J Hosp Infect, 1995 Dec, 31(4), 247 - 52 Sequential emergence of multi-resistant Klebsiella pneumoniae in Bahrain; Wallace MR et al.; During the mid-1980s, nosocomial infections due to aminoglycoside-resistant Klebsiella pneumonia were prevalent in the intensive care unit (ICU) of the Salamanya Medical Centre, Bahrain . In an attempt to control the spread of such organisms, the third-generation cephalosporins were introduced in early 1987 . Subsequently there was a marked increase in the incidence of cephalosporin resistance among Klebsiella spp . isolated in the ICU . In 1990, over 60% of Klebsiella isolates were resistant to both cephalosporins and aminoglycosides . Cephalosporin resistance was due to production of extended-spectrum beta-lactamases encoded on the same plasmid as aminoglycoside resistance . The incidence of cephalosporin resistance declined during 1991-1992, which was coincident with severe restrictions on the use of third-generation cephalosporins and the preferential use of ciprofloxacin and imipenem for nosocomial klebsiella infections . Sequential overuse of aminoglycosides and cephalosporins for nosocomial klebsiella infection may select for organisms resistant to both classes of antibiotics. Clin Infect Dis, 1995 Dec, 21(6), 1501 - 3 An outbreak of infusion-related Klebsiella pneumoniae bacteremia in a liver transplantation unit; Goetz AM et al.; Over a 1-month period, there were five episodes of infusion-related Klebsiella pneumoniae bacteremia in four liver transplantation patients housed in the same ward . Investigation of nursing practices revealed that a common normal-saline bag, to which intravenous (iv) tubing and a stopcock were attached, was used to flush iv catheters . The iv tubing and stopcock were changed at sporadic intervals . Cultures of the normal saline and iv equipment yielded K . pneumoniae, which had the same susceptibility pattern as the patients' isolates . Isolates recovered during the outbreak from the patients and from the iv saline/equipment were of the same strain, as determined by pulsed-field electrophoresis of Xba I-digested genomic DNA . Termination of the practice of flushing iv catheters with a common normal-saline bag halted the outbreak. Eur J Nucl Med, 1995 Dec, 22(12), 1411 - 5 Imaging of pulmonary disease in rheumatoid arthritis using J001X scintigraphy: preliminary results; Goupille P et al.; The purpose of this study was to determine the ability of technetium-99m J001X scintigraphy to image active pulmonary involvement in patients suffering from rheumatoid arthritis (RA) . J001X is a fully characterized acylated poly(1,3)galactoside, isolated from Klebsiella membranes, which is able to bind recruited macrophages after aerosol administration . J001X scintigraphy was compared with high-resolution computed tomography (HRCT), pulmonary function tests (PFTs) and bronchoalveolar lavage (BAL) in 15 patients suffering from RA . Patients were considered to have pulmonary involvement when they had an interstitial syndrome on HRCT and a decrease of 20% in TCO/VE (transfer coefficient) on PFTs and/or an abnormal BAL (lymphocytosis higher than 20% and/or percentage of neutrophils higher than 10%) . Pulmonary involvement was present in eight patients, and absent in seven . Of the eight patients with pulmonary involvement, all had abnormal BAL, two had an interstitial syndrome on HRCT, two had decreased TCO/VE and three had positive J001X scintigraphy . Of the seven patients without pulmonary involvement, six had normal BAL (not available in one), two had an interstitial syndrome on HRCT, one had decreased TCO/VE and two had positive J001X scintigraphy . According to our gold standard of pulmonary involvement, the sensitivity of J001X scintigraphy for the detection of pulmonary involvement in RA was 37.5%, the specificity was 71.4% and the positive predictive value was 60% . The ability of J001X scintigraphy to detect active pulmonary involvement during RA appears unclear in this study but it may detect processes unnoticed by the other modalities . These patients will be followed 12 and 24 months later and the changes in J001X scintigraphy, HRCT and PFTs will be compared to demonstrate whether J001X scintigraphy is able to assess an active process in the pulmonary involvement during RA and to specify its predictive value. J Formos Med Assoc, 1995 Dec, 94(12), 713 - 8 Bacteremia in acute pancreatitis of different etiologies; Chang KK et al.; To evaluate the rationale of using antibiotics in acute pancreatitis and to determine whether the indication for their use depends upon the etiology of the pancreatitis, the records of 202 patients with acute pancreatitis were retrospectively reviewed . The incidence of abnormal body temperature, leukocytosis, bacteremia and the results of biochemistry tests in different etiologies of the disease were investigated . Pancreatitis was found to be alcohol-related (47 patients), gallstone-related (105 patients), idiopathic (26 patients) and miscellaneous (24 patients) . On admission, 83 patients had abnormal body temperature and 146 patients showed leukocytosis . Bacteremia occurred in 20 patients . Of these, 15 had gallstone-related pancreatitis, two had pancreatic cancers and one developed bacteremia after endoscopic retrograde cholangio-pancreatography (ERCP) . These 18 patients had abnormal biochemistry results (including high serum levels of direct bilirubin, alkaline phosphatase and gamma-glutamyltransferase) and dilated bile ducts on imaging studies, indicating biliary infections . The remaining two patients with bacteremia included one alcoholic patient and one patient with idiopathic pancreatitis . The most commonly involved pathogens were Escherichia coli and Klebsiella pneumoniae . In addition, eight patients (4%) developed secondary pancreatic infections during hospitalization; the blood cultures of seven of these patients were negative on admission . Although fever and leukocytosis are not good predictors of infection in acute pancreatitis our results showed that bacteremia is common in patients whose pancreatitis is related to gallstones, ERCP or pancreatic malignancy with obstructive jaundice . We recommend that antibiotics be used only in this subset of acute pancreatitis patients. Appl Environ Microbiol, 1995 Dec, 61(12), 4167 - 71 Polymer production by Klebsiella pneumoniae 4-hydroxyphenylacetic acid hydroxylase genes cloned in Escherichia coli; Gibello A et al.; The expression of Klebsiella pneumoniae hpaA and hpaH genes, which code for 4-hydroxyphenylacetic acid hydroxylase in Escherichia coli K-12 derivative strains, is associated with the production of a dark brown pigment in the cultures . This pigment has been identified as a polymer which shows several of the characteristics reported for microbial melanins and results from the oxidative activity of 4-hydroxyphenylacetic acid hydroxylase on some dihydroxylated compounds to form o-quinones . A dibenzoquinone is formed from the oxidation of different mono- or dihydroxylated aromatic compounds by the enzyme prior to polymerization . We report a hydroxylase activity, other than tyrosinase, that is associated with the synthesis of a bacterial melanin. Gene, 1995 Dec 1, 166(1), 11 - 7 A new gene, cbl, encoding a member of the LysR family of transcriptional regulators belongs to Escherichia coli cys regulon; Iwanicka-Nowicka R et al.; The cbl (cysB-like) gene has been identified in Escherichia coli . The analysis of the cloned cbl sequence revealed strict homology to an ORF of unknown function found initially in Klebsiella aerogenes {Schwacha and Bender, J . Bacteriol . 175 (1993) 2107-2115} . The predicted Cbl protein has structural features of the LysR family of transcriptional activators . It is also strongly similar to the CysB protein, the activator of the cys regulon . The position of cbl on the Ec physical map has been established at a 2070-kb (43.5 min) region between asnU and asnV . The gene is expressed in vivo as a 1-kb monocistronic transcript starting from one major transcription start point . Unexpectedly, the in vivo expression of cbl has shown dependence on CysB, belonging to the same family of proteins . The promoter region of cbl binds purified CysB protein in a manner similar to other CysB-responsive promoters . A cbl disruption mutant was constructed by insertion of a KmR gene cartridge into the ORF on the chromosome . Phenotypes related to cbl expression suggest the involvement of the gene in an accessory regulatory circuit within the cys regulon engaging, in the last step, the function of the cysM gene encoding O-acetylserine (thiol)-lyase B. Carbohydr Res, 1995 Nov 30, 278(1), 103 - 11 Synthesis of the tetrasaccharide repeating unit of the antigen from Klebsiella type 2; Misra AK et al.; The disaccharide ethyl 2,4,6-tri-O-acetyl-3-O-(2,3,4,6-tetra-O-benzyl-alpha- D-glucopyranosyl)-1-thio-beta-D-glucopyranoside (6) and methyl 2,6-di-O-benzyl-3-O-(methyl 2,3,4-tri-O-benzyl-alpha-D- glucopyranosyluronate)-beta-D-mannopyranoside (21) have been synthesized and condensed in the presence of methyl triflate to afford a tetrasaccharide derivative . Removal of protecting groups gave methyl 3-O-(methyl alpha-D-glucopyranosyluronate)-4-O-(3-O-alpha-D- glucopyranosyl-beta-D-glucopyranosyl)-beta-D-mannopyranoside (23), the repeating unit of the antigen from Klebsiella type 2, in the form of its methyl ester methyl glycoside. Mol Microbiol, 1995 Nov, 18(3), 579 - 91 nasST, two genes involved in the induction of the assimilatory nitrite-nitrate reductase operon (nasAB) of Azotobacter vinelandii; Gutierrez JC et al.; An operon including two new genes (nasS and nasT) has been defined, cloned and sequenced . The deduced NASS protein is homologous to NRTA from Synechococcus sp . and to NASF from Klebsiella pneumoniae, two proteins involved in nitrate uptake . The predicted NAST polypeptide is homologous to the regulator proteins of the two-component regulatory systems . NASS plays a negative regulatory role in the synthesis of the nitrate and nitrite reductase . NAST is required for the expression of the nitrite-nitrate reductase operon (nasAB) . Expression of the nasST operon is not under the control of the NTR system and is not regulated by the nitrogen source . A Phi(nasA-lacZ) fusion has been used to analyse expression of the nasAB operon in three different genetic backgrounds with altered nitrate reductase activity . Beta-galactosidase activity in two of them was independent of nitrate but in a mutant unable to reduce nitrate, nas-4, it was normally induced by nitrate. Mol Microbiol, 1995 Nov, 18(3), 533 - 46 Regulation of anaerobic citrate metabolism in Klebsiella pneumoniae; Bott M et al.; Three enzymes are specifically required for uptake and catabolism of citrate by Klebsiella pneumoniae under anaerobic conditions: a Na+ -dependent citrate carrier (CitS), citrate lyase (CitDEF), and the Na+ pump oxaloacetate decarboxylase (OadGAB) . The corresponding genes are clustered on the chromosome, with the citCDEFG genes located upstream and divergent to the citS-oadGAB genes . We found that expression of citS from its native promoter in Escherichia coli requires the DNA region downstream of oadB . Nucleotide sequence analysis of this region revealed the presence of two adjacent genes, citA and citB . By sequence similarity, the predicted CitA and CitB proteins were identified as members of the two-component regulatory systems . The sensor kinase CitA contained, in the N-terminal half, two putative transmembrane helices which enclosed a presumably periplasmic domain of about 130 amino acids . The C-terminal half of the response regulator CitB harboured a helix-turn-helix motif typical of DNA-binding proteins . K . pneumoniae citB null mutants were unable to grow anaerobically with citrate as the sole carbon and energy source (Cit- phenotype) . When cultivated anaerobically with citrate plus glycerol, all of the citrate-specific fermentation enzymes were synthesized in the wild type, but not in the citB mutants . This showed that citS, oadGAB and citDEF required the CitB protein for expression and therefore are part of a regulon . In the wild type, synthesis of CitS, oxaloacetate decarboxylase and citrate lyase was dependent on the presence of citrate, sodium ions and a low oxygen tension . In a citA null mutant which expressed citB constitutively at high levels, none of these signals was required for the formation of the citrate fermentation enzymes . This result suggested that citrate, Na+, and oxygen exerted their regulatory effects via the CitA/CitB system . In the presence of these signals, the citAB gene products induced their own synthesis . The positive autoregulation occurred via co-transcription of citAB with citS and oadGAB. Proteins, 1995 Nov, 23(3), 370 - 5 Assessment of a protein fold recognition method that takes into account four physicochemical properties: side-chain packing, solvation, hydrogen-bonding, and local conformation; Matsuo Y et al.; A protein fold recognition method was tested by the blind prediction of the structures of a set of proteins . The method evaluates the compatibility of an amino acid sequence with a three-dimensional structure using the four evaluation functions: side-chain packing, solvation, hydrogen-bonding, and local conformation functions . The structures of 14 proteins containing 19 sequences were predicted . The predictions were compared with the experimental structures . The experimental results showed that 9 of the 19 target sequences have known folds or portions of known folds . Among them, the folds of Klebsiella aerogenes urease beta subunit (KAUB) and pyruvate phosphate dikinase domain 4 (PPDK4) were successfully recognized; our method predicted that KAUB and PPDK4 would adopt the folds of macromomycin (Ig-fold) and phosphoribosylanthranilate isomerase:indoleglycerol-phosphate