitro sensitivity of clinical isolates of Klebsiella pneumoniae to oral cephalosporins}; Bedenic B et al.; Extended-spectrum beta-lactamases which confer resistance to oxymino cephalosporins, are derived from common TEM-1, TEM-2 and SHV-1 . beta-lactamases by mutations which alter the configuration of active site and expand their spectrum of activity . The aim of this investigations was to determine the effect of these enzymes on in vitro susceptibility of clinical isolates of Klebsiella pneumoniae to oral cephalosporins . beta-lactamases were characterised by isoelectric focusing and polymerase chain reaction . The susceptibility of K . pneumoniae isolates harboring these enzymes to oral cephalosporins was determined by broth-microdilution method and the stability of these antibiotics to enzyme degradation was tested by macroiodometric method . Based on the isoelectric point beta-lactamases were assigned into three groups: SHV-2 type (pI 7.6), SHV-5 type (pI 8.2) and TEM-1 derivates (pI 5.4) . All types of beta-lactamases described in this investigation were associated with high level of resistance to older cephalosporins like cephalexin, cefaclor, cefuroxime, cefotiam, cefprozil and cefadroxil and to amoxycillin too . Third generation oral cephalosporins like ceftibuten and cefetamet were more stable in the presence of beta-lactamases but some of the strains were resistant to these antibiotics too . SHV-5 beta-lactamases producers displayed higher level of resistance to most of oral cephalosporins compared to SHV-2 and TEM producing strains . According to the results of this investigation oral cephalosporins cannot be considered as a therapeutic option for the treatment of infections caused by ESBL producing K . pneumoniae isolates.

Biochem Biophys Res Commun, 2000 Aug 2, 274(2), 496 - 9
Membrane permeability modifications are involved in antibiotic resistance in Klebsiella pneumoniae; Chevalier J et al.; Two Klebsiella pneumoniae strains selected according to their high cross-resistance pattern to cephalosporins were characterized by (i) outer membrane protein content such as OmpA or nonspecific porins, (ii) MICs of various cephalosporins and unrelated antibiotics, (iii) beta-lactamase production, and (iv) active efflux of fluoroquinolones . An association of porin deficiency and beta-lactamase production induced a noticeable cephalosporin resistance . In addition to these mechanisms, the presence of an active efflux participating in high-level fluoroquinolone resistance was identified in one strain . The decrease of antibiotic uptake associated with efflux explains the Klebsiella adaptation against the drugs present in the environment .

Biochemistry, 2000 Jul 25, 39(29), 8575 - 84
Kinetic and structural characterization of urease active site variants; Pearson MA et al.; Klebsiella aerogenes urease uses a dinuclear nickel active site to catalyze urea hydrolysis at >10(14)-fold the spontaneous rate . To better define the enzyme mechanism, we examined the kinetics and structures for a suite of site-directed variants involving four residues at the active site: His320, His219, Asp221, and Arg336 . Compared to wild-type urease, the H320A, H320N, and H320Q variants exhibit similar approximately 10(-)(5)-fold deficiencies in rates, modest K(m) changes, and disorders in the peptide flap covering their active sites . The pH profiles for these mutant enzymes are anomalous with optima near 6 and shoulders that extend to pH 9 . H219A urease exhibits 10(3)-fold increased K(m) over that of native enzyme, whereas the increase is less marked ( approximately 10(2)-fold) in the H219N and H219Q variants that retain hydrogen bonding capability . Structures for these variants show clearly resolved active site water molecules covered by well-ordered peptide flaps . Whereas the D221N variant is only moderately affected compared to wild-type enzyme, D221A urease possesses low activity ( approximately 10(-)(3) that of native enzyme), a small increase in K(m), and a pH 5 optimum . The crystal structure for D221A urease is reminiscent of the His320 variants . The R336Q enzyme has a approximately 10(-)(4)-fold decreased catalytic rate with near-normal pH dependence and an unaffected K(m) . Phenylglyoxal inactivates the R336Q variant at over half the rate observed for native enzyme, demonstrating that modification of non-active-site arginines can eliminate activity, perhaps by affecting the peptide flap . Our data favor a mechanism in which His219 helps to polarize the substrate carbonyl group, a metal-bound terminal hydroxide or bridging oxo-dianion attacks urea to form a tetrahedral intermediate, and protonation occurs via the general acid His320 with Asp221 and Arg336 orienting and influencing the acidity of this residue . Furthermore, we conclude that the simple bell-shaped pH dependence of k(cat) and k(cat)/K(m) for the native enzyme masks a more complex underlying pH dependence involving at least four pK(a)s.

Chang Gung Med J, 2000 Apr, 23(4), 240 - 5
Endogenous Klebsiella pneumoniae endophthalmitis associated with prostate abscess: case report; Chuang LH et al.; A young alcoholic and heavy smoker was admitted because of Klebsiella pneumoniae bacteremia . The abdominal and pelvic computed tomography with enhancement revealed an abscess in the right prostate . He experienced blurred vision of his left eye on the third day during admission . Initial ocular examination revealed mild reaction in the anterior chamber and vitreous as well as multiple Roth's spots in the retina . The administration of empirical antibiotics including cefazolin and gentamicin intravenously was changed to ceftriaxone and amikacin after ophthalmologic consultation . After parenteral antibiotics for 3 weeks, the systemic condition was controlled, and the vision remained stable . During the past decades, many cases of endogenous Klebsiella pneumoniae endophthalmitis associated with liver abscess were reported in Taiwan . However, documented reports of this condition associated with a prostate abscess are rare . This case, classified as posterior focal endogenous endophthalmitis, had good prognosis under appropriate antibiotic treatment . Early diagnosis and prompt therapy are important for control of sepsis and restoration of vision.

Afr J Med Med Sci, 1997 Mar-Jun, 26(1-2), 51 - 3
Outbreak of neonatal Klebsiella septicaemia: a review of antimicrobial sensitivities; Akindele JA et al.; A 10-week prospective study was undertaken to document the antibiotic susceptibilities of klebsiella organisms which were responsible for an outbreak of septicaemia on the neonatal units of the University College Hospital, Ibadan, Nigeria . The thirty-nine isolates obtained comprised K . pneumoniae, 18 (46.2%), K . aerogenes, 17 (43.6%), K . edwardsii, 3 (7.7%), and K . oxytoca, 1(2.5%) . All the strains were sensitive to ciprofloxacin and ofloxacin, but resistant to ampicillin . The percentage of qualitative sensitivities of the klebsiella species to other available drugs were 41% for ceftazidime, 36% for cefotaxime, 31% for ceftriazone, 23% for cefuroxime, 21% for gentamycin, and 15% for kanamycin . Quantitative sensitivities of the three most commonly isolated sub-types to netilmycin were 63%, 36%, and 33%, respectively . A comparison with a previous antibiotic susceptibility study still showed persistent resistance to the available aminoglycosides.

Microbiol Immunol, 2000, 44(5), 395 - 400
Hexagonal assembly of the magnesium salt of an R-form lipopolysaccharide from Klebsiella pneumoniae: its lowered stability compared with original non-electrodialyzed preparation; Ito H et al.; The magnesium salt of R-form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN-111 (O3-:K1-) that was prepared after the removal of cationic materials by electrodialysis formed essentially the same ordered hexagonal lattice structure with a lattice constant of 14 to 15 nm as the original non-electrodialyzed preparation of the R-form LPS . When the magnesium salt was suspended in 50 mM glycine buffer or Tris buffer at pH 1.4 to 9.5 and kept at 4 C for 24 hr, its content of Mg was markedly decreased, and its hexagonal lattice structure was changed to a swollen hexagonal lattice structure with extended lattice constants at pH 1.4 and to a loose mesh-like structure at pH 3.0 or higher . In the original non-electrodialyzed preparation of the R-form LPS, the release of Mg and disintegration of the hexagonal lattice structure did not occur by suspending in buffers at pH 1.4 to 8.5 at 4 C for 24 hr, but occurred only at pH 9.0 or higher . The results suggest that organic cations that can be removed by electrodialysis play some part in tight binding to Mg2+ and in stabilizing the ordered hexagonal assembly of the R-form LPS.

Rev Esp Quimioter, 1999 Sep, 12(3), 229 - 33
{In vitro activity of piperacillin-tazobactam against Klebsiella pneumoniae clinical isolates, producers or not of extended spectrum beta-lactamases}; Alarcon T et al.; The aim of this study was to determine the in vitro activity of piperacillin-tazobactam against 81 clinical isolates of Klebsiella pneumoniae . The clinical specimens were processed according to standard microbiological procedures and 81 K . pneumoniae isolates were identified using MicroScan Panels following the manufacturer's recommendations . A double disk diffusion method was applied to detect extended spectrum betalactamases (ESBL) (43 isolates were positive and 38 were negative) . Minimum inhibitory concentrations (MIC) were determined by an agar dilution technique using Mueller-Hinton . The following antibiotics were studied: piperacillin with 4 mg/l of tazobactam, amoxicillin-clavulanic acid in a 2:1 proportion, cefotaxime, ceftriaxone, cefepime, imipenem and meropenem . The MIC(90) were 16/4 mg/l for piperacillin-tazobactam, 16/8 for amoxicillin-clavulanic acid, 16 for ceftriaxone, 16 for cefotaxime, 4 for cefepime, 0.25 for imipenem and 0.032 for meropenem in ESBL-positive strains . In ESBL-negative strains the MIC90 were as follows: 4/4 mg/l for piperacillin-tazobactam, 8/4 for amoxicillin-clavulanic acid, 0.064 for ceftriaxone, 0.125 for cefotaxime, 0.125 for cefepime, 0.125 for imipenem and 0.016 for meropenem . All betalactams showed excellent in vitro activity against ESBL non-producer K . pneumoniae . Moreover, piperacillin-tazobactam and both carbapenems showed good in vitro activity against EBSL-producer K . pneumoniae.

Appl Environ Microbiol, 2000 Jul, 66(7), 3083 - 7
A new Klebsiella planticola strain (Cd-1) grows anaerobically at high cadmium concentrations and precipitates cadmium sulfide; Sharma PK et al.; Heavy metal resistance by bacteria is a topic of much importance to the bioremediation of contaminated soils and sediments . We report here the isolation of a highly cadmium-resistant Klebsiella planticola strain, Cd-1, from reducing salt marsh sediments . The strain grows in up to 15 mM CdCl(2) under a wide range of NaCl concentrations and at acidic or neutral pH . In growth medium amended with thiosulfate, it precipitated significant amounts of cadmium sulfide (CdS), as confirmed by x-absorption spectroscopy . In comparison with various other strains tested, Cd-1 is superior for precipitating CdS in cultures containing thiosulfate . Thus, our results suggest that Cd-1 is a good candidate for the accelerated bioremediation of systems contaminated by high levels of cadmium.

J Chemother, 2000 Jun, 12(3), 204 - 7
Diversity of beta-lactam resistance levels among related Klebsiella pneumoniae strains isolated in an intensive care unit; Paniara et al.; Seven multiresistant Klebsiella pneumoniae strains isolated in an intensive care unit were studied . Susceptibility to beta-lactams was determined with the E test . Molecular-typing of the isolates was performed by a PCR-based technique (ERIC2-PCR) . Sonic cell-extracts were used as beta-lactamase preparations . Beta-lactamase quantities were evaluated measuring nitrocefin hydrolysis . The similarity of the ERIC2-PCR patterns indicated that the seven isolates constituted a single clone . The levels of resistance to oximino-beta-lactams, however, were different . There were indications that the differences in susceptibilities were due, at least partly, to differences in the levels of expression of an extended-spectrum beta-lactamase (most likely SHV-5) . Related K . pneumoniae isolates may exhibit different levels of resistance to beta-lactams . Therefore, comparison of resistance phenotypes is of limited usefulness in epidemiological investigations of nosocomial infections caused by resistant K . pneumoniae.

West Afr J Med, 1999 Jan-Mar, 18(1), 1 - 5
Acute laryngotracheobronchitis in Nigerian children; Osinusi K et al.; In a prospective study of cases of severe acute respiratory infections seen at the University College Hospital, Ibadan over a 30-month period, 39 cases were found to have acute upper airway obstruction; out of this number, acute laryngotracheobronchitis (croup) accounted for 35 (90%) . The ages of the 35 ranged from 2 to 53 months with 86% being less than 3 years old, while there was a male preponderance (M:F ratio, 1.7:1) . Croup was associated with bronchopneumonia in 29%, measles in 2.9%, and with both in 40% of the 35 cases . The mean respiratory rate in patients in whom croup co-existed with measles and pneumonia was significantly higher than that in patients with croup alone (P < 0.001) . Six (55%) of the eleven viral identifications made were Parainfluenza virus types 1-3, while Respiratory syncytial virus was identified in three patients . Of the 18 blood cultures done, only one was positive, yielding Klebsiella pneumoniae in a two-year old undernourished child with associated measles and bronchopneumonia . Mortality was 11%; all the four children who died had associated bronchopneumonia, which complicated measles infection in three . The findings in this study support the view that antibiotic therapy is not routinely indicated in the management of croup . Furthermore, measles and bronchopneumonia were identified as indicators of poor prognosis requiring extra care in management.

