J Bacteriol, 1990 Feb, 172(2), 818 - 23 Role of the promoter in activation of transcription by nitrogen regulator I phosphate in Escherichia coli; Ray L et al.; The protein nitrogen regulator I (NRI)-phosphate is known to activate the initiation of transcription of the Escherichia coli glnA gene . This activation is facilitated by the binding of the protein to NRI-specific sites located upstream of the sigma 54-dependent glnA promoter . To determine whether binding of NRI-phosphate to upstream sites is sufficient for activation, we placed several promoters not normally activated by NRI-phosphate downstream of NRI binding sites and measured activation in intact cells and in an in vitro transcription system . We found that the sigma 70-dependent lac promoter was not activated, that the sigma 54-dependent Klebsiella pneumoniae nifH promoter was weakly activated, and that a nifH promoter altered in the RNA polymerase binding site was almost as well activated as the glnA promoter . We conclude that the sensitivity of the susceptible promoter depends on the presence of NRI binding sites, but that the presence of bound NRI-phosphate upstream of a promoter is not sufficient for activation of transcription by RNA polymerase . This activation is determined by the structure of the RNA polymerase binding site . We suggest that sigma 54-but not sigma 70-dependent promoters are susceptible to activation by NRI-phosphate and that the nucleotide sequence of the sigma 54-RNA polymerase binding site is an important determinant of the efficiency of activation. Pneumonol Pol, 1990 Feb-Mar, 58(2-3), 112 - 7 {Etiological factors in secondary bacterial infections from the authors' own clinical material}; Misiewicz A et al.; An analysis of etiological factors of secondary bacterial infections was carried out in 64 patients with pneumonias . In most of the patients gram negative rods were cultured . Most often the pneumonias were caused by Klebsiella pneumoniae and Escherichia coli . A single causative agent was found in the majority of the patients (60.7%), less frequently two, least three. Wei Sheng Wu Xue Bao, 1990 Feb, 30(1), 54 - 7 {Study on identification for the motility and flagella antigen of four international reference strains of Escherichia coli}; Yang Z et al.; Authors identified four strains of Escherichia coli from Collaborative Centre for reference and research on Escherichia coli-Klebsiella (WHO) in Denmark . Our results were different from original record and reference . It was shown that strain H311b and H5 were motile strains, and its H antigen respectively is H11 and H27 . The strain W27 is nonmotile . H antigen of the strain H511 is H40, not H8 . Antigen formula of four strains respectively is 26:60:11 (strain H311b), 81:97:27 (strain H5), 115:-:- (strain W27), 102:-:40 (strain H511). J Dairy Sci, 1990 Feb, 73(2), 363 - 72 Growth of gram-negative bacteria in dry cow secretion; Todhunter D et al.; Gram-negative bacteria (n = 192) isolated from infected bovine mammary glands were tested for growth in a pooled source of dry cow secretion . Growth of Klebsiella pneumoniae in dry cow secretion was greater than growth of Escherichia coli and Klebsiella oxytoca . Escherichia coli originating during the early dry period exhibited greater growth in dry cow secretion than those originating around calving or during lactation . Klebsiella pneumoniae growth did not differ with time of origin of intramammary infection . Escherichia coli, K . oxytoca, and K . pneumoniae growth in a synthetic medium was reduced by apolactoferrin plus Ig . Growth reduction was greatest for E . coli . Citrate reversed growth inhibition . The inhibitory properties of dry cow secretion for E . coli may contribute to the low number of naturally occurring intramammary infections originating during the early part of the dry period . Inhibitory properties of dry cow secretion are partially explained by lactoferrin acting in conjunction with antibody to prevent iron acquisition by many gram-negative bacteria. Antimicrob Agents Chemother, 1990 Feb, 34(2), 326 - 31 Application of mathematical model to experimental chemotherapy of fatal murine pneumonia; Hishikawa T et al.; Two beta-lactam antibiotics, cefazolin and cefmenoxime, were administered in an experimental model of murine pneumonia caused by Klebsiella pneumoniae in a way which enabled us to approximate the serum antibiotic concentration time course in humans . Bacterial counts during the experiments were subjected to nonlinear least-squares analyses by using a mathematical model that explained the bacterial killing by the antibiotic concentration time course and other factors associated with antimicrobial potency and bacterial growth . Cefazolin gave a killing curve that changed synchronously with the drug levels in serum; in contrast, cefmenoxime gave a curve that was prolonged as compared with the change in the drug levels in serum . Multiple correlation coefficients were about 0.9, and the model worked well for bacterial count data . Parameters relating to antimicrobial potency of the drugs, bacterial growth rate, and drug distribution into the tissue were estimated numerically. Mol Gen Genet, 1990 Feb, 220(3), 481 - 4 A general method for the transfer of plasmid-borne mutant alleles to the chromosome of Klebsiella pneumoniae using bacteriophage lambda: transfer of pqq genes; Meulenberg JJ et al.; A generally applicable method is described for reintroduction of mutant plasmid-borne alleles to the chromosome of Klebsiella pneumoniae using bacteriophage lambda . We used this method to make stable chromosomal transposon insertions in genes for biosynthesis of pyrroloquinoline quinone in K . pneumoniae. J Autoimmun, 1990 Feb, 3(1), 37 - 42 The antibody repertoire of early human B cells . II: Expression of anti-DNA-related idiotypes; Lydyard PM et al.; Cord blood B cells were immortalized in vitro with Epstein-Barr virus (EBV) . Supernatants containing greater than 500 ng/ml IgM from clones/lines were tested for expression of anti-DNA-associated 16/6 and PR4 idiotypes (id) by ELISA . Four of 70 lines, but no clones, were positive for 16/6 id and none expressed the PR4 id . The presence of 16/6 id on four cell lines was associated with specificity for ssDNA, cardiolipin and Fc of IgG . No association was seen with binding to the K30 polysaccharide of Klebsiella . One clone binding this antigen also had anti-ssDNA, anti-Fc and anti-cardiolipin activity but did not express 16/6 id . Our data support the germ-line nature of the 16/6 id and are consistent with the notion that IgM autoantibody-producing B cells use VH genes which are part of the normal B cell repertoire. Eur J Biochem, 1990 Jan 26, 187(2), 353 - 60 Characterisation of the Klebsiella pneumoniae nitrogen-fixation regulatory proteins NIFA and NIFL in vitro; Austin S et al.; Transcriptional activation by the Klebsiella pneumoniae nitrogen-fixation-specific positive control protein, NIFA, (nifA gene product) has been demonstrated in vitro in S30 extracts from cells which overproduce this protein . The activity of NIFA was dramatically reduced in vitro in the presence of the negative regulatory protein NIFL (nifL gene product) but was not inhibited by the presence of a mutant NIFL protein, NIFL2404 . Transcriptional activation from the nifH promoter by NIFA was dependent on the alternative sigma factor, sigma 54, and also required the presence of an upstream activator sequence . NIFA activity was temperature-sensitive in vitro (as it is in vivo) which is due, at least in part, to the intrinsic lability of the protein itself . The majority of overproduced NIFA and NIFL was insoluble after low-speed centrifugation and was inactive in vitro . A low level of less aggregated NIFA protein present in cell extracts was responsible for in vitro activity and this fraction was partially purified. FEMS Microbiol Lett, 1990 Jan 15, 55(1-2), 33 - 8 Two novel transferable extended-spectrum beta-lactamases from Klebsiella pneumoniae in Tunisia; Ben Redjeb S et al.; Two novel beta-lactamases conferring multiresistance to antibiotics including oxyimino beta-lactams have been identified in two nosocomial K . pneumoniae strains isolated in Tunis in 1986 and 1988 . Both enzymes were encoded by ca . 150-kilobase plasmids . Donor and transconjugant strains producing these enzymes exhibited highly similar pattern of resistance (CTX phenotype) to beta-lactams including penicillins and oxyimino beta-lactams e.g . cefotaxime, ceftriaxone, ceftazidime, and aztreonam . High and variable synergy (16 to 1066-fold) was obtained when combined to 0.1 microgram/ml of clavulanate (beta-lactamase inhibitor) . The isoelectric points of these two enzymes were 5.4 and 6.4 . These beta-lactamases differed from TEM types by hydrolysis for cefotaxime or ceftriaxone but were inhibited by clavulanate and cloxacillin . DNA hybridization studies suggested that that the genes of these enzymes may be derived from genes encoding TEM-type enzymes. J Biol Chem, 1990 Jan 15, 265(2), 831 - 7 Selective killing of Klebsiella pneumoniae by 5-trifluoromethylthioribose . Chemotherapeutic exploitation of the enzyme 5-methylthioribose kinase; Gianotti AJ et al.; 5'-Deoxy-5'-methylthioadenosine (MTA), an important intermediate in methionine recycling, can be metabolized by one of two mechanisms that appear to be mutually exclusive . In human cells, MTA is degraded in one step to adenine and 5-methylthioribose 1-phosphate (MTR-1-P) via MTA phosphorylase . In contrast, certain microbes metabolize MTA in two steps: first to 5-methylthioribose (MTR) followed by conversion to MTR-1-P . The enzymes involved in this two-step conversion are MTA nucleosidase and MTR kinase . In both cases, MTR-1-P is subsequently recycled to methionine . Because MTR kinase is "unique" to microbes (it is also found in plant tissue) and since it is essential to microbial methionine salvage, we hypothesized that MTR kinase is a promising target for chemotherapeutic exploitation . We demonstrate that 5-trifluoromethylthioribose (TFMTR), a structural analog of MTR, is a potent inhibitor of the MTR kinase-containing organism Klebsiella pneumoniae . TFMTR not only inhibits the growth of K . pneumoniae in a dose-dependent manner (50% inhibition at approximately 40 nM) but also competitively inhibits MTR kinase activity (Ki approximately 7 microM) . Furthermore, TFMTR is shown to be a substrate for MTR kinase (Km = 1.7 microM), suggesting that the drug could be converted to toxic products (e.g . trifluoromethionine or carbonothionic difluoride) in enzyme-containing organisms . Structural analogs of MTR represent a new class of compounds with the potential for treating diseases caused by MTR kinase-containing microorganisms. Nucleic Acids Res, 1990 Jan 11, 18(1), 151 - 6 Cloning and sequence analysis of gyrA gene of Klebsiella pneumoniae; Dimri GP et al.; The gene gyrA encoding the DNA gyrase A subunit of Klebsiella pneumoniae has been cloned in the plasmid pBR322 . Bases of about 3.5 Kb DNA have been sequenced to locate the gyrA gene . An open reading frame of 2628 nucleotides coding for a 97 KD protein has been identified . Homology to the extent of about 85% was detected at the nucleotide level and about 90% at the amino acid level, when the sequences were compared with that of Escherichia coli gyrA . Some very interesting differences have, however, been found in the promoter region. J Biol Chem, 1990 Jan 5, 265(1), 42 - 6 Hydrogen peroxide mediates the oxidative inactivation of enzymes following the switch from anaerobic to aerobic metabolism in Klebsiella pneumoniae; Chevalier M et al.; Klebsiella pneumoniae utilizes distinct pathways for the anaerobic and aerobic metabolism of glycerol . During anaerobic growth, glycerol is first converted to dihydroxyacetone by glycerol dehydrogenase; subsequent phosphorylation yields dihydroxyacetone phosphate . During aerobic growth, glycerol is initially phosphorylated to yield glycerol 3-phosphate; subsequent reduction then gives dihydroxyacetone phosphate . A coordinated response occurs when anaerobically growing cells are switched to aerobic conditions . Synthesis of glycerol dehydrogenase is repressed, glycerol dehydrogenase is inactivated, and the protein is degraded . Ethanol dehydrogenase and propanediol oxidoreductase are also inactivated when cells are exposed to oxygen (Johnson, E . A., Levine, R . L., and Lin, E . C . C . (1985) J . Bacteriol . 