synthase (TIM barrel), respectively, and the experimental structure revealed that they actually adopt the predicted folds . The predictions for the other targets were not successful, but they often gave secondary structural patterns similar to those of the experimental structures. Infection, 1995 Nov-Dec, 23(6), 371 - 7 Salicylate-enhanced exposure of Klebsiella pneumoniae subcapsular components; Salo RJ et al.; The capsular polysaccharide (CPS) of Klebsiella pneumoniae is an important virulence factor . Salicylate, which inhibits CPS production, was used to expose subcapsular antigens and components that may play an important role in host defense . Salicylate treatment greatly increased phagocytosis of five O1 serotypes by human polymorphonuclear leukocytes with normal rabbit serum and rabbit antisera against purified O1 lipopolysaccharide (O1LPS) as opsonins (p < 0.01 or < 0.05) . Similar results were obtained with rabbit antiserum against a non-encapsulated isogenic strain . To further determine how salicylate increases susceptibility to phagocytosis, the binding of monoclonal antibodies against O1LPS or the LPS core and the binding of complement component C3b were measured by ELISA . The data indicate that salicylate reduced the barrier of CPS in serotypes O1:K1, O1:K10, and O1:K16 and unmasked subcapsular antigenic components in serotypes O1:K2 and O1:K66 so that bound opsonins could react with receptors on phagocytes . Serum bactericidal assays supported this conclusion . Therefore, decapsulating agents such as salicylate accentuate phagocytosis of K . pneumoniae by making subcapsular antigens and components accessible to immune and nonimmune host defences and vaccination with subcapsular antigens may exhibit optimal protection against lethal infection when combined with salicylate therapy. Vet Microbiol, 1995 Nov, 47(1-2), 9 - 15 Plasmid profiles of Klebsiella pneumoniae isolated from bovine mastitis; Kikuchi N et al.; Plasmids of Klebsiella pneumoniae strains isolated from mastitic milk of cows were detected, and the epidemiological significance of their profiles was investigated . Of 47 K . pneumoniae isolates, 41 (87.2%) possessed plasmid(s) . Although the molecular size of the plasmids ranged from 1.9 to 140 megadaltons (Md), the 125 Md plasmid was predominant (31/41, 75.6%) . There was a great diversity in the plasmid profiles, however, no correlation was found between the plasmid profiles and capsular types of Klebsiella or the source of strains (dairy farms), except for one farm . All isolates obtained from cows on that farm possessed the 125 Md plasmid and their capsular type was 9. Appl Microbiol Biotechnol, 1995 Nov, 43(6), 985 - 8 Microbial production of specifically ring-13C-labelled 4-hydroxybenzoic acid; Muller R et al.; When transformed with a recombinant vector carrying the ubiC gene (encoding chorismate pyruvate-lyase, EC 4.1.3.27) the triple mutant (Phe-, Trp-, Tyr-) Klebsiella pneumoniae 62-1 excretes 4-hydroxybenzoic acid instead of chorismic acid . The recombinant strain can be used to produce in high yield specifically ring-labelled 4-hydroxybenzoic acid from isotopically labelled glucose. J Hosp Infect, 1995 Nov, 31(3), 177 - 87 Klebsiella pneumoniae infections in Greek hospitals . Dissemination of plasmids encoding an SHV-5 type beta-lactamase; Legakis NJ et al.; A total of 160 Klebsiella pneumoniae clinical strains consecutively isolated in 14 Greek hospitals in a three-month period was examined . Application of capsular typing using 72 monovalent antisera combined with phage-typing using a set of 15 Klebsiella-specific phages showed the absence of epidemic strains . However, 41% of the isolates examined displayed high level resistance to ceftazidime and aztreonam and, in most of the cases, to more than one aminoglycoside as well as to other antibacterial drugs . Nearly all of these multi-resistant strains were epidemiologically distinct on the basis of their capsular serotype and phage reactivity . After examination of 14 distinct strains, it was found that in nine cases, the resistance characters were readily transferred to Escherichia coli recipients through large self-transmissible plasmids (15-100 MDa) . Six of the nine plasmids had equal molecular weight (60 MDa) and displayed similar fragment profiles upon digestion with restriction endonuclease EcoRI . Isoelectric focusing and hydrolytic studies showed that the prominent beta-lactamase produced by the transconjugants harbouring the 60 MDa plasmids and the respective K . pneumoniae parent strains, was an extended-spectrum beta-lactamase of the SHV-5 type . It appears that among K . pneumoniae strains isolated in Greek hospitals a significant resistance rate to both newer beta-lactams and amino-glycosides has been established through the acquisition of promiscuous multi-resistant plasmids which share a high degree of similarity. J Bacteriol, 1995 Nov, 177(22), 6352 - 61 Lethality and survival of Klebsiella oxytoca evoked by conjugative IncN group plasmids; Rodriguez M et al.; The transmission of plasmid pCU1 (or other IncN group plasmid) into a population of Klebsiella oxytoca cells reduces the viability of the population . A 2,400-bp region adjacent to traA is responsible for this phenotype and includes two regions, called kikA and kikC . Klebsiella cells which received this region and survived were found to acquire a chromosomal mutation which renders them immune to killing even after the plasmid is cured from the cells . To obtain insight into the mode of this apparent lethality, an appropriate pCU1lacZ derivative was constructed . It could be introduced with high efficiency into Klebsiella cells . Analyses of the resultant colonies indicate that the loss of viability is not a consequence of the death of plasmid-free segregants . On the contrary and unlike postsegregational killing by plasmids, cells survived by losing the plasmid or by acquiring, secondarily, a chromosomal mutation which confers immunity to killing. Infect Immun, 1995 Nov, 63(11), 4336 - 44 Molecular characterization and adhesive properties of CF29K, an adhesin of Klebsiella pneumoniae strains involved in nosocomial infections; Di Martino P et al.; We previously described a CS31A-related protein, CF29K, expressed by Klebsiella pneumoniae strains involved in nosocomial infections . In this study, we cloned and sequenced cf29A, the structural gene of the CF29K protein, and showed that CF29K is an antigenic subtype of CS31A . The CF29K protein was found to be identical to the CS31A-L protein on the basis of biochemical and immunological properties . In contrast, the CS31A-H protein presented a different apparent molecular mass during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a different limited degradation pattern with endopeptidase V8, and a specific conformational epitope . We cloned and sequenced the CS31A-L structural gene and confirmed that CF29K and CS31A-L are identical, but their major subunits differ from ClpG (the CS31A-H subtype major subunit) by one amino acid at position 89 of the mature protein, which is a lysine in CF29K instead of the asparagine in ClpG . Site-directed mutagenesis experiments demonstrated that the biochemical and immunological differences between CS31A-H and CF29K or CS31A-L were dependent only on the amino acid at position 89 of the mature protein . To study the adhesive properties of CS31A-H and CF29K in the same Escherichia coli reference strain, we performed transcomplementation experiments with the cloned CS31A major-subunit structural genes or cloned cf29A gene and the clp accessory genes of the CS31A operon . We showed that CS31A-L, CF29K, and CS31A-H were involved in adhesion to Caco-2 and Int-407 cells but not to HEp-2 cells . Nevertheless, K . pneumoniae strains and corresponding E . coli transconjugants producing CF29K adhered to cultured Caco-2, Int-407, and HEp-2 cells, indicating the expression of another R-plasmid-encoded adhesin that mediated adhesion to HEp-2 cells . The carbohydrate part of the eucaryotic receptor of CF29K and CS31A-H adhesins was investigated by adhesion inhibition experiments with Int-407 cells . Although CS31A and CF29K belong to the K88 adhesin family, the receptor does not contain N-acetyl-D-galactosamine residues but contains N-acetylneuraminic acid and N-acetyl-D-glucosamine. Gene, 1995 Oct 16, 164(1), 89 - 94 A Klebsiella aerogenes moaEF operon is controlled by the positive MoaR regulator of the monoamine regulon; Azakami H et al.; A 30-kDa protein accumulated upon induction by a high concentration of tyramine or dopamine in cells of Klebsiella aerogenes (Ka) . These cells carried a plasmid (pAS123) that included the arylsulfatase operon (atsBA) . Deletion analysis showed that the region essential for induction of the 30-kDa protein was located within a 2.0-kb cloned segment downstream of the atsBA operon . The nucleotide (nt) sequence of the 2.0-kb fragment revealed two open reading frames (ORFs), moaE and moaF . Transcription from a putative promoter of moaE was induced by the addition of tyramine, and the moaF gene was co-transcribed from this monoamine-inducible Ka promoter . The deduced Ka MoaE protein was homologous to insect-type alcohol dehydrogenase . The sequence of the 18 amino acids from the N-terminus of the purified 30-kDa protein agreed with that deduced from the nt sequence of moaF . Using a Ka strain with a mutant moaR gene, we found that MoaR, that acts as the positive regulator of the monoamine regulon, also acts as the positive regulator of the moaEF operon. Carbohydr Res, 1995 Oct 2, 275(2), 371 - 9 The structure of the acidic exopolysaccharide produced by Pseudomonas "gingeri" strain Pf9; Cescutti P et al.; The structure of the acidic exopolysaccharide produced by the mushroom pathogen Pseudomonas "gingeri" strain Pf9, a bacterium which causes ginger blotch, was investigated by chemical analysis, mass spectrometry and 1D and 2D NMR spectroscopy . The polysaccharide consists of the linear trisaccharide repeating unit {formula: see text} where the cyclic pyruvic acetal groups at O-4 and O-6 of the mannopyranosyl residues have the S-configuration . Methylation analysis under neutral conditions and NMR data showed that the mannose residues are acetylated at O-2 . This exopolysaccharide has the same structure as the E . coli K55 capsular polysaccharide and differs from the Klebsiella K5 capsular polysaccharide only in the position of acetylation (C-2 of the glucopyranose residue). J Chemother, 1995 Oct, 7(5), 406 - 13 Effects of sub-minimal inhibitory concentrations of antimicrobial agents on the cell surface of Klebsiella pneumoniae and phagocytic killing activity; Nomura S et al.; Changes in the phagocytic killing activity, capsule structure, and physicochemical properties such as the hydrophobicity and charge of the cell surface were studied in Klebsiella pneumoniae treated with sub-minimal inhibitory concentrations (MICs) of various antimicrobial agents . The phagocytic killing activity of macrophages was enhanced by penicillins, cephems, and monobactam in the absence of antibodies specific to the capsule or complement . No enhancement was observed with new quinolones, aminoglycosides, macrolide, or carbapenem . The thickness of the capsule structure was considerably reduced after the treatment with penicillins, cephems, and monobactam compared with the untreated control, and it was slightly reduced by new quinolones . No changes were observed in the capsule structure with aminoglycosides, macrolide, and carbapenem . The hydrophobicity on the cell surface of the bacteria was considerably increased after the treatment with penicillins, cephems, and monobactam compared with the control, slightly increased with new quinolones and carbapenem, and not changed with aminoglycosides and macrolide . The negative charge of the cell surface of the bacteria was reduced by penicillins, cephems, and monobactam compared with the control . It was slightly reduced by new quinolones and carbapenem but was not reduced by aminoglycosides and macrolide . These findings suggest that sub-MIC beta-lactam drugs such as penicillins, cephems, and monobactams cause thinning of the capsule of K . pneumoniae with increases in the hydrophobicity and decreases in the negative charge of the cell surface, which reduces the physical repulsion between the K . pneumoniae and phagocytes and enhances the sensitivity of the bacteria to phagocytic killing activity. Lupus, 1995 Oct, 4(5), 375 - 89 Analysis of three new idiotypes on human monoclonal autoantibodies; Kalsi JK et al.; We have identified and characterised three new idiotypes on human IgM McAbs generated from the splenocytes of a SLE patient with active disease . RT-6, which binds H1 and Sm/RNP, expresses essentially a private Id . Its expression is limited to a small number of human McAbs and the sera from patients with infectious diseases . In contrast RT-72Id and RT-84Id, expressed on McAbs which are polyreactive for two or more antigens, have a public distribution . RT-72Id and RT-84Id are found on McAbs from murine and human adult, and foetal tissues . In sera, significant numbers of SLE, RA and patients with other autoimmune diseases are positive for both Ids . RT-84Id is also elevated in SLE relatives and spouses, and in patients with Klebsiella infection . No correlation with disease activity, IgM or IgG levels was observed with either Id . However, RT-72Id was significantly associated with anti-ssDNA antibodies and RhF . RT-6Id and RT-72Id are located on the framework regions of the mu heavy chain, whereas RT-84Id is present on the kappa light chain, within the binding site . The McAbs are encoded by mainly germline genes: heavy chains of RT-6, RT-72 and RT-84 are encoded by the genes VH26, VH4.22 and VH4.21, respectively, and the light chain sequences of RT-6 and