Am J Surg, 2000 Feb 1, 179(2 Suppl 1), 41 - 44
Immunologic responses to pneumonia; Polk HC et al.; Pneumonia in the critically ill surgical patient often results from the bombardment of a previously normal pulmonary system with therapeutic foreign bodies, hospital pathogens, and impairment of the host defenses . Despite its long history as a significant clinical problem, a woefully inadequate amount of study has been directed toward therapy . We created an experimental model of a differential pulmonary infection using a strain of Klebsiella pneumoniae . We then compared the progressively affected pneumonic process versus the normal parenchyma . We measured neutrophil and monocyte complement antibody receptor expression and monocyte and macrophage class II major histocompatibility antigens (HLA-DR) via percent of cells and mean fluorescent intensity outcomes from flow cytometry . The main difference between infected versus noninfected tissues was monocyte DR expression, which was consistently depressed in cells from infected parenchyma . What follows is a discussion of the implications of this work as well as other work in the immunology of pneumonia and cytokine expression . Possible therapeutic modalities are included.

Nucleic Acids Res, 2000 Jul 1, 28(13), 2563 - 70
The role of region II in the RNA polymerase sigma factor sigma(N) (sigma(54)); Southern E et al.; Bacterial RNA polymerase holoenzymes containing the sigma subunit sigma(N) (sigma(54)) can form a stable closed complex with promoter DNA but only undergo transition to an open complex and transcription initiation when acted on by an activator protein . Proteins of the sigma(N) family have a conserved N-terminal region of 50 amino acids (Region I) that is separated from a conserved C-terminal region of around 360 amino acids (Region III) by a much more variable sequence of between 30 and 110 residues (Region II) . We have investigated the role of Region II in Klebsiella pneumoniae sigma(N) by studying the properties of deletions of all or part of the region both in vivo and in vitro . We found that whilst Region II is not essential, deletion of all or part of it can significantly impair sigma(N) activity . Deletions have effects on DNA binding by the isolated sigma factor and on holoenzyme formation, but the most marked effects are on transition of the holoenzyme from the closed to the open complex in the presence of the activator protein.

Am J Hematol, 2000 Jul, 64(3), 218 - 20
Ruptured Klebsiella pneumoniae liver abscess after high-dose cyclophosphamide for severe aplastic anemia; Lee JJ et al.; A 19-year-old woman with severe aplastic anemia who had previously failed antithymocyte globulin/cyclosporine A received high-dose cyclophosphamide without bone marrow rescue . On day +14, she complained of right upper quadrant abdominal pain and fever . A CT scan of the abdomen showed multiple liver abscesses with rupture and Klebsiella pneumoniae was isolated from blood . In spite of aggressive antibiotic therapy, she rapidly deteriorated and died of overwhelming sepsis . To our knowledge, our patient is the first case of fatal ruptured liver abscess after high-dose cyclophosphamide in a patient with severe aplastic anemia .

J Nephrol, 2000 Mar =Apr, 13(2), 155 - 8
Bilateral emphysematous pyelonephritis with perirenal abscess cured by conservative therapy; Lim CS et al.; Emphysematous pyelonephritis is a rare life-threatening infection of the renal parenchyma . It usually affects unilateral kidney and occurs mostly in diabetic patients . It is characterized by the presence of gas within the renal parenchyma and requires prompt diagnosis and early aggressive therapy . Bilateral emphysematous pyelonephritis is even more rare and is associated with high mortality . We describe a case of a 62-year-old diabetic woman who presented with nonketotic hyperosmolar coma and bilateral emphysematous pyelonephritis caused by Klebsiella pneumoniae . Diagnosis of bilateral emphysematous pyelonephritis was confirmed by an abdominal computed tomographic scan and microbiologic studies . Our patient was successfully treated using percutaneous catheter drainage and long-term antibiotic therapy.

Sex Transm Infect, 2000 Apr, 76(2), 134 - 6
A colorimetric detection system for Calymmatobacterium granulomatis; Carter JS et al.; OBJECTIVE: To incorporate the first polymerase chain reaction (PCR) assay for Calymmatobacterium granulomatis into a colorimetric detection system for use in routine diagnostic laboratories . METHODS: A capture oligonucleotide specific for the Klebsiella phoE gene was covalently linked to tosyl activated magnetic beads . Biotinylated phoE PCR products obtained from 14 positive specimens from patients with donovanosis and isolates of Klebsiella pneumoniae, K rhinoscleromatis, and K ozaenae were cleaved with HaeIII for the purpose of differentiation, captured by the prepared beads, and subjected to standard EIA detection methodology . Eight samples from unrelated genital conditions underwent the same procedure . It was anticipated from the sequence data that the biotinylated fragment would be cleaved from the capture oligonucleotide target region in the three Klebsiella phoE products (that is, a negative colorimetric result) while the entire fragment of interest would remain intact in the positive C granulomatis phoE products (that is, a positive colorimetric result) . RESULTS: All 14 positive specimens from patients with donovanosis gave strong colorimetric readings with this detection system . Isolates of K pneumoniae, K rhinoscleromatis, K ozaenae, and the eight specimens from unrelated genital conditions were negative . CONCLUSION: The successful development of a colorimetric detection system for C granulomatis incorporating two levels of specificity enables the molecular diagnosis of this condition to be undertaken by routine diagnostic laboratories . This should have an important role in the Australian government's campaign to eradicate donovanosis by 2003 though the test still needs to undergo trials and be validated using a larger number of samples from geographically diverse parts of the world in order to ascertain the generalisability of the methodology.

Chest, 2000 Jun, 117(6), 1795 - 8
Laryngotracheobronchial involvement in a patient with nonendemic rhinoscleroma; Yigla M et al.; We report the first case of rhinoscleroma in an Israeli citizen, a former sailor with a transatlantic shipping company . Characteristic histologic changes from a tracheal biopsy and isolation of Klebsiella rhinoscleromatis from a blood culture after diagnostic bronchoscopy confirmed the diagnosis . Extreme delay in the diagnosis, a not uncommon feature in nonendemic areas, was associated with severe advanced laryngotracheobronchial disease . Treatment with quinolones was followed by significant improvement, but the patient died 1 month after presentation, apparently from upper airway obstruction.

Int J Antimicrob Agents, 2000 Jun, 15(1), 37 - 42
Molecular basis of extended-spectrum beta-lactamase production in nosocomial isolates of Klebsiella pneumoniae from M¿erida, Venezuela; Araque M et al.; Twelve Klebsiella pneumoniae isolates resistant to expanded-spectrum cephalosporins and aztreonam, from patients with nosocomial septicaemia at the intensive care unit of the Andes University Hospital, M inverted question markerida, Venezuela, were studied for production of extended-spectrum beta-lactamase (ESbetaL) activity . All were also resistant to gentamicin, kanamycin, tetracycline and chloramphenicol but sensitive to cefoxitin, imipenem, amikacin and tobramycin . Production of ESbetaL activity was confirmed by restoring susceptibility to ceftazidime in the presence of clavulanic acid . All isolates carried an identical plasmid of approximately 87 kb . Resistance to beta-lactams, aminoglycosides, tetracycline and chloramphenicol was lost en bloc after plasmid curing by treatment with acridine orange and was transferable en bloc to Escherichia coli by conjugation . Transconjugants always showed the same plasmid profile as that of Klebsiella donors . Isoelectric focusing analysis of the crude extracts of transconjugants showed in all cases, the presence of two beta-lactamases of pI 5.4 and 8.2 . Analysis of the plasmid carried by one of the transconjugants by means of hybridization assays, revealed the presence of both bla(TEM) and bla(SHV) determinants . Cloning and sequencing of each determinant identified them as bla(TEM-1) and bla(SHV-5), respectively, the latter being responsible for the ESbetaL activity . Results of this study indicate that ESbetaL determinants of the SHV-type carried by transferable elements, are spreading among nosocomial isolates of K . pneumoniae in Merida, Venezuela.

J Postgrad Med, 2000 Jan-Mar, 46(1), 18 - 22
Bacterial nosocomial pneumonia in Paediatric Intensive Care Unit; Tullu MS et al.; AIMS: To determine the incidence, risk factors, mortality and organisms causing nosocomial pneumonia (NP) in intubated patients in Paediatric Intensive Care Unit (PICU) . MATERIALS & METHODS: All patients with endotracheal (ET) tube with or without mechanical ventilation (MV) in a PICU of a tertiary care teaching hospital were included in this prospective study . Clinical parameters and investigations were evaluated in patients who developed nosocomial pneumonia (NP) . Colonisation of the ET tube tip was studied by culture and the antibiotic susceptibility pattern of the isolates was determined . RESULTS: Sixty-nine patients had an ET tube inserted and fifty-nine of these underwent MV . ET tube tip colonisation was seen in 70 out of 88 ET tubes inserted . The incidence of NP in patients with ET tube was 27.54% (7.96/100 days of ET intubation) . NP developed only in patients undergoing MV . The main risk factors for developing NP were - duration of MV and duration of stay in the PICU . Age, sex, immunocompromised status and altered sensorium did not increase the risk of NP . The mortality in cases with NP was 47 . 37% . E . coli and Klebsiella were the commonest organisms isolated from the ET tube tip cultures with maximum susceptibility to amikacin and cefotaxime . CONCLUSIONS: NP developed only in patients undergoing MV . Duration of MV and duration of stay in the PICU increased the risk of developing NP.

J Rheumatol, 2000 Jun, 27(6), 1453 - 60
Cellular immune response to Klebsiella pneumoniae antigens in patients with HLA-B27+ ankylosing spondylitis; Dominguez-Lopez ML et al.; OBJECTIVE: To study the reactivity of peripheral blood mononuclear cells (PBMC) of patients with ankylosing spondylitis (AS) and rheumatoid arthritis (RA) and healthy controls to Klebsiella pneumoniae antigens and to the GroEL-like proteins from K . pneumoniae (HSP60Kp) and Mycobacterium leprae recombinant heat shock protein 65 (rHSP65Ml) . METHODS: PBMC of 13 patients with AS and 9 with RA and 10 controls were stimulated in vitro by heat shock induced K . pneumoniae antigens in a cell blot assay, by insolubilized HSP60Kp, by cytosolic proteins (CP) from K . pneumoniae cultivated at 37 degrees C or 45 degrees C, by soluble HSP60Kp, or by rHSP65Ml . RESULTS: In the cell blot assay 7/13 AS and 3/9 RA patients responded to fraction 4, which contains mainly HSP60Kp, and no controls responded (AS vs . controls: p = 0.007) . The response to the insolubilized HSP60Kp was positive in 6/13 AS patients but negative in RA patients and controls (p = 0.004) . The response to CP45 degrees C was positive in 7/13 AS, in 2/9 RA, and no controls (AS vs controls: p<0.015) . Response to the soluble HSP60Kp was found in 7/13 AS and 5/9 RA patients, but no controls (AS vs . controls: p = 0.0075) . Response to rHSP65Ml was positive in 3/13 AS, 7/9 RA patients, and 1/10 controls (AS vs RA: p = 0.027; RA vs . controls: p = 0.005; AS vs . controls: nonsignificant) . CONCLUSION: In PBMC of the majority of patients with AS and in some with RA, but not in healthy controls, there are cells that proliferate in the presence of HSP60 of K . pneumoniae.

Zh Mikrobiol Epidemiol Immunobiol, 1999 May-Jun, (3), 20 - 6
{The use of computer technologies in the operational analysis of outbreaks of hospital infections}; Bondarenko VM et al.; The description of the mathematical model for the study of outbreaks of hospital infections caused by Klebsiella pneumoniae in intensive care units and the example of its use are presented . Calculations in connection with the evaluation and prognostication of the number of sick and deceased children on account of Klebsiella hospital infection were carried out with the use of the proposed computer program . The importance creating an effective hospital information and analytical system, suitable for the treatment and analysis of data on the epidemiological monitoring of hospital infections, with the elements permitting the registration and prognostication of the cases of the appearance and development of hospital infections, the evaluation of the morbidity level, as well as the effectiveness of using different prophylactic measures.

J Bacteriol, 2000 Jul, 182(13), 3681 - 7
Effects of perturbations of the nitrogenase electron transfer chain on reversible ADP-ribosylation of nitrogenase Fe protein in Klebsiella pneumoniae strains bearing the Rhodospirillum rubrum dra operon; Halbleib CM et al.; The redox state of nitrogenase Fe protein is shown to affect regulation of ADP-ribosylation in Klebsiella pneumoniae strains transformed by plasmids carrying dra genes from Rhodospirillum rubrum . The dra operon encodes dinitrogenase reductase ADP-ribosyltransferase and dinitrogenase reductase-activating glycohydrolase, enzymes responsible for the reversible inactivation, via ADP-ribosylation, of nitrogenase Fe protein in R . rubrum . In bacteria containing the dra operon in their chromosomes, inactivation occurs in response to energy limitation or nitrogen sufficiency . The dra gene products, expressed at a low level in K . pneumoniae, enable transformants to reversibly ADP-ribosylate nitrogenase Fe protein in response to the presence of fixed nitrogen . The activities of both regulatory enzymes are regulated in vivo as described in R . rubrum . Genetic perturbations of the nitrogenase electron transport chain were found to affect the rate of inactivation of Fe protein . Strains lacking the electron donors to Fe protein (NifF or NifJ) were found to inactivate Fe protein more quickly than a strain with wild-type background . Deletion of nifD, which encodes a subunit of nitrogenase MoFe protein, was found to result in a slower inactivation response . No variation was found in the reactivation responses of these strains . It is concluded that the redox state of the Fe protein contributes to the regulation of the ADP-ribosylation of Fe protein.