164, 479-483) . Exposure of anaerobically growing cells to low concentrations of hydrogen peroxide also inactivated these three enzymes and led to rapid degradation of glycerol dehydrogenase . Glycerol dehydrogenase was purified and characterized after in vivo oxidative modification initiated by hydrogen peroxide . No differences in molecular weight, amino acid composition, or Km were detected between the native and oxidatively modified forms, although the modified enzyme had only 10% of the catalytic activity of the native form . The oxidatively modified enzyme was very susceptible to degradation by subtilisin while the native enzyme was resistant . Chloramphenicol prevented the inactivation and degradation of glycerol dehydrogenase caused by exposure to oxygen but did not block that caused by hydrogen peroxide . Thus, protein synthesis appears necessary for in vivo oxidative modification caused by exposure to oxygen but is not necessary when the process is initiated by exposure to hydrogen peroxide . The newly synthesized protein(s) presumably catalyzes the production of hydrogen peroxide which is required for the metal-catalyzed oxidative modification of susceptible enzymes. Radiat Res, 1990 Jan, 121(1), 71 - 5 Trehalose dimycolate enhances survival of fission neutron-irradiated mice and Klebsiella pneumoniae-challenged irradiated mice; McChesney DG et al.; The survival of B6D2F1 female mice exposed to lethal doses of fission neutron radiation is increased when trehalose dimycolate (TDM) preparations are given either 1 h after exposure or 1 day before exposure to radiation . TDM in an emulsion of squalene, Tween 80, and saline was the most effective formulation for increasing the 30-day survival of mice when given 1 day before (90%) or 1 h after (88%) exposure to radiation . An aqueous suspension of a synthetic analog of TDM was less effective at increasing 30-day survival (60%) when given 1 day prior to radiation exposure and not effective when given 1 h after radiation . Mice receiving a sublethal dose (3.5 Gy) of fission neutron radiation and either the TDM emulsion or synthetic TDM 1 h after irradiation were substantially more resistant to challenge with 10, 100, 1000, or 5000 times the LD50/30 dose of Klebsiella pneumoniae than untreated mice. J Infect, 1990 Jan, 20(1), 21 - 31 Klebsiella pneumoniae bacteraemia at an urban general hospital; Feldman C et al.; Of 47 patients with Klebsiella pneumoniae bacteraemia admitted to the Hillbrow Hospital, Johannesburg during a period of 18 months, 31 were males and 16 were females . Features predisposing to illness were found in 89.4% patients, chronic alcoholism, neoplastic disease and diabetes mellitus being the most common . Twenty-five infections were acquired in hospital and 22 in the community . Most patients (59.6%) had pneumonia . All isolates of K . pneumoniae were resistant to ampicillin (100%); several (42.6%) were resistant to other antibiotics also . The overall mortality rate was 55.3% . A higher mean initial blood pressure and lower concentrations of serum urea and bilirubin were found in survivors . None of the 28 patients, surviving more than 48 h who received combined therapy with an aminoglycoside and a beta-lactam antibiotic (to which the organism was susceptible) died . Among the remaining patients treated with either an appropriate beta-lactam agent alone, an appropriate aminoglycoside alone or ciprofloxacin the combined mortality rate was 83.3% (P = 0.007). Crit Care Med, 1990 Jan, 18(1), 67 - 71 Detoxification of plasma containing lipopolysaccharide by adsorption; Bysani GK et al.; We compared the ability of 13 materials, all with known affinity for lipopolysaccharide (LPS), to remove LPS from water, pooled normal human plasma, and plasma from a patient with Gram-negative bacterial sepsis . Escherichia coli O111:B4 LPS was added to water and pooled normal human plasma to a concentration of 200 ng/ml and aliquots were adsorbed in parallel with each of the materials . Plasma containing 25 ng/ml of LPS was obtained by plasmapheresis from a patient with fatal Klebsiella pneumoniae sepsis and was similarly adsorbed . Ethanolamine bound to Sepharose 4B served as a control adsorbent . LPS was most readily removed from water and least readily removed from plasma obtained from the patient with sepsis . Nonetheless, some materials removed substantial quantities of LPS from the patient's plasma . Upon ip injection of control-adsorbed patient's plasma into LPS-sensitized mice, death occurred in 13 of 13 mice within 12 h . In contrast, when this plasma was first adsorbed with activated charcoal, Kaopectate, or polymyxin B bound to Sepharose 4B, death occurred 12 h after challenge in 0/13, 0/13, and 2/13 mice, respectively (p less than .0001 by actuarial life table analysis of survival distributions) . Thus, plasma that contains bacterial toxins produced during Gram-negative bacterial sepsis can be detoxified by adsorption. Indian Pediatr, 1990 Jan, 27(1), 21 - 6 Bacterial contamination of reconstituted oral rehydration solution; Nagarajan L et al.; The extent and nature of bacterial contamination in oral rehydration solution reconstituted for use by individuals and for group of patients was studied . Twenty three volunteers (all qualified doctors) were asked to reconstitute a packet of prepackaged salt in half litre of clean unboiled water obtained from taps at their residence . Five ml aliquots of ORS were collected at 6, 12 and 24 hours after reconstitution for bacteriologic study . The water used by volunteers to reconstitute the ORS as well as throat swabs, peri-anal swabs and nail clippings of volunteers yielded pathogenic bacteria in all the subjects/samples . All the 23 specimens of ORS prepared by volunteers when cultured at 6 hours after reconstitution yielded pathogenic bacteria . The bacterial colony counts were found to be unacceptably high at 12 hours . Five ml samples of reconstituted ORS prepared in bulk in the children ward of PGIMER, Chandigarh were cultured at 12, and again at 24 hours after reconstitution on 10 different days . These yielded Klebsiella pneumoniae in 8 specimens (80%) and E . coli in 2 (20%) . The bacterial colony count was unacceptably high, 12 hours after reconstitution. Microbiol Immunol, 1990, 34(2), 147 - 51 Cytotoxic component(s) of Klebsiella oxytoca on HEp-2 cells; Higaki M et al.; A cytotoxic component(s) was detected in culture filtrates of Klebsiella oxytoca isolated from patients with antibiotic-associated hemorrhagic colitis . Eleven of 12 isolates exhibited cytotoxicity on HEp-2 cells . The cytotoxic activity of K . oxytoca strain MH43-1 was stable for the treatment of 60 C for 30 min, but inactivated by the treatment of 100 C for 15 min . This cytotoxicity was not destroyed by the treatment with trypsin or pronase, and the component was filtrable through a membrane filter which cut off molecular weight 5,000. J Urol (Paris), 1990, 96(1), 60 - 1 {Emphysematous pyelonephritis}; Heritier P et al.; The authors report on a case of a emphysematous pyelonephritis due to Klebsiella pneumoniae, occurring in a diabetic patient and successfully treated by nephrectomy . They compare it to the literature. Scand J Rheumatol Suppl, 1990, 87, 74 - 9; discussion 79-80 Trial and error in producing ankylosing-spondylitis-selective antisera according to Andrew Geczy; Beukelman CJ et al.; Geczy found that rabbit sera raised against Klebsiella strain K43 cross-reacted with the cells from HLA-B27 positive patients with ankylosing spondylitis (AS) . Other laboratories failed to reproduce these results . After a series of unsuccessful attempts, however, we managed to prepare one selective antiserum, using E . coli, isolated from a Dutch Bechterew patient, in offspring of rabbits Geczy sent us . Ever since we obtained irreproducible results only . This paper reports about the many attempts we have made to produce a discriminating antiserum for use in a combined vital stain and dye-exclusion assay. Arch Microbiol, 1990, 154(5), 489 - 95 Catabolism of 3-hydroxybenzoate by the gentisate pathway in Klebsiella pneumoniae M5a1; Jones DC et al.; Growth of Klebsiella pneumoniae M5a1 on 3-hydroxybenzoate leads to the induction of 3-hydroxybenzoate monooxygenase, 2,5-dihydroxybenzoate dioxygenase, maleylpyruvate isomerase and fumaryl-pyruvate hydrolase . Growth in the presence of 2,5-dihydroxybenzoate also induces all of these enzymes including the 3-hydroxybenzoate monooxygenase which is not required for 2,5-dihydroxybenzoate catabolism . Mutants defective in 3-hydroxybenzoate monooxygenase fail to grow on 3-hydroxybenzoate but grow normally on 2,5-dihydroxybenzoate . Mutants lacking maleylpyruvate isomerase fail to grow on 3-hydroxybenzoate and 2,5-dihydroxybenzoate . Both kinds of mutants grow normally on 3,4-dihydroxybenzoate . Mutants defective in maleylpyruvate isomerase accumulate maleylpyruvate when exposed to 3-hydroxybenzoate and growth is inhibited . Secondary mutants that have additionally lost 3-hydroxybenzoate monooxygenase are no longer inhibited by the presence of 3-hydroxybenzoate . The 3-hydroxybenzoate monooxygenase gene (mhbM) and the maleylpyruvate isomerase gene (mhbI) are 100% co-transducible by P1 phage. Int J Immunopharmacol, 1990, 12(5), 491 - 6 Effect of RU 41740 on autologous hemopoietic reconstitution of sublethally irradiated mice; Rezzoug F et al.; RU 41740, a glyco-protein extract of Klebsiellae pneumoniae 01 K2 strain, is known to have immunomodulator activities . It acts on macrophages as well as on B and T-cells, and enhances their cytokine production, particularly interleukin 1 (IL-1) . The fact that cytokines have an effect on hemopoiesis led us to suspect an effect of RU 41740 on hemopoietic reconstitution . In this study, a model of autologous reconstitution of a hemopoietic system after sublethal irradiation in mice was used . C57 BL/6 mice were treated orally with RU 41740 (10 mg/kg/day) before or after irradiation (6.5 Gy) . LPS of Klebsiella pneumoniae was used as a positive control . The hemopoietic reconstitution occurred more rapidly in treated animals especially when RU 41740 was given before irradiation. Chemotherapy, 1990, 36(5), 337 - 44 In vitro and in vivo effects of subinhibitory concentrations of clindamycin on experimental Klebsiella pneumoniae sepsis; Arbo A et al.; We studied the effect of subinhibitory doses of clindamycin on the course of experimental Klebsiella pneumoniae sepsis . Wistar rats were injected intraperitoneally with an inoculum containing 5 x 10(6) colony-forming units of K . pneumoniae resistant to clindamycin (minimum inhibitory concentration greater than 128 micrograms/ml) and then distributed to receive clindamycin 10 mg/kg/day or placebo for 10 days . All animals were bacteremic at 3 h . When the magnitude of bacteremia was compared, no difference was seen during the first 24 h; however, by 72 h the clindamycin-treated group had a significant decrease in the number of colony-forming units per milliliter blood (p less than 0.01) . The mortality rate showed a tendency to decrease in the treated group (0%) as compared with the control group (30%) . By 120 h, 3 of the 9 (33%) surviving animals from the control group were still bacteremic versus 0 of 11 (0%) in the clindamycin-treated group . These results suggest that subinhibitory clindamycin therapy can improve bacterial clearance and survival during the course of experimental K . pneumoniae sepsis. Arch Microbiol, 1990, 154(2), 162 - 7 Construction of a new catabolic pathway for D-fructose in Escherichia coli K12 using an L-sorbose-specific enzyme from Klebsiella pneumoniae; Wohrl BM et al.; Starting with a fruK (formerly fpk) mutant of Escherichia coli K12 lacking D-fructose-1-phosphate kinase (E.C . 2.7.1.3.), fructose positive derivatives were isolated after introduction of the cloned gene sorE from Klebsiella pneumoniae coding for an L-sorbose-1-phosphate reductase . The new pathway was shwon to proceed from D-fructose via D-fructose-1-phosphate and D-mannitol-1-phosphate to D-fructose 6-phosphate . It involves a transport system and enzymes encoded in the fru and the mtl operons from E . coli K12 as well as in the sor operon from K . pneumoniae respectively. Indian J Pathol Microbiol, 1990 Jan, 33(1), 53 - 6 R plasmids from clinical and environmental Klebsiella pneumoniae; Gadre DV; A total of 200 strains of K . pneumoniae, 150 from clinical and 50 from hospital environmental source were studied for antibiotic resistance pattern . The strains from clinical source showed higher incidence of drug resistance as compared to environmental strains . 50 clinical and 32 environmental multidrug resistant strains were tested for R plasmids . The biogroup 1 showed higher incidence of autotransferring and non autotransferring R plasmids. Microbiol Immunol, 1990, 34(5), 427 - 38 Inhibitory effect of Ca2+ on formation of Mg2(+)-mediated two-dimensional hexagonal lattice structure by an R-form lipopolysaccharide from Klebsiella pneumoniae; Kato N et al.; When the R-form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN-111 (O3-:K1-), from which cationic material had been removed by electrodialysis, was suspended in 50 mM Tris buffer at pH 8.5 containing 0.1 mM or higher concentrations of MgCl2, it formed an ordered two-dimensional hexagonal lattice structure and its center-to-center distance (lattice constant) depended upon the concentration of MgCl2 and reached the shortest value (14 nm) at 10 mM . In contrast, in the presence of 0.1 to 10 mM CaCl2 in place of MgCl2, the electrodialyzed LPS did not form such an ordered hexagonal lattice structure but formed an irregular network structure with a center-to-center distance of 19 to 20 nm . We investigated interaction of Mg2+ and Ca2+ in formation of the hexagonal lattice structure by the electrodialyzed LPS suspended in 50 mM Tris buffer at pH 8.5 . When 0.1 mM or higher concentrations of CaCl2 were mixed with 1 mM MgCl2 or when 1 mM or higher concentrations of CaCl2 was mixed with 10 mM MgCl2, the electrodialyzed LPS did not form the hexagonal lattice structure of the magnesium salt type but formed the irregular network structure of the calcium salt type . In the coexistence of equimolar or higher concentrations of CaCl2 together with 1 or 10 mM MgCl2, the binding of Mg to the electrodialyzed LPS was significantly inhibited and, conversely, the binding of Ca was enhanced as compared with when MgCl2 or CaCl2 was present alone . However, the coexistence of 10 times less molar concentrations of CaCl2 did not significantly inhibit the binding of Mg to the electrodialyzed LPS . Therefore, the inhibition of formation of the Mg2(+)-mediated hexagonal lattice structure of the electrodialyzed LPS by equimolar or higher concentrations of CaCl2 accompanied the inhibition of binding of Mg but that by 10 times less molar concentrations of CaCl2 did not accompany it. Int J Immunopharmacol, 1990, 12(4), 397 - 402 Interleukin-6 gene expression and production induced in human monocytes by membrane proteoglycans from Klebsiella pneumoniae; Sironi M et al.; The present study was designed to investigate