RT-72 are derived from DPL11 and HK102 . Immunofluorescent staining revealed the presence of RT-72Id and RT-84Id positive immunoglobulin deposits in 18% and 45%, respectively, of the lupus renal sections compared with none in the disease control group, suggesting that these Ids may contribute to the pathology of the disease. Br J Haematol, 1995 Oct, 91(2), 471 - 3 The use of an anti-beta 2-glycoprotein-I assay for discrimination between anticardiolipin antibodies associated with infection and increased risk of thrombosis; McNally T et al.; Antiphospholipid antibodies (aPAs), occurring in association with infection, are not generally associated with an increased risk of thrombosis . Anticardiolipin antibodies (aCL) from patients with infection, unlike those from patients with SLE, do not have the beta 2GPI cofactor requirements . Antibodies to beta 2GPI (alpha beta 2GPI) are more closely associated with a previous history of thrombosis than aCL in patients with SLE . In the present study we have investigated the reactivity of the alpha beta 2GPI assay for aPAs associated with infection . Serum from 114 patients with infections including syphilis (n = 11), tuberculosis (n = 63) and Klebsiella (n = 42) were assayed for alpha beta 2GPI and aCL antibodies . The incidence of aCL in serum of patients with tuberculosis . Klebsiella infection and syphilis was 6.0%, 5.0% and 64.0%, respectively, but all patients were negative for alpha beta 2GPI . These results indicate that the alpha beta 2GPI assay is negative in patients with transiently positive aCL assays associated with infection. Protein Sci, 1995 Oct, 4(10), 2234 - 6 Urease activity in the crystalline state; Moncrief MB et al.; Crystalline Klebsiella aerogenes urease was found to have less than 0.05% of the activity observed for the soluble enzyme under standard assay conditions . Li2SO4, present in the crystal storage buffer at 2 M concentration, was shown to inhibit soluble urease by a mixed inhibition mechanism (Ki's of 0.38 +/- 0.05 M for the free enzyme and 0.13 +/- 0.02 M for the enzyme-urea complex) . However, the activity of crystals was less than 0.5% of the expected value, suggesting that salt inhibition does not account for the near absence of crystalline activity . Dissolution of crystals resulted in approximately 43% recovery of the soluble enzyme activity, demonstrating that protein denaturation during crystal growth does not cause the dramatic diminishment in the catalytic rate . Finally, crushed crystals exhibited only a three-fold increase in activity over that of intact crystals, indicating that the rate of substrate diffusion into the crystals does not significantly limit the enzyme activity . We conclude that urease is effectively inactive in this crystal form, possibly due to conformational restrictions associated with a lid covering the active site, and propose that the small amounts of activity observed arise from limited enzyme activity at the crystal surfaces or trace levels of enzyme dissolution into the crystal storage buffer. Biosci Biotechnol Biochem, 1995 Oct, 59(10), 1987 - 90 Molecular cloning and nucleotide sequence of purine nucleoside phosphorylase and uridine phosphorylase genes from Klebsiella sp; Takehara M et al.; Klebsiella sp . LF 1202 was isolated as a bacterium that can assimilate adenosine as a sole source of carbon and nitrogen {F . Ling et al., Agric . Biol . Chem., 55, 573-575 (1991)} from a soil sample . Both the purine nucleoside phosphorylase (PNPase) and uridine phosphorylase (UPase) of this bacterium were induced simultaneously when the bacterium was cultured in a medium containing adenosine or uridine as a sole source of carbon and nitrogen . This induction profile is different from that of Escherichia coli . Here we cloned and sequenced the gene corresponding to each enzyme . The open reading frame (ORF) of the PNPase gene consisted of 717 bp that encoded a polypeptide of 239 amino acids with a molecular weight of 26,198 . The ORF of the UPase gene consisted of 834 bp that encoded a polypeptide of 278 amino acids with a molecular weight of 28,912. J Bacteriol, 1995 Oct, 177(19), 5535 - 8 Repression of the Klebsiella aerogenes nac promoter; Feng J et al.; Transcription of the nitrogen-regulated nac promoter of Klebsiella aerogenes requires sigma54 RNA polymerase, is activated by the phosphorylated form of the transcription factor nitrogen regulator I (NRI) (NtrC), and is repressed by the product of the nac gene, Nac . Nac protects a large portion of the nac control region, extending from positions -130 to -70, from digestion by DNase I . This site(s) lies immediately upstream from the site at which sigma 54 RNA polymerase binds, is downstream of a high-affinity binding site for the transcriptional activator NRI approximately P, and partially overlaps a low-affinity NRI approximately P-binding site . Binding of Nac to the DNA resulted in bending of the DNA but did not interfere with the binding of sigma 54 RNA polymerase to the promoter or with the binding of NRI approximately P to either the high-affinity site or low-affinity site . Furthermore, transcription assays with various wild-type and mutant templates suggested that Nac did not exclude NRI approximately P from either the low- or high-affinity sites, nor did Nac interfere with the ability of the polymerase to form the open complex when the binding sites for NRI approximately P were moved to different locations upstream from the promoter . Rather, Nac seemed to repress by an antiactivation mechanism in which the interaction of the NRI approximately P, bound at its normal sites, with sigma 54 RNA polymerase, bound to the promoter, was prevented. J Bacteriol, 1995 Oct, 177(19), 5523 - 34 Activation of transcription initiation from the nac promoter of Klebsiella aerogenes; Feng J et al.; The nac gene of Klebsiella aerogenes encodes a bifunctional transcription factor that activates or represses the expression of several operons under conditions of nitrogen limitation . In experiments with purified components, transcription from the nac promoter was initiated by sigma 54 RNA polymerase and was activated by the phosphorylated form of nitrogen regulator I (NRI) (NtrC) . The activation of the nac promoter required a higher concentration of NRI approximately P than did the activation of the Escherichia coli glnAp2 promoter, and both the promoter and upstream enhancer element contributed to this difference . The nac promoter had a lower affinity for sigma 54 RNA polymerase than did glnAp2, and uninitiated competitor-resistant transcription complexes formed at the nac promoter decayed to competitor-sensitive complexes at a greater rate than did similar complexes formed at the glnAp2 promoter . The nac enhancer, consisting of a single high-affinity NRI-binding site and an adjacent site with low affinity for NRI, was less efficient in stimulating transcription than was the glnA enhancer, which consists of two adjacent high-affinity NRI-binding sites . When these binding sites were exchanged, transcription from the nac promoter was increased and transcription from the glnAp2 promoter was decreased at low concentrations of NRI approximately P . Another indication of the difference in the efficiency of these enhancers is that although activation of a nac promoter construct containing the glnA enhancer was relatively insensitive to subtle alterations in the position of these sites relative to the position of the promoter, activation of the natural nac promoter or a nac promoter construct containing only a single high-affinity NRI approximately P binding site was strongly affected by subtle alterations in the position of the NRI approximately P binding site(s), indicating a face-of-the-helix dependency for activation. Biol Pharm Bull, 1995 Sep, 18(9), 1259 - 63 Decrease in surface charge density of Klebsiella pneumoniae treated with cefodizime and enhancement of the phagocytic function of human polymorphonuclear leucocytes stimulated by the drug-treated bacteria; Muratsugu M et al.; Treatment of Klebsiella pneumoniae 109 with cefodizime, ceftazidime, cefbuperazone, cefotaxime or flomoxef at a sub-minimum inhibitory concentration (sub-MIC) (1/4 MIC) for 1 h altered its morphology . The bacteria treated with cefodizime at sub-MICs (1/8, 1/4 MIC) enhanced the chemiluminescencence (CL) of human polymorphonuclear leucocytes (PMNs), implying an increase in the production of active oxygen species in association with phagocytosis, whereas the cells treated with other cephalosporins at the same sub-MICs did not . Furthermore, a significant decrease in electrophoretic mobility was induced by the bacteria treated with cefodizime at sub-MICs (1/8, 1/4 MIC), but other cephalosporins neither increased nor decreased the electrophoretic mobility significantly . These findings suggested that cefodizime caused a morphological change, with a decrease in negative surface charge density of K . pneumoniae, more easily than ceftazidime, cefbuperazone, cefotaxime or flomoxef, followed by an increase in the phagocytic activity of PMNs. Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi, 1995 Sep-Oct, 36(5), 336 - 41 Ultrasound-guided percutaneous drainage of liver abscess in children; Ni YH et al.; This study's purpose was to investigate the effectiveness of ultrasound-guided percutaneous drainage (US-PD) as treatment modality for pediatric pyogenic liver abscess when compared with other modalities . Ten consecutive patients, aged from 2 months to 22 years, were enrolled in this study . In addition to antibiotics, US-PD was attempted in each case unless the procedure was judged unsuitable . Six patients were treated with US-PD while four were not, for various reasons . US-PD was performed under an intravenous anesthesia and with the aid of real-time sonography . An indwelling pigtail catheter was usually inserted during US-PD . Fever subsided within one to six days, but the abscess cavity closed later . Failure to respond to US-PD may relate to the huge size of abscess cavity; surgical drainage provides an option . A trend toward a shorter hospital stay and defervescence day was found in the US-PD group than in the non-US-PD group . Prognosis was generally good and none in this series died of this disease or the procedure . Klebsiella pneumoniae was the pathogen most frequently encountered . Seven of the ten patients had underlying disease, and hemoglobinopathy was frequently associated . In conclusion, adequate drainage is recommended as the most effective treatment modality for pediatric liver abscess, and US-PD is the first choice. Fundam Appl Toxicol, 1995 Sep, 27(2), 177 - 84 Local versus systemic immunotoxicity of isobutyl nitrite following subchronic inhalation exposure of female B6C3F1 mice; Ratajczak HV et al.; Female B6C3F1 mice were exposed to isobutyl nitrite (IBN) by inhalation at 0, 37.5, 75, or 150 ppm for 6 hr per day, 5 days per week for 15 weeks . The potential of this compound to induce immunotoxicity was assessed during the 3rd, 13th, 14th, and 15th week of exposure and after 2 weeks of recovery following the 15 weeks of exposure . Both systemic and lung immune functions were examined, including body and lymphoid organs weights, pulmonary macrophage function and host defense, expression of splenic lymphocyte cell-surface markers, natural killer cell function, mixed lymphocyte reaction, and induction of specific antibody to a T-cell-dependent antigen . There was a dose-related suppression of T-cell-dependent antibody-forming cell responses in the spleen following IBN exposure; however, other measures of T-cell and nonspecific immunity were not significantly affected . A dose-related increase of H202 production by alveolar macrophages was present after 12 but not after 68 exposures to IBN . In contrast, pulmonary host defense mechanisms against Klebsiella pneumoniae were unaffected . These results suggest that in the absence of changes in host resistance, IBN may have selective and partially reversible effects on the immune system. J Bacteriol, 1995 Sep, 177(18), 5261 - 9 Identification of the transcriptional activator controlling the butanediol fermentation pathway in Klebsiella terrigena; Mayer D et al.; The gene budR, whose product is responsible for induction of the butanediol formation pathway under fermentative growth conditions in Klebsiella terrigena, has been cloned and sequenced . This gene is separated from the budABC operon by a nontranslated region of 106 bp and transcribed in the opposite direction . budR codes for a protein of molecular weight 32,124, the sequence of which exhibits characteristics of regulators belonging to the LysR family . When transferred into the heterologous host Escherichia coli, budR activates expression of budA'-lacZ transcriptional and translational fusions with a regulatory pattern identical to that in K . terrigena, namely, induction by acetate, low pH, and anaerobiosis . Induction by acetate was specific, indicating that it is the physiological inducer . Primer extension analysis located the start site of transcription to two positions, 23 and 24 bp upstream of the budR initiation codon, and also showed that BudR strongly autoregulates its own expression . The products of fhlA, arcA, hip, ntrA, and katF did not influence expression of the bud operon . A mutation in fnr, however, led to a threefold increase in expression, indicating that Fnr acts as a repressor . The results support the notion that BudR coordinates the activity of the energy-conserving, nonreductive, but acidifying acetate formation pathway with the expression of the non-energy-conserving, reductive, but nonacidifying butanediol pathway. J Bacteriol, 1995 Sep, 177(17), 5088 - 98 Synthesis of pyrroloquinoline quinone in vivo and in vitro and detection of an intermediate in the biosynthetic pathway; Velterop JS et al.; In