J Appl Microbiol, 2000 Jun, 88(6), 1038 - 48
A novel large plasmid carrying multiple beta-lactam resistance genes isolated from a Klebsiella pneumoniae strain; Li L et al.; Klebsiella pneumoniae clinical isolates were selected according to the results of antibiotic susceptibility tests . Most of them were resistant to multiple antibiotics, including ampicillin, ceftazidime, cefotaxime and aminoglycosides . Large plasmids were observed in these Kl . pneumoniae strains by pulsed-field gel electrophoresis with S1 nuclease digestion . The Kl . pneumoniae strains investigated produced one to two extrachromosomal bands with a mobility corresponding to 97 approximately 145 kbp linear DNA molecules . A 100 kbp plasmid, designated pK1, was observed in the multiply resistant strain K250 . pK1 had sequences homologous to both the TEM-1 and the aphD probe which were associated with beta-lactam and aminoglycoside resistance . pK1 was transformed into Escherichia coli strain DH5alpha and was found to confer resistance to ampicillin, ceftazidime, cefotaxime and kanamycin . A 8 kbp BamHI DNA fragment of pK1 that carried the ampicillin resistance gene (minimum inhibitory concentration > 1000 microgram ml-1) was cloned into the BamHI site of pACYC184 . Sequence determination showed that this cloned fragment carried a TEM-1 gene . These findings suggest that pK1 is novel in that it appears to carry genes for resistance to ampicillin, cefotaxime and ceftazidime, as well as kanamycin.

Mol Microbiol, 2000 May, 36(3), 697 - 709
The 5'-proximal hairpin loop of lbi RNA is a key structural element in repression of D-galactan II biosynthesis in Klebsiella pneumoniae serotype O1; Warnecke JM et al.; The lbi (lipopolysaccharide biosynthesis interfering) RNA of phage Acm1, an untranslated RNA transcript of 97 nucleotides, previously shown to affect O-polysaccharide biosynthesis in various Escherichia coli strains, was found to downregulate the synthesis of the D-galactan II component of the O-specific polysaccharide in Klebsiella pneumoniae serotype O1 . Enzymatic and Pb2+ probing experiments revealed that lbi RNA consists of two consecutive stem-loop structures, the 5'-proximal hairpin loop of 15 nucleotides being particularly accessible to single strand-specific probes . Based on the assumption that the 5'-proximal hairpin loop may be involved in an antisense interaction with cellular target RNAs, we randomly mutagenized one or two of its central nucleotides . Expression of mutated lbi RNA variants in K . pneumoniae serotype O1 relieved at least partly the repression of D-galactan II formation . In addition, a truncated version of lbi RNA lacking the 3'-proximal hairpin loop was almost as efficient as the wild-type RNA in downregulating D-galactan II synthesis . The results obtained indicate that the 5'-proximal hairpin loop of lbi RNA functions as a key structural element in the mechanism leading to the inhibition of D-galactan II biosynthesis in K . pneumoniae serotype O1.

Indian J Pediatr, 2000 Mar, 67(3), 169 - 74
Clinico-bacteriological study of neonatal septicemia in Hubli; Tallur SS et al.; Septicemia is a leading cause of neonatal morbidity and mortality in India . In a study of 242 infants with septicemia conducted between March 1996 & June 1997 at Hubli, Karnataka, 43.39% infants had 'very early onset' sepsis (VOS), 40.08%, had 'early onset' sepsis (EOS), and 16.53% 'late onset' sepsis (LOS) . 54.55% neonates had birth weight below 2000 g and 39.67% were born before 37 weeks of gestation . The cardiorespiratory signs and jaundice were the most frequent clinical features . The blood culture positivity rate was 64.87% . Klebsiella species was the commonest causative pathogen found and multidrug resistance was frequent . The overall mortality rate was 47.52% and the case fatality rate in LOS was higher than in VOS and EOS (p < 0.001) . The mortality was significantly higher in neonates with lower birth weight and lower gestational age (p < 0.001) . The study underlines the importance of monitoring the various features of neonatal septicemia, as well as the drug resistance of the pathogens from the nurseries.

J Infect Dis, 2000 Jun, 181(6), 2075 - 9 Epub 2000 May 23.
A 5-year study of the seroepidemiology of Klebsiella pneumoniae: high prevalence of capsular serotype K1 in Taiwan and implication for vaccine efficacy; Fung CP et al.; Seroepidemiology of Klebsiella pneumoniae was determined for 1000 nonrepetitive K . pneumoniae isolates collected by a medical center in Taiwan during 1993-1997 . Of these, 630 isolates (63%) were from community-acquired infections; the rest were from hospital-acquired infections . The isolates were serotyped according to capsular antigen by countercurrent immunoelectrophoresis . About 77% were typeable . Serotypes K1 and K2 accounted for 21.7% and 9.3% of the isolates, respectively, followed by K57 (5.1%), K54 (4.2%), K21 (3 . 3%), and K16 (3%) . The frequency of serotype K1 among bacteremic isolates (30.8%) far exceeded that reported by other investigators worldwide . Molecular typing of random K1 isolates by pulsed-field gel electrophoresis revealed several different pulsotypes, suggesting a nonclonal spread . This study indicates that a Klebsiella vaccine developed in Europe is not optimal for use in Taiwan because it does not contain the most predominant serotypes-K1, K54, and K57.

Indian J Pediatr, 1996 Sep-Oct, 63(5), 704 - 6
Neonatal empyema thoracis; Gupta R et al.; Empyema thoracis, a serious complication of pneumonia, fortunately remains a less common cause of respiratory distress in neonates . Only 14 cases of neonatal empyema thoracis have been described in the world literature . The condition is characterized by its rarity, inability to identify any consistent predisposing factors, uncertain pathogenesis, rapid course, lack of consensus on management and a high mortality . We describe here two cases of empyema aged 6 and 8 days caused by E . Coli and Klebsiella respectively . Out of them one survived . A brief review of literature follows the above account.

Indian J Pediatr, 1995 Mar-Apr, 62(2), 237 - 9
Is vernix caseosa a protective material to the newborn? A biochemical approach; Baker SM et al.; Twenty random samples of vernix caseosa were collected from immediately born neonates, in Jamahiriya Hospital, Benghazi . Biochemical studies of these samples revealed presence of lipids (62.5%), proteins (36%) and carbohydrate (1.5%) . Also we could observe inhibition of staph . aureus and klebsiella growth on nutrient agar by this vernix . This observation could be explained either by its higher asparagine content or by its elevated lipid component . In addition tripalmitin was found to be the major lipid constituent, responsible for its hydrophobic property . So we recommend leaving this vernix layer on newborn skin until spontaneous drying.

Biochim Biophys Acta, 2000 Jun 1, 1466(1-2), 328 - 38
Membrane topology of the Na(+)/citrate transporter CitS of Klebsiella pneumoniae by insertion mutagenesis; van Geest M et al.; The sodium ion dependent citrate transporter of Klebsiella pneumoniae (CitS) is a member of the bacterial 2-hydroxycarboxylate transporter family . Membrane topology models of the protein, largely based on reporter molecule fusions to C-terminally truncated CitS molecules, indicate that the protein traverses the membrane 11 times with the NH(2)-terminus in the cytoplasm and the COOH-terminus in the periplasm . Furthermore, the structure is characterized by unusual long loops in the COOH-terminal half of the protein: one hydrophobic segment between transmembrane segments V and VI in the periplasm and three long loops connecting transmembrane segments VI and VII, VIII and IX and X and XI in the cytoplasm . The 10 kDa biotin acceptor domain and six consecutive His residues (His-tag) were inserted at different positions in the four long loops and the effect on transport activity and protein stability was analyzed . Six out of seven insertion mutants were stably expressed and three of these had retained significant transport activity . The sidedness of the tags in the mutants that tolerated the insertion was determined by proteolysis experiments . The results support the 11 transmembrane segment model that was based upon truncated CitS proteins.

Biosens Bioelectron, 2000 Mar, 15(1-2), 7 - 11
Application of a luminescence-based pH optrode to monitoring of fermentation by Klebsiella pneumoniae; Chan CM et al.; The application of an optical sensor based on immobilization of the ruthenium complex {Ru(bpy)2(dhphen)}2+ (bpy = 2,2'-bipyridine, dhphen = 4,7-dihydroxy-1,10-phenanthroline) in Nafion to pH monitoring of fermentation by Klebsiella pneumoniae is described . Interference from the culture medium can be eliminated by addition of a black microporous filter membrane (pore size: 0.45 microm) on top of the sensing film . The response of this pH optrode was found to show good correlation with the conventional pH electrode.

J Trop Pediatr, 2000 Apr, 46(2), 86 - 91
Emergence of multidrug-resistant gram-negative organisms in a neonatal unit and the therapeutic implications; Musoke RN et al.; Multidrug-resistant organisms are increasing worldwide . Over the years we have noted increasing resistance of organisms isolated in our neonatal unit . There is a need therefore to scrutinize the problem so as to be able to plan for the future . Over a 5-month period, 716 infants were admitted of which 192 were screened for sepsis . Overall, 121 (16.7 per cent) had positive blood cultures . The predominant organisms were Gram negative (73.6 per cent of isolates) with Klebsiella species topping the list at 31 per cent . Case fatality for infants infected with Gram negative organisms was 41 per cent . Resistance to gentamicin was 20 per cent chloramphenicol 23.6 per cent, and amoxicillin/ampicillin 66.3 per cent . Of worry is the resistance to ceftazidime 19.1 per cent, and cefuroxime 21.3 per cent, with the figures rising to 27 per cent when more specialized tests are done (disc approximation and potentiation tests) . If these drugs cannot be used in 20-27 per cent of cases then the situation is serious . The contributory factors to increased resistance include: non-investigation of infants put on antibiotics (50 per cent of cases); prolonged (73 per cent) and sometimes unjustified (41.7 per cent) use of antibiotics; and non-utilization of investigations when these are done (52 per cent) together with the delay in getting results back in the ward (6 days).

Biochemistry, 2000 May 9, 39(18), 5389 - 96
Fluoride inhibition of Klebsiella aerogenes urease: mechanistic implications of a pseudo-uncompetitive, slow-binding inhibitor; Todd MJ et al.; Klebsiella aerogenes urease uses a dinuclear nickel active site to catalyze the hydrolysis of urea . Here, we describe the steady-state and pre-steady-state kinetics of urease inhibition by fluoride . Urease is slowly inhibited by fluoride in both the presence and absence of substrate . Steady-state rate studies yield parallel double-reciprocal plots; however, we show that fluoride interaction with urease is not compatible with classical uncompetitive inhibition . Rather, we propose that fluoride binds to an enzyme state (E) that is in equilibrium with resting enzyme (E) and produced during catalysis . Fluoride binding rates are directly proportional to inhibitor concentration . Substrate reduces both the rate of fluoride binding to urease and the rate of fluoride dissociation from the complex, consistent with urea binding to E and E.F in addition to E . Fluoride inhibition is pH-dependent due to a protonation event linked to fluoride dissociation . Fluoride binding is pH-independent, suggesting that fluoride anion, not HF, is the actual inhibitor . We assess the kinetic results in terms of the known protein crystal structure and evaluate possible molecular interpretations for the structure of the E state, the site of fluoride binding, and the factors associated with fluoride release . Finally, we note that the apparent uncompetitive inhibition by fluoride as reported for several other metalloenzymes may need to be reinterpreted in terms of fluoride interaction with the corresponding E states.

Antimicrob Agents Chemother, 2000 Jun, 44(6), 1499 - 505
A novel complex mutant beta-lactamase, TEM-68, identified in a Klebsiella pneumoniae isolate from an outbreak of extended-spectrum beta-lactamase-producing Klebsiellae; Fiett J et al.; Twenty-two Klebsiella pneumoniae and two K . oxytoca extended-spectrum beta-lactamase (ESBL)-producing isolates were collected in 1996 from patients in two pediatric wards of the University Hospital in Wroclaw, Poland . Molecular typing has revealed that the K . pneumoniae isolates represented four different epidemic strains . Three kinds of enzymes with ESBL activity (pI values of 5.7, 6.0, and 8.2) were identified . The pI 6.0 beta-lactamases belonged to the TEM family, and sequencing of the bla(TEM) genes amplified from representative isolates revealed that these enzymes were TEM-47, previously identified in K . pneumoniae isolates from pediatric hospitals in Lodz and Warsaw . One of the TEM-47-producing strains from Wroclaw was very closely related to the isolates from the other cities, and this indicated countrywide spread of the epidemic strain . The pI 5.7 beta-lactamase was produced by a single K . pneumoniae isolate for which, apart from oxyimino-beta-lactams, the MICs of beta-lactam-inhibitor combinations were also remarkably high . Sequencing revealed that this was a novel TEM beta-lactamase variant, TEM-68, specified by the following combination of mutations: Gly238Ser, Glu240Lys, Thr265Met, and Arg275Leu . The new enzyme has most probably evolved from TEM-47 by acquiring the single substitution of Arg275, which before was identified only twice in enzymes with inhibitor resistance (IR) activity . TEM-68 was shown to be a novel complex mutant TEM beta-lactamase (CMT-2) which combines strong ESBL activity with relatively weak IR activity and, when expressed in K . pneumoniae, is able to confer high-level resistance to a wide variety of beta-lactams, including inhibitor combinations . This data confirms the role of the Arg275Leu mutation in determining IR activity and documents the first isolation of K . pneumoniae producing the complex mutant enzyme.

