J Biol Chem, 1988 Jul 15, 263(20), 9598 - 606 Intermediates in the conversion of 5'-S-methylthioadenosine to methionine in Klebsiella pneumoniae; Furfine ES et al.; Extracts of Klebsiella pneumoniae oxidatively convert 1-phospho-5-S-methylthioribose (1-PMTR) to alpha-keto-gamma-methylthiobutyrate, a precursor of methionine, and to S-methylthiopropionate and formate . One equivalent of formate is produced per equivalent of alpha-keto-gamma-methylthiobutyrate and two equivalents of formate per equivalent of methylthiopropionate . Two compounds were identified as intermediates in this reaction sequence: 1-phospho-5-S-methylthioribulose (1-PMT-ribulose) and 1-phospho-2,3-diketo-5-S-methylpentane . The enzyme, 1-PMTR isomerase, which converts 1-PMTR to 1-PMT-ribulose was highly purified . In addition, a protein fraction was isolated which converts 1-PMT-ribulose to the phosphodiketone . A second protein fraction was isolated that converts the phosphodiketone to an intermediate which has not been isolated so far . This intermediate is oxidatively converted to alpha-keto-gamma-methylthiobutyrate and S-methylthiopropionate by a third protein fraction . Methylthiopropionate is not derived from free alpha-keto-gamma-methylthiobutyrate. J Biol Chem, 1988 Jul 15, 263(20), 9640 - 5 The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae . Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes; Schwarz E et al.; The gene encoding the alpha-subunit of the Na+ pump oxalacetate decarboxylase of Klebsiella pneumoniae was cloned and sequenced . The deduced primary structure of the protein was confirmed by protein sequencing of about 30% of the polypeptide chain . The gene has a GC content of 67% and codes for 596 amino acids . The N-terminal methionine is removed in the mature protein which has a calculated molecular mass of 63,600 daltons . The protein consists of two different domains that are connected by a stretch of amino acid residues susceptible to proteolytic cleavage . Limited proteolysis of the native enzyme with trypsin produced fragments of about 51 kDa and 10.2 kDa, the latter of which started with valine 491 and contained the biotin prosthetic group . Peptide sequencing indicated binding of the biotin prosthetic group to lysine 561, 35 residues from the C terminus . The decarboxylase contains an extended alanine- and proline-rich region (positions 502-532) on the N-terminal side of the 10.2-kDa biotin domain . This sequence includes a total of 16 alanine and 9 proline residues. J Clin Immunol, 1988 Jul, 8(4), 319 - 31 Anti-Klebsiella K30 phospholipid antibodies in systemic lupus erythematosus: antigen cross-reactions and idiotypic sharing with antibodies to DNA and Klebsiella K30 polysaccharide; Harkiss GD et al.; Patients with systemic lupus erythematosus were examined for the presence of serum antibodies reactive with phospholipids (PL) extracted from the membranes of Klebsiella K30 (K30PL) . Affinity-purified anti-K30PL antibodies were tested for their ability to cross-react with other PL antigens or DNA and for the presence of idiotypic markers known to be associated with anti-DNA antibodies or antibodies to Klebsiella K30 polysaccharide (K30p) . Affinity-purified antibodies to K30PL, phosphatidylserine (PS), or phosphatidylinositol (PL) uniformly cross-reacted with each other . Analysis of the PL preparations by thin-layer chromatography and reversed-phase high-performance liquid chromatography (HPLC) showed the presence of several components . Lupus sera reacted with one component mainly in the HPLC-fractionated K30PL preparation, although this component appeared to be present in the other PL preparations . Direct-binding and inhibition studies showed that affinity-purified antibodies to the K30PL extract, PS, or PI reacted poorly with DNA . However, the anti-K30PL antibodies possessed a prominent anti-DNA idiotypic marker (AM Id) in 61% of the patients and an anti-K30p idiotypic marker (SP Id) in 94% of the patients . The results thus show that anti-K30PL antibodies are idiotypically related to anti-DNA and anti-K30p antibody subpopulations, although they do not share the same antigen-binding characteristics. Mol Microbiol, 1988 Jul, 2(4), 433 - 42 The role of activator binding sites in transcriptional control of the divergently transcribed nifF and nifLA promoters from Klebsiella pneumoniae; Minchin SD et al.; The regulatory region spanning the divergently transcribed nifF and nifLA promoters contains a NIFA-specific upstream activator sequence (UAS) located around +59, and two NTRC binding sites centred at -142 and -163 with respect to the nifLA transcription start site . We have constructed mutations in each of these binding sites and examined their role in transcriptional activation of the divergently transcribed promoters . Analysis of a mutation at +60 confirms that the UAS is required for efficient NIFA-mediated activation of nifF transcription . This sequence is also required for maximal activation of the nifLA promoter . Mutations at -169 and -148, within the two NTRC binding sites, reduce activation of the nifLA promoter by NTRC in vivo and lower the affinity of the activator for these sites in vitro . Phosphorylation of NTRC by NTRB is required for efficient binding of NTRC to these sites. J Bacteriol, 1988 Jul, 170(7), 3297 - 300 Cloning of nifHD from Nostoc commune UTEX 584 and of a flanking region homologous to part of the Azotobacter vinelandii nifU gene; Defrancesco N et al.; The heterocystous cyanobacterium Nostoc commune UTEX 584 contains two nifH-like sequences (nifH1 and nifH2) in addition to nifHD . A region of DNA 1 kilobase upstream from the 5' end of nifH showed considerable sequence similarity to part of the published nifU sequences of Azotobacter vinelandii and Klebsiella pneumoniae. J Gen Microbiol, 1988 Jul, 134 ( Pt 7), 1779 - 84 Construction of multicopy expression vectors for regulated over-production of proteins in Klebsiella pneumoniae and other enteric bacteria; Kleiner D et al.; A number of expression vectors have been constructed to allow over-production of selected gene products in Klebsiella pneumoniae and other enteric bacteria . The plasmids use the strong hybrid trp-lac (tac) promoter for gene expression, which is regulated by the lacIQ allele of the lac repressor carried on the vector . This provides very tight regulation of gene expression, which is important for over-production of proteins which may be detrimental to cell growth . The vectors carry the standard mp18 cloning nest in which all the restriction sites are unique to the plasmid (with the exception of EcoRI in pDK7) . Derivatives were constructed carrying kanamycin, chloramphenicol or ampicillin resistance as selectable markers, the first two of which are advantageous in K . pneumoniae due to the high inherent beta-lactamase activity of this organism. Can J Microbiol, 1988 Jul, 34(7), 918 - 21 A high molecular weight lipopolysaccharide specific bacteriophage for Klebsiella pneumoniae; Benedi VJ et al.; High-molecular weight lipopolysaccharide (O antigen enriched fraction) from Klebsiella pneumoniae was determined to be the receptor for bacteriophage FC3-1 . A methodology for the identification of the lipopolysaccharide component involved in FC3-1 bacteriophage reception was used that is suitable for other phages and host bacteria. Rev Infect Dis, 1988 Jul-Aug, 10(4), 885 - 91 Interactions of new plasmid-mediated beta-lactamases with third-generation cephalosporins; Labia R et al.; The kinetic constants of three recently identified plasmid-mediated beta-lactamases--SHV-2, CTX-1, and CAZ-1--markedly active against third-generation cephalosporins were analyzed in comparison with three better-characterized beta-lactamases--two plasmid-mediated enzymes, TEM-2 and PIT-2/SHV-1, and R-30, a beta-lactamase from Klebsiella oxytoca that has few similarities to the newer enzymes . All of these enzymes are synthesized constitutively, demonstrate efficient hydrolysis of penicillins, are highly susceptible to the action of clavulanic acid and sulbactam, and have no detectable activity against the cephamycins and imipenem . With the methoxyimino cephalosporins, including those of the third generation, the rates of hydrolysis observed for the SHV-2, CTX-1, and CAZ-1 enzymes are high and show no relation to those observed for the other presently known beta-lactamases . Structure-activity relations suggest that the oxime substituent of these cephalosporins is a major structural factor in the catalytic process observed with the three new beta-lactamases. Ann Inst Pasteur Immunol, 1988 Jul-Aug, 139(4), 401 - 7 Protection against fatal Klebsiella pneumoniae sepsis in the squirrel monkey Saimiri sciureus after immunization with a capsular polysaccharide vaccine; Postal JM et al.; An anti-Klebsiella pneumoniae K5a capsular polysaccharide vaccine was evaluated as a preventive approach for protecting squirrel monkeys, Saimiri sciureus, in our breeding colony . Based on an 8-month vaccination schedule over a period of more than two years, this vaccine, regardless of the animal's age, resulted in a reduction of this particular bacterial infection and its generally associated fatal outcome . IgG antibody responses in naive and vaccinated animals were monitored over an extended period by radioimmunoassay and showed a marked increase above the initial naturally occurring antibody titres . No side-effects were observed after repeated vaccination of several hundred animals during a two-year period. J Wildl Dis, 1988 Jul, 24(3), 585 - 6 Klebsiella pneumoniae as a cause of pneumonia and septicemia in a civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria; Enurah LU et al.; A fatal case of acute pneumonia and septicemia that occurred in a captive civet kitten (Civettictis civetta) in the Jos Zoo, Nigeria is reported . Diagnosis was based on clinical signs, necropsy findings, and the isolation of Klebsiella pneumoniae from the lung, intestine, liver and heart blood of the animal . This is the first report of clinical K . pneumoniae infection in a wild or captive animal in Nigeria. Ann Otol Rhinol Laryngol, 1988 Jul-Aug, 97(4 Pt 1), 422 - 6 Functional disorder of eustachian tube in experimental otitis media with effusion following inoculation of bacterial endotoxin; Ohashi Y et al.; A 10-micrograms/mL solution of lipopolysaccharide derived from Klebsiella pneumoniae was inoculated into the middle ears of guinea pigs . The animals were killed painlessly on the first, third, or seventh day after inoculation, and the mucosal samples from the bony portion of the eustachian tube were examined for ciliary activity and epithelial morphology . On the first and third days, when middle ear effusions were present, deterioration of ciliary activity and morphologic changes in the mucociliary system were observed . On the seventh day, when middle ear effusions were absent, the ciliary activity had recovered to normal . Our data show that endotoxin extracted from K pneumoniae can produce otitis media with effusion and that dysfunction of cilia caused by endotoxin is a factor responsible for the manifestation of otitis media. Mol Gen Genet, 1988 Jul, 213(1), 175 - 8 Complementation of a truncated membrane-bound Enzyme IINag from Klebsiella pneumoniae with a soluble Enzyme III in Escherichia coli K12; Vogler AP et al.; Cloning and analysis of the gene nagE encoding Enzyme IINag (EIINag) from Klebsiella pneumoniae revealed strong similarities with the corresponding gene from Escherichia coli K12 . Truncated EIINag proteins were generated by inserting a series of Tn1725 transposons into the structural gene; the positions of the insertions were mapped by restriction enzyme analysis, and the activity of the polypeptides determined by in vitro and in vivo tests . Insertions in the region encoding the amino-terminal half of the protein invariably abolished transport and phosphorylation activity, while truncated proteins lacking a C-terminal domain homologous to the soluble Enzyme III (crr gene) could be complemented by this molecule to nearly wild-type activity. Mol Microbiol, 1988 Jul, 2(4), 443 - 54 Cloning and characterization of an albicidin resistance gene from Klebsiella oxytoca; Walker MJ et al.; A DNA fragment containing a gene for resistance to the antibiotic albicidin was isolated from Klebsiella oxytoca and shown to be expressed in Escherichia coli, where it also protected bacteriophage T7 replication from inhibition by albicidin . In vivo translation analysis demonstrated that the cloned 2.2kb DNA