Klebsiella pneumoniae, six genes, constituting the pqqABCDEF operon, which are required for the synthesis of the cofactor pyrroloquinoline quinone (PQQ) have been identified . The role of each of these K . pneumoniae Pqq proteins was examined by expression of the cloned pqq genes in Escherichia coli, which cannot synthesize PQQ . All six pqq genes were required for PQQ biosynthesis and excretion into the medium in sufficient amounts to allow growth of E . coli on glucose via the PQQ-dependent glucose dehydrogenase . Mutants lacking the PqqB or PqqF protein synthesized small amounts of PQQ, however . PQQ synthesis was also studied in cell extracts . Extracts made from cells containing all Pqq proteins contained PQQ . Lack of each of the Pqq proteins except PqqB resulted in the absence of PQQ . Extracts lacking PqqB synthesized PQQ slowly . Complementation studies with extracts containing different Pqq proteins showed that an extract lacking PqqC synthesized an intermediate which was also detected in the culture medium of pqqC mutants . It is proposed that PqqC catalyzes the last step in PQQ biosynthesis . Studies with cells lacking PqqB suggest that the same intermediate might be accumulated in these mutants . By using pqq-lacZ protein fusions, it was shown that the expression of the putative precursor of PQQ, the small PqqA polypeptide, was much higher than that of the other Pqq proteins . Synthesis of PQQ most likely requires molecular oxygen, since PQQ was not synthesized under anaerobic conditions, although the pqq genes were expressed. J Bacteriol, 1995 Sep, 177(17), 5078 - 87 The C-terminal domain of NifL is sufficient to inhibit NifA activity; Narberhaus F et al.; In Klebsiella pneumoniae, transcription of all nif (nitrogen fixation) operons except the regulatory nifLA operon itself is regulated by the proteins NifA and NifL . NifA, an enhancer-binding protein, activates transcription by RNA polymerase containing the alternative sigma factor sigma 54 . The central catalytic domain of NifA is sufficient for transcriptional activation, which can occur from solution . In vivo, NifL antagonizes the action of NifA in the presence of molecular oxygen or combined nitrogen . Inhibition has also been shown in vitro, but it was not responsive to environmental signals . Assuming a two-domain structure of NifL, we localized inhibition by NifL to its carboxy (C)-terminal domain, which is more soluble than the intact protein . The first line of evidence for this is that internal deletions of NifL containing an intact C-terminal domain were able to inhibit transcriptional activation by NifA in a coupled transcription-translation system . The second line of evidence is that the isolated C-terminal domain of NifL (assayed as a fusion to the soluble maltose-binding protein {MBP}) was sufficient to inhibit transcriptional activation by the central domain of NifA in a purified transcription system . The final line of evidence is that an MBP fusion to the C-terminal domain of NifL inhibited transcriptional activation by NifA in vivo . On the basis of these data, we postulate that the inhibitory function of NifL lies in its C-terminal domain and hence infer that this domain is responsible for interaction with NifA . Gel filtration experiments with MBP-NifL fusion derivatives lacking portions of the N- or C-terminal domain of the protein revealed that the C-terminal domain is the most soluble part of NifL . Up to 50% of two MBP-NifL truncations containing only the C-terminal domain appeared to be in a defined dimeric state. Kansenshogaku Zasshi, 1995 Sep, 69(9), 1017 - 20 {A case of gas-containing liver abscess with multiple metastatic lesions}; Higashi T et al.; A 63-year-old male was admitted to our hospital because of high fever and delirium . He had been diagnosed as diabetes mellitus five years before but not treated at all . An abdominal CT scan showed gas-containing abscess in the right lobe of the liver . Klebsiella pneumoniae and Bacteroides distasonis were cultured both from the punctured specimen of the abscess and from arterial blood . Catheter drainage was carried out percutaneously under guidance with ultrasonography and antibiotics was administered intravenously . He was diagnosed as purulent meningitis by lumbar puncture on admission and as endophthalmitis because of swelling of the left eyeball on hospital day 4 . CT scan also showed multiple metastatic lesions in the cerebrum and in the lung . After three months, he was discharged from the hospital in good condition, except for loss of vision of the left eye. Arthritis Rheum, 1995 Sep, 38(9), 1277 - 82 Klebsiella pneumoniae-reactive T cells in blood and synovial fluid of patients with ankylosing spondylitis . Comparison with HLA-B27+ healthy control subjects in a limiting dilution study and determination of the specificity of synovial fluid T cell clones; Hermann E et al.; OBJECTIVE . To study the frequency of Klebsiella pneumoniae-responsive T cells in the peripheral blood (PB) of ankylosing spondylitis (AS) patients compared with that in healthy HLA-B27+ donors, and to examine T lymphocyte clones (TLC) derived from AS patient synovial fluid (SF) for the presence of Klebsiella reactivity . METHODS . Limiting dilution analysis of PB T cells in 8 patients with active AS and in 8 HLA-B27+ healthy subjects was used to determine the frequency of PB T cells responsive to K pneumoniae and Escherichia coli GroEL . SF T cells from a patient with active AS were cloned, and 125 TLC were characterized in proliferation assays . RESULTS . There were fewer T cells in the PB of AS patients that reacted with K pneumoniae than in the PB of healthy HLA-B27+ subjects . The frequencies of E coli GroEL-responsive T cells were approximately 5-10 times lower in all subjects tested (healthy donors and AS patients), but without significant differences between the 2 groups . Two CD4+ TLC that recognized K pneumoniae (1 cross-reactive with E coli) as well as 3 TLC that recognized GroEL (2 CD4+, 1 T cell receptor gamma/delta+) were isolated from the SF of a patient with actige AS . CONCLUSION . Our results indicate that there is a quantitative reduction of K pneumoniae-responsive T cells in the PB of AS patients as compared with healthy controls . This may reflect a defective peripheral T cell defense in the immune response to Klebsiella and may allow bacterial antigens to reach the synovium, where they initiate specific T cell responses. Epidemiol Mikrobiol Imunol, 1995 Sep, 44(3), 111 - 4 {Occurrence of Klebsiella pneumonia strains resistant to third-generation cephalosporins at the Pediatric Clinic of the Medical School Hospital in Olomouc}; Kolar M et al.; The authors describe the high incidence of strains of Klebsiella pneumoniae resistant to cephalosporins of the IIIrd generation at the department of pathological neonates of the Paediatric Clinic of the Faculty Hospital in Olomouc . They discuss possible reasons for this adverse situation and the way how to resolve it . They emphasize the necessity of rational administration of antibiotics and continuous monitoring of resistance in the most frequent causal agents of bacterial infections. Am J Vet Res, 1995 Sep, 56(9), 1213 - 8 Effect of phenylbutazone and flunixin meglumine on acute toxic mastitis in dairy cows; Dascanio JJ et al.; A double-blinded randomized prospective clinical trial was designed to evaluate the effectiveness of flunixin meglumine and phenylbutazone for treatment of acute toxic mastitis in dairy cows . All cows were treated 4 times at 12-hour intervals by intramammary infusion of gentamicin (150 mg) . A total of 45 dairy cows with toxic mastitis were randomly assigned to 1 of 3 treatment groups: group 1 (control), saline solution, IV; group 2, 1 g of flunixin meglumine, IV; or group 3, 4 g of phenylbutazone, IV . Physical examination and udder variables were assessed at initial examination and 24 hours later . Milk production was recorded at regular intervals from 1 week before until 10 weeks after development of mastitis . Rear quarters (34/45) were more commonly affected than front quarters . Thirty-five cows returned to the herd, 9 cows were culled, and 1 cow died . There were no significant differences among treatment groups in the need for further treatment or outcome . Klebsiella spp (18/45) and Escherichia coli (16/45) were the most common pathogens isolated by culture of milk from affected quarters . The overall bacteriologic cure rate on days 7 and 14 was 64 and 75%, respectively . At the time of initial examination, cows of the control group had higher rectal temperature than did cows of the flunixin group . At the examination 24 hours later, the rectal temperature of cows in all treatment groups was lower than the temperature at initial examination; at that time (24 hours), however, there were no significant differences in temperature among the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS) Carbohydr Res, 1995 Aug 25, 273(2), 197 - 205 Structure of the capsular polysaccharide from the Klebsiella K8 reference strain 1015; Erbing B et al.; The structure of the capsular polysaccharide from the Klebsiella K8 reference strain 1015 has been elucidated . The structure was deduced from sugar analysis, different methylation analyses, a uronic acid degradation, and NMR spectroscopy . It is concluded that the polysaccharide is composed of pentasaccharide repeating units with the structure: {formula: see text} The structure differs from that of the previously published structure of the capsular polysaccharide from Klebsiella K8, which originates from another strain and has the following structure: {formula: see text} The serological similarity between the two strains is most likely derived from a common tetrasaccharide which is substituted in different ways in the two strains . Since the strain in the present investigation originates from the Klebsiella K reference strain collection of the International Escherichia and Klebsiella Centre, Copenhagen, Denmark, it is suggested that it should keep the designation K8 . The other polysaccharide with Klebsiella K8 specificity should be renamed as K8,52,59 based on the cross-reactivity of the strain (I . Orskov, unpublished). Carbohydr Res, 1995 Aug 22, 273(1), 53 - 62 Structural studies of the capsular polysaccharide from Klebsiella type 7; Kjellberg A et al.; The structure of the capsular polysaccharide elaborated by Klebsiella type 7 has been investigated . NMR spectroscopy together with sugar and methylation analysis have been the main methods used . A uronic acid degradation was also employed . The polysaccharide consists of hexasaccharide repeating units having the following structure . {formula: see text} FEBS Lett, 1995 Aug 7, 369(2-3), 243 - 8 Molecular mimicry and ankylosing spondylitis: possible role of a novel sequence in pullulanase of Klebsiella pneumoniae; Fielder M et al.; Molecular mimicry has been shown between two sequences of Klebsiella pneumoniae pulD secretion protein (DRDE) with HLA-B27 (DRED) and pulA (pullulanase) enzyme (Gly-X-Pro) with types I, III and IV collagen respectively . IgG antibody levels in AS patients were elevated against 16mer synthetic peptides of HLA-B27 and pulD by enzyme immunosorbent assay (ELISA) compared to controls (P < 0.001) . ELISA assays against K . pneumoniae grown in the absence and presence of pullulan demonstrated significant levels of IgA antibody in AS patients compared to controls (P < 0.001) . Increased IgA and IgG antibody levels to pulA and types I and IV collagen were observed in AS patients compared to controls (P < 0.001) . These observations could be relevant in the sequence of molecular events in AS. Indian J Med Res, 1995 Aug, 102, 53 - 5 Ceftazidime resistance among Klebsiella pneumoniae in south India; Abigail S et al.; Fifteen strains of K . pneumoniae resistant to ceftazidime were tested for extended spectrum beta lactamase (ESBL) production by double disc diffusion synergy test using ceftazidime and augmentin, and by testing for reduction in minimum inhibitory concentration of ceftazidime in combination with clavulanic acid . All strains were found to produce ESBL . We document the presence of ESBL producing strains in south India . There is a possibility of the spread of this resistance to other bacteria because the resistance could be transferred to recipient Escherichia coli from two strains. J Chir (Paris), 1995 Aug-Sep, 132(8-9), 342 - 5 {Chronic osteomyelitis caused by Klebsiella pneumoniae with a 50 year course . Medicosurgical management}; Sarfati F et al.; The 50-year clinical course of a case of osteomyelitis of the left tibia is reported . Surgical specimens were positive for Klebsiella pneumoniae allowing treatment with certriaxone and ciprofloxacine . After antibiotic therapy, regional skin expansion was possible with excision and simultaneous closure of a wide area of damaged skin resulting from the long-term infection. Biol Pharm Bull, 1995 Aug, 18(8), 1089 - 93 Time-dependent changes in the pharmacokinetics and renal excretion of xanthine derivative enprofylline induced by bacterial endotoxin in rats; Nadai M et al.; Time-dependent changes in the pharmacokinetics and renal handling of enprofylline induced by bacterial endotoxin (Klebsiella pneumoniae LPS) were investigated in rats . To evaluate the early effect of LPS on kidney functions and the renal excretion of enprofylline, which is an organic anion drug excreted primarily by an active tubular secretion, LPS (250 micrograms/kg) was infused for 5 min under constant infusion at rates of 2.3 and 23 micrograms/min/kg for inulin and enprofylline, respectively . LPS caused a drop in the glomerular filtration rate (GFR), estimated as the renal clearance of inulin, to 65-75% of that observed in the control rats within 30 min after the LPS treatment . The renal clearance (CLr) of