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