the effect of membrane proteoglycans (MPG) from Klebsiella pneumoniae on IL-6 production by human peripheral blood monocytes . Exposure in vitro to MPG induced release of IL-6 activity from human monocytes, as assessed by the 7TD1 hybridoma assay . MPG-induced hybridoma growth factor activity was blocked by anti-IL-6 antibodies . MPG induced expression in human monocytes of IL-6 mRNA transcripts as assessed by Northern blot analysis . Induction of IL-6 in mononuclear phagocytes may play a role in the immunomodulatory activity of MPG. Infection, 1990, 18 Suppl 1, S5 - 9 Review of available trials of selective decontamination of the digestive tract (SDD); van Saene HK et al.; Eighteen studies on Selective Decontamination of the Digestive Tract (SDD) have been published up to now . A statistically significant reduction of infection rate was found in fourteen out of the fifteen controlled studies . Although all the studies were designed to evaluate infection-related morbidity as the end point, ten centres have reported fatality rates . Six centres out of the ten showed a statistically significant reduction in mortality in patients receiving SDD versus control . A recent French study describes the eradication of an outbreak of a multi-resistant Klebsiella with SDD . The Paris trial suggests a major impact of the SDD maneuver on the ICU ecology . Emergence of resistance to the SDD agents among gram-positive cocci has been described, although the clinical impact of this antibiotic side effect has not been reported so far . There are three indications for SDD, as follows: (i) trauma patients; (ii) liver transplant recipients and (iii) outbreaks of multi-resistant organisms. Mikrobiol Zh, 1990 Jan-Feb, 52(1), 20 - 2 {The antilysozyme activity of bacteria in the genus Klebsiella}; Turianitsa AI et al.; The ability to inactivate lysozyme was found in representatives of three species of the genus Klebsiella bacteria: K . pneumoniae (117 strains), K . rhinoscleromatis (104 strains), K . ozaenae (90 cultures) . The test cultures displayed a different antilysozyme activity, inactivating from 2 to 30 micrograms/ml of the enzyme . Taking into account the lysozyme role in the immunity and chitin synthesis processes in the organism of insects, the inactivation of the enzyme by Klebsiella may be considered as one of possible mechanisms of the entomopathogenic action of these bacteria. J Gen Microbiol, 1990 Jan, 136 ( Pt 1), 171 - 80 The purification, characterization and role of the d-type cytochrome oxidase of Klebsiella pneumoniae during nitrogen fixation; Smith A et al.; Klebsiella pneumoniae synthesized only b-type and d-type cytochromes under the wide range of growth conditions tested, and reaction with CO revealed two potential oxidases . The o-type oxidase was produced only in the presence of O2 and appeared to be repressed by glucose . The d-type oxidase was, by contrast, produced only in the absence of measurable O2 (less than 1 microM), and was the only oxidase expressed in nitrogen-fixing conditions . It was extracted from the membrane, purified and shown to be similar to that from E . coli in being a heterodimer (subunits of Mr 52,000 and 35,000), in containing two distinguishable b haems and haem d (one or two molecules per molecule of oxidase), and in being able to react with O2 to give a stable oxygenated intermediate . The purified d-type cytochrome oxidase had a very high affinity for O2 (Km 20 nM; measured by the spectral properties of leghaemoglobin) . It is proposed that this provides a role for this oxidase in lowering the O2 concentration to allow nitrogenase synthesis and function, and to provide a terminal oxidase to permit electron-transport-coupled ATP synthesis which supports the increase in efficiency of nitrogen fixation observed under microaerobic conditions. Am J Reprod Immunol, 1990 Jan-Feb, 22(1-2), 42 - 8 Characterization of autoantigens relevant to experimental autoimmune orchitis (EAO) in mice immunized with a mixture of syngeneic testis homogenate and Klebsiella O3 lipopolysaccharide; Yokochi T et al.; Experimental autoimmune orchitis (EAO) in mice has been induced by repeated injection of a mixture of syngeneic testis homogenate and Klebsiella O3 lipopolysaccharide (KO3 LPS) as a potent adjuvant . The antisera obtained from mice with EAO lesions defined several antigens with apparent molecular weights (MW) of 38,000 (38 kd), 86 kd, 100 kd, and greater than 200 kd by the immunoblotting method . These antigens were organ-specific and exclusively present on the acrosome of spermatozoa, suggesting that these acrosomal antigens were highly relevant to EAO . It was found that the antigen with a fairly high MW (greater than 200 kd) was expressed on spermatozoa from the epididymis . Furthermore, the acrosomal 86 kd antigen was predominantly expressed in the testis, while the 100 kd antigen was dominant in the spermatozoa from the epididymis . It was therefore suggested that the 86 kd and 100 kd antigens in the acrosome were differentially expressed on the process of maturation of spermatozoa. Microbiol Immunol, 1990, 34(2), 185 - 95 Protective effect of recombinant human interleukin-2 against lethal infection caused by Klebsiella pneumoniae; Iizawa Y et al.; The effects of recombinant human interleukin-2 (rIL-2) administered prophylactically on the death of CBA/J mice challenged with Klebsiella pneumoniae 27 intraperitoneally were examined . rIL-2 administered subcutaneously at 20 micrograms per mouse for 7 days enhanced survival after a lethal challenge . The injection of anti-asialo GM1 antibody did not influence the effect of rIL-2 . In mice given rIL-2, the number of peritoneal macrophages increased, and the infiltration of polymorphonuclear leukocytes (PMN) into the peritoneal cavity after the bacterial challenge was enhanced . In addition, adoptive transfer of sera and peritoneal exudate cells (PEC), consisting of an approximately equal number of macrophages and PMN, obtained from mice given rIL-2 enhanced resistance to a K . pneumoniae infection, compared with adoptive transfer of sera and PEC obtained from mice not given rIL-2 . These results indicate that rIL-2 protects mice from a lethal challenge with K . pneumoniae, and suggest that the protective effect is due to an increase in the number of phagocytic cells and in the cooperative activity of the sera and the phagocytic cells. Arch Microbiol, 1990, 153(5), 478 - 84 Quantification of multiple-substrate controlled growth--simultaneous ammonium and glucose limitation in chemostat cultures of Klebsiella pneumoniae; Rutgers M et al.; In chemostat cultures of Klebsiella pneumoniae (K . aerogenes) NCTC 418 we measured the concentrations of glucose and ammonium and we varied the ratio of the (limiting) concentrations of glucose and ammonium in the feed medium . By doing this at different dilution rates we found a range where growth rate varies with either concentration in the culture when the other concentration in the culture is held constant . This proves that within this range, dual-substrate controlled growth occurs . Dual substrate-controlled growth was accompanied by yield coefficients for glucose and for ammonium that were intermediate between the yield coefficients obtained for single glucose or single ammonium limitation . We quantified the control by either substrate in terms of the flux control coefficient with respect to that substrate, where flux refers to growth rate . Dual-substrate controlled growth is reflected by the finding that both flux control coefficients exceed zero, simultaneously . In the transition of glucose to ammonium limitation, the control gradually shifts from glucose to ammonium. Arch Microbiol, 1990, 153(3), 272 - 5 Recovery of exponentially growing cultures of Klebsiella pneumoniae NCIB 418 after heat shocks; Heitzer A et al.; Exponentially growing cultures of Klebsiella pneumoniae were subjected to heat shocks in the superoptimal and supermaximal temperature ranges for growth on glucose in a defined mineral salts medium . Transitory changes in the specific growth rate constant during recovery were evident . The response was heat shock temperature and exposure time dependent . Cell viability determinations, based on colony counts, indicated complete recovery from heat treatments at superoptimal temperatures . In contrast, at supermaximal temperatures, discrepancies in colony counts on different agars were observed . The kinetic response of the specific growth rate constant after a heat shock at supermaximal temperatures is explained by segregation within the bacterial population. Adv Exp Med Biol, 1990, 256, 247 - 55 Interaction of Mg2+ and Ca2+ in in vitro hexagonal assembly of R-form lipopolysaccharides; Kato N et al.; The R-form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN-111 (03-:Kl-), from which cationic material had been removed by electrodialysis, formed an orderly hexagonal lattice structure when suspended in 50 mM Tris buffer at pH 8.5 containing MgCl2 . The center-to-center distance (lattice constant) of the hexagonal lattice structure depended upon the concentration of MgCl2 and reached the shortest value (15 nm) at 10 mM . In contrast, CaCl2 could not produce the orderly hexagonal lattice structure but produced an irregular network structure with a center to center distance of 19 to 20 nm . When the LPS was suspended in Tris buffer containing 10 mM MgCl2 mixed with 1 or 10 mM CaCl2, formation of the orderly hexagonal lattice structure of the magnesium salt type was inhibited and the LPS showed the structure of the calcium salt type . When 1 or 10 mM CaCl2 was mixed with 10 mM MgCl2, the binding of Mg to the LPS was significantly inhibited compared with when 10 mM MgCl2 was added alone . On the contrary, when 10 mM CaCl2 was mixed with 10 mM MgCl2, the binding of Ca to the LPS was enhanced compared with when 10 mM CaCl2 was added alone . It was therefore concluded that the inhibition of formation of the hexagonal lattice structure of the magnesium salt type by addition of CaCl2 is due to the inhibition of the binding of Mg to the LPS . Such a competitive interaction of Mg2+ and Ca2+ was also observed with the electrodialyzed LPS of Escherichia coli K-12. Mol Microbiol, 1990 Jan, 4(1), 73 - 85 Molecular characterization of pulA and its product, pullulanase, a secreted enzyme of Klebsiella pneumoniae UNF5023; Kornacker MG et al.; The determined nucleotide sequence of the Klebsiella pneumoniae UNF5023 gene pulA comprises a single open reading frame coding for a 1090-residue precursor of the secreted protein pullulanase . The predicted sequence of this protein is highly homologous to that of pullulanase of Klebsiella aerogenes strain W70 . However, the UNF5023 pullulanase lacks a collagen-like sequence present at the N-terminus of the mature W70 enzyme and differs further from the W70 pullulanase around residue 300 and at the C-terminus . Pullulanases with or without the collagen-like sequence could not be separated by gel electrophoresis under denaturing or non-denaturing conditions, and were unaffected by collagenase . A large central domain which is highly conserved in both UNF5023 and W70 polypeptides contains eight short sequences that are also found in amylases and iso-amylases . Linker mutations in the region of the UNF5023 pulA gene coding for this domain abolished catalytic activity without affecting transport of the polypeptide across the outer membrane . Hybrid proteins comprising at least the amino-terminal 656 residues of prepullulanase fused to alkaline phosphatase were partially localized to the cell surface, as judged by their accessibility to anti-pullulanase serum in immuno-fluorescence tests . On the basis of these results, we tentatively propose that secretion signals required for recognition and translocation across the outer membrane via the pullulanase-specific extension of the secretion pathway are located near the N-terminus of the pullulanase polypeptide. Mol Microbiol, 1990 Jan, 4(1), 59 - 72 Analysis of the subcellular location of pullulanase produced by Escherichia coli carrying the pulA gene from Klebsiella pneumoniae strain UNF5023; Pugsley AP et al.; Three different techniques, protease accessibility, cell fractionation and in situ immunocytochemistry, were used to study the location of the lipoprotein pullulanase produced by Escherichia coli K12 carrying the cloned pullulanase structural gene (pulA) from Klebsiella pneumoniae, with or without the K . pneumoniae genes required to transport pullulanase to the cell surface (secretion-competent and secretion-incompetent, respectively) . Pullulanase produced by secretion-competent strains could be slowly but quantitatively released into the medium by growing the cells in medium containing pronase . The released pullulanase lacked the N-terminal fatty-acylated cysteine residue (and probably also a short N-terminal segment of the pullulanase polypeptide), confirming that the N-terminus is the sole membrane anchor in the protein . Pullulanase produced by secretion-incompetent strains was not affected by proteases, confirming that it is not exposed on the cell surface . Pullulanase cofractionated with both outer and inner membrane vesicles upon isopycnic sucrose gradient centrifugation, irrespective of the secretion competence of the strain . Examination by electronmicroscopy of vesicles labelled with antipullulanase serum and protein A-gold confirmed that pullulanase was associated with both types of vesicles . When thin-sectioned cells were examined by the same technique, pullulanase was found to be located mainly on the cell surface of the secretion-competent cells and mainly in the proximity of the inner membrane in the secretion-incompetent cells . Thus, while the results from three independent techniques (substrate accessibility, protease accessibility and in situ immunocytochemistry) show that pullulanase is transported to the cell surface of secretion-competent cells, this could not be confirmed by cell-fractionation techniques . Possible explanations for this discrepancy are discussed. Mol Microbiol, 1990 Jan, 4(1), 29 - 37 The function of isolated domains and chimaeric proteins constructed from the transcriptional activators NifA and NtrC of Klebsiella pneumoniae; Drummond MH et al.; A model for the domain structure of sigma 54-dependent transcriptional activators, based on sequence data, has been tested by examining the function of truncated and chimaeric proteins . Removal of the N-terminal domain of NtrC abolishes transcriptional activation, indicating that this domain is positively required for activator function . Over-expression of this domain as a separate peptide appears to titrate out the phosphorylating activity of NtrB . Removal of the N-terminal domain of NifA reduces activation 3-4-fold . The residual activity is particularly sensitive to inhibition by NifL, suggesting that the role of the N-terminal domain is to block the action of NifL in derepressing conditions . The C-terminal domain of NtrC showed repressor activity when expressed as a separate peptide . This domain is necessary for activator function even when NtrC binding sites are deleted from promoters . A point mutation in the ATP-binding motif of the NtrC central domain, Ser169 to Ala, also abolished activator function . Exchanging the N-terminal domains of Klebsiella pneumoniae NtrC, NifA and Escherichia coli OmpR, did not produce any hybrid activity, suggesting that N-terminal domains in the native proteins specifically recognize the rest of the molecule. J Antimicrob Chemother, 1990 Jan, 25(1), 127 - 32 Survival after intramuscular or inhalation administration of gentamicin in neutropenic guinea pigs infected by Klebsiella pneumoniae; Trnovec T et al.; Guinea pigs were infected by intranasal administration of Klebsiella pneumoniae on day 7 after irradiation with a dose of 4 Gy and treated on days 8-10 by gentamicin 4 mg/kg bd im or by inhalation leading to estimated systemically absorbed doses of 5.9 mg/kg/day . All animals died; however, gentamicin treatment significantly (P less than 0.05, Fisher's exact test) delayed death in the specific time intervals: 7-13 days (with the exception of day 11) and 12-26 days after im and inhalation administration respectively . Comparison of the two treatments by Fisher's exact test showed a significant advantage (P less than 0.05) for im vs inhalation treatment on days 9 and 10 only . The log rank test gave somewhat different results in that the delay of death after im administration was highly significantly (P less than 0.002) different from control whereas results after inhalation were not significantly different from control (P = 0.06) . Moreover, according to the log rank test, im administration delayed death significantly (P = 0.04) better than inhalation . In conclusion these data do not show any advantage of inhalation over im administration for treatment of experimental pneumonia, indeed they are compatible with im administration being marginally more effective. Vestn Otorinolaringol, 1990 Jan-Feb, (1), 31 - 8 {Ultrastructural studies of Klebsiella rhinoscleromatis}; Karchev T et al.; In 1870, F . Hebra described a slowly progressing granulomatous disease of the nasal mucous membrane he called rhinoscleroma . Later, N . M . Volkovich and A . Frisch isolated from patients suffering from this disease a microorganism they termed Klebsiella rhinoscleromatis . In 1932, S . Belinov proposed to name the disease Scleroma respiratorium because the pathological process developing in rhinosclerosis may involve not only upper but also lower airways . In 1961, T . Steffen and J . Smith demonstrated that Klebsiella rhinoscleromatis conformed to Koch's postulates and was an etiological factor of inflammatory changes typical of scleroma . This paper presents results of ultrastructural examinations of Klebsiella rhinoscleromatis isolated from two patients with Scleroma respiratorium . It was shown that as the degenerative process developed, the amount of electron-dense material around the bacterial cell wall decreased progressively and only fragments of the cellular membrane remained. Infection, 1990 Jan-Feb, 18(1), 48 - 52 Perspectives of beta-lactamases inhibitors in therapy of infections caused by Escherichia coli or Klebsiella with plasmidic resistance to third generation cephalosporins; Bauernfeind A; New plasmidic beta-lactamases inactivating so far stable cephalosporins, aztreonam and cephamycins restrict the use of these antibiotics in therapy of infections, e.g., by Escherichia coli and Klebsiella . Thus, combinations of beta-lactamase inhibitors and beta-lactam antibiotics were investigated in vitro with regard to their therapeutic perspectives . Minimal inhibitory concentrations and the kinetics of killing in a pharmacodynamic model were determined . Extended broad spectrum beta-lactamases (EBS-beta-lactamases) representative both for the TEM- and SHV-type were included . None of the available fixed combinations of penicillins and beta-lactamase inhibitors appears useful for therapy of infections caused by producers of EBS-beta-lactamases . In contrast, combinations of piperacillin and tazobactam or sulbactam plus cephalosporins (cefoperazone, cefotaxime, ceftazidime) or aztreonam are highly active (both by their MICs and bactericidal activity) against TEM-type EBS-beta-lactamases, but less promising for the SHV-type EBS-beta-lactamases, and plasmidic cephamycinase . Of the beta-lactams available, the monobactam carumonam and the carbapenems (imipenem, meropenem) remain safe in infections caused by E . coli and Klebsiella EBSBase producers. Rev Paul Med, 1990 Jan-Feb, 108(1), 26 - 8 {Rotavirus in newborns less than 24 hours of age}; Tavares Junior Pde A; During 22 months, 687 samples, including feces and meconium of patients from the baby nursery (group A--236 samples) and from newborns seen in the outpatient clinic (group B--451 samples), were submitted to ELISA, PAGE, and ME, searching for rotavirus and were also cultivated for bacteria, at the HUAP (UFF) . Nonsymptomatic newborns were the only positive cases for rotavirus, a rate of 2.11% . Contamination at delivery was suspected, but there was no correlation between either cesarean or vaginal labor and maternal diarrhea with newborn diarrhea after delivery . Klebsiella pneumoniae was the main bacteria in the symptomatic babies from the ward. Plasmid, 1990 Jan, 23(1), 27 - 34 Direct sequencing of the amplified structural gene and promoter for the extended-broad-spectrum beta-lactamase TEM-9 (RHH-1) of Klebsiella pneumoniae; Mabilat C et al.; Extended-broad-spectrum beta-lactamase TEM-9, detected in a clinical isolate of Klebsiella pneumoniae, confers high-level resistance to recent cephalosporins, in particular ceftazidime, and to the monobactam aztreonam . Using oligonucleotide probes, we found that the plasmid gene blaT-9 encoding TEM-9 differs from characterized blaT genes by a new combination of already known mutations . Gene blaT-9 was further studied by direct sequencing of an amplified 1.1-kb DNA fragment which contained the open reading frame and its promoter . Analysis of the nucleotide and of the deduced amino acid sequence confirmed the hybridization results and indicated that TEM-9 differs from TEM-1 by four amino acid substitutions: Phe at position 19 and Met at position 261, which have been found in TEM-4 and are known not to expand the enzyme substrate range; Lys 102, detected in TEM-3 and TEM-4, and Ser 162, present in TEM-5 and TEM-7 . Each of the latter substitutions enlarges the substrate spectrum of the enzymes and they are found associated for the first time in TEM-9. Arch Microbiol, 1990, 153(5), 502 - 5 The role of magnesium and calcium ions in the glucose dehydrogenase activity of Klebsiella pneumoniae NCTC 418; Buurman ET et al.; Magnesium-limited chemostat cultures of Klebsiella pneumoniae NCTC 418 with 20 microM CaCl2 in the medium showed a low rate of gluconate plus 2-ketogluconate production relative to potassium- or phosphate-limited cultures . However, when the medium concentration of CaCl2 was increased to 1 mM, the glucose dehydrogenase (GDH) activities also increased and became similar to those observed in potassium- or phosphate limited cultures . It is concluded that this is due to Mg2+ and Ca2+ ions being involved in the binding of pyrroloquinoline quinone (PQQ) to the GDH apoenzyme . There seems to be an absolute requirement of divalent cations for proper enzyme functioning and in this respect Ca2+ ions could replace Mg2+ ions . The high GDH activity which has been found in cells grown under Mg2(+)-limited conditions in the presence of higher concentrations of Ca2+ ions, is compatible with the earlier proposal that GDH functions as an auxiliary energy generating system involved in the maintenance of high transmembrane ion gradients. Ter Arkh, 1990, 62(12), 74 - 6 {Microbiocenosis and the status of the intestinal mucosa in alkylosing spondyloarthritis}; Guseinov NI et al.; Intestinal microbiocenosis was examined in 53 patients with ankylosing spondyloarthritis (AS) . Of these, 23 had the central and 30 presented with the peripheral form . 34 patients underwent endoscopic examinations (rectoromanoscopy, ileocolonoscopy) . As a result, in 74% of cases with the central form of AS and in 80% of cases with the peripheral form of AS, alterations in the intestinal microflora were revealed, which can be regarded as intestinal dysbacteriosis (ID), with dysbacteriosis being more pronounced in patients with the peripheral form . In such patients it manifested by a considerable decrease of the amount of bifido- and lactoflora, which promoted the increment of the amount of opportunistic microorganisms, particularly Klebsiella . Macroscopic signs of nonspecific inflammation of the small and large intestines were identified in 63% of patients with the central and in 67% of those with the peripheral form of AS . Mean-while histomorphological study of biopsy specimens of the intestinal mucosa showed that all the examinees had chronic inflammation of varying degree. Medicina (B Aires), 1990, 50(6), 543 - 7 Dissemination of plasmid-mediated amikacin resistance among pathogenic Klebsiella pneumoniae; Chamorro RM et al.; Plasmids isolated from Klebsiella pneumoniae strains that caused outbreaks in pediatric units in various geographical regions of Argentina harbored genetic determinants for resistance to amikacin . By using restriction endonuclease and Southern blot hybridization analysis it was determined that all of the strains carried plasmids with Tn1331-related elements indicating that transposition of these elements may have played an important role in the dissemination process of resistance to amikacin. Drugs Exp Clin Res, 1990, 16(12), 615 - 9 Post-antibiotic effect of azithromycin on respiratory tract pathogens; Debbia EA et al.; Azithromycin is a new azalide antibiotic with structural modifications that confer to the molecule acid stability, extension of antibacterial spectrum that includes important Gram-negative pathogens, long elimination half-life, and tendency to concentrate into various tissues where it persists for extended periods of time . The existence and length of a post-antibiotic effect (PAE), an important parameter for the characterization of new antibiotic molecules, has not yet been evaluated for this agent . In this study the PAE of azithromycin was assessed against representative respiratory pathogens included in the in vitro antimicrobial activity of the drug . The results obtained indicate that azithromycin produce a significant PAE on all Gram-positive and Gram-negative bacteria tested, resulting in an average value of 3.5 h for both S . pyogenes and S . pneumoniae, 3 h for B . catarrhalis and H . influenzae, and 2 h for Klebsiella spp . These findings support previous reports underlining the remarkable in vitro activity of azithromycin against H . influenzae, a pathogen poorly susceptible to the classical macrolides . Furthermore, the present demonstration of the existence of a long PAE of azithromycin against other Gram-positive and Gram-negative bacteria extends the pharmacokinetic advantages of the drug and strongly supports the application of this azalide in the therapy of respiratory infections. Scand J Rheumatol, 1990, 19(5), 341 - 9 The use of enzyme immunoassay (EIA) and radiobinding assay to investigate the cross-reactivity of Klebsiella antigens and HLA B27 in ankylosing spondylitis patients and healthy controls; Baines M et al.; The binding of rabbit anti-Klebsiella antibodies to tissue-typed lymphocytes obtained from 30 ankylosing spondylitis (AS) patients and 54 healthy subjects has been measured by an enzyme immunoassay method . HLA B27 positive lymphocytes obtained from either AS patients (t = 3.60; p less than 0.001) or healthy subjects (t = 3.77; p less than 0.001) were found to bind Klebsiella antibodies to a significantly greater extent than non-B27 lymphocytes obtained from healthy controls . Absorption studies demonstrated that HLA B27 positive lymphocytes absorbed out significantly more anti-Klebsiella antibodies than HLA B27-negative lymphocytes (t = 6.76; p less than 0.005) . These results are compatible with cross-reactivity or molecular mimicry between HLA B27 and epitopes on some Gram-negative bacteria such as Klebsiella. Gene, 1989 Dec 21, 85(1), 153 - 60 Functional expression of a Rhodospirillum rubrum gene encoding dinitrogenase reductase ADP-ribosyltransferase in enteric bacteria; Fu HA et al.; The function of the cloned draT gene of Rhodospirillum rubrum was studied by placing it under the control of the tac promoter in the vector, pKK223-3 . After induction with isopropyl-beta-D-thiogalactopyranoside, dinitrogenase reductase ADP-ribosyltransferase (DRAT) activity was detected in crude extracts of the heterologous hosts Escherichia coli and Klebsiella pneumoniae . In addition, the expression of draT produced a Nif- phenotype in the otherwise wild-type K . pneumoniae strains, the result of the ADP-ribosylation of accumulated dinitrogenase reductase (DR) . DR from a nifF- background was also susceptible to ADP-ribosylation, indicating that the oxidized form of DR will serve as a substrate for DRAT in vivo . A mutation that changes the Arg-101 residue of DR, the ADP-ribose attaching site, eliminates the ADP-ribosylation of DR in vivo, confirming the necessity of this residue for modification. Biochem J, 1989 Dec 15, 264(3), 657 - 61 A transient-kinetic study of the nitrogenase of Klebsiella pneumoniae by stopped-flow calorimetry . Comparison with the myosin ATPase; Thorneley RN et al.; The pre-steady-state kinetics of MgATP hydrolysis by nitrogenase from Klebsiella pneumoniae were studied by stopped-flow calorimetry at 6 degrees C and at pH 7.0 . An endothermic reaction (delta Hobs . = +36 kJ.mol of ATP-1; kobs . = 9.4 s-1) in which 0.5 proton.mol of ATP-1 was released, has been assigned to the on-enzyme cleavage of MgATP to yield bound MgADP + Pi . The assignment is based on the similarity of these parameters to those of the corresponding reaction that occurs with rabbit muscle myosin subfragment-1 (delta Hobs . = +32 kJ.mol of ATP-1; kobs . = 7.1 s-1; 0.2 proton released.mol of ATP-1) {Millar, Howarth & Gutfreund (1987) Biochem . J . 