enprofylline decreased in conjunction with GFR, while the percentage of decrease in the CLr was slightly greater than that in GFR . LPS-induced decreases in the CLr for enprofylline and GFR continued over the testing period of 120 min . The time-dependent effect of LPS on the pharmacokinetics of enprofylline was examined by a single injection of enprofylline (2.5 mg/kg) to rats pretreated 2, 10 or 24 h earlier with or without LPS . The pharmacokinetic parameters of enprofylline were determined by a model-independent method . Significant changes in the systemic clearance for enprofylline were observed in rats pretreated 2 and 10 h earlier with LPS, but no such changes were observed in rats pretreated 24 h earlier with LPS . These findings indicate the existence of a time-dependent effect of LPS on the pharmacokinetics of enprofylline, and suggest that LPS at a dose of 250 micrograms/kg, at least, does not induce cytotoxicity to kidney cells. Cent Eur J Public Health, 1995 Aug, 3(3), 129 - 31 An outbreak of gentamicin-resistant Klebsiella pneumoniae in a neonatal ward; Shamseldin el Shafie S et al.; Gentamicin-resistant Klebsiella pneumoniae (GRKP) was isolated from nineteen patients in the neonatal ward of Sultan Qaboos University Hospital (SQUH) in Oman . Thirteen cases were infected and six were colonized . Two infected patients died due to septicaemia, eight cases occurred within SQUH and eleven cases were imported from other hospitals . Sixteen isolates were of the same serotype (K25), three were non typeable . Referred patients from peripheral hospitals were the source of the organism and hand carriage the probable vehicle of transmission . The first strain of gentamicin-sensitive Kleb . pneumoniae was isolated five months after combating the outbreak. Ann Rheum Dis, 1995 Aug, 54(8), 631 - 5 IgA1 and IgA2 subclass antibodies against Klebsiella pneumoniae in the sera of patients with peripheral and axial types of ankylosing spondylitis; Maki-Ikola O et al.; OBJECTIVE--To study further the Klebsiella specific serum antibody response in patients with axial and peripheral types of ankylosing spondylitis (AS) . METHODS--IgA1 and IgA2 subclass antibodies to Klebsiella pneumoniae were measured by enzyme linked immunosorbent assay in the sera of 171 patients with axial or peripheral type AS, and in sera of 100 healthy controls . The effect of 26 weeks of sulphasalazine treatment on the antibody levels in the two types of AS was also analysed . RESULTS--K pneumoniae specific antibody levels of both IgA1 and IgA2 subclasses were increased in the sera of patients with AS compared with healthy controls . The increased levels were present in patients with axial and with peripheral AS, and there were no statistically significant differences in the antibody levels between these two groups . Sulphasalazine treatment decreased the Klebsiella specific antibody level of IgA1 subclass in patients with axial AS, but there were no statistically significant changes in the IgA2 subclass, or in the patients with peripheral type AS . CONCLUSIONS--These results agree with earlier published findings suggesting that IgA (especially Klebsiella specific IgA) may have a role in the pathogenetic mechanisms of both peripheral and axial types of AS . In addition, it seems that both IgA1 and IgA2 subclasses are involved in the disease process. Eur J Biochem, 1995 Aug 1, 231(3), 790 - 801 Synthesis of the oxaloacetate decarboxylase Na+ pump and its individual subunits in Escherichia coli and analysis of their function; Di Berardino M et al.; The oadGAB genes encoding the gamma, alpha and beta-subunits of the oxaloacetate decarboxylase Na+ pump in Klebsiella pneumoniae have been cloned on plasmid pSK-GAB and expressed in Escherichia coli . The membranes of the recombinant E . coli clone contained about three times as much catalytically active oxaloacetate decarboxylase (3 mg protein/2 g wet cells) as those of the K . pneumoniae strain from which the genes were derived . The enzyme was solubilised from the membranes with Triton X-100 and purified . Its Na+ transport function was demonstrated after reconstitution into proteoliposomes . Proteoliposomes containing only the membrane-bound subunits beta and gamma (not the peripheral alpha-subunit) were unable to catalyse Na+ translocation in response to a transmembrane Na+ (delta pNa+) or electrical gradient (delta psi) . Individual subunits of oxaloacetate decarboxylase and combinations of two subunits were expressed from appropriate derivatives of plasmid pSK-GAB . The hydrophobic subunits beta and beta gamma were membrane-bound as expected . Interestingly, the alpha-subunit was located in the cytoplasm if expressed separately or together with beta, but became membrane-bound if expressed together with gamma . A gamma alpha complex was isolated from such membranes by avidin-Sepharose affinity chromatography . Interactions of the gamma-subunit with the water-soluble alpha-subunit and with the membrane-bound beta-subunit are therefore required to form the oxaloacetate decarboxylase complex . The combinations of separately expressed subunits gamma alpha + beta and beta gamma+alpha were shown to yield the catalytically active enzyme . The alpha or the beta-subunit and the combinations of these subunits with the gamma-subunit were therefore expressed in E . coli in a catalytically competent state . Functional expression of the separate gamma-subunit, however, could not be demonstrated . The alpha-subunit was strongly overexpressed from a pT7-7 derived plasmid, but was only partially biotinylated under these conditions . On coexpression of the birA gene encoding biotin ligase the major part (80-100%) of the overexpressed alpha-subunit was biotinylated . Highly purified alpha-subunit was obtained by fractionated precipitation of the soluble cell fraction with ammonium sulfate . Incubation of the alpha-subunit with oxaloacetate led to a CO2 transfer to its prosthetic biotin group with the formation of stoichiometric amounts of pyruvate . The velocity of the CO2 transfer to the biotin on the alpha-subunit was about three orders of magnitude too low to account for the rate of the overall reaction . The carboxyltransfer reaction was significantly accelerated if the gamma-subunit was additionally present.(ABSTRACT TRUNCATED AT 400 WORDS) J Bacteriol, 1995 Aug, 177(16), 4820 - 4 Activation of the Escherichia coli lacZ promoter by the Klebsiella aerogenes nitrogen assimilation control protein (NAC), a LysR family transcription factor; Pomposiello PJ et al.; A chimeric promoter with the nitrogen assimilation control protein binding site from hutUp of Klebsiella aerogenes fused to the lacZ core promoter from Escherichia coli was built and cloned in a lacZ reporter plasmid . This construct showed a 14-fold increase of beta-galactosidase activity upon nitrogen limitation . Primer extension experiments showed that the nitrogen assimilation control protein activates lacZp1 in a position-dependent manner. Laryngoscope, 1995 Aug, 105(8 Pt 1), 854 - 6 Rhinoscleroma treated with ciprofloxacin: a case report; Avery RK et al.; Rhinoscleroma, a chronic progressive infection of the nose and associated structures caused by Klebsiella rhinoscleromatis, has posed a therapeutic dilemma since its identification in the late 1800s . Although a number of antibiotics have been found to be effective in this relapsing disorder, the lengthy duration of treatment can lead to problems with adverse effects and compliance, especially with the traditional therapies of streptomycin and tetracycline . We report on a patient with extensive nasal rhinoscleroma who achieved pathologic and bacteriologic resolution during treatment with oral ciprofloxacin after previous courses of tetracycline and trimethoprim-sulfamethoxazole . Ciprofloxacin may prove to be useful in the therapy of rhinoscleroma because it is convenient for oral administration, achieves good tissue levels, is concentrated in macrophages, and is generally well tolerated as long-term therapy . As mentioned in a recent review of patients with rhinoscleroma at the Mayo Clinic, the fluoroquinolones deserve further study as potentially highly effective agents for this uncommon but significant infectious condition. Dtsch Med Wochenschr, 1995 Jul 28, 120(30), 1040 - 4 {Vaso-occlusive crises in sickle-cell anemia}; Lang SM et al.; HISTORY AND FINDINGS: Severe pain suddenly occurred in the shaft of the right thigh in a 21-year-old patient of Turkish descent with known homozygotic sickle cell anaemia . He also had marked scleral jaundice and anaemia (haemoglobin 9.1 g/dl) . An X-ray film of the lower leg pointed to a bone infarct . Type of pain, partial pressure of oxygen in blood (70 mm Hg), concentration of lactate dehydrogenase and the bilirubin level (4,5 mg/dl) as signs of haemolysis, as well as the fall in haemoglobin, indicated a vascular occlusive crisis in sickle cell anaemia . TREATMENT AND COURSE: The symptoms regressed within 8 days of the patient receiving fluids, analgesics and oxygenation by nasal tube . A few weeks later he developed a fever, productive cough and severe pain in joints and abdomen . Blood p02 was 54 mm Hg . Pneumonia in the right lung base was the cause of this renewed life-threatening crisis, which was again associated with haemolysis . Klebsiella having been found in sputum he was treated with cefotaxim (1 g twice daily intravenously) and oxygen insufflation via nasal tube (7 l/min) . His condition quickly improved . CONCLUSIONS: In patients with sickle cell anaemia it is important (1) to regard crises as potentially life-threatening conditions, (2) to detect and treat infections as possible causes early and (3) to provide sufficient oxygen insufflation quickly, especially in pulmonary infections. J Immunol, 1995 Jul 15, 155(2), 722 - 9 Neutralization of IL-10 increases survival in a murine model of Klebsiella pneumonia; Greenberger MJ et al.; Effective host defense against bacterial infection is dependent upon the vigorous recruitment and activation of neutrophils and macrophages . We hypothesized that IL-10 is produced in the setting of bacterial pneumonia, and this cytokine may attenuate host defense by inhibiting the expression of important activating and chemotactic cytokines . CD-1 mice were challenged with either 30 microliters of saline or saline containing 10(3) CFUs of Klebsiella pneumoniae intratracheally (i.t.) and lungs were harvested at 8, 24, and 48 h . The i.t . inoculation with K . pneumoniae resulted in a 13-, 14-, and 8-fold increase in lung homogenate TNF, macrophage inflammatory protein-2 (MIP-2), and macrophage inflammatory protein-1 alpha (MIP-1 alpha) levels, respectively, as compared with control animals . In addition, we observed an increase in IL-10 mRNA and protein levels in lung homogenates, maximal at 48 h postinoculation . To establish the biologic relevance of IL-10 in Klebsiella pneumonia, we passively immunized CD-1 mice with 0.5 ml of rabbit anti-murine IL-10 serum or preimmune serum i.p . 2 h before i.t . administration of K . pneumoniae . Treatment of animals with anti-IL-10 serum resulted in increased levels of TNF, MIP-2, and MIP-1 alpha, respectively, within lung homogenates at 24 and 48 h, as compared with preimmune-treated animals . Furthermore, neutralization of IL-10 resulted in a significant decrease in K . pneumoniae CFU in both lung homogenates and plasma harvested at 48 h, as well as a significant increase in survival in these animals . Our studies indicate that 1) IL-10 is produced during Klebsiella pneumonia; and 2) inhibition of IL-10 bioactivity in vivo results in enhanced bacterial clearance, increased expression of proinflammatory cytokines, and prolonged survival. FEMS Microbiol Lett, 1995 Jul 15, 130(1), 51 - 7 Virulence factors (aerobactin and mucoid phenotype) in Klebsiella pneumoniae and Escherichia coli blood culture isolates; Vernet V et al.; We examined the presence of two virulence factors in 241 blood isolates of Klebsiella pneumoniae from patients hospitalized during 1989 and 1990 in 7 French hospitals, and 125 blood isolates of Escherichia coli from one hospital . Aerobactin was scored phenotypically and genotypically with an intragenic DNA probe of 2 kb . The mucoid phenotype was assessed by culture on trypticase soy agar and by genotypic analysis (intragenic DNA probe of 235 bp) . Only 6% K . pneumoniae isolates were aerobactin-positive with no significant variation according to geographical location while 20% of K . pneumoniae isolates displayed the mucoid phenotype, with a significant variation according to hospital . Aerobactin was always associated with the mucoid phenotype . The frequency of aerobactin production but not mucoid phenotype (14%) was higher among E . coli isolates (48%) . They harbored two types of large plasmids . Intraperitoneal injection into mice of 10(3) cfu of K . pneumoniae producing both virulence factors demonstrated that capsular serotype K2 was the more virulent K23 and K28. Carbohydr Res, 1995 Jul 11, 271(2), 177 - 83 Synthesis of the tetrasaccharide repeating unit of the antigen from Klebsiella type 20; Das SK et al.; Starting from D-mannose, D-galactose and D-glucuronolactone, two disaccharide blocks, namely methyl 4,6-di-O-benzyl-2-O-(2,3,4,6-tetra-O-benzyl-beta-D-galactopyranosyl)- alpha-D-mannopyranoside, acting as acceptor, and ethyl 4,6-di-O-acetyl-2-O-allyl-3-O-(methyl 2,3,4-tri-O-acetyl-beta-D-gluco-pyranosyluronate)-1-thio-beta-D- galactopyranoside, acting as donor, were synthesised . The two disaccharides were then allowed to react to give, after deprotection, methyl 2-O-beta-D-galactopyranosyl-3-O-(3-O-beta-D-glucopyranosyluronic acid-alpha-D-galactopyranosyl)-alpha-D-mannopyranoside which is the methyl glycoside of the tetrasaccharide repeating unit of the said antigen. Orv Hetil, 1995 Jul 2, 136(27), 1447 - 9 {Vertical transmission causing hepatitis C in a newborn infant}; Grasselly M et al.; There is a case report of an intrauterine growth retarded infant who was suffering of sepsis caused by Klebsiella pneumoniae . An exchange transfusion was carried out because of hyperbilirubinaemia on the 5th day of life . 