Antimicrob Agents Chemother, 2000 Jun, 44(6), 1438 - 42
Prevalence of SHV-12 among clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamases and identification of a novel AmpC enzyme (CMY-8) in Southern Taiwan; Yan JJ et al.; Twenty (8.5%) of 234 nonrepetitive clinical isolates of Klebsiella pneumoniae from southern Taiwan were found to produce extended-spectrum beta-lactamases (ESBLs): 10 strains produced SHV-12, 4 produced SHV-5, 2 produced a non-TEM non-SHV ESBL with a pI of 8.3, 3 produced a novel AmpC beta-lactamase designated CMY-8 with a pI of 8.25, and 1 produced SHV-12 and an unidentified AmpC enzyme with a pI of 8.2 . The CMY-8 enzyme confers a resistance phenotype similar to CMY-1 and MOX-1, and sequence comparisons showed high homologies (>95%) of nucleotide and amino acid sequences among these three enzymes . Plasmid and pulse-field gel electrophoresis analyses revealed that all isolates harboring an SHV-derived ESBL were genetically unrelated, indicating that dissemination of resistance plasmids is responsible for the spread of SHV ESBLs among K . pneumoniae in this area . All three isolates carrying CMY-8 had identical genotypic patterns, suggesting the presence of an epidemic strain.

EMBO J, 2000 May 15, 19(10), 2229 - 36
Domain structure of secretin PulD revealed by limited proteolysis and electron microscopy; Nouwen N et al.; Secretins, a superfamily of multimeric outer membrane proteins, mediate the transport of large macromolecules across the outer membrane of Gram-negative bacteria . Limited proteolysis of secretin PulD from the Klebsiella oxytoca pullulanase secretion pathway showed that it consists of an N-terminal domain and a protease-resistant C-terminal domain that remains multimeric after proteolysis . The stable C-terminal domain starts just before the region in PulD that is highly conserved in the secretin superfamily and apparently lacks the region at the C-terminal end to which the secretin-specific pilot protein PulS binds . Electron microscopy showed that the stable fragment produced by proteolysis is composed of two stacked rings that encircle a central channel and that it lacks the peripheral radial spokes that are seen in the native complex . Moreover, the electron microscopic images suggest that the N-terminal domain folds back into the large cavity of the channel that is formed by the C-terminal domain of the native complex, thereby occluding the channel, consistent with previous electrophysiological studies showing that the channel is normally closed.

EMBO J, 2000 May 15, 19(10), 2221 - 8
Pilus formation and protein secretion by the same machinery in Escherichia coli; Sauvonnet N et al.; The secreton (type II secretion) and type IV pilus biogenesis branches of the general secretory pathway in Gram-negative bacteria share many features that suggest a common evolutionary origin . Five components of the secreton, the pseudopilins, are similar to subunits of type IV pili . Here, we report that when the 15 genes encoding the pullulanase secreton of Klebsiella oxytoca were expressed on a high copy number plasmid in Escherichia coli, one pseudopilin, PulG, was assembled into pilus-like bundles . Assembly of the 'secreton pilus' required most but not all of the secreton components that are essential for pullulanase secretion, including some with no known homologues in type IV piliation machineries . Two other pseudopilins, pullulanase and two outer membrane-associated secreton components were not associated with pili . Thus, PulG is probably the major component of the pilus . Expression of a type IV pilin gene, the E.coli K-12 gene ppdD, led to secreton-dependent incorporation of PpdD pilin into pili without diminishing pullulanase secretion . This is the first demonstration that pseudopilins can be assembled into pilus-like structures.

J Biol Chem, 2000 May 19, 275(20), 14903 - 9
Involvement of the acute phase protein alpha 1-acid glycoprotein in nonspecific resistance to a lethal gram-negative infection; Hochepied T et al.; Resistance to gram-negative infection can be induced by pretreating animals with several agents such as turpentine and interleukin (IL)-1 . Because these agents are powerful inducers of acute phase proteins, we wondered whether these proteins, more particularly alpha(1)-acid glycoprotein (alpha(1)-AGP), are involved in nonspecific resistance to infection . Turpentine and IL-1 protect completely against a lethal challenge of Klebsiella pneumoniae when given 48 and 12-48 h before the challenge, respectively . alpha(1)-AGP induction in the serum reached peak values 48 h after turpentine and 12-48 h after IL-1 injection . Administration of alpha(1)-AGP, 2 h before a challenge of K . pneumoniae, significantly increased the survival . Numbers of bacteria cultured from blood and organs were significantly lower in mice pretreated with a protective dose of turpentine, IL-1, or alpha(1)-AGP . These data suggest that alpha(1)-AGP is a possible mediator in turpentine- or IL-1-induced protection because time points of maximal induction of alpha(1)-AGP by turpentine or IL-1 and of optimal protection by alpha(1)-AGP coincide . Transgenic overexpression of rat alpha(1)-AGP protected mice from a K . pneumoniae infection . Bacterial counts in blood and organs were significantly lower in transgenic mice, and only in control mice were large necrotic areas, apoptosis, and blood clots observed in the spleen . Our data suggest that alpha(1)-AGP prevents gram-negative infections and may be an essential component in nonspecific resistance to infection.

Jpn J Infect Dis, 1999 Feb, 52(1), 16 - 7
Enterotoxigenicity of Klebsiella pneumoniae associated with childhood gastroenteritis in Madras, India; Ananthan et al.; Klebsiella pneumoniae was isolated as the predominant growth from 12 of 100 children under 3 years of age suffering from acute diarrhea . Of the 12 isolates, SA1, SA2, SA4, SA5, SA6 produced a secretogenic response in the ligated rabbit ileal loop, and one isolate, SA3, induced a diarrheagenic response in suckling mice . Two isolates, SA7 and SA8, were diarrheagenic in both assays . Strains SA9, SA10, SA11, and SA12 were found to be non-enterotoxigenic . These isolates belonged to serotypes K6, 16, 25, 30, 39, 46, 49, 53, 66, and 81 . All eight enterotoxigenic strains were resistant to ampicillin, streptomycin, ceftazidime, cefuroxime, and cotrimoxazole . Only quinolones such as ciprofloxacin and norfloxacin appear to be effective against enterotoxigenic K . pneumoniae.

Am J Surg, 2000 Feb, 179(2A Suppl), 41S - 44S
Immunologic responses to pneumonia; Polk HC et al.; Pneumonia in the critically ill surgical patient often results from the bombardment of a previously normal pulmonary system with therapeutic foreign bodies, hospital pathogens, and impairment of the host defenses . Despite its long history as a significant clinical problem, a woefully inadequate amount of study has been directed toward therapy . We created an experimental model of a differential pulmonary infection using a strain of Klebsiella pneumoniae . We then compared the progressively affected pneumonic process versus the normal parenchyma . We measured neutrophil and monocyte complement antibody receptor expression and monocyte and macrophage class II major histocompatibility antigens (HLA-DR) via percent of cells and mean fluorescent intensity outcomes from flow cytometry . The main difference between infected versus noninfected tissues was monocyte DR expression, which was consistently depressed in cells from infected parenchyma . What follows is a discussion of the implications of this work as well as other work in the immunology of pneumonia and cytokine expression . Possible therapeutic modalities are included.

J Antimicrob Chemother, 2000 May, 45(5), 611 - 6
Survey of Klebsiella pneumoniae producing extended-spectrum beta-lactamases at a Portuguese hospital: TEM-10 as the endemic enzyme; Barroso H et al.; One hundred and thirty-eight isolates of Klebsiella pneumoniae showing resistance to ceftazidime were isolated from different wards of the Hospital de Santa Maria, Lisbon . The genomic DNA of the isolates was analysed by pulsed-field gel electrophoresis (PFGE) and two patterns were predominant . In all isolates the presence of a single large plasmid of about 50 kb suggested that propagation of the outbreak prominently involved plasmid spread . The deduced amino acid sequence indicated the presence of a TEM-10 beta-lactamase . This extended-spectrum beta-lactamase was present among K . pneumoniae isolates, was widely disseminated in different wards and remained persistent as a result of an outbreak involving the dissemination of both the multi-resistance plasmids harbouring the bla gene and the isolates themselves.

J Chemother, 2000 Apr, 12(2), 145 - 52
High prevalence of extended spectrum beta-lactamase production among Klebsiella pneumoniae strains isolated at a University Hospital in Turkey; Gulay Z et al.; Beta-lactam susceptibility and beta-lactamase patterns of a random sample of 44 Klebsiella pneumoniae strains that had been isolated from nosocomial infections at Dokuz Eylul University Hospital in Izmir, were investigated . All strains were amoxycillin resistant but in the presence of clavulanic acid 26 became sensitive . Similarly 39 of the strains were resistant to ceftazidime and cefotaxime; clavulanic acid restored sensitivity to ceftazidime in 28 and to cefotaxime in 25 of these resistant strains . Extended spectrum beta-lactamase (ESBL) production was positive in 84% of the isolates as determined by the double disk synergy test . Isoelectric focusing revealed that each strain produced one to four beta-lactamases, pI 7.6 enzymes being the most prevalent . Other enzymes with pIs of 8.4, 8.2, 5.4, 7.8 were also detected . Resistance to ceftazidime was transferred from 18 of the 44 isolates to the recipient Escherichia coli K-12 at 37 degrees C . The transconjugants were examined for their plasmid content and the plasmids were characterized by their size and resistance profile . Fourteen different restriction pattern groups were identified with Eco R1 . The results indicate a high prevalence of ESBL production in nosocomial K . pneumoniae isolates in Izmir and have major implications concerning the clinical use of later generation cephalosporins.

Microbiol Immunol, 2000, 44(3), 193 - 6
Disintegration of Mg2+ -induced hexagonal assembly of an R-form lipopolysaccharide from Klebsiella pneumoniae by treatment with CaCl2; Ito H et al.; R-form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN-111 (O3-: K1-), which was precipitated by the addition of 2 volumes of ethanol containing 10 mM MgCl2 for the purification process, ultrastructurally exhibited membrane pieces consisting of an ordered hexagonal lattice structure with a lattice constant of 14 to 15 nm . When the R-form LPS was suspended in 50 mM tris (hydroxymethyl) aminomethane buffer (at pH 8.5) containing 1 mM or higher concentrations of CaCl2 and kept at 4 C for 10 hr, the ordered hexagonal lattice structure of the R-form LPS was disintegrated and changed to an irregular rough, mesh-like structure . By treatment with CaCl2, the content of Mg in the LPS was markedly decreased, and conversely, the content of Ca was increased to a level depending upon the concentration of CaCl2 . Results indicate that the addition of CaCl2 to suspensions of the Mg-bound R-form LPS result in a tighter binding of Ca2+ to the R-form LPS and the release of Mg2+ from the R-form LPS, and as a consequence, destroys the Mg2+ -induced ordered hexagonal lattice structure of the R-form LPS.

J Bacteriol, 2000 May, 182(10), 2879 - 85
Contribution of cysteine desulfurase (NifS protein) to the biotin synthase reaction of Escherichia coli; Kiyasu T et al.; The contribution of cysteine desulfurase, the NifS protein of Klebsiella pneumoniae and the IscS protein of Escherichia coli, to the biotin synthase reaction was investigated in in vitro and in vivo reaction systems with E . coli . When the nifS and nifU genes of K . pneumoniae were coexpressed in E . coli, NifS and NifU proteins in complex (NifU/S complex) and NifU monomer forms were observed . Both the NifU/S complex and the NifU monomer stimulated the biotin synthase reaction in the presence of L-cysteine in an in vitro reaction system . The NifU/S complex enhanced the production of biotin from dethiobiotin by the cells growing in an in vivo reaction system . Moreover, the IscS protein of E . coli stimulated the biotin synthase reaction in the presence of L-cysteine in the cell-free system . These results strongly suggest that cysteine desulfurase participates in the biotin synthase reaction, probably by supplying sulfur to the iron-sulfur cluster of biotin synthase.