248, 683-690} . MgATP-dependent electron transfer from the nitrogenase Fe-protein to the MoFe-protein was monitored by stopped-flow spectrophotometry at 430 nm and occurred with kobs . value of 3.0 s-1 at 6 degrees C . Thus, under these conditions, hydrolysis of MgATP precedes electron transfer within the protein complex . Evidence is presented that suggests that MgATP cleavage and subsequent electron transfer are reversible at 6 degrees C with an overall equilibrium constant close to unity, but that, at 23 degrees C, the reactions are essentially irreversible, with an overall equilibrium constant greater than or equal to 10. Cancer, 1989 Dec 1, 64(11), 2368 - 76 Klebsiella bacteremia . A 10-year review in a cancer institution; Bodey GP et al.; A total of 368 episodes of Klebsiella bacteremia occurred in 330 cancer patients, representing a rate of four episodes per 1000 hospital admissions . Eighty-eight percent of these infections were acquired nosocomically and 58% of the patients received antibiotics during the preceding 10-day period . There was pulmonary infection in 24% of the patients, shock in 25%, and disseminated intravascular coagulation in 7% . The overall response rate was 69% . Response rates were significantly lower among patients with shock (25% versus 83%), hemorrhage (29% versus 76%), and pneumonia (37% versus 79%) . The combination of a cephalosporin plus an aminoglycoside produced the highest response rate (79%) . Klebsiella sp continue to be an important cause of infection in patients with cancer. Antimicrob Agents Chemother, 1989 Dec, 33(12), 2096 - 100 Molecular characterization of the gene encoding SHV-3 beta-lactamase responsible for transferable cefotaxime resistance in clinical isolates of Klebsiella pneumoniae; Nicolas MH et al.; In Klebsiella pneumoniae 86-4, cefotaxime resistance was due to a transferable broad-spectrum beta-lactamase, SHV-3 . The plasmid-borne gene encoding SHV-3 has been cloned, and the primary structure of the enzyme was deduced from its nucleotide sequence . SHV-3 differs from SHV-1 in two positions . The extended substrate profile of SHV-3 probably results from the substitution of Ser-213 for Gly, as in SHV-2, whereas replacement of Arg-180 by Leu resulted in a decrease in the pI from 7.6 to 7.0 . The blashv-3 gene is highly homologous (92% DNA sequence identity) with the chromosomal gene coding for LEN-1 beta-lactamase of K . pneumoniae, suggesting that the origin of the SHV-encoding genes now present on many plasmids may be chromosomal. Clin Rheumatol, 1989 Dec, 8(4), 517 - 21 Infectious arthritis caused by Klebsiella . A report of two cases; Markusse HM et al.; Klebsiella arthritis is uncommon . This report describes 2 cases seen in one year in one hospital and reviews the previous 9 well-documented cases . Despite a long delay in diagnosis, antibiotic treatment without drainage procedures resulted in complete cure in one patient and cure with some impairment in the other. Arch Pathol Lab Med, 1989 Dec, 113(12), 1381 - 3 Disseminated Klebsiella rhinoscleromatis infection; Porto R et al.; A 35-year-old obese black American woman presented with nausea, vomiting, diarrhea, fever, cough, and chest pain of 2 weeks duration . She was pancytopenic and acidotic, with respiratory failure and hypotension . A diagnosis of septic shock was made, and the patient died 48 hours after admission . Blood cultures were positive for organisms that were reported to be Klebsiella rhinoscleromatis . At autopsy she had massive hepatic necrosis with numerous Mikulicz's cells . The lungs, spleen, and bone marrow were also involved . To our knowledge, this is the first report of a case of systemic infection with K rhinoscleromatis. Infect Immun, 1989 Dec, 57(12), 3778 - 82 Reduction of capsular polysaccharide production in Klebsiella pneumoniae by sodium salicylate; Domenico P et al.; Heavily encapsulated Klebsiella pneumoniae (serotypes 1 and 2) was cultured in the presence of sodium salicylate . The addition of salicylate (2 to 30 micrograms/ml) progressively decreased the amount of capsular polysaccharide produced by all strains without significantly inhibiting cell growth . Further addition of salicylate (50 to 200 micrograms/ml) was progressively inhibitory to cell growth and decreased the production of polysaccharide only slightly . The optimal concentration of salicylate that could reduce the polysaccharide production of heavily encapsulated, virulent strains by 50% or more was 30 micrograms/ml . Mutants of these bacteria that produced less capsule were affected by salicylate to a lesser degree . All concentrations of salicylate tested were physiologically achievable in humans and within the therapeutic range of aspirin . The addition of calcium and magnesium partially reversed the effects of salicylate on polysaccharide production . Chelating agents, particularly EGTA {ethylene-bis(oxyethylenenitrile)tetraacetic acid}, reduce capsule production as salicylate did . Thus, the chelation of calcium and magnesium by salicylate could account, at least in part, for the reduction of capsule . Optical density measurements allowed for rapid monitoring of capsule production in various culture media because a large part of culture turbidity was apparently due to the capsule . Decreased production of the primary K . pneumoniae virulence factor with salicylate may have therapeutic potential. J Vasc Surg, 1989 Dec, 10(6), 635 - 41 Mycotic aneurysms of the suprarenal abdominal aorta: prolonged survival after in situ aortic and visceral reconstruction; Atnip RG; Necrotizing infection of the arterial wall causes rupture and false ("mycotic") aneurysm formation, with a very poor prognosis if untreated . Cure can be achieved by surgical drainage and debridement, with restoration of arterial continuity through uncontaminated tissues . The dilemma of applying these principles to the treatment of mycotic aneurysms of the suprarenal aorta is that no remote or extraanatomic routes are available to maintain perfusion to the viscera . We report the first case of Klebsiella suprarenal mycotic aortic aneurysm successfully treated with in situ prosthetic reconstruction of the aorta and visceral arteries, and we have reviewed the 21 other suprarenal mycotic aortic aneurysms reported in the English-language literature . Repair was performed in 20 of the 22 cases, with in situ prosthetic reconstruction performed in 18 . Prolonged survival has been achieved in 16 patients after in situ repair . No long-term survival has been reported after extraanatomic reconstruction of the aorta and visceral arteries in patients with such aneurysms . We conclude that in situ prosthetic reconstruction, accompanied by thorough drainage and debridement, prolonged parenteral antibiotic therapy, and permanent suppressive oral antibiotics, offers the best chance for survival in these patients. J Nutr Sci Vitaminol (Tokyo), 1989 Dec, 35(6), 661 - 5 Effects of the yeast extract components pyrroloquinoline quinone and aspartic acid on vitamin B12 production in Klebsiella pneumoniae IFO 13541; Ohsugi M et al.; The production of vitamin B12 from carbohydrates, peptone, casamino acid, etc., by intestinal bacteria was investigated . Klebsiella pneumoniae IFO 13541 was the most efficient strain for vitamin B12 production, which depended exclusively on the concentration of yeast extract added to the medium . A concentrated solution of yeast extract (1 ml) was chromatographed on a Sephadex G-25 column (1 x 180 cm) and eluted with H2O (eighty fractions of 3 ml each were collected) . It was found that fractions in which bacterial growth was most prevalent also exhibited the highest amount of vitamin production . The effectiveness of yeast extract was shown by the participation of pyrroloquinoline quinone and aspartic acid in the growth stimulation and in the vitamin B12 production in this strain. J Biol Response Mod, 1989 Dec, 8(6), 656 - 64 Comparative effects of F1 and P1 fractions obtained from a Klebsiella pneumoniae glycoproteic extract (RU 41740) on polymorphonuclear leukocytes; el Abbouyi A et al.; RU 41740 is a glycoprotein extract from Klebsiella pneumoniae described as a macromolecular aggregation of a lipopolysaccharide (LPS)-associated protein (F1 fraction) and a glycoproteic complex (P1 fraction) . The human polymorphonuclear (PMN) response was studied after incubation of the cells in the presence of RU 41740, F1 and P1 fractions, or F1-P1 complex . Oxidative metabolism was assessed by chemiluminescence, O2 consumption, O2- generation, and degranulation by beta-glucuronidase release . Results were compared to data obtained with a homologous LPS . RU 41740, F1 fraction, and F1-P1 complex increased the respiratory burst of PMNs stimulated by opsonized zymosan (OZ) . N-formylmethionylleucylphenylalanine (fMLP), phorbol myristate acetate, or the calcium ionophore A23187 . The beta-glucuronidase release was stimulated by the same compounds when OZ or fMLP were used as stimuli . These effects were dose-dependent . In contrast, P1 fraction was inactive . Addition of polymyxin B resulted in a profound inhibition of both the F1 fraction and LPS activities but only in a partial inhibition of RU 41740 effects . These results strengthen the hypothesis that different biochemical pathways are involved in the enhancement of stimulated neutrophil functions by RU 41740. J Gen Microbiol, 1989 Dec, 135 ( Pt 12), 3173 - 81 Novel aerobactin receptor in Klebsiella pneumoniae; Williams P et al.; Several Klebsiella pneumoniae strains which produced enterochelin but not aerobactin were nevertheless sensitive to cloacin DF13 . In contrast, a strain of serotype K1:O1 which produced both siderophores was cloacin-resistant . Loss by mutation of the O1 but not K1 antigen rendered this strain cloacin-sensitive, indicating that the O1 antigen prevented access of cloacin to the cloacin/aerobactin receptor . Unlike the K1:O1 strain, the aerobactin-negative strains failed to hybridize in a colony blot assay with an aerobactin receptor gene probe prepared from pColV-K30 . However, antisera raised against the 74 kDa pColV-K30 aerobactin receptor cross-reacted with a 76 kDa outer-membrane protein in each K . pneumoniae strain . In addition to the 76 kDa protein, the K1:O1 strain also produced a strongly cross-reacting 74 kDa protein . To determine whether these aerobactin-negative strains could use aerobactin, mutants unable to synthesize siderophores were isolated . Aerobactin promoted the growth of these mutants in iron-deficient media . The evidence presented suggests that some K . pneumoniae strains produce an aerobactin iron-uptake system without apparent production of aerobactin and which is probably based on a 76 kDa receptor, the gene for which does not hybridize with aerobactin receptor gene encoded on pColV-K30. Burns, 1989 Dec, 15(6), 403 - 6 Long-term experience with 1 per cent topical silver sulphadiazine cream in the management of burn wounds; Sawhney CP et al.; Three hundred and forty-two patients with 10-50 per cent body surface area burns were studied prospectively over the 5-year period from 1982 to 1986 for the effectiveness of topical 1 per cent silver sulphadiazine . Various parameters were studied including: (i) healing time of deep partial skin thickness burns, (ii) eschar separation time, (iii) conversion rate of deep dermal burns to full skin thickness burns, (iv) burn wound surface bacterial flora and their changing pattern over the years, (v) incidence of invasive sepsis and (vi) overall mortality . There was a remarkable decrease in the time taken for the healing of deep dermal burns, and the conversion rate of deep dermal burns to full skin thickness was significantly reduced . Eschar separation was delayed considerably . There was a total change in the predominent surface micro-organisms from Staph . aureus, which was predominant in 1982, to pseudomonas species and klebsiella in 1986 . Moreover, there was the emergence of a new variety of micro-organism within the last 2 years . The incidence of invasive infection and overall mortality