10 days later high bilirubin concentration was observed again parallel to the liver enzyme abnormalities . HCV-related antibodies were detectable in the sera of the newborn and his mother as well . The results of analysis of HCV RNA with the polymerase chain reaction in the infant and his mother suggest vertical transmission of HCV infection . This is the first case of HCV infection in the newborn caused by vertical transmission in Hungary. J Antimicrob Chemother, 1995 Jul, 36(1), 241 - 6 Bactericidal effect of beta-lactams and amikacin alone or in association against Klebsiella pneumoniae producing extended spectrum beta-lactamase; Roussel-Delvallez M et al.; Ten extended spectrum beta-lactamases producing strains of Klebsiella pneumoniae characterized by analytical isoelectric focusing and studied for their susceptibility to beta-lactam antibiotics, either alone or in combination with a beta-lactamase inhibitor (clavulanic acid and sulbactam) and in association with amikacin . The extended spectrum beta-lactamases were derived from either TEM (CTX-1 = TEM-3) or SHV (CAZ-4 = SHV-5) . Killing curves were studied with antibiotics at clinical by achievable concentrations, at MIC and MIC x 4 . At MIC, cefotetan, cefotaxime and ceftazidime lacked bactericidal activity . Imipenem was more rapidly bactericidal than meropenem or co-amoxiclav . At MIC x 4, cefotetan and cefotaxime exhibited bactericidal effect but this was less than for imipenem which gave a reduction of 4 log10 of the inoculum . Cefotaxime plus sulbactam gave no bactericidal effect compared with cefotaxime plus co-amoxiclav . A bactericidal effect with cefotaxime plus sulbactam was seen with the addition of amikacin . At clinical concentrations cefotaxime plus co-amoxiclav +/- amikacin was as efficient as imipenem +/- amikacin with a rapid bactericidal effect (5-6 log10 in 30-60 min) . We proposed that cefotaxime+co-amoxiclav might be considered as an alternative to imipenem for the treatment of extended spectrum beta-lactamase associated K . pneumoniae injections. Chemotherapy, 1995 Jul-Aug, 41(4), 267 - 75 Mechanism of enhancement of bactericidal activity of phagocytes against Klebsiella pneumoniae treated with subminimal inhibitory concentrations of cefodizime; Nomura S et al.; The effects of a sub-MIC of cefodizime on the morphology of the capsular structures and on the surface physicochemical properties, such as hydrophobicity and charge, of encapsulated Klebsiella pneumoniae were studied . The enhancement of bactericidal activity of macrophages against bacteria treated with sub-MICs of antibiotics was evaluated as the killing index . Cefodizime treatment gave the highest value of 32 . Electron microscope observations revealed that the capsular material layer of cefodizime-treated K . pneumoniae was markedly thinner (32 nm) than that of untreated bacteria (160 nm) or bacteria treated with other antibiotics (75-90 nm) . Contact angle measurement revealed that the surface of cefodizime-treated K . pneumoniae was more hydrophobic than that of untreated bacteria or bacteria treated with other antibiotics . Furthermore, the negative charge of the surface of K . pneumoniae decreased significantly with cefodizime treatment compared with the surface of untreated bacteria . These findings suggest that the treatment of K . pneumoniae with a sub-MIC of cefodizime reduced the thickness of the capsular material layer and that these changes increased the surface hydrophobicity of the bacteria and decreased the negative charge of the bacterial surface to render K . pneumoniae more susceptible to phagocytic activity by reducing the physical repulsion between the bacteria and phagocytes. Infect Immun, 1995 Jul, 63(7), 2537 - 40 Binding of mannose-binding protein to Klebsiella O3 lipopolysaccharide possessing the mannose homopolysaccharide as the O-specific polysaccharide and its relation to complement activation; Jiang GZ et al.; Lipopolysaccharide from Klebsiella pneumoniae O3, which possesses the mannose homopolysaccharide as the O-specific polysaccharide, exhibits an extraordinarily high ability to activate the human complement system . We isolated the mannose-binding protein with a Klebsiella O3 lipopolysaccharide affinity column . The protein isolated had a molecular mass of much higher than 200 kDa, and it consisted of subunits with an apparent molecular mass of 32 kDa . The NH2-terminal sequence of the 32-kDa subunits was completely consistent with a part of the amino acid sequence of human serum mannose-binding protein . In immunoblotting, an anti-mannose-binding protein monoclonal antibody was definitely reactive with the isolated protein with the higher molecular mass . The protein isolated was bound exclusively to lipopolysaccharides possessing the mannose homopolysaccharide, not to lipopolysaccharide possessing the heteropolysaccharides . Klebsiella O3 lipopolysaccharide did not exhibit a high anticomplement activity in the serum from which the mannose-binding protein was depleted . It was concluded that the serum factor that bound to Klebsiella O3 lipopolysaccharide may be mannose-binding protein and that it may play a crucial role in the strong complement activation by Klebsiella O3 lipopolysaccharide. Rinsho Shinkeigaku, 1995 Jun, 35(6), 667 - 9 {A case of thrombocytopenia due to heparin therapy for a progressing ischemic stroke}; Arai M et al.; A 44-year-old man was admitted, complaining of weakness of his left limbs . On admission, he was oriented and cooperative . He showed positive Barre sign only on the left leg . Deep tendon reflexes were unremarkable; pathological reflexes were negative . There was no cerebellar ataxia or sensory deficit . Immediately after admission, monoplegia of the left leg progressed . He was started on heparin infusion therapy . His paralysis had been resolved by the 5th day . MRI of the brain demonstrated an infarction involving the body of the corpus callosum, the right cingulate gyrus, and the right paracentral lobule . Neuropsychologic examination demonstrated a disturbance of interhemispheric transfer of position . On the 10th day he developed Klebsiella ozaenae bacteremia . On the 11th day, platelet count was 9.2 x 10(4)/microliter . Antithrombin III, thrombin antithrombin III complex, D-dimer, and FDP levels were unremarkable, which is quite atypical for DIC associated with bacterial infection . Platelet factor 4 and beta-thromboglobulin levels were remarkably high, suggesting activated platelet aggregation . We made a diagnosis of heparin-induced thrombocytopenia and discontinued heparin infusion . Three days later, platelet count began to increase and creatinine levels returned normal . All patients receiving heparin should be monitored for platelet count. J Bacteriol, 1995 Jun, 177(12), 3546 - 55 The nitrogen assimilation control protein, NAC, is a DNA binding transcription activator in Klebsiella aerogenes; Goss TJ et al.; A 32-kDa polypeptide corresponding to NAC, the product of the Klebsiella aerogenes nac gene, was overexpressed from a plasmid carrying a tac'-'nac operon fusion and purified to near homogeneity by taking advantage of its unusual solubility properties . NAC was able to shift the electrophoretic migration of DNA fragments carrying the NAC-sensitive promoters hutUp, putPp1, and ureDp . The interaction between NAC and hutUp was localized to a 26-bp region centered approximately 64 bp upstream of the hutUp transcription initiation site . Moreover, NAC protected this region from DNase I digestion . Mobility shift and DNase I protection studies utilizing the putP and ureD promoter regions identified NAC-binding regions of sizes and locations similar to those found in hutUp . Comparison of the DNA sequences which were protected from DNase I digestion by NAC suggests a minimal NAC-binding consensus sequence: 5'-ATA-N9-TAT-3' . In vitro transcription assays demonstrated that NAC was capable of activating the transcription of hutUp by sigma 70-RNA polymerase holoenzyme when this promoter was presented as either a linear or supercoiled DNA molecule . Thus, NAC displays the in vitro DNA-binding and transcription activation properties which have been predicted for the product of the nac gene. Yakugaku Zasshi, 1995 Jun, 115(6), 446 - 59 {A stereoselective synthesis of tilivalline and its analogs utilizing a new Mannich type intramolecular cyclization}; Aoyama T et al.; Tilivalline (1a), a metabolite isolated from Klebsiella pneumoniae var . oxytoca, belongs to a group of pyrrolo{2,1-c}{1,4}benzodiazepines, a characteristic skeleton of anthramycin-type antitumor antibiotics . We have accomplished a completely stereoselective, efficient and convenient synthesis of 1a utilizing a new Mannich type intramolecular cyclization as a key step . Further, a computational chemical analysis clarified the effect of zinc chloride on the high stereoselectivity in the tilivalline synthesis . To aim both the extension of the scope of the new Mannich type intramolecular cyclization and the studies on the structure-biological activity relationship, we further extended the method to the synthesis of tilivalline derivatives and 2-(3'-indolyl)-1,4-benzodiazepines (50) . Investigation on the cytotoxicity of 1a and its analogs has revealed that 1a shows the strong cytotoxicity toward mouse leukemia L 1210 cells and the replacement of the indole function of 1a with cyano one increases the cytotoxicity of 1a about 100 times (IC50 = 0.05 microgram/ml). Antimicrob Agents Chemother, 1995 Jun, 39(6), 1365 - 8 Point mutation in the pribnow box, the molecular basis of beta-lactamase overproduction in Klebsiella oxytoca; Fournier B et al.; Klebsiella oxytoca mutants resistant to a variety of beta-lactams were obtained in vitro on aztreonam . Constitutive beta-lactamase production was much higher in the mutants than in the susceptible strains (75-fold) . The only difference observed in these mutants compared with the susceptible strains were point mutations in the Pribnow box: a transversion (G-->T) in the first base for one mutant or a transition (G-->A) in the fifth base of the -10 consensus sequence for the other three mutants . The transcriptional output of the beta-lactamase gene (blaOXY) from the mutants was significantly higher than that of the blaOXY gene from the susceptible strains. Rhinology, 1995 Jun, 33(2), 57 - 60 Successful treatment of ozena with ciprofloxacin; Nielsen BC et al.; Rhinitis chronica foetida, or ozena, is a rare chronic inflammatory disease . The aetiology and pathogenesis are still not satisfactory explained . For many years various medical and surgical methods for the treatment of this slowly progressive and disabling disease have been tried without permanent success so far . The new fluoroquinolones with excellent effect on gram-negative bacteria and high suitability for oral use offer a potentially attractive treatment for ozena . We review our experience in the treatment of 10 patients with ciprofloxacin in a daily dose of 500-750 mg b.i.d . for 1-3 months . The patients have been followed regularly for up to 26-74 months after treatment and in all of them we registered permanent disappearance of odour, crusting, and growth of Klebsiella ozenae . We conclude that ciprofloxacin provides a step towards better conservative therapy for patients with ozena. Intern Med, 1995 Jun, 34(6), 554 - 8 Magnetic resonance imaging of multiple brain abscesses of the bilateral basal ganglia; Nagase T et al.; A 64-year-old woman developed multiple brain abscesses of the basal ganglia associated with Klebsiella pneumoniae septicemia . Magnetic resonance (MR) images showed three different stages of the brain abscesses . The images of early cerebritis of this site mimicked lacunar infarctions or dilated Virchow-Robin spaces . The differentiation of the brain abscess from lacunae and dilated Virchow-Robin spaces is discussed, together with the evolution of the brain abscesses on MR images. Int J Cancer, 1995 May 29, 61(5), 716 - 21 Liposomal doxorubicin-induced toxicity: depletion and impairment of phagocytic activity of liver macrophages; Daemen T et al.; Doxorubicin entrapped within conventional liposomes (200 nm in diameter; lip-Dox) has major toxic effects on liver macrophages of the rat for a considerable period of time following i.v . administration, with respect to both specific phagocytic capacity and cell numbers . At different time-points after injection of lip-Dox or free doxorubicin, radiolabeled, negatively charged, "empty" test liposomes were injected . Phagocytic capacity was determined by isolating the liver macrophages and measuring the amount of macrophage-associated radioactivity . Four subfractions of liver macrophages of different cell-size and with intrinsically different phagocytic capacity were isolated . Twenty-four hours after injection of lip-Dox, the phagocytic capacity of the larger-sized liver macrophages was strongly decreased . The relatively low intrinsic phagocytic capacity of the smaller-sized macrophages was only slightly impaired . Phagocytic capacity after injection of lip-Dox was nearly restored to control values after 14 days . Blood clearance of Klebsiella pneumoniae bacteria after pre-treatment with lip-Dox was strongly decreased . Pre-treatment with the free drug and/or placebo liposomes had no effect on phagocytic and bacterial blood-clearance capacity . A major depletion of the liver macrophage population was observed, as revealed by both macrophage isolation and histology . Only 2 weeks after injection of lip-Dox, the number of cells