Indian J Pediatr, 1997 Jan-Feb, 64(1), 116 - 8
Perianal necrotizing fasciitis in a neonate; Ozkan H et al.; A two-week-old term male infant, weighing 1,600 grams was transferred to the neonatology unit of Dokuz Eylul University hospital with sharply demarcated cutaneous gangrene surrounding the perianal region . He did well at birth . In his history, on the 10th postnatal day, a red, painful, warm cutaneous lesion was observed which was thought to be secondary to repeated and inappropriate rectal temperature measurements . Besides an ill-appearing child, a nontender frank cutaneous gangrene developed within several days . Klebsiella pneumoniae was cultured from the involved area . Blood cultures were negative . A frozen section of soft tissue biopsy could not be performed because of the localization of the lesion . The patient was successfully treated by surgical debridement and high doses of parenteral antibiotics.

Hinyokika Kiyo, 2000 Feb, 46(2), 105 - 7
{A case of infected renal cyst extending to leg abscess}; Ueno Y et al.; A 45-year-old woman was referred to our hospital with the chief complaint of left flank pain, left leg pain and loss of appetite . Computed tomography scan and magnetic resonance imaging demonstrated a large cystic mass in the left kidney, which we diagnosed as an infected renal cyst . Under ultrasonic guidance, percutaneous puncture and drainage of the renal cyst were performed . After her leg pain worsened, computed tomography revealed abscesses in the left leg, suggesting an infected renal cyst extending to the leg through the obturator foramen . Under general anesthesia, incision and drainage were performed . Cultures from the cyst and abscess fluid showed Klebsiella pneumoniae . Our case is the 82nd case of an infected renal cyst in the Japanese literature.

Infect Immun, 2000 May, 68(5), 2402 - 9
Binding to and opsonophagocytic activity of O-antigen-specific monoclonal antibodies against encapsulated and nonencapsulated Klebsiella pneumoniae serotype O1 strains; Held TK et al.; The high mortality of nosocomial infections caused by Klebsiella spp . has acted as a stimulus to develop immunotherapeutic approaches targeted against surface molecules of these bacteria . Since O-antigen-specific antibodies may add to the protective effect of K antisera, we tested the functional and binding capacity of O-antigen-specific monoclonal antibodies (MAbs) raised against different Klebsiella O antigens . The MAbs tested were specific for the O-polysaccharide partial antigens D-galactan II (MAb Ru-O1), D-galactan I (MAb IV/4-5), or core oligosaccharide (MAb V/9-5) of the Klebsiella serogroup O1 antigen . In enzyme-linked immunosorbent assay binding experiments, we found that all MAbs recognized their epitopes on intact capsule-free bacteria; however, binding to encapsulated wild-type strains belonging to different K-antigen serotypes was significantly reduced . The K2 antigen acted as the strongest penetration barrier, while the K7 and K21 antigens allowed some, though diminished, antibody binding . In vitro phagocytic killing experiments showed that MAb Ru-O1 possessed significant opsonizing activity for nonencapsulated O1 serogroup strains and also, to a much lesser extent, for encapsulated strains belonging to the O1:K7 and O1:K21 serotypes . MAbs or antisera specific for the D-galactan II antigen may thus be the most promising agents for further efforts to develop a second-generation Klebsiella hyperimmune globulin comprising both K- and O-antigen specificities.

Arch Med Res, 2000 Jan-Feb, 31(1), 28 - 36
Klebsiella pneumoniae 35 and 36 kDa porins are common antigens in different serotypes and induce opsonizing antibodies; Alcantar-Curiel MD et al.; BACKGROUND: Klebsiella pneumoniae is a major cause of neonatal sepsis and nosocomial infections in Mexico . Antibiotic therapy is the first choice for treatment but the increase in multiple resistance strains has forced scientists to look for alternative treatments, such as immunotherapy . In this work, we propose that porins could be a common antigen among four different capsular serotypes of Klebsiella pneumoniae for the production of immune sera with opsonizing capacity . METHODS: The 35 and 36 kDa porins from four different serotypes of the bacteria were isolated by the Nikaido method followed by purification in Sephacryl column chromatography . The 36 kDa of serotype K8 was further purified by electroelution . The 35 and 36 kDa porins were used to obtain rabbit polyclonal antibodies (PolyAb) to the four serotypes and the 36 kDa from K8 for the production of monoclonal antibodies (MoAb) . Antigenic reactivity of PolyAb and MoAb were analyzed by ELISA and WB and their opsonizing capacity for human PMN was measured by chemiluminescence (CL) using capsulated and non-capsulated bacteria . RESULTS: Porins from the four strains showe electrophoretic homology and cross reaction by ELISA and WB . CL assays indicated that PolyAb opsonized heterologous strains and that MoAb perform this in the absence of capsule . CONCLUSIONS: K.pneumoniae 35 and 36 kDa porins are common antigens for the four serotypes studied and induce opsonizing antibodies.

Microb Pathog, 2000 Apr, 28(4), 211 - 20
Recognition of B cells epitopes of the Klebsiella pneumoniae GroEL-like protein by HLA-B27 positive subjects; Cancino-Diaz M et al.; The presence of antibodies against antigens of K . pneumoniae in HLA-B27 positive patients with ankylosing spondylitis (AS), has been well documented . We have previously reported that sera from HLA-B27 positive subjects react with the K . pneumoniae GroEL-like protein (HSP60Kp) and have higher titers than HLA-B27 negative individuals . We cloned the gene that codes for this protein, determined hydrophilic regions by computer analysis of the predicted amino acid sequence and found that residues 389-397, 360-368 and 282-290, were possible B cell epitopes . To test this prediction, and to determine if the HLA-B27 positive and negative AS patients recognize the same or different epitopes, we truncated the hsp60Kp gene, from the 3; terminal nucleotide, to obtain fragments having or not the predicted epitopes . Four polypeptides of 40, 37, 30 and 18 kDa were obtained and analysed, by ELISA and inhibition of ELISA, for their reactivity with IgG antibodies from three high responders HLA-B27 positive AS patients and three HLA-B27 negative subjects who recognized the rHSP60Kp . Sera from both HLA-B27 positive and negative subjects reacted equally well with rHSP60Kp or with the 40 and 37 kDa peptides, which do not have residues 389-397 and 360-368, respectively, but reactivity was lost with the 30 kDa peptide, which also lacks residues 282-290 . Contrary to what we expected, antibodies from HLA-B27 negative and positive individuals recognized the same epitope of the HSP60Kp . Our results indicate that the important epitope for B cells could be the 282-290 region and that the contribution of the two other predicted regions is minimal . We also conclude that the differences in response to the HSP60Kp in HLA-B27 positive AS patients and HLA-B27 negative individuals is not qualitative, but only quantitative .

J Bacteriol, 2000 May, 182(9), 2567 - 73
A single amino acid substitution in a mannosyltransferase, WbdA, converts the Escherichia coli O9 polysaccharide into O9a: generation of a new O-serotype group; Kido N et al.; wbdA is a mannosyltransferase gene that is involved in synthesis of the Escherichia coli O9a polysaccharide, a mannose homopolymer with a repeating unit of 2-alphaMan-1,2-alphaMan-1,3-alphaMan-1, 3-alphaMan-1 . The equivalent structural O polysaccharide in the E . coli O9 and Klebsiella O3 strains is 2-alphaMan-1,2-alphaMan-1, 2-alphaMan-1,3-alphaMan-1,3-alphaMan-1, with an excess of one mannose in the 1,2 linkage . We have cloned wbdA genes from these O9 and O3 strains and shown by genetic and functional studies that wbdA is the only gene determining the O-polysaccharide structure of O9 or O9a . Based on functional analysis of chimeric genes and site-directed mutagenesis, we showed that a single amino acid substitution, C55R, in WbdA of E . coli O9 converts the O9 polysaccharide into O9a . DNA sequencing revealed the substitution to be conserved in other E . coli O9a strains . The reverse substitution, R55C, in WbdA of E . coli O9a resulted in lipopolysaccharide synthesis showing no ladder profile instead of the conversion of O9a to O9 . This suggests that more than one amino acid substitution in WbdA is required for conversion from O9a to O9.

Biochemistry, 2000 Apr 18, 39(15), 4320 - 6
Essential role of tyrosine 229 of the oxaloacetate decarboxylase beta-subunit in the energy coupling mechanism of the Na(+) pump; Jockel P et al.; The membrane-bound beta-subunit of oxaloacetate decarboxylase from Klebsiella pneumoniae catalyzes the decarboxylation of carboxybiotin, which is coupled to Na(+) translocation and consumes a periplasmically derived proton . Upon site-directed mutagenesis of 20 polar and/or conserved residues within putative membrane-integral regions, the specific oxaloacetate decarboxylase activities were reduced to various extents, but only the enzyme with a Y229F mutation was completely inactive . We propose that Y229 is part of the network by which the proton of S382 is delivered to carboxybiotin, where it is consumed upon catalyzing the immediate decarboxylation of this acid-labile compound . Unlike S382 or D203, Y229 appears to be not involved in Na(+) binding, because in the Y229F orY229A mutants, the beta-subunit was protected from tryptic digestion by 50 mM NaCl like in the wild-type enzyme . Oxaloacetate decarboxylase with a betaC291E mutation was unstable in the absence of Na(+) and dissociated into an alpha-gamma subcomplex and the beta-subunit . The enzyme could only be isolated in the presence of 0 . 5 M NaCl . These results are consistent with the notion that the beta-subunit changes its conformation upon Na(+) binding.

J Antimicrob Chemother, 2000 Apr, 45(4), 467 - 73
Molecular epidemiology of an outbreak due to IRT-2 beta-lactamase-producing strains of Klebsiella pneumoniae in a geriatric department; Girlich D et al.; In February 1998, 195 patients in the geriatric department of a French hospital were screened for the presence of co-amoxiclav-resistant Klebsiella pneumoniae . Eleven co-amoxiclav-resistant isolates obtained all produced an identical IRT-2 beta-lactamase . These K . pneumoniae isolates were clonally related and harboured a c . 55 kb non-conjugative plasmid encoding a non-class-1 integron-located blaIRT-2 gene . This study underlines that geriatric departments may be a reservoir for antibiotic-resistant strains and that IRT beta-lactamase-producing strains may be nosocomial pathogens.

J Clin Microbiol, 2000 Apr, 38(4), 1495 - 7
Incidence and identification of Klebsiella planticola in clinical isolates with emphasis on newborns; Westbrook GL et al.; Studies conducted in France and Germany suggest that up to 19% of clinically identified Klebsiella sp . are actually Klebsiella planticola, an environmental species that has been attributed to two cases of septicemia, with a rare isolate of Klebsiella terrigena (0 . 4%) being identified . A 1-year survey of newborns on a neonatal ward, also conducted in Germany, reported that 72% of Klebsiella sp . were Klebsiella oxytoca and 8.7% were K . planticola . The tests necessary to identify these species are not found in most clinical identification schemes or in the database matrices of most commercial identification products . To determine the incidence of unrecognized K . planticola among the Klebsiella sp . isolates in our collection, we used the battery of seven supplemental tests amended from the work of Monnet and Freney to test 352 stock isolates and 84 fresh clinical isolates from four local hospitals . After testing 436 strains of Klebsiella, only one strain was identified as a possible K . planticola and none was identified as K . terrigena . We tested an additional 43 stock strains of K . oxytoca isolated from newborns by using eight biochemical tests and found one additional strain of K . planticola . The occurrence of K . planticola in our collection is far less frequent than that observed in other countries.

Jpn J Antibiot, 1999 Sep, 52(9), 585 - 94
{Antibacterial activity of beta-lactam antibiotics against extended-spectrum beta-lactamase producing bacteria}; Kuga A et al.; MICs of various beta-lactam antibiotics by themselves and in combination with beta-lactamase inhibitor (clavulanic acid) against extended spectrum beta-lactamase (ESBL) producing strains of Escherichia coli and Klebsiella pneumoniae which were isolated from clinical materials were investigated . Furthermore, based on the results obtained, a procedure to detect ESBL producing strains was proposed . The MICs of beta-lactam antibiotics against beta-lactamase producing strains were investigated . At first, beta-lactamase was investigated by the drug sensitivity pattern (MIC) to beta-lactam antibiotics and by the substrate profiles of beta-lactamase extracted from the transconjugant of E . coli K-12 strains . After that, we classified the beta-lactamase producing gene by PCR method . Furthermore, a proposal was made for an antibiotic to be used in the confirmation of mixed type beta-lactamase . The data obtained by the above investigations were compiled and used to determine the limit concentration of each beta-lactam against beta-lactamase producing strains including ESBL . By using beta-lactam antibiotics at the following concentrations, it is considered possible to classify beta-lactamase; ampicillin (64 micrograms/ml), ampicillin/clavulanic acid (32/5 micrograms/ml), piperacillin (64 micrograms/ml), cefotaxime (1 microgram/ml), cefpodoxime (2 micrograms/ml), ceftazidime (1 microgram/ml), cefmetazole (4 micrograms/ml), cefminox (2 micrograms/ml), cefepime (0.5 microgram/ml), aztreonam (1 microgram/ml) and imipeneme (1 microgram/ml) . This method may be used as a reference in investigating the prevalence of beta-lactam resistant isolates by ESBL producing E . coli and K . pneumoniae.