was significantly reduced. Genetics, 1989 Dec, 123(4), 635 - 48 DNA sequence analysis of artificially evolved ebg enzyme and ebg repressor genes; Hall BG et al.; The ebg system has been used as a model to study the artificial selection of new catalytic functions of enzymes and of inducer specificities of repressors . A series of mutant enzymes with altered catalytic specificities were previously characterized biochemically as were the changes in inducer specificities of mutant, but fully functional, repressors . The wild type ebg operon has been sequenced, and the sequence differences of the mutant enzymes and repressors have been determined . We now report that, contrary to our previous understanding, ebg enzyme contains 180-kD alpha-subunits and 20-kD beta-subunits, both of which are required for full activity . Mutations that dramatically affect substrate specificity and catalytic efficiency lie in two distinct regions, both well outside of the active site region . Mutations that affect inducer specificity of the ebg repressor lie within predicted sugar binding domains . Comparisons of the ebg beta-galactosidase and repressor with homologous proteins of the Escherichia coli and Klebsiella pneumoniae lac operons, and with the galactose operon repressor, suggest that the ebg and lac operons diverged prior to the divergence of E . coli from Klebsiella . One case of a triple substitution as the consequence of a single event is reported, and the implications of that observation for mechanisms of spontaneous mutagenesis are discussed. Br J Surg, 1989 Dec, 76(12), 1282 - 6 Protective effects of recombinant human tumour necrosis factor alpha and interferon gamma against surgically simulated wound infection in mice; Hershman MJ et al.; Tumour necrosis factor alpha and interferon gamma have both been shown to have immunoregulatory properties, and to be able to influence, several microbial infections . This study showed that tumour necrosis factor was effective in modifying surgically simulated wound infections when administered both as prophylaxis and as therapy . Two models were used; one was an intramuscular bacterial challenge, and the other involved the use of a bacteria-laden thigh suture . The test bacterium for both models was Klebsiella pneumoniae, a common surgical pathogen in our surgical service . Interferon gamma was an effective biological response modifier in these models . Tumour necrosis factor was more potent than interferon gamma and there was no additive or synergistic effect with interferon gamma . This indicates potentially different mechanisms of action for these two cytokines. Biochim Biophys Acta, 1989 Nov 23, 977(2), 142 - 9 Effect of concentration of substrates and products on the growth of Klebsiella pneumoniae in chemostat cultures; Rutgers M et al.; Non-equilibrium thermodynamics (NET) can be used to describe microbial growth . In this description, the concentrations of products contribute to the driving forces of the metabolic processes (anabolism and catabolism) . Thus, in contrast to the model of bacterial growth of Monod (Recherches sur la Croissance les Cultures Bacteriennes (1942) Herman et Cie, Paris), it is predicted that the growth rate of a bacterial chemostat culture is, in principle, dependent on the concentration of the catabolic product (for instance HCO3-) during catabolite limitation and on the concentration of the anabolic product (for instance biomass) during anabolite limitation . In order to test this prediction, Klebsiella pneumoniae was grown in aerobic citrate-limited, glucose-limited or ammonia-limited chemostat cultures . Ammonia-limited cultures were considered to be essentially anabolite-limited, whereas citrate limitation was used as a representative for catabolite limitation . In ammonia-limited or in glucose-limited cultures it was found that the growth rate was independent of the biomass concentration present . In the NET description this means that the 'back' reaction (i.e., in the direction from biomass to substrates) is saturated with respect to biomass . On the other hand, in citrate-limited cultures, the steady-state concentration of citrate increased with the concentration of the catabolic product HCO3- . At relatively low concentrations of HCO3-, 'thermodynamic back-pressure' of growth (i.e., increase in product concentration was compensated by an increase in substrate concentration so that the driving force for growth remained almost constant) was demonstrated as predicted by the NET model . At concentrations above 40 mM, a kinetic (allosteric) effect of HCO3- was detected . This was concluded from a reduced growth yield on citrate, and from a significant decrease in the maximal growth rate and the maximal oxygen consumption rate after relief of the citrate limitation. Biochem J, 1989 Nov 15, 264(1), 257 - 64 Site-directed mutagenesis of the Klebsiella pneumoniae nitrogenase . Effects of modifying conserved cysteine residues in the alpha- and beta-subunits; Kent HM et al.; The five conserved cysteine residues present in the alpha-subunit and the three conserved cysteine residues present in the beta-subunit of nitrogenase component 1 were individually changed to alanine . Mutations in the alpha-subunit at positions 63, 89, 155 and 275 and in the beta-subunit at positions 69, 94 and 152 all resulted in a loss of diazotrophic growth and component 1 activity and loss of the normal e.p.r . signal of the component 1 protein . Component 2 activity was retained . Replacement of cysteine-184 in the alpha-subunit with alanine greatly diminished, but did not eliminate, diazotrophic growth and component 1 activity . Substitution of serine for cysteine at position 152 in the beta-subunit, in contrast with the substitution of alanine at this position, resulted in the formation of active component 1 . Replacement of the non-conserved cysteine-112 in the beta-subunit with alanine did not greatly perturb diazotrophic growth or the activity of component 1 . Extracts prepared from a mutant, with cysteine-275 of the alpha-subunit replaced by alanine, complemented extracts of a mutant unable to synthesize the iron-molybdenum cofactor of nitrogenase, indicating that the alanine-275 substitution increases the availability of cofactor . Furthermore extracts of this mutant exhibited an e.p.r . signal similar to that of extracted iron-molybdenum cofactor . These data suggest a role for cysteine-275 as a ligand to the cofactor. Mikrobiol Zh, 1989 Nov-Dec, 51(6), 25 - 30 {The compositional characteristics of the cellular fatty acids in strains of Klebsiella pneumoniae serovar K 1}; Vasiurenko ZP et al.; Klebsiella pneumoniae, serovar K1 strains, differ essentially in the composition of cellular fatty acids from serovar strains K 2, K 3, K 7-K 72, K 74-K 80 of this species and are analogous to K . rhinoscleromatis and K . ozaenae by the given trait . The differences concern the concentration of cyclopropane fatty acids and their biosynthetic precursors--monounsaturated fatty acids, that is typical of a number of other bacteria belonging to the related species and, evidently, may be a marker of phenetic differences at the level of related taxonomic groups . The data obtained evidence for taxonomic separation of K . pneumoniae serovar K1 strains from representatives of other serovars of the given species. FEMS Microbiol Lett, 1989 Nov, 53(1-2), 5 - 9 Biochemical, immunological and physicochemical comparisons between OHIO-1 and four SHV-type beta-lactamases; Vedel G et al.; The biochemical, immunological and physicochemical properties of the beta-lactamase OHIO-1 were compared to those of four beta-lactamases commonly found in Klebsiella pneumoniae: SHV-1, SHV-3 and the beta-lactamases of strains GN 11-03 and GN 422 . The substrate profile of SHV-1, OHIO-1 and of the beta-lactamases GN 11-03 and GN 422 were similar, while that of SHV-3 appeared comparable to that of the extended spectrum SHV-2 . Moreover, anti-TEM-1 serum inactivated OHIO-1 as well as SHV-1 and the beta-lactamases of strains GN 11-03 and GN 422 . Analysis of the electrophoretic mobilities, isoelectric points and titration curves demonstrated that OHIO-1 and the 4 other beta-lactamases examined were closely related variants . From these findings it appears that OHIO-1 could be classified among the SHV-type beta-lactamases. Zentralbl Hyg Umweltmed, 1989 Nov, 189(2), 147 - 50 An improved broth-microdilution procedure for testing metal ion resistance of Klebsiella pneumoniae; Podbielski A et al.; A new broth-microdilution method for testing metal ion resistance was designed in order to circumvent the formation of insoluble complexes between cations and compounds of the test medium . High-level cefotaxime resistant K . pneumoniae isolates tested by this procedure proved to be susceptible to Cd, Hg and Zn but resistant to Pb, Mo and W . The metal ion resistance was not linked to the beta-lactam resistance gene. EMBO J, 1989 Nov, 8(11), 3491 - 9 Transcriptional activation of the Klebsiella pneumoniae nifLA promoter by NTRC is face-of-the-helix dependent and the activator stabilizes the interaction of sigma 54-RNA polymerase with the promoter; Minchin SD et al.; Activation of transcription at the Klebsiella pneumoniae nifLA promoter requires the phosphorylated form of the positive control protein NTRC, together with RNA polymerase modified by the alternative sigma factor sigma 54 . Dimethylsulphate and potassium permanganate were used as probes to analyse the interaction of NTRC and sigma 54-RNA polymerase with supercoiled nifLA promoter DNA in vitro . In contrast to the glnAp2 promoter, sigma 54 holoenzyme did not protect guanine residues in the nifLA promoter from methylation in the absence of the activator . We propose that NTRC stabilizes the interaction of sigma 54-RNA polymerase with the -24, -12 region, in addition to its role in catalysing open complex formation . Phosphorylated NTRC binds to two sites located greater than 100 nucleotides upstream of the -24, -12 region; it also induces hyper-methylation of a G residue at -23 . Enhanced methylation at -23 is not co-operative with the binding of activator to the upstream sites and may account for the ability of NTRC, when present at high concentration, to activate transcription in the absence of the upstream binding sites . The insertion of spacer mutations at -86 indicates that transcriptional activation of the nifLA promoter at low NTRC concentrations is face-of-the-helix dependent, both in vivo and in vitro . We propose that correct positioning of activator molecules at the upstream binding sites stabilizes the interaction of sigma 54-RNA polymerase with the downstream region via the formation of a DNA loop. J Clin Microbiol, 1989 Nov, 27(11), 2539 - 43 Colonization of neonates in a nursery ward with enteropathogenic Escherichia coli and correlation to the clinical histories of the children; Senerwa D et al.; Stool samples were examined from 30 preterm neonates admitted to a nursery ward; 16 neonates had diarrhea, 12 constituted an age-matched control group without diarrhea, and 2 had an unknown history regarding diarrhea . Variable numbers of enteropathogenic Escherichia coli serotype O111:HNT strains possessing the gene coding for the enteroadherence factor (EAF) were found in stool samples from 13 of the neonates . No other microbiological enteropathogen was found . A total of 294 strains (9 or 10 from each neonate, comprising 229 E . coli and 65 Klebsiella pneumoniae strains) were characterized with respect to plasmid content and grouped into 37 plasmid profile groups . Diarrhea was found not to be correlated with any specific plasmid profile or with the presence of the EAF-positive strains but rather with the number of strains with one specific plasmid profile or with the number of EAF-positive strains (of the 9 or 10 strains) isolated from each stool sample . All the neonates who died had diarrhea (5 died of 16 with diarrhea); all five of the neonates who died possessed strains with one specific plasmid profile group, and EAF-positive strains were isolated from four of them . Of the seven neonates from whom seven or more EAF-positive isolates were isolated, three died, compared with only one of five of those from whom only a few (1 to 3 of 10) EAF-positive strains were isolated . Both plasmid profiling and genetic probing with the EAF probe were found to be good alternatives when serotyping is not available for identification of O111:HNT enteropathogenic E . coli strains. Antimicrob Agents Chemother, 1989 Nov, 33(11), 1915 - 20 Multiplicity of TEM-derived beta-lactamases from Klebsiella pneumoniae strains isolated at the same hospital and relationships between the responsible plasmids; Chanal CM et al.; Five plasmid-mediated beta-lactamases conferring high-level resistance to ceftazidime were isolated from Klebsiella pneumoniae strains in the same hospital . These enzymes had isoelectric points ranging from 5.3 to 6.5 (CAZ-1, 5.55; CAZ-2, 6.0; CAZ-3, 5.3; CAZ-6, 6.5; and CAZ-7, 6.3) . All isolates and their Escherichia coli transconjugants were highly resistant to amoxicillin (MICs, greater than 4,096 micrograms/ml), piperacillin (64 to 256 micrograms/ml), cephalothin (32 to 256 micrograms/ml), and ceftazidime (32 to 512 micrograms/ml) but remained moderately susceptible to cefotaxime (0.5 to 8 micrograms/ml) . Only CAZ-6- and CAZ-7-producing strains were highly resistant to aztreonam (64 to 128 micrograms/ml) . All the isolates remained susceptible to moxalactam and imipenem . The reduced activity of piperacillin, cefotaxime, ceftazidime, or aztreonam was restored by 2 micrograms of clavulanate, sulbactam, tazobactam, or brobactam per ml for E . coli producing CAZ-2, CAZ-3, and CAZ-7 . Sulbactam had a lower protective effect than other