had returned to that seen in control animals . In view of the important host-defense functions of the liver macrophages, especially in the control of tumor growth and infection, the findings reported here should be taken into consideration when lip-Dox is to be administered in anti-tumor therapy. Science, 1995 May 19, 268(5213), 998 - 1004 The crystal structure of urease from Klebsiella aerogenes; Jabri E et al.; The crystal structure of urease from Klebsiella aerogenes has been determined at 2.2 A resolution and refined to an R factor of 18.2 percent . The enzyme contains four structural domains: three with novel folds playing structural roles, and an (alpha beta)8 barrel domain, which contains the bi-nickel center . The two active site nickels are 3.5 A apart . One nickel ion is coordinated by three ligands (with low occupancy of a fourth ligand) and the second is coordinated by five ligands . A carbamylated lysine provides an oxygen ligand to each nickel, explaining why carbon dioxide is required for the activation of urease apoenzyme . The structure is compatible with a catalytic mechanism whereby urea ligates Ni-1 to complete its tetrahedral coordination and a hydroxide ligand of Ni-2 attacks the carbonyl carbon . A surprisingly high structural similarity between the urease catalytic domain and that of the zinc-dependent adenosine deaminase reveals a remarkable example of active site divergence. J Mol Biol, 1995 May 12, 248(4), 781 - 803 Core RNA polymerase and promoter DNA interactions of purified domains of sigma N: bipartite functions; Cannon W et al.; The sigma N class of sigma factors confer upon RNA polymerase the requirement for enhancer-binding activator proteins . The sigma-N (sigma N) protein of Klebsiella pneumoniae was analysed by the assay of purified peptides comprising domains or regions of sigma N defined by proteolysis or by homology alignment, respectively . The NH2-terminal Region I is required for the correct interaction of holoenzyme with the promoter, and promoter complexes forming with a truncated sigma N lacking Region I are not activatable . The complexes lack the DNA structure believed to represent nucleated strand separation but still make close contacts with this promoter part . Determinants of specific DNA recognition by sigma N were shown to reside in a C-terminal 16 kDa peptide, and core RNA polymerase binding determinants in an adjacent peptide . The latter contacts and appears to pack against the DNA-binding domain . Thus the DNA-binding and core-binding domains are bipartite in function, consistent with core functioning as an allosteric effector of the sigma DNA-binding activity . The DNA-binding and core-binding domains together include Region III of sigma N . Although not the primary determinant of core or DNA recognition, the acidic Region II of sigma N influenced both activities . Regions I and II in combination with core RNA polymerase thus appear to control the activity of C-terminal DNA contacting surfaces to allow formation of a closed promoter complex that is susceptible to activation. Lett Appl Microbiol, 1995 May, 20(5), 303 - 7 Comparison of selective agars for the isolation and identification of Klebsiella oxytoca and Escherichia coli from environmental drinking water samples; Packer PJ et al.; Various selective media were assessed for their ability to detect and differentiate Klebsiella oxytoca and Escherichia coli in environmental water samples . Only two, Membrane Lauryl Sulphate agar and Deoxycholate Agar, could differentiate the two coliforms from each other and from the 'background' heterotrophs in water and this was a consequence of E . coli's ability to grow at 44 degrees C and 37 degrees C whereas Kl . oxytoca could only grow at 37 degrees C . Modified M-FC medium effectively differentiated Kl . oxytoca but not E . coli in environmental samples . Other media characterized the different coliforms in pure culture but failed to do likewise in environmental samples . For example, pure cultures of E . coli fluoresced when MUG was added to the medium but single colonies on a mixed species plate failed to do so . MT7 agar distinguished the two coliforms from water heterotrophs but not from each other. Am J Gastroenterol, 1995 May, 90(5), 767 - 70 Rupture of pyogenic liver abscess; Chou FF et al.; OBJECTIVE: Our objective was to study the clinical manifestations, course, treatment, and results obtained in 23 patients with ruptured pyogenic liver abscess and compare these findings with those of nonruptured cases . METHODS: Four hundred twenty-four patients with clinical diagnoses of pyogenic liver abscess were enrolled in the study . Among these, 23 patients had ruptured pyogenic liver abscess . The clinical manifestations, incidence of septic shock, laboratory findings, concurrent diabetes mellitus, etiology of abscess, and results of the treatment were recorded . Qualitative data were analyzed by chi 2 test, and quantitative data were analyzed by Student's t test . RESULTS: Except for abdominal pain and septic shock, other symptoms, such as fever, chills, and jaundice, were similar in ruptured and nonruptured groups . Laboratory findings indicated that the group with ruptured liver abscess had higher levels of bilirubin, blood glucose, and aspartate aminotransferase than the non-ruptured group . Of the patients with ruptured abscess, 14 (60.9%) had diabetes mellitus and 15 (65.2%) were cryptogenic . Klebsiella pneumoniae was the bacteria most often isolated in both blood cultures and liver aspirates . Surgical intervention--draining the abscess and cleaning the abdominal cavity--was the only means of saving the patients' lives . The overall mortality rate was higher in this group (43.5%) than in the nonruptured group (15.5%) . CONCLUSIONS: Ruptured pyogenic liver abscess should be suspected if septic shock and diffuse abdominal pain are found in a patient with pyogenic liver abscess, concurrent with high levels of bilirubin, aspartate aminotransferase, and blood glucose . Surgery is the only treatment for this condition. Infect Immun, 1995 May, 63(5), 2026 - 32 Adherence properties of an mrkD-negative mutant of Klebsiella pneumoniae; Hornick DB et al.; The role of the mrkD gene in attachment by a type 3 fimbriate Klebsiella pneumoniae strain was further characterized . A clinical isolate, K . pneumoniae IA565, was found to contain two copies of the gene encoding the fimbrial subunit, mrkA, and one copy of the gene encoding the adhesin subunit, mrkD . One copy of mrkA was located on the bacterial chromosome, and the other copy was associated with mrkD and located on a plasmid . The plasmid-borne mrk gene cluster was lost when K . pneumoniae IA565 was subcultured serially in broth at 44 degrees C . The resulting mrkD-negative strain, designated K . pneumoniae IApc35, did not exhibit the following adherence characteristics associated with K . pneumoniae possessing MrkD-positive fimbriae: agglutination of tannic acid-treated human erythrocytes and attachment to trypsinized human buccal cells . However, K . pneumoniae IApc35 produced type 3 fimbriae that were composed of the characteristic 21.5-kDa major fimbrial subunit, were reactive with specific serum, and were visualized specifically by immunoelectron microscopy . K . pneumoniae IApc35 retained a copy of the mrkA gene on its chromosome . This mrkA-containing gene cluster could be complemented by a recombinant plasmid carrying only the mrkD gene, resulting in restoration of the K . pneumoniae IA565-like adhesive phenotype and demonstration of type 3 filament-associated MrkD subunits by using colloidal gold labeling and immunoelectron microscopy . These data indicate that K . pneumoniae may contain multiple copies of the mrk genes which may be present simultaneously on both plasmid and chromosomal DNAs and which may encode fimbriae with different binding specificities. Chemotherapy, 1995 May-Jun, 41(3), 178 - 86 In vitro and in vivo enhancement of bactericidal activity of phagocytes against Klebsiella pneumoniae treated with subminimal inhibitory concentrations of cefodizime; Nomura S et al.; The effects of a subminimal inhibitory concentration (sub-MIC) of cefodizime on the bactericidal activity of phagocytes against encapsulated Klebsiella pneumoniae were studied in an in vitro system using an established mouse macrophage cell line, and in an in vivo system using mice in which many phagocytes were induced in the peritoneal cavity . In the in vitro system, the bactericidal activity of mouse macrophages against K . pneumoniae treated with a sub-MIC of cefodizime was significantly enhanced, and was greater than that of cefotaxime or cefoperazone . Significantly more bacteria treated with a sub-MIC of cefodizime were killed by serum complement than those treated with cefotaxime or cefoperazone . In the in vivo system, cefodizime-treated bacteria were phagocytosed and killed by phagocytes in the mouse peritoneal cavity, whereas, cefotaxime- and cefoperazone-treated and untreated bacteria were hardly phagocytosed at all or killed by phagocytes in the mouse peritoneal cavity, and bacterial regrowth was observed 24 h after bacterial challenge . Furthermore, the virulence of K . pneumoniae in mice was reduced more by treatment with cefodizime than with cefotaxime or cefoperazone . These findings indicate that K . pneumoniae treated with a sub-MIC of cefodizime become more susceptible to the bactericidal activity of phagocytes both in vitro and in vivo . This provides evidence that cefodizime at a sub-MIC may act together with the phagocytes against bacterial infections. Antimicrob Agents Chemother, 1995 May, 39(5), 1187 - 90 In vitro activities of various beta-lactam antimicrobial agents against clinical isolates of Escherichia coli and Klebsiella spp . resistant to oxyimino cephalosporins; Jett BD et al.; Broth microdilution testing was used to study the activity of several beta-lactam antimicrobial agents, including piperacillin-tazobactam and cefepime, against 108 clinically derived Escherichia coli and Klebsiella sp . strains resistant to oxyimino cephalosporins (i.e., putative extended-spectrum beta-lactamase producers) . On the basis of the percentage of susceptible strains, imipenem (100%), cefotetan (> or = 92%), and piperacillin-tazobactam (> or = 86%) were the most active agents . Cefepime activity (52 to 64% susceptible) was comparable to that of cefotaxime (40 to 63% susceptible) and aztreonam (20 to 63% susceptible) . Among all beta-lactams tested, imipenem and cefotetan demonstrated the highest and most consistent level of activity and were the least affected by challenges with increased sizes of inocula of these resistant organisms. Antimicrob Agents Chemother, 1995 May, 39(5), 1093 - 6 Effects of MICs and sub-MICs of antibiotics on production of capsular polysaccharide of Klebsiella pneumoniae; Held TK et al.; In the present study, we examined whether MICs and sub-MICs of antimicrobial agents belonging to two different classes, ciprofloxacin and ceftazidime, were able to influence the production and release of cell-associated and soluble (extracellular) capsular polysaccharide (CPS), respectively, in a heavily encapsulated strain of Klebsiella pneumoniae (B5055) . Using a CPS-specific enzyme-linked immunosorbent assay, we found that the amount of cell-associated CPS increased in a dose-dependent manner by more than 10-fold under the influence of the MIC of ceftazidime and by more than 100-fold under the influence of the MIC of ciprofloxacin . The largest amounts of CPS were measured by using the MIC of either antibiotic substance . Electron microscopic studies showed that the diameter of the capsule was significantly increased compared with the diameter for untreated controls . Thus, both antimicrobial agents genuinely stimulated CPS production. J Formos Med Assoc, 1995 May, 94(5), 232 - 7 Comparison of pyogenic liver abscesses caused by Klebsiella pneumoniae and non-K . pneumoniae pathogens; Chang FY et al.; A retrospective study on patients with pyogenic liver abscesses was conducted to clarify the different clinical presentations among patients with Klebsiella pneumoniae abscesses and those with non-K . pneumoniae abscesses . From 1981 to 1993, the medical records of 146 adults with culture-confirmed pyogenic liver abscesses who attended Tri-Service General Hospital in Taipei were studied . Abscesses due to K . pneumoniae accounted for 114 (78%) of pyogenic liver abscesses . When compared to patients with non-K . pneumoniae abscesses, patients with K . pneumoniae liver abscesses had significantly higher proportions of monomicrobial infections, unknown sources of infection and solitary abscesses . Patients with K . pneumoniae liver abscesses were found to have diabetes mellitus more often than patients with non-K . pneumoniae liver abscesses (66% vs 19%) . Septicemia was found more frequently in patients with K . pneumoniae liver abscesses than in patients with non-K . pneumoniae liver abscesses (50% vs 27%) . The clinical presentations among the two groups were, otherwise, not significantly different . Regardless of the microbial etiology, patients with diabetes mellitus had longer periods of fever after treatment and hospitalization than patients without diabetes . The reason for the high relative frequency of liver abscesses in Taiwan and its more frequent occurrence in diabetes mellitus remains unclear. Microbiol Res, 1995 May, 150(2), 167 - 72 Antibacterial, antidermatophytic and antitoxigenic activities of onion (Allium cepa L.) oil; Zohri AN et al.; The inhibitory effect of onion oil against the growth of various isolates of bacteria representing Gram-positive (4 isolates) and Gram-negative (4 isolates) species were studied . Results show that onion oil was highly active against all Gram-positive bacteria tested and only one isolate (Klebsiella