J Postgrad Med, 1997 Oct-Dec, 43(4), 98 - 101
Klebocin typing of Klebsiella species isolated from nosocomial infection in intensive care unit; Pal RB et al.; Klebocin typing and antibiotic resistance have been studied for 518 strains of Klebsiella pneumoniae, {106 from intensive care unit (ICU) sites, 182 from ICU staff flora, 192 from patient flora and 38 from clinical specimens} . The overall typability was 71.62% . The most common mnemonic types among various sources were 111, 211, and 112 . Of the total strains tested, 28.37% strains were found to be untypable . These strains are labelled as "444" . When klebocin typing was used in association with antibiogram, in 86.84% cases of clinical infection probable source of infection could be detected . Thus a combination of two typing methods poses a significant contribution in epidemiological studies.

Biochemistry (Mosc), 2000 Mar, 65(3), 304 - 8
Molecular analysis of polyphosphate accumulation in bacteria; Kuroda A et al.; The dynamic behavior of inorganic polyphosphate (polyP), its accumulation and disappearance, is the most striking aspect of polyP metabolism in bacteria . Imbalance between polyP synthesis and degradation results in fluctuations of polyP by 100- to 1000-fold . We here review recent results with respect to this polyP metabolism in bacteria . PolyP accumulation in response to amino acid starvation, accompanied by increased levels of stringent factors, has been observed in Escherichia coli . Inhibition by stringent factors of polyphosphatase interrupts the dynamic balance between the synthesis and degradation of polyP, accounting for polyP accumulation . Polyphosphate kinase is required for activation of intracellular protein degradation, which is required for adaptation at the onset of amino acid starvation . The adaptation to amino acid starvation is mediated by the network of stringent response and polyP metabolism . PolyP accumulation independent of stringent response has also been observed . Novobiocin, an inhibitor for DNA gyrase, stimulated accumulation of polyP but not that of stringent factors . However, a temperature-sensitive DNA gyrase mutant did not exhibit polyP accumulation at the non-permissive temperature . Antagonistic relationship of polyP to nucleic acid synthesis, explored by Harold, appears to be more complicated . We discuss relationship of Pi regulation to polyP accumulation in E . coli and Klebsiella aerogenes . A function of polyP as an in vivo phosphagen affecting polyP accumulation is also discussed.

Protein Sci, 2000 Jan, 9(1), 121 - 8
Interaction with magnesium and ADP stabilizes both components of nitrogenase from Klebsiella pneumoniae against urea denaturation; Zou L et al.; The nitrogenase enzyme of Klebsiella pneumoniae consists of two separable proteins, each with multiple subunits and one or more oxygen sensitive metallocenters . The wild-type nitrogenase proteins are stable to electrophoresis in high concentrations of urea under anaerobic conditions . Addition of Mg+2 and ADP greatly increases the stability of the smaller Fe protein (from <4 to >6 M for full unfolding), an effect directly analogous to stabilization in p21ras induced by Mg+2 and GDP . Stabilization by Mg+2 is slight for the holo MoFe protein (from approximately 1.5 to approximately 2.4 M) but more dramatic for the apo protein form of the MoFe protein accumulated by certain Fe protein (nifH gene) mutants . The potent product inhibitor of nitrogenase function, MgADP, increases stability of the MoFe protein more than Mg+2 alone, to approximately 3.6 M, showing that nucleotides interact with the MoFe protein . Mutations of the nifM gene result in slower accumulation of less stable Fe protein, indicating that NifM is involved in correct folding of the Fe protein . Mutationally altered proteins are often difficult to purify for study because of their inherent instability, low expression level, or oxygen lability . Crude extracts of 11 different mutants of Fe protein (nifH gene) were examined by transverse urea gradient gels to rapidly screen for stabilizing interactions in the presence or absence of substrate or inhibitor analogs . Amino acid alterations D44N and R188C, at the interface of the dimer, in the vicinity of the nucleotide binding site(s), have significantly lower stability than the wild-type enzyme in the absence of Mg+2 but comparable stability in its presence, showing the importance of Mg+2 in the subunit interactions . Mutations N163S and E266K, in which residues normally involved in hydrogen bonding far from the active site were altered, are more labile than the wild-type even with Mg+2 added . Seven other mutants, though nonfunctional, did not appear altered in stability compared to the wild-type.

Biosci Biotechnol Biochem, 2000 Feb, 64(2), 428 - 31
Cloning and sequencing of the maltohexaose-producing amylase gene of Klebsiella pneumoniae; Momma M; The molecular characterization of the maltohexaose-producing amylase gene of Klebsiella pneumoniae revealed an open reading frame in which 2,031 base pairs encode a protein of 677 amino acids with a calculated molecular weight of 75,921 . The amylase gene had high similarities of 73.6% in DNA sequence and 79.3% in deduced amino acid sequence with the periplasmic alpha-amylase MalS gene of Escherichia coli.

J Med Entomol, 2000 Mar, 37(2), 246 - 9
Bactericidal effects of sugar-fed antibiotics on resident midgut bacteria of newly emerged anopheline mosquitoes (Diptera: Culicidae); Toure AM et al.; A protocol was developed for significantly reducing resident midgut bacteria in newly emerged anopheline mosquitoes using a combination of antibiotics . Pupa harvested from colony-reared Anopheles gambiae s.l . Giles and Anopheles stephensi (Liston) were placed in cages wiped previously with 70% alcohol and kept under UV light for 24 h . Emerging adult mosquitoes were fed for 3 consecutive days on antibiotic solution, consisting of 0.4% gentamicin sulfate and 1% penicillin-streptomycin solution in a 10% sterile sucrose solution . Bacterial suspensions of Escherichia coli, Klebsiella pneumoniae (Schroeter, 1886), and Pseudomonas stutzeri (Lehmann & Neumann, 1896) isolated from wild-caught anophelines were fed to antibiotic-treated mosquitoes starved for 24 h via either sugar or membrane-feeding . Mosquitoes dissected 1 and 24 h after blood-feeding or sugar-feeding, and plated on trypticase soy agar plates, yielded the same type of bacteria fed originally without evidence of contaminants . There was no residual effect of the antibiotics on introduced single bacteria strains as judged by the presence of bacteria in antibiotic-treated mosquitoes . This experimental reduction of resident midgut bacteria and their replacement with single strains in newly emerged anopheline mosquitoes should facilitate further investigations of the interactions between malaria parasites and bacteria found in the midguts of mosquitoes.

Antimicrob Agents Chemother, 2000 Apr, 44(4), 1081 - 4
SHV-13, a novel extended-spectrum beta-lactamase, in Klebsiella pneumoniae isolates from patients in an intensive care unit in Amsterdam; Yuan M et al.; Eleven clonally related Klebsiella pneumoniae isolates were examined . These had been isolated at an intensive care unit in Amsterdam in 1994 . Their resistance was associated with a conjugative 170-kb plasmid which encoded a novel SHV beta-lactamase designated SHV-13 . The SHV-13 enzyme had two substitutions compared with SHV-1: Leu35Gln and Gly238Ala . It hydrolyzed cefotaxime much more rapidly than ceftazidime or aztreonam.

Clin Infect Dis, 2000 Mar, 30(3), 473 - 8
Epidemiology of ciprofloxacin resistance and its relationship to extended-spectrum beta-lactamase production in Klebsiella pneumoniae isolates causing bacteremia; Paterson DL et al.; A prospective study of Klebsiella pneumoniae bacteremia was performed in 12 hospitals in 7 countries . Of 452 episodes of bacteremia, 25 (5.5%) were caused by K . pneumoniae that was resistant in vitro to ciprofloxacin . Extended-spectrum beta-lactamase (ESBL) production was detected in 15 (60%) of 25 ciprofloxacin-resistant isolates, compared with 68 (16%) of 427 ciprofloxacin-susceptible strains (P=.0001) . Multivariate analysis revealed that risk factors for ciprofloxacin resistance in K . pneumoniae included prior receipt of a quinolone (P=.0065) and an ESBL-producing strain (P=.012) . In all, 18% of ESBL-producing isolates were also ciprofloxacin-resistant . Pulsed-field gel electrophoresis showed that 11 of the 15 ciprofloxacin-resistant ESBL-producing strains belonged to just 4 genotypes, suggesting that patient-to-patient transmission of such strains occurred . The close relationship between ESBL production and ciprofloxacin resistance is particularly worrisome because the first reported instance of plasmid-mediated ciprofloxacin resistance has been in an isolate of K . pneumoniae also possessing an ESBL.

Int J Radiat Biol, 2000 Feb, 76(2), 261 - 71
WR-151327 increases resistance to Klebsiella pneumoniae infection in mixed-field- and gamma-photon-irradiated mice; Ledney GD et al.; PURPOSE: To determine the efficacy of WR-151327 (WR) {S-3-(3-methylaminopropylamino) propylphosphorothioic acid; (CH3-HN-(CH2)3-NH-(CH2)3-S-PO3H2)} in increasing resistance to bacterial infection after a sublethal dose of gamma-photons or mixed-field neutrons plus gamma-photons . MATERIALS AND METHODS: B6D2F1/J female mice received 200 mg/kg WR i.p . or saline vehicle 20-30 min before or after sham (0 Gy) or 7.0 Gy 60Co gamma-photon irradiation . WR or saline vehicle was given only before 3.5 Gy TRIGA-reactor-produced mixed-field {n/(n+y) = 0.67} irradiation . Four days after drug treatment or drug treatment and irradiation, graded doses of Klebsiella pneumoniae were injected s.c . into mice, and 30-day survival was recorded . To assess haemopoietic changes other unirradiated and irradiated mice not injected with bacteria were given WR or saline . Peripheral blood (PB) and femoral bone marrow (BM) cells were measured 1, 3 or 4, 7, 10 and 14 or 15 days later . RESULTS: WR pretreatment increased resistance to infection in irradiated but not in unirradiated mice . Bacterial CFU-LD50/30 values for 0 Gy saline-treated mice were 1.20x10(6); for 0 Gy WR-treated mice 1.16x10(6); for gamma-photon-irradiated saline-treated mice 3.02x10(1); for gamma-photon-irradiated WR-treated mice 1.24x10(4); for mixed-field-irradiated saline-treated mice 1.94x10(2); and for mixed-field-irradiated WR-treated mice 6.13x10(3) . WR-induced resistance to infection paralleled increased numbers of PB white cells, neutrophils, platelets, femoral BM cells and granulocyte macrophage colony-forming cells (GM-CFC) in irradiated mice not given bacteria . CONCLUSIONS: These studies quantify the resistance to bacterial infection in mice treated with WR before sublethal irradiation . The findings suggest that WR treatment increases resistance to infection in immunocompromised hosts.

Burns, 2000 Mar, 26(2), 178 - 80
Bacteriology of burn wounds in Enugu, Nigeria; Ozumba UC et al.; A retrospective study of bacterial infection in 71 burned patients over a 5-year period (1993-1997) was carried out . The commonest colonizing organism was Klebsiella species (26.7%) followed by Staph aureus (25.6%) . There was a very high degree of resistance by these organisms to commonly available antibiotics in Nigeria, with the result that more expensive antibiotics such as the cephalosporins were required . The poor socioeconomic condition of most of the patients was a very important pre-disposing factor to burn wound infection, as only 25% of patients were able to afford the cost of wound microscopy and culture, thus leading to limited numbers of cultures being performed, the result being their prescription of antibiotics was made generally on an empirical basis . Restriction in the misuse of antibiotics and establishment of an infection control until will help to lower the incidence of infection.

Eur J Clin Microbiol Infect Dis, 2000 Jan, 19(1), 64 - 8
Epidemiology of quinolone resistance of Klebsiella pneumoniae and Klebsiella oxytoca in Europe; Brisse S et al.; The epidemiology of quinolone resistance and the concomitant resistance to other antibiotic classes was investigated in 445 Klebsiella pneumoniae and 238 Klebsiella oxytoca isolates . Decreased susceptibility to ciprofloxacin was found in 7.2% and 3.4% of these two species, respectively . Ciprofloxacin resistance was significantly linked to ceftazidime resistance, the hallmark of extended-spectrum beta-lactamase production, as well as to resistance to all antibiotic classes tested . Using automated ribotyping, seven intrahospital- and interhospital-transmitted clones of ciprofloxacin-resistant isolates were found . The newer fluoroquinolones sitafloxacin and clinafloxacin may become increasingly valuable, since they proved to be active also against ciprofloxacin-resistant isolates.

Diagn Microbiol Infect Dis, 2000 Feb, 36(2), 113 - 7
Screening and confirmatory testing for extended spectrum beta-lactamases (ESBL) in Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca clinical isolates; Hadziyannis E et al.; Escherichia coli and Klebsiella spp . were screened for ESBL based on routine susceptibility testing results . Isolates with intermediate or resistant susceptibilities for extended spectrum cephalosporins or aztreonam were reported as probable ESBL producers . By using the NCCLS proposed ESBL confirmatory method, we tested 61 screen-positive isolates from 42 patients, 30 randomly selected susceptible isolates, and 12 isolates with previously characterized beta-lactamases . Ceftazidime contributed to 97% of screen-positive isolates, whereas aztreonam added a single patient isolate . An ESBL was confirmed in 86% of K . pneumoniae, 100% of K . oxytoca, and 20% of E . coli screen-positive single patient isolates . None of the susceptible isolates were shown to produce ESBL . Based on these findings a comment regarding the presence of ESBL seems sufficient for Klebsiella spp . but confirmatory testing is indicated for E . coli . 0.25 microg/mL was used to indicate the presence of ESBL, the specificity of the assay increased to 100% . The NCCLS ESBL phenotypic confirmatory method was reproducible and accurate enough to be used in the clinical laboratory.

J Postgrad Med, 1998 Oct-Dec, 44(4), 93 - 6
Effect of honey on multidrug resistant organisms and its synergistic action with three common antibiotics; Karayil S et al.; A total of 15 bacterial strains (7 Pseudomonas & 8 Klebsiella species) isolated from various samples which showed multi-drug resistance were studied to verify in vitro antibacterial action of honey on the principle of Minimum Inhibitory Concentration (MIC) & its synergism with 3 common antibiotics--Gentamicin, Amikacin & Ceftazidime . The MIC of honey with saline for both organisms was found to be 1:2 . The synergistic action was seen in the case of Pseudomonas spp . and not with Klebsiella spp.

Kansenshogaku Zasshi, 2000 Jan, 74(1), 24 - 9
{Extended-spectrum beta-lactamase (ESBL) produced by Escherichia coli and Klebsiella pneumoniae isolated from Teikyo University Hospital--the second report}; Kawakami S et al.; We studied the high-level resistant to cefotaxime (CTX, MIC > or = 512 micrograms/ml) clinical isolates of Escherichia coli and Klebsiella pneumoniae in Teikyo University Hospital . The CTX-resistance could be transferred to E . coli K-12 chi 1037 or ML4903 strains from 30 of the 33 isolates by conjugation at a frequency of 10(-4) . When the hydrolysis rate of benzylpenicillin was 100%, the beta-lactamases which were extracted from the transconjugants hydrolyzed CTX, CAZ and AZT at the rate of 38-95%, 0-8.6% and 0-56%, respectively . These results demonstrate that these enzymes should be categorized into ESBL . The nucleotide sequence of CTX-resistant gene was identified to that of the CTX-M2 gene which was first described in Argentina . It was found to have 99.9% homology to Toho-1 gene in Japan and 99.6% homology to CMY-2 gene . Using a PCR methods for the detection of one of ESBL gene such as CTX-M2, Toho-1 or CMY-2, the DNA was amplified from all strains (11 isolates of E . coli and 21 isolates of K . pneumoniae).

Int J Infect Dis, 2000, 4(1), 21 - 5
Ceftazidime-resistant Klebsiella pneumoniae bloodstream infection in children with febrile neutropenia; Ariffin H et al.; OBJECTIVES: To evaluate prevalence of ceftazidime-resistant Klebsiella pneumoniae (CRKP) in the pediatric oncology unit of University Hospital, Kuala, Lumpur, and to identify differences between febrile neutropenic pediatric patients with CRKP and ceftazidime-sensitive K . pneumoniae (CSKP) bacteremia . MATERIALS AND METHODS: Febrile neutropenic patients treated between January 1996 and December 1997 at the pediatric oncology unit of University Hospital, Kuala Lumpur, were prospectively studied . Empirical antibiotic therapy consisted of ceftazidime and amikacin . Those who developed K . pneumoniae bacteremia were identified, and clinical features analyzed . Ceftazidime-resistance was documented via disk-diffusion testing . Production of extended-spectrum beta-lactamase (ESBL) was inferred on the basis of synergy between ceftazidime and amoxicillin-clavulanic acid . The different features between the two groups and variables associated with the development of CRKP bacteremia were analyzed using chi-square and t-tests and calculation of odds ratios . A multivariate analysis was used to identify independent factors for CRKP development . RESULTS: Ceftazidime-resistance was seen in 51.6% of all K . pneumoniae isolates, and all these isolates were inferred to be ESBL producers . All isolates were sensitive to imipenem . Susceptibility to gentamicin was 90.5% . The mean continuous hospital stay prior to the detection of bacteremia was 13.7 days overall, but significantly longer in the CRKP group (21.9 d) compared to the CSKP group (4.3 d) (P = 0.003) . Children with CRKP were more likely to have received antibiotics in the 2 weeks prior to detection of bacteremia (87.5% of cases) than the CSKP group (20.0% of cases) (P = 0.0008) . Sepsis-related mortality was higher in those with CRKP (50.0%) than in the CSKP group (13.3%) (P = 0.02) . Patients who did not receive CRKP-directed antibiotics within 48 hours of admission were more likely to have a fatal outcome than those who did (P = 0.009) . Logistic regression analysis identified use of third-generation cephalosporins 2 weeks prior to presentation and a hospital stay of 2 weeks or more as independent risk factors for development of CRKP . CONCLUSIONS: More than half of total K . pneumoniae isolated from blood cultures in the unit were ceftazidime-resistant . Children with febrile neutropenia with prolonged hospital stay and recent prior antibiotic exposure are at high risk of developing CRKP bacteremia . Mortality was significantly higher in this group . Early commencement of appropriate antibiotics (e.g., imipenem with or without gentamicin), according to susceptibility study results, may be beneficial in such circumstances.

FEMS Microbiol Lett, 2000 Mar 1, 184(1), 53 - 6
A preliminary survey of extended-spectrum beta-lactamases (ESBLs) in clinical isolates of Klebsiella pneumoniae and Escherichia coli in Japan; Yagi T et al.; We conducted a survey of extended-spectrum beta-lactamases (ESBLs) among 16805 Escherichia coli and 9794 Klebsiella pneumoniae clinical isolates recovered from 196 separate medical institutions during the period January 1997 to January 1998 . Using the criteria for minimal inhibitory concentrations (MICs) of oxyimino-cephalosporins of >/=8 microg ml(-1) and confirmation by double-disk test, we detected 15 E . coli and 34 K . pneumoniae isolates producing ESBLs . Genotypes of ESBLs determined by PCR with type-specific primers included one TEM-derived and 24 SHV-derived ESBLs, in addition to 24 Toho-1-type ESBLs, one of the major types of ESBLs reported in Japan . Nucleotide sequence analysis of SHV-specific PCR products revealed that SHV-12 was the dominant type of SHV-derived ESBL . In addition, we also identified TEM-26 and SHV-2 . This is the first report characterizing TEM- and SHV-derived ESBLs in Japan.

Infect Immun, 2000 Mar, 68(3), 1265 - 70
Febrile core temperature is essential for optimal host defense in bacterial peritonitis; Jiang Q et al.; Fever, a nonspecific acute-phase response, has been associated with improved survival and shortened disease duration in infections, but the mechanisms of these beneficial responses are poorly understood . We previously reported that increasing core temperature of bacterial endotoxin (LPS)-challenged mice to the normal febrile range modified expression of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), and IL-6, three cytokines critical to mounting an initial defense against microbial pathogens, but survival was not improved in the warmer animals . We speculated that our inability to show a survival benefit of optimized cytokine expression in the warmer animals reflected our use of LPS, a nonreplicating agonist, rather than an infection with viable pathogens . The objective of this study was to determine if increasing murine core temperature altered cytokine expression and improved survival in an experimental bacterial peritonitis model . We showed that housing mice at 35.5 degrees C rather than 23 degrees C increased core temperature from 36.5 to 37.5 degrees C to 39.2 to 39.7 degrees C, suppressed plasma TNF-alpha expression for the initial 48 h, delayed gamma interferon expression, improved survival, and reduced the bacterial load in mice infected with Klebsiella pneumoniae peritonitis . We showed that the reduced bacterial load was not caused by a direct effect on bacterial proliferation and probably reflected enhanced host defense . These data suggest that the increase in core temperature that occurs during bacterial infections is essential for optimal antimicrobial host defense.

J Perinatol, 1999 Jun, 19(4), 311 - 4
Successful treatment of late-onset infection due to resistant Klebsiella pneumoniae in an extremely low birth weight infant using ciprofloxacin; Khaneja M et al.; OBJECTIVE: This paper presents a case in which an extremely low birth weight infant with multidrug-resistant Klebsiella pneumoniae infection was successfully treated with ciprofloxacin and gentamicin . STUDY DESIGN: A clinical case report of a neonate who received broad spectrum antibiotics for possible infection despite negative cultures . The infant developed sepsis and meningitis resulting from multidrug-resistant K . pneumoniae, which was treated with ciprofloxacin and gentamicin . The literature for the use of ciprofloxacin in pediatric patients was reviewed . RESULTS: The infant responded to the antibiotic regimen with sterilization of blood and cerebrospinal fluid; no adverse effects were attributable to the ciprofloxacin . Although ciprofloxacin has been found to cause irreversible injury to cartilage in juvenile laboratory animals, a review of the literature found that this complication occurs rarely if at all in pediatric patients . Ciprofloxacin has been found to be effective in the treatment of multidrug-resistant Gram-negative infections in pediatric patients, including premature infants . CONCLUSION: Ciprofloxacin should be considered in the treatment of neonatal infection caused by multidrug-resistant Gram-negative organisms . Although the published experience with this drug suggests that it is effective and that significant toxicity is not common, its use should be restricted to the treatment of serious infections for which an alternative antibiotics is not available.

Int J Immunopharmacol, 2000 Jan, 22(1), 1 - 14
Androstenediol stimulates myelopoiesis and enhances resistance to infection in gamma-irradiated mice; Whitnall MH et al.; The ionizing radiation-induced hemopoietic syndrome is characterized by defects in immune function and increased mortality due to infections and hemorrhage . Since the steroid 5-androstene-3beta, 17beta-diol (5-androstenediol, AED) modulates cytokine expression and increases resistance to bacterial and viral infections in rodents, we tested its ability to promote survival after whole-body ionizing radiation in mice . In unirradiated female B6D2F1 mice, sc AED elevated numbers of circulating neutrophils and platelets and induced proliferation of neutrophil progenitors in bone marrow . In mice exposed to whole-body (60)Co gamma-radiation (3 Gy), AED injected 1 h later ameliorated radiation-induced decreases in circulating neutrophils and platelets and marrow granulocyte-macrophage colony-forming cells, but had no effect on total numbers of circulating lymphocytes or erythrocytes . In mice irradiated (0, 1 or 3 Gy) and inoculated four days later with Klebsiella pneumoniae, AED injected 2 h after irradiation enhanced 30-d survival . Injecting AED 24 h before irradiation or 2 h after irradiation increased survival to approximately the same extent . In K . pneumoniae-inoculated mice (irradiated at 3-7 Gy) and uninoculated mice (irradiated at 8-12 Gy), AED (160 mg/kg) injected 24 h before irradiation significantly promoted survival with dose reduction factors (DRFs) of 1.18 and 1.26, respectively . 5-Androstene-3beta-ol-17-one (dehydroepiandrosterone, DHEA) was markedly less efficacious than AED in augmenting survival, indicating specificity . These results demonstrate for the first time that a DHEA-related steroid stimulates myelopoiesis, and ameliorates neutropenia and thrombocytopenia and enhances resistance to infection after exposure of animals to ionizing radiation.

J Med Microbiol, 2000 Feb, 49(2), 115 - 9
Expression of putative virulence factors by clinical isolates of Klebsiella planticola; Podschun R et al.; A total of 92 clinical isolates of Klebsiella planticola from man was examined with respect to the production of haemagglutinins and siderophores, serum resistance and distribution of capsular types . For comparison, a group of 207 clinical isolates of K . pneumoniae was also studied . The percentages of K . planticola strains able to express mannose-sensitive haemagglutination, indicating type 1 fimbriae (83%) and mannose-resistant and Klebsiella-like agglutination, indicating type 3 fimbriae (69%), as well as to produce the siderophores enterobactin (100%) and aerobactin (2.2%) were almost identical to those of the K . pneumoniae strains . Similarly, the proportion of serum-resistant strains (30%) was comparable to that of K . pneumoniae (25%) . The capsule types most often detected in K . planticola were K14 (13%), K2 (9%) and K70 (9%) . The incidence of K2, which is the predominant capsular type in K . pneumoniae, was similar in both species . These findings show that K . planticola, which is being detected with increasing frequency in clinical specimens from man, has the ability to express similar putative virulence factors to K . pneumoniae, suggesting that they may have similar pathogenicity.

Immunopharmacology, 2000 Jan, 46(1), 1 - 13
The immunostimulating activities of anti-tumor polysaccharide from K1 capsular (polysaccharide) antigen isolated from Klebsiella pneumoniae; Ho CY et al.; We have previously shown that K1 capsular polysaccharide antigen (K1CPS) of Klebsiella exhibits anti-tumor activities . In the present study, we examined the effect of K1CPS on cytotoxic effector cells . We found that K1CPS could activate many cytotoxic effector cells including alloreactive cytotoxic T cells and tumor-infiltrating lymphocytes (TILs) . Moreover, K1CPS could increase the anti-tumor activity of lymphokine-activated killer (LAK) cells, both in vitro and in vivo . The i.p . injection of K1CPS in low dose could enhance the LAK cytotoxicity and the effect was further potentiated by coculture of LAK cells with K1CPS and low concentration of murine rIL-2 in vitro . The phenotypic characterization revealed that K1CPS might contribute to the increase in CD3+ LAK cell subpopulation by its in vivo priming effect . In addition, the K1CPS-treated LAK cells were able to inhibit the growth of WEHI-164 tumor cells in vivo in Winn-type inhibition assay . Subcutaneous (s.c.) and intraperitoneal (i.p.) adoptive infusion of LAK cells (splenocytes from K1CPS-treated WEHI-164-bearing mice cultured with K1CPS-plus-rIL-2) into WEHI-164 sarcoma-bearing mice could slightly cause regression in terms of tumor diameter, and more significantly in sarcoma weight.

Biosci Biotechnol Biochem, 1999 Dec, 63(12), 2064 - 8
Production of an extracellular polysaccharide bioflocculant by Klebsiella pneumoniae; Nakata K et al.; Klebsiella pneumoniae H12 produced a newly identified extracellular polysaccharide in an ethanol medium with a yield of 3.0 g/l . The molar composition of the polysaccharide was 56.04% galactose, 25.92% glucose, 10.92% galacturonic acid, 3.71% mannose, and 3.37% glucuronic acid . The addition of 0.5%-1.5% NaCl increased production . The polysaccharide flocculated with kaolin clay in suspension at the concentration of 1 ppm in a 300-ppm solution of CaCl2 . Almost all bacterial species cells aggregated in the polysaccharide solution . The ability to flocculate with kaolin clay changed with the pH and with the concentrations of coexisting cation and anion species . The polysaccharide flocculant may participate in in vivo bacterial aggregation or adherence to host organisms.

J Antimicrob Chemother, 2000 Feb, 45(2), 183 - 9
Antimicrobial resistance amongst Klebsiella spp . collected from intensive care units in Southern and Western Europe in 1997-1998; Babini GS et al.; A 1994 survey of 35 intensive care units (ICUs) in Western and Southern Europe found extended-spectrum beta-lactamases (ESBLs) in 220/966 (23%) klebsiellae . A follow-up survey from May 1997 to October 1998 collected klebsiellae from 24 ICUs, including 23 that participated in 1994 . Twenty-one ICUs sent 433 eligible isolates, of which 110 (25%) had ESBLs . The prevalence of ESBLs had not changed significantly from 1994 but the proportion of ESBL-producers resistant to piperacillin/tazobactam had risen from 31% to 63% (P < 0.001), and most of this resistance was high level (MICs >/= 128 + 4 mg/L) . The proportion of Klebsiella oxytoca isolates hyperproducing K1 beta-lactamase rose from 8% in 1994 to 21% in 1997-1998 (P < 0 . 001) . Most klebsiellae (99%) were very susceptible to meropenem (mode MIC 0.03 mg/L) but three had decreased susceptibility (MICs 2-4 mg/L) . These could not hydrolyse carbapenems . Aminoglycoside resistance was not significantly changed in prevalence from 1994; ciprofloxacin resistance occurred in 31% of ESBL-producers in both years, but had increased among non-producers (2% in 1994 versus 7% in 1997-1998, P < 0.001).

Appl Environ Microbiol, 2000 Feb, 66(2), 783 - 7
Immunolocalization of dinitrogenase reductase produced by Klebsiella pneumoniae in association with Zea mays L; Chelius MK et al.; The endophytic lifestyle of Klebsiella pneumoniae is described, including the production of dinitrogenase reductase by bacteria residing in maize root tissue . The green fluorescent protein (GFP) was used to detect the colonization of maize by K . pneumoniae strains 2028 and 342 . These strains were found to reside in intercortical layers of the stem and within the region of maturation in the root . The production of dinitrogenase reductase by GFP-tagged bacteria was visualized using immunolocalization . This activity was only apparent when bacteria were supplied with an exogenous carbon source . The results suggest that maize provides a suitable habitat for K . pneumoniae and that this species is capable of producing nitrogenase under the appropriate plant cultivation conditions.

Mol Microbiol, 2000 Jan, 35(2), 428 - 34
Na+ translocation by complex I (NADH:quinone oxidoreductase) of Escherichia coli; Steuber J et al.; Following on from our previous discovery of Na+ pumping by the NADH:ubiquinone oxidoreductase (complex I) of Klebsiella pneumoniae, we show here that complex I from Escherichia coli is a Na+ pump as well . Our study object was the Escherichia coli mutant EP432, which lacks the Na+/H+ antiporter genes nhaA and nhaB and is therefore unable to grow on LB medium at elevated Na+ concentrations . During growth on mineral medium, the Na+ tolerance of E . coli EP432 was influenced by the organic substrate . NaCl up to 450 mM did not affect growth on glycerol and fumarate, but growth on glucose was inhibited . Correlated to the Na+ tolerance was an increased synthesis of complex I in the glycerol/fumarate medium . Inverted membrane vesicles catalysed respiratory Na+ uptake with NADH as electron donor . The sodium ion transport activity of vesicles from glycerol/fumarate-grown cells was 40 nmol mg-1 min-1 and was resistant to the uncoupler carbonyl-cyanide m-chlorophenylhydrazone (CCCP), but was inhibited by the complex I-specific inhibitor rotenone . With an E . coli mutant deficient in complex I, the Na+ transport activity was low (1-3 nmol mg-1 min-1), and rotenone was without effect.

Adv Perit Dial, 1998, 14, 80 - 2
Temporary stoppage of peritoneal dialysis when laparoscopic procedures are performed on patients undergoing CAPD/CCPD: a change in policy; Goel S et al.; Laparoscopic surgery has been gaining in popularity . Prior to 1995, whenever our patients had a laparoscopic procedure it was carried out with the standard precautions, and the patient stayed on peritoneal dialysis (PD) . We noticed a high incidence (71.4%) of peritonitis post laparoscopic surgery . Four of the peritonitis patients had cholecystectomies and 1 had a gastrostomy for placement of a feeding tube laparoscopically . The patients' ages ranged from 31 to 74 years; 4 (80%) were females . They had been on PD for a mean of 24 months (range 6-48 months) . Peritonitis was diagnosed within a mean of 22.2 hours (r: 12-36 h) post procedure . The causative organisms were Klebsiella in two cases, Staph . aureus in one, Pseudomonas in one, with no growth observed in one . Following this, we instituted a deliberate policy of temporary cessation of PD for 2 weeks post laparoscopic procedure, maintaining the patient on hemodialysis during this period . Since the adoption of this policy, we have had no peritonitis in 7 patients who have undergone laparoscopic procedures.

J Nurs Care Qual, 2000 Jan, 14(2), 57 - 62
Reevaluating practice changes: return to standard precautions after an outbreak of multidrug-resistant Klebsiella oxytoca; Giramonti KM et al.; Change in medical practice is usually made to solve immediate problems . The continued use of these changes should then be reevaluated . Following an outbreak of K . oxytoca in our Neonatal Intensive Care Unit (NICU), glove use for all patient contact was mandated . We evaluated the need for continued glove use after resolution of the outbreak . No change in colonization patterns was seen after returning to standard precautions, and, as a secondary benefit, financial savings resulting from decreased glove use was realized . Following implementation of any practice change, routine reevaluation will help to determine when that change is no longer needed or beneficial.

Antimicrob Agents Chemother, 2000 Feb, 44(2), 453 - 5
Characterization of TEM-56, a novel beta-lactamase produced by a Klebsiella pneumoniae clinical isolate; Neuwirth C et al.; TEM-56 produced by a Klebsiella pneumoniae clinical isolate is a novel beta-lactamase of isoelectric point 6.4 that confers a moderate resistance level to expanded-spectrum cephalosporins . The amino acid sequence deduced from the corresponding bla gene showed two amino acid replacements with respect to the TEM-2 sequence: Glu-104 to Lys and His-153 to Arg . This enzyme showed catalytic properties close to those of TEM-18 . Thus, TEM-56 appears as a new TEM mutant, an intermediary between TEM-18 and the extended-spectrum beta-lactamase TEM-21.

Antimicrob Agents Chemother, 2000 Feb, 44(2), 362 - 7
High-level expression of chromosomally encoded SHV-1 beta-lactamase and an outer membrane protein change confer resistance to ceftazidime and piperacillin-tazobactam in a clinical isolate of Klebsiella pneumoniae; Rice LB et al.; We describe Klebsiella pneumoniae 15571, a clinical isolate resistant to ceftazidime MIC = 32 microg/ml) and piperacillin-tazobactam (MICs = 1,024 and 128 microg/ml) . K . pneumoniae 15571 expresses a single beta-lactamase with a pI of 7.6 . However, when cloned in a high-copy-number vector in Escherichia coli, this bla(SHV-1) gene did not confer resistance to ceftazidime, a spectrum consistent with the nucleotide sequence, which was nearly identical to those of previously described bla(SHV-1) genes . Outer membrane protein (OMP) analysis of K . pneumoniae 15571 revealed a decrease in the quantity of a minor 45-kDa OMP in comparison to that in K . pneumoniae 44NR, a low-level ampicillin-resistant strain that also expresses a chromosomally determined bla(SHV-1) . Crude beta-lactamase enzyme extracts from K . pneumoniae 15571 produced roughly 200-fold more beta-lactamase activity than K . pneumoniae 44NR . Northern hybridization analysis revealed that this difference was explainable by quantifiable differences in transcription of the bla(SHV-1) gene in the two strains . Primer extension analysis of bla(SHV-1) mRNA from K . pneumoniae 15571 and 44NR indicated that the transcriptional start sites were identical in the two strains . DNA sequencing of the promoter regions upstream of the of bla(SHV-1) open reading frames in the two K . pneumoniae strains revealed an A-->C change in the second position of the -10 region in K . pneumoniae 44NR compared to that in 15571 . Site-directed mutagenesis of the cloned K . pneumoniae 15571 bla(SHV-1), in which the A in the second position of the 15571 -10 region was changed to a C, resulted in a substantial lowering of the MIC of ampicillin . When the levels of beta-lactamase enzyme expression in E . coli were compared, the bla(SHV-1) downstream of the altered -10 region produced 17-fold less beta-lactamase enzyme . These results indicate that elevated levels of ceftazidime resistance can result from a combination of increased enzyme production and minor OMP changes and that levels of chromosomally encoded SHV-1 beta-lactamase production can vary substantially with a single-base-pair change in promoter sequence.

Infect Immun, 2000 Feb, 68(2), 953 - 5
Capsular polysaccharide is a major complement resistance factor in lipopolysaccharide O side chain-deficient Klebsiella pneumoniae clinical isolates; Alvarez D et al.; We have previously demonstrated the existence of Klebsiella pneumoniae clinical isolates deficient in the lipopolysaccharide O side chain, the major factor for resistance to complement-mediated killing in this bacterial species . These isolates are complement resistant, and their mechanisms to resist complement were investigated by selecting transposon-generated complement-sensitive mutants . One mutant with a drastically reduced capacity to grow in nonimmune human serum carried the transposon inserted in an open reading frame of a gene cluster involved in capsule synthesis . This mutant produced less capsule, bound more molecules of the complement component C3, and was more sensitive to complement-mediated and opsonophagocytic killings than was the parent strain . Four additional clinical isolates representing four different K serotypes were studied, and results showed that capsular polysaccharide is a major complement resistance factor in these O side chain-deficient isolates.

Eur J Surg, 1999 Dec, 165(12), 1159 - 61
Choledochal cysts in children: a review of 28 years of treatment in a Dutch children's hospital; Postema RR et al.; OBJECTIVE: To analyse the results of surgical treatment of choledochal cysts . DESIGN: Retrospective study . SETTING: Children's hospital, The Netherlands . PATIENTS: 14 children presenting with choledochal cysts . INTERVENTIONS: Choledochoduodenostomy, Roux-en-Y choledochojejunostomy or Roux-en-Y hepaticojejunostomy . MAIN OUTCOME MEASURES: Morbidity and mortality RESULTS: The mean age of the patients was 20 months (2 weeks to 7 years) . 10 patients had a type I choledochal cyst; three a type IV, and one a type V . Mean follow-up period was 6 years (18 months to 16 years) . One patient with a type I cyst died of Klebsiella pneumoniae that was resistant to treatment . One patient with a type I cyst treated by choledochojejunostomy had two episodes of cholangitis . Another patient with a type I cyst, treated by choledochoduodenostomy, had one episode of cholangitis . Both could be treated with antibiotics . The other patients had had no complications up to 1997 . CONCLUSION: This rare anomaly may lead to severe complications when left untreated or after late treatment . It is easy to manage with low associated morbidity.

What Is Genetic Engineering?, What Is Bioreactor?, What Is Antibiotic?, What Is Protein?, What Is Molecular Microbiology?, a, Microbe, i, Microorganism, c, Bacteria, n, Bacteriology, i, Microorganisms, a, Denitrifying, r, Yeasts, o, Antimicrobial, s, Pseudomonas aeruginosa, a, Neisseria, e, Streptococci, a, Microbiological, s, Fermentations, e, Yeasts, i, Escherichia coli, o, Rhodococci, n, Neisseria, s, Shigella, s, Escherichia coli, c, Shigella, n, Escherichia coli, o, Pichia, a, Staphylococcus aureus, r, Escherichia coli, a, Microbial, n, Antimicrobial

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005