inhibitors for E . coli harboring CAZ-1 and especially CAZ-6 . Except for CAZ-1, which was mediated by a 150-kilobase (kb) plasmid (pCFF14), the other ceftazidimases were mediated by plasmids of 85 kb with EcoRI digestion patterns similar to that of pCFF04 encoding CTX-1 beta-lactamase . A TEM probe hybridized with a 19-kb EcoRI fragment of all these closely related plasmids. Can J Microbiol, 1989 Nov, 35(11), 994 - 9 Use of a bacteriophage-encoded glycanase enzyme in the generation of lipopolysaccharide O side chain deficient mutants of Escherichia coli O9:K30 and Klebsiella O1:K20: role of O and K antigens in resistance to complement-mediated serum killing; McCallum KL et al.; Coliphage K30 lysates contain free and phage-associated forms of a bacteriophage-encoded capsule depolymerase (glycanase) enzyme, active against the serotype K30 capsular polysaccharide of Escherichia coli . The free glycanase has been purified to apparent homogeneity . The molecular weight of the enzyme was estimated at 450,000, and when heated in SDS at 100 degrees C, the enzyme dissociated into two subunits of 90,000 and 52,000 . The glycanase enzyme was used as a reagent to reversibly degrade the capsular layers on cells of Escherichia coli O9:K30 and Klebsiella O1:K20 . This treatment rendered these bacteria sensitive to their respective lipopolysaccharide-specific bacteriophages, coliphage O9-1 and Klebsiella phage O1-3 . This novel approach facilitated isolation of lipopolysaccharide O antigen side chain deficient mutants which retained the ability to synthesize the capsule . The response of defined mutants, O+:K-, O-:K+, and O-:K-, to exposure to nonimmune rabbit serum was measured . Results showed that the primary barrier against complement-mediated serum killing in both Escherichia coli O9:K30 and Klebsiella O1:K20 was the O antigen side chains of the lipopolysaccharide molecules . In both strains, the capsule played no role in the determination of serum resistance. Carbohydr Res, 1989 Oct 31, 193, 147 - 55 A re-investigation of the structure of the capsular polysaccharide of Klebsiella K10; Dutton GG et al.; The capsular polysaccharide of Klebsiella K10 was investigated by methylation analysis and 1H-n.m.r . spectroscopy, and the deacetylated bacteriophage-degraded polysaccharide by 1D- and 2D-n.m.r . spectroscopy . The repeating unit was shown to be a branched hexasaccharide (see text) . OAc substituents were located on the terminal and 2-linked galactopyranosyl residues by Prehm methylation of a low-molecular-weight fraction obtained by bacteriophage degradation. J Biol Chem, 1989 Oct 15, 264(29), 17524 - 31 Biosynthesis and secretion of pullulanase, a lipoprotein from Klebsiella aerogenes; Murooka Y et al.; We constructed, by site-directed mutagenesis, a mutant pullulanase gene in which the cysteine residue in a pentapeptide sequence, Leu16-Leu-Ser-Gly-Cys20 within the NH2-terminal region of pullulanase from Klebsiella aerogenes, is replaced by serine (Ser20) . The modification, processing, and subcellular localization of the mutant pullulanase were studied . Labeling studies with {3H}palmitate and immunoprecipitation with mouse antiserum raised against pullulanase showed that the wild form of both the extracellular and intracellular pullulanases contained lipids, whereas the mutant enzyme was not modified with lipids . Only the Cys20 was modified with glyceryl lipids . The bulk of the mutant pullulanase was located in the periplasm, but a portion of the unmodified, mutant pullulanase was secreted into the medium . Mutant pullulanases from the extracellular and the periplasm were purified and their NH2-terminal sequences were determined . Both the mutant pullulanases were cleaved between residues of Ser13 and Leu14 which is 6-amino acid residues upstream of the lipid modified pullulanase cleavage site . This new cleavage was resistant to globomycin, an inhibitor of the prolipoprotein signal peptidase of Escherichia coli . These results indicate that the pentapeptide sequence plays an important role in maturation and translocation of pullulanase in K . aerogenes . However, the modification of pullulanase with lipids seems to be not essential for export of the enzyme across the outer membrane. J Bacteriol, 1989 Oct, 171(10), 5638 - 45 Bradyrhizobium japonicum glnB, a putative nitrogen-regulatory gene, is regulated by NtrC at tandem promoters; Martin GB et al.; The glnB gene from Bradyrhizobium japonicum, the endosymbiont of soybeans (Glycine max), was isolated and sequenced, and its expression was examined under various culture conditions and in soybean nodules . The B . japonicum glnB gene encodes a 12,237-dalton polypeptide that is highly homologous to the glnB gene products from Klebsiella pneumoniae and Escherichia coli . The gene is located directly upstream from glnA (encoding glutamine synthetase), a linkage not observed in enteric bacteria . The glnB gene from B . japonicum is expressed from tandem promoters, which are differentially regulated in response to the nitrogen status of the medium . Expression from the downstream promoter involves the B . japonicum ntrC gene product (NtrC) in both free-living and symbiotic cells . Thus, glnB, a putative nitrogen-regulatory gene in B . japonicum, is itself Ntr regulated, and NtrC is active in B . japonicum cells in their symbiotic state. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1989 Oct-Dec, 34(4), 313 - 24 {An episode of hospital infection due to Klebsiella pneumoniae in a neonatal department}; Paraschivescu I et al.; Enterotoxin-producing Klebsiella pneumoniae was implicated in the induction of intrahospital infections in new-born babies . A total of 46 children and 4 adults (hospital personnel) were involved . Most of the subjects (82.6%) had median and light forms of gastroenterocolitis, and recovered following biological re-equilibration . In 17.39% of the cases the evolution was more severe due to advanced dehydration and secondary dissemination of the infection . Two children (approximately 4%) died . Factors that favored the dissemination of the infection were hygiene deficiencies and ignorance of functioning rules of materno-infantile units, and these included: admission to the hospital of working personnel with acute phenomena of enterocolitis; administration of sweetened solutions that were prepared without control and stored at room temperature; the "critical" point represented by the special room for "the accommodation" of the newborns, a "key-point" where infection was disseminated to other wards following dispersion of "adapted babies". Mol Gen Genet, 1989 Oct, 219(1-2), 97 - 105 Analysis of the nag regulon from Escherichia coli K12 and Klebsiella pneumoniae and of its regulation; Vogler AP et al.; Four genes, nagR, A, B and E, clustered in the nag locus of Escherichia coli K12 and Klebsiella pneumoniae, were cloned and physically mapped, and the corresponding gene products involved in amino sugar metabolism identified . Expression of the nag genes was also analysed using a series of lacZ fusions . In both bacteria, the genes are arranged in two divergent operons and controlled by a common NagR repressor . The corresponding gene nagR was found to map in the first operon together with the promoter proximal gene nagB, encoding the enzyme D-glucosamine isomerase (deaminase) (NagB) and the middle gene nagA, coding for N-acetyl-glucosamine deacetylase (NagA) . Polar mutations in nagB and nagA prevent the efficient expression of nagR and cause constitutive expression of all nag genes . This includes the gene nagE encoding Enzyme IINag of the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS), encoded in the second divergently transcribed operon . No further gene is found in this operon which in both organisms is directly adjacent to the gene glnS . It is interesting that the NagR repressor also affects the mannose PTS (genes manX, Y, Z), the second transport system involved in amino sugar uptake and phosphorylation. Mol Microbiol, 1989 Oct, 3(10), 1349 - 59 Positive control of colanic acid synthesis in Escherichia coli by rmpA and rmpB, two virulence-plasmid genes of Klebsiella pneumoniae; Nassif X et al.; In Klebsiella pneumoniae, the mucoid phenotype, which is a virulence factor, is distinct from capsule production . It is positively controlled by a plasmid gene, designated rmpA . When introduced into certain Escherichia coli strains, rmpA induces expression of a mucoid phenotype, which results from overproduction of colanic acid at 30 degrees C but not at 37 degrees C . In E . coli, production of colanic acid is regulated by three genes: rcsA and rcsB which act as positive regulators, and rcsC which is a negative effector . In this work we present evidence that the rmpA gene complemented an rcsA, Ion double mutant of E . coli, but not an rcsA, Ion+ isolate . This leads to the suggestion that rmpA expressed an rcsA-like activity and like rcsA, was negatively controlled at post-transcriptional level by the Lon protease . The nucleotide sequence of rmpA is reported . No homology could be found between the 27 kiloDalton RcsA protein and the deduced amino acid sequence of the 15.5 kiloDalton RmpA protein . Another gene, rmpB, which was required in E . coli recA isolates for full expression of rmpA at 30 degrees C, has been identified on the K . pneumoniae virulence plasmid and shown to encode a 37 kiloDalton protein . Although rmpB was closely linked to rmpA, it was not present on the same transcriptional unit . These results suggested that induction of colanic acid synthesis by the K . pneumoniae virulence gene rmpA, was, at least in E . coli, under the control of the RecA network via rmpB, which may act as a positive regulator of rmpA . We conclude that these plasmid genes may function in K . pneumoniae as regulatory genes controlling the mucoid phenotype, which is itself encoded by the chromosome. J Antimicrob Chemother, 1989 Oct, 24(4), 509 - 21 Comparative study of five plasmid-mediated ceftazidimases isolated in Klebsiella pneumoniae; Sirot D et al.; Five plasmid-mediated beta-lactamases conferring a high level of resistance to ceftazidime were isolated from Klebsiella pneumoniae strains . These ceftazidimases (CAZ) differed in their isoelectric point (from 5.3 to 8.2) and were encoded by large self-transferable plasmids of 85 kb (CAZ-2, CAZ-3) or greater than or equal to 150 kb (CAZ-1, CAZ-4, CAZ-5) . The 85 kb plasmids seemed closely related to pCFF04 encoding CTX-1 enzyme and belonged to the same incompatibility group 7 or M . These beta-lactamases hydrolysed all beta-lactams with the exception of cephamycins and carbapenems . For CAZ-1, CAZ-2 and CAZ-3 producers, MICs of ceftazidime (32-256 mg/l) were higher than MICs of cefotaxime (0.12-2 mg/l) and aztreonam (1-16 mg/l) . For the strains producing the beta-lactamases CAZ-4 and CAZ-5, MICs of aztreonam were the highest (greater than or equal to 256 mg/l) . The impaired activities of cephalosporins and monobactams were restored equally well by 2 mg/l of clavulanate, sulbactam and CL-298741 for CAZ-2 producing strains (wild type and transconjugant) . Sulbactam (2 mg/l) had a lower protective effect than other inhibitors on ceftazidime for CAZ-1 and CAZ-3 producing K . pneumoniae . The protective effect of sulbactam (2 mg/l) was lower than that of the other inhibitors on all beta-lactams for CAZ-4 and CAZ-5 producers . The enzymes CAZ-1, CAZ-2 and CAZ-3 derived from TEM beta-lactamase whereas CAZ-4 and CAZ-5 derived from SHV-1 enzyme. Jpn J Antibiot, 1989 Oct, 42(10), 2135 - 40 {Klebsiella pneumoniae meningitis associated with liver abscess: a case report}; Yanagawa T et al.; We report a rare case of Klebsiella pneumoniae meningitis associated with liver abscess, which was successfully treated with cefotaxime (CTX), one of the third-generation cephalosporins . A 53-year-old man was admitted to Keio University Hospital on June 13, 1988, because of a fever and a headache . On June 3, he suddenly started shivering and his temperature rose to 39 degrees C . He then began to complain of polydipsia, polyuria, and a weight loss of 4 kg a week . On June 11, he developed a severe headache . Four years prior to this incident, he had been diagnosed as having diabetes after a routine medical examination, but had neglected to undergo medical treatment . On admission, laboratory data showed leukocytosis, hyperglycemia (394 mg/dl) and ketonuria (4+) . A lumbar puncture yielded cloudy cerebrospinal fluid (CSF) containing 500/3 cells/mm8, of which about 70% were neutrophils . A diagnosis of diabetic ketoacidosis and purulent meningitis was made . A treatment with ampicillin (ABPC) and CTX, (12 g/day, each) was begun . On the third day, cultures of a blood specimen and CSF yielded both K . pneumoniae . The MICs of CTX to K . pneumoniae isolated from blood and CSF were both 0.05 microgram/ml . ABPC was discontinued, gentamicin was administered for 2 days, CTX was continued at the same dosage level and an administration of prednisolone 40 mg daily was begun.