pneumoniae) of Gram-negative bacteria . The inhibitory effect of onion oil against nine different species of dermatophytic fungi were also studied . Onion oil (200 ppm) completely inhibited the growth of Microsporum canis, M . gypseum and Trichophyton simii while the growth of both, Chrysosporium queenslandicum and Trichophyton mentagrophytes was completely inhibited by 500 ppm of onion oil . The growth of four other species of dermatophytic fungi was gradually reduced by increasing the concentrations of onion oil . The inhibitory effect of onion oil was also tested against four toxigenic isolates of fungi . Onion oil at different concentrations (100, 200 and 500 ppm) tested gradually reduced fungal growth and aflatoxin production by Aspergillus flavus IMI 89,717 and A . parasiticus var . globosus IMI 120,920 . Fungal growth and production of sterigmatocystin and rubratoxin A by A . versicolor IMI 16,139 and Penicillium rubrum IMI 136,127 were completely inhibited by the addition of 200 ppm onion oil. Infection, 1995 May-Jun, 23(3), 163 - 7 Klebsiella oxytoca meningitis: frequent association with neurosurgical procedures; Tang LM et al.; Klebsiella oxytoca meningitis is a rare condition . Nine patients were diagnosed between 1981 and 1993 at our institution . These accounted for 2.3% of 393 patients with blood and/or cerebrospinal fluid culture-proven bacterial meningitis . K . oxytoca was noted in both community-acquired meningitis and nosocomial meningitis . Eight of the nine cases were patients who had undergone neurosurgical procedures . Four were mixed bacterial meningitis . All K . oxytoca isolates were susceptible to third-generation cephalosporins and all but one to chloramphenicol . Antibiotic therapy was successful in eight patients but failed in one. Mol Gen Genet, 1995 Apr 20, 247(2), 189 - 98 The role of uridylyltransferase in the control of Klebsiella pneumoniae nif gene regulation; Edwards R et al.; The glnD gene in enteric bacteria encodes a uridylyltransferase/uridylyl-removing enzyme which acts as the primary nitrogen sensor in the nitrogen regulation (Ntr) system . We have investigated the role of this enzyme in transcriptional regulation of nitrogen fixation genes in Klebsiella pneumoniae by cloning glnD from this organism and constructing a null mutant by insertional inactivation of the chromosomal gene using the omega interposon . K . pneumoniae glnD encodes a 102.3 kDa polypeptide which is highly homologous to the predicted products of both Escherichia coli glnD and Azotobacter vinelandii nfrX . The glnD-omega mutant was unable to uridylylate PII and was altered in adenylylation/deadenylylation of glutamine synthetase . Uridylyltransferase was required for derepression of ntr-regulated promoters such as glnAp2 and pnifL but was not involved in the nif-specific response to changes in nitrogen status mediated by the nifL product . We conclude that a separate, as yet uncharacterised, nitrogen control system may be responsible for nitrogen sensing by NifL. Infect Immun, 1995 Apr, 63(4), 1318 - 28 Aggregative adherence of Klebsiella pneumoniae to human intestine-407 cells; Favre-Bonte S et al.; Aggregative adhesion of Klebsiella pneumoniae LM3 to Intestine-407 (Int-407) cells was studied . Adhesive capacities were affected by the bacterial growth phase (with a maximum of adherence obtained during the exponential phase), temperature, multiplicity of infection, and length of incubation with Int-407 cells . Adhesion occurred through a cytochalasin D-sensitive process and was greatly reduced after treatment of Int-407 with cycloheximide, indicating that aggregative adhesion requires active participation of Int-407 cells . Transmission electron microscopy revealed that adherent bacteria were surrounded by a capsule-like material, apparently involved in both bacterium-Int-407 cell and bacterium-bacterium adherence . Examination with a scanning electron microscope showed interactions of intestinal cell microvilli with bacteria and formation in 3 h of a fibrous network within and around the bacterial clusters . We speculate that aggregative adhesion of K . pneumoniae mediated by a capsule-like extracellular material might explain the persistence of these strains inside the host gastrointestinal tract. J Am Vet Med Assoc, 1995 Apr 1, 206(7), 1018 - 21 Septic cholangiohepatitis and cholangiocarcinoma in a horse; Durando MM et al.; Septic cholangiohepatitis was diagnosed in an 11-year-old Warmblood gelding with a history of intermittent colic and fever . Klebsiella pneumoniae, susceptible to gentamicin, was cultured from the biopsy specimen . However, treatment with gentamicin was unsuccessful, and histologic examination and bacteriologic culture of a biopsy specimen obtained 3 weeks later revealed progression of the hepatic inflammation and yielded growth of gentamicin-resistant K pneumoniae . At this time, several discrete hyperechoic structures, suggestive of biliary calculi, were seen ultrasonographically . A change in antibiotic treatment was associated with gradual resolution of clinical signs . Five months after initial examination, the horse had a sudden onset of severe right forelimb lameness . The horse responded to treatment with antibiotics and phenylbutazone, but lameness and fever that was unresponsive to treatment recurred 7 months later, and the horse was euthanatized . Necropsy revealed nodules throughout the liver and a mass associated with the right metacarpophalangeal joint . Histologic and immunohistochemical examination revealed carcinomatous infiltration of the liver and metacarpophalangeal joint . The tumor was probably of biliary origin . Carcinoma should be considered in cases of septic cholangiohepatitis unresponsive to antibiotic treatment. J Bacteriol, 1995 Apr, 177(8), 1947 - 51 Evidence for the presence of urease apoprotein complexes containing UreD, UreF, and UreG in cells that are competent for in vivo enzyme activation; Park IS et al.; In vivo activation of Klebsiella aerogenes urease, a nickel-containing enzyme, requires the presence of functional UreD, UreF, and UreG accessory proteins and is further facilitated by UreE . These accessory proteins are proposed to be involved in metallocenter assembly (M . H . Lee, S . B . Mulrooney, M . J . Renner, Y . Markowicz, and R . P . Hausinger, J . Bacteriol . 174:4324-4330, 1992) . A series of three UreD-urease apoprotein complexes are present in cells that express ureD at high levels, and these complexes are thought to be essential for in vivo activation of the enzyme (I.-S . Park, M . B . Carr, and R . P . Hausinger, Proc . Natl . Acad . Sci . USA 91:3233-3237, 1994) . In this study, we describe the effect of accessory gene deletions on urease complex formation . The ureE, ureF, and ureG gene products were found not to be required for formation of the UreD-urease complexes; however, the complexes from the ureF deletion mutant exhibited delayed elution during size exclusion chromatography . Because these last complexes were of typical UreD-urease sizes according to native gel electrophoretic analysis, we propose that UreF alters the conformation of the UreD-urease complexes . The same studies revealed the presence of an additional series of urease apoprotein complexes present only in cells containing ureD, ureF, and ureG, along with the urease subunit genes . These new complexes were shown to contain urease, UreD, UreF, and UreG . We propose that the UreD-UreF-UreG-urease apoprotein complexes represent the activation-competent form of urease apoprotein in the cell. Arch Surg, 1995 Apr, 130(4), 401 - 5; discussion 406 The comparison of clinical course and results of treatment between gas-forming and non-gas-forming pyogenic liver abscess; Chou FF et al.; OBJECTIVES: To study and review the clinical manifestations, courses, and results of treatment in 83 cases of verified gas-forming pyogenic liver abscess . DESIGN: Case series . SETTING: Both primary and referral hospital care . PATIENTS: Four hundred twenty-four patients with clinical diagnosis of pyogenic liver abscess were enrolled in the study . Eighty-three patients had gas-forming abscesses and 341 had non-gas-forming abscesses . The clinical manifestations, duration of symptoms, incidence of septic shock, laboratory findings, concurrent diabetes mellitus, cause of abscess, size of abscess, and results of treatment were recorded . MAIN OUTCOME MEASURES: A chi 2 test for qualitative data and Student's test for quantitative data . RESULTS: Duration of symptoms were shorter (mean +/- SD, 5.2 +/- 5.3 vs 7.6 +/- 10 days) (P < .005) and the incidence of septic shock was higher in the gas-forming than in the non-gas-forming group (32.5% vs 11.7%) (P < .01) . Laboratory findings revealed high levels of blood glucose, aspartate aminotransferase, alkaline phosphatase, and serum urea nitrogen in the gas-forming group . The size of abscess was usually bigger (> 5 cm) in this group . In the gas-forming group, 71 patients (85.5%) had diabetes mellitus and 65 patients (78.3%) had conditions of cryptogenic origin . Klebsiella pneumoniae was the main bacteria, in blood culture and liver aspirates, especially in gas-forming liver abscess . Medical treatment and/or aspiration carried a high mortality rate (44.4%) in the gas-forming group; also, the overall mortality rate was higher in this group than in the non-gas-forming group (27.7% vs 14.4%) (P < .01) . CONCLUSIONS: The gas-forming liver abscess may be a disease of wide spectrum of severity and may run a fulminating course . Strong antibiotics with early adequate drainage are mandatory . Surgery should not be delayed if necessary. J Infect Dis, 1995 Apr, 171(4), 938 - 47 Efficacy of gentamicin or ceftazidime entrapped in liposomes with prolonged blood circulation and enhanced localization in Klebsiella pneumoniae-infected lung tissue; Bakker-Woudenberg IA et al.; Polymer (PEG-PE)-coated liposomes exhibit prolonged circulation time in blood and substantial localization in Klebsiella pneumoniae-infected lung tissue in rats . Therefore, to determine the therapeutic effect, gentamicin and ceftazidime were entrapped in these liposomes and administered to rats experimentally infected with pneumonia: Relatively high and sustained concentrations of liposome-associated antibiotic in blood were observed . Compared with antibiotics alone, one dose of liposome-entrapped gentamicin or ceftazidime increased the therapeutic effect of the drugs, survival of rats, and bacterial killing in lungs . One dose of liposome-entrapped ceftazidime was as effective as a continuous 2-day infusion of nonentrapped ceftazidime . Since antibiotic-containing liposomes are stable during circulation and liposome-entrapped ceftazidime and gentamicin have low bactericidal activity in vitro, the superior therapeutic effect of the liposome-encapsulated antibiotics results from localization and subsequent degradation of liposomes and the resulting release of entrapped antibiotic at the infection site. J Med Assoc Thai, 1995 Apr, 78(4), 210 - 6 Liver abscess in polycystic liver disease; Assantachai P et al.; A case of an elderly female whose clinical manifestations were atypical in view of physiologic aging changes and multiple pathology commonly found in the geriatric group . The liver abscess had presented clinically and radiologically like a hepatic malignancy and pus was obtained during the process of liver biopsy . Even though anchovy-like pus suggested amebic in origin, the microscopic exam showed numerous polymorphonuclear leucocyte and the culture both from the blood and pus grew Klebsiella spp . as well as the study for E . histolytica antibody was also negative . Pyogenic liver abscess was the working diagnosis and indicated for surgical drainage . Adult polycystic liver disease was found intraoperatively and the liver pathology did reveal the presence of many amebae in the cavity of the right lobe of the liver . After the administration of ceftriaxone and metronidazole as well as the surgical drainage, the patient recovered uneventfully within sixteen days of admission. Nippon Ronen Igakkai Zasshi, 1995 Apr, 32(4), 266 - 9 {Evaluation of respiratory tract disease accompanying malignant lymphoma in the elderly}; Niitsu N et al.; We studied the usefulness of bronchoscopy in the diagnosis of pulmonary complications in elderly patients with non-Hodgkin's lymphoma (NHL) . The subjects were 12 patients 65 or older with NHL . Two of these patients underwent transbronchial lung biopsies (TBLB), all of them underwent bronchiolavage (BAL) and 2 with tumors underwent transbronchial biopsies of the tumors . Diagnosis was obtained in 8 out of 12 patients (66.7%) . Two patients showed infiltration of lymphoma cells, 2 had cytomegalovirus (CMV) infection, 1 had pneumocystosis (P . carinii), 2 had fungal infections and 1 had a bacterial infection . TBLB was performed in 2 patients with platelet counts of over 5 x 10(4)/microliters . Lymphoma cell infiltration was diagnosed in one of these patients and a fungal infection in the other . BAL was performed on all patients . CMV was detected in 2 cases by the PCR method, P . carinii was detected in 1 case and aspergillus in 1 case . Klebsiella pneumonia was diagnosed in 1 patient . Since respiratory tract complications are often serious in NHL and the prognosis depends on early diagnosis and treatment, it is important to perform bronchoscopy on elderly patients and obtain a definitive diagnosis. Eur J Pediatr, 1995 Apr, 154(4), 299 - 303 Effect of nutritional and HIV status on bacteraemia in Zimbabwean children who died at home; Wolf BH et al.; From July 1992 to May 1993 a study was performed of the relationship between bacteraemia, nutritional status and HIV status in 212 out of 334 consecutive infants and children aged 0-5 years, who had died at home in Bulawayo, Zimbabwe . The remaining 122 c