(ABSTRACT TRUNCATED AT 250 WORDS) J Natl Med Assoc, 1989 Oct, 81(10), 1033 - 40 Preservation of myocardial ultrastructure after 24 hours of Klebsiella sepsis: histologic, functional, and biochemical correlations; Hoover EL et al.; Myocardial function with ultrastructure and high energy phosphate levels in dogs was correlated after 24 hours of sepsis using live Klebsiella aerogenes . All animals developed progressive hemodynamic deterioration over a 24 hour period . Mean arterial pressure decreased from 148 +/- 7 mmHg to 85 (P less than 0.01) and cardiac output decreased from 3.43 +/- .31 to 1.6 +/- 0.5 L/min . Left ventricular stroke work decreased from 48.2 +/- 5 to 18.1 +/- 6 gm-meters (P less than 0.001) . Systemic and pulmonary vascular resistances were increased at 24 hours (3,538 +/- 27 to 7,404 +/- 1,400 dyne/sec/cm-5 (P less than 0.01), and 185 +/- 20 and 619 +/- 90 dyne/sec/cm-5 (P less than 0.001), respectively . Left ventricular function curves at 24 hours showed a fixed low output . However, myocardial ultrastructure was preserved and high energy phosphate levels remained normal . These observations correlate well with the changes seen clinically in early gram negative sepsis in hypovolemic patients . Thus, this appears to be a suitable model for further investigation of the effects of gram negative sepsis on myocardial performance, ultrastructure, and maintenance of energy stores. Proc Natl Acad Sci U S A, 1989 Oct, 86(19), 7346 - 50 In vitro activity of the nitrogen fixation regulatory protein NIFA; Santero E et al.; We have detected activity of the nitrogen fixation regulatory protein NIFA of Klebsiella pneumoniae in vitro . To do so we directed synthesis of NIFA in a coupled transcription-translation system and detected its ability to activate expression of a translational fusion between the nifH and lacZ genes . We infer that NIFA stimulates initiation of transcription by sigma 54 holoenzyme from the nifHDK promoter . The activity of NIFA was lost rapidly under both aerobic and anaerobic conditions at 30 degrees C and was lost somewhat less rapidly at 0 degrees C . Loss of activity was not accompanied by degradation of NIFA polypeptide . Loss of activity was approximately exponential and was not affected by NIFA concentration over a 5-fold range . Therefore, NIFA inactivation does not appear to be due to self-association . We found that the factor in crude extracts previously demonstrated to bind to the nifHDK promoter-regulatory region {Beynon, J., Cannon, M., Buchanan-Wollaston, V., and Cannon, F . (1983) Cell 34, 665-671} is the integration host factor, which is known to bend DNA . Since the binding site for integration host factor lies between the upstream binding site for NIFA and the nifHDK promoter, integration host factor may bend the DNA between these two sites to facilitate productive interactions between NIFA and sigma 54 holoenzyme. J Bacteriol, 1989 Oct, 171(10), 5729 - 35 Structure-function relationships in the alpha subunit of Klebsiella pneumoniae nitrogenase MoFe protein from analysis of nifD mutants; Govezensky D et al.; Crude extracts of wild-type, nitrogenase-derepressed Klebsiella pneumoniae fractionated by nondenaturing gel electrophoresis contain, in addition to the major form of the MoFe protein, two minor variants of lower electrophoretic mobility . Of seven Nif- mutants of K . pneumoniae with nonpolar point mutations in nifD (encoding the alpha subunit of Kp1), three exhibit a wild-type-like electrophoretic pattern, whereas in the remaining four, the slowest-migrating form becomes the predominant species . Amino acid substitutions in mutants of the first type are located in the N terminus of NifD and include Gly-85 to Arg (UN1661), Glu-121 to Lys (UN1649), and Gly-161 to Asp (UN1683) . Mutations of the second type are Gly-186 to Asp (UN1648), Gly-195 to Glu (UN1680), Ser-443 to Pro (UN1793), and Gly-455 to Asp (UN1650) . Six of the mutated residues show interspecies conservation, three are close to conserved cysteines, and two are located next to conserved histidines . Based on evidence pointing to the possibility that the lowest-mobility form lacks the iron-molybdenum cofactor, these results provide insights into the functional significance of specific sites in the alpha subunit of the MoFe protein. Ann Intern Med, 1989 Oct 1, 111(7), 581 - 91 Molecular mimicry in HLA-B27-related arthritis; Yu DT et al.; A unique feature of patients with ankylosing spondylitis and reactive arthritis is that almost all share the HLA type B27 . The primary structures of the HLA-B27 antigens have been determined . At least six variants exist . However, disease predisposition does not appear to be restricted to a particular variant . One hypothesis about the pathogenesis of arthritis is that the bacteria that cause the arthritis carry components that are cross-reactive with HLA-B27 antigens . Several reactive bacterial components have indeed been identified using monoclonal anti-HLA-B27 antibodies . Even more striking is the identification, through a computerized search, of a Klebsiella protein . This protein carries a stretch of six amino acids identical to residues 72 to 77 of two of the HLA-B27 variants . A synthetic peptide carrying these six amino acids of HLA-B27 protein is reactive with serum antibodies in some patients with arthritis . With this knowledge, investigators will be able to formulate new approaches for examining the pathogenesis of HLA-B27-associated arthritis. Mol Gen Genet, 1989 Oct, 219(1-2), 235 - 40 Identification of a regulatory nifA type gene and physical mapping of cloned new nif regions of Azospirillum brasilense; Singh M et al.; Three new Tn5-mutagenized nif genes of Azospirillum brasilense were characterized . The sizes of the restriction fragments and the restriction maps of the cloned nif DNA regions showed that these nif genes are distinct from those reported earlier, e.g . nifHDK, nifE, nifUS, fixABC . The Nif27 mutant was identified as a nifA type regulatory gene of A . brasilense (a) by genetic complementation with nifA of Klebsiella pneumoniae, (b) by the absence of nitrogenase iron protein in western protein blots and (c) by its inability to activate expression of a nifH-lacZ fusion . The growth characteristics of the three mutants showed that none of them is defective in general nitrogen regulatory (ntr) genes . Also, no homology was detected between the three nif DNA regions of the mutants, cloned in pMS188, pMS189 and pMS197, and the K . pneumoniae nif, glnA or ntr genes . In addition, the fixABC genes of Bradyrhizobium japonicum did not show any hybridization with the cloned Azospirillum genes . Unlike the situation in enteric bacteria, the nif genes in A . brasilense are scattered and span a region of about 65 kb. J Antimicrob Chemother, 1989 Oct, 24(4), 561 - 72 Comparison of the in-vitro effect of several macrolides on the oxidative burst of human neutrophils; Labro MT et al.; We have compared the in-vitro interaction of five macrolides (roxithromycin, erythromycin, spiramycin, oleandomycin and josamycin) with human neutrophils (PMN) . Only roxithromycin strongly impaired the oxidative burst of PMN assessed by luminol amplified chemiluminescence, superoxide anion generation, and myeloperoxidase-mediated iodination of proteins . This effect was observed only for high concentrations of this drug (100 and 50 mg/l) . Furthermore, the sensitivity of PMN to the depressive effect of roxithromycin permitted the definition of two kinds of PMN: in Highly Sensitive (HS)-PMN, the oxidative response was completely abolished while in Moderately Sensitive (MS)-PMN, a decreased, but yet measurable (20-50% of the control), response was obtained . The roxithromycin-induced depression of PMN was time-dependent and partly reversed by washing . Chemotaxis was also impaired by roxithromycin (100 mg/l) but phagocytosis of Klebsiella pneumoniae was unaltered even at high concentrations of the drug . Since roxithromycin displays the highest intracellular uptake, compared with the other macrolides assessed in this study, this could explain the results observed here . The relevance to the clinical situation needs further study . This effect of roxithromycin could be useful to control the inflammatory process associated in certain infectious diseases, in particular if high concentrations of the drug are obtained in tissues. Eur J Clin Microbiol Infect Dis, 1989 Oct, 8(10), 878 - 87 Impact of the dosage schedule on the efficacy of ceftazidime, gentamicin and ciprofloxacin in Klebsiella pneumoniae pneumonia and septicemia in leukopenic rats; Roosendaal R et al.; The impact of the dosage schedule on the therapeutic efficacy of antibiotics was investigated in experimental Klebsiella pneumoniae pneumonia and septicemia in leukopenic rats . The daily doses (mg/kg) that protected 50% of the animals from death, when calculated for administration at 6 h intervals or by continuous infusion, were as follows: ceftazidime 24.4 and 1.5 (p less than 0.001), gentamicin 2.8 and 3.8 (p greater than 0.05), and ciprofloxacin 3.3 and 6.5 (p less than 0.05), respectively . This correlates with the observation that ceftazidime killed Klebsiella pneumoniae slowly but constantly, and relatively independently of concentration, whereas killing by gentamicin or ciprofloxacin was rapid, and markedly dependent on antibiotic concentration . Exposure of bacteria for 1 h to concentrations of fivefold the MBC did not give rise to a postantibiotic effect for any of the drugs . In our model ceftazidime was far more effective when given continuously than when administered at 6 h intervals . On the other hand, the activity of gentamicin was not influenced by the mode of administration, whereas ciprofloxacin was slightly more effective when given intermittently . However, to avoid misleading conclusions regarding the use of antibiotics in humans, the pharmacokinetic differences between rats and man must be considered when interpreting these results. An Med Interna, 1989 Oct, 6(10), 531 - 3 {Liver abscesses due to Klebsiella pneumoniae and acute anterior uveitis}; Cordero M et al.; Liver abscesses are not very frequent and those produced by Klebsiella spp . are quite exceptional . These germs have been related to the development of acute anterior uveitis (AAU) in patients whose susceptibility has been linked to HLA B27 . We present a case of a female, HLA B27 negative, who developed AAU during a course of liver abscesses caused by Klebsiella pneumoniae. Arch Invest Med (Mex), 1989 Oct-Dec, 20(4), 287 - 95 {Usefulness of 5 microbiological markers for the differentiation of strains of Klebsiella ozaenae causing an outbreak of nosocomial infection}; Garcia-Perez M et al.; Five different microbiological markers (biotyping, antimicrobial susceptibility, heavy-metal susceptibility, klebocin susceptibility and number of extrachromosomal DNA fragments) were utilized in order to distinguish between Klebsiella ozaenae strains arising from a nosocomial infection outbreak and strains of some species isolated from hospital environment . Antimicrobial susceptibility pattern to 13 different antibiotics was the most reliable maker, because 85% of the outbreak strains shown resistance to 8-10 antibiotics and only two strains of hospital environment had similar pattern . Sensitivity for this marker was 85.0% specificity 93.5% and positive predictive value of 89.5% . Other markers were less efficient with a positive predictive value of 79.2%, for biotyping, 70.4% for klebocin susceptibility, 69.2% for number of extrachromosomal DNA fragments and 60.6% for heavy-metals susceptibility . We discuss the applicability of each one of these markers in different circumstances. J Biol Chem, 1989 Sep 25, 264(27), 15835 - 42 Competitive inhibitors of Klebsiella aerogenes urease . Mechanisms of interaction with the nickel active site; Todd MJ et al.; We examined several compounds for their mechanisms of inhibition with the nickel-containing active site of homogeneous Klebsiella aerogenes urease . Thiolate anions competitively inhibit urease and directly interact with the metallocenter, as shown by the pH dependence of inhibition and by UV-visible absorbance spectroscopic studies . Cysteamine, which possesses a cationic beta-amino group, exhibited a high affinity for urease (Ki = 5 microM), whereas thiolates containing anionic carboxyl groups were uniformly poor inhibitors . Phosphate monoanion competitively inhibits a protonated form of urease with a pKa of less than 5 . Both the thiolate and phosphate inhibition results are consistent with charge repulsion by an anionic group in the urease active site . Acetohydroxamic acid (AHA) was shown to be a slow-binding competitive inhibitor of urease . This compound forms an initial E.AHA complex which then undergoes a slow transformation to yield an E.AHA* complex; the overall dissociation constant of AHA is 2.6 microM . Phenylphosphorodiamidate, also shown to be a slow-binding competitive inhibitor, possesses an overall dissociation constant of 94 pM . The tight binding of phenylphosphorodiamidate was exploited to demonstrate the presence of two active sites per enzyme molecule . Urease contains 4 mol of nickel/mol enzyme, hence there are two nickel ions/catalytic unit . Each of the two slow-binding inhibitors are proposed to form complexes in which the inhibitor bridges the two active site nickel ions . The inhibition results obtained for K . aerogenes urease are compared with inhibition studies of other ureases and are interpreted in terms of a model for catalysis proposed for the jack bean enzyme (Dixon, N.E., Riddles, P.W., Gazzola, C., Blakely, R.L., and Zerner, B . (1980) Can . J . Biochem . 58, 1335-1344). J Biol Chem, 1989 Sep 5, 264(25), 14710 - 5 The sodium ion translocating oxaloacetate decarboxylase of Klebsiella pneumoniae . Sequence of the integral membrane-bound subunits beta and gamma; Laussermair E et al.; The sequences upstream and downstream of the cloned gene for the alpha-subunit of the Na+ pump oxaloacetate decarboxylase of Klebsiella pneumonia were determined . An open reading frame in the upstream region was identified as the gene for the gamma-subunit, and an open reading frame in the downstream region represents the gene for the beta-subunit . The deduced primary structure of the gamma- and beta-subunit was confirmed by protein sequencing of about 37 and 22%, respectively, of each polypeptide chain . The gene for the gamma-subunit has a GC content of 64% and codes for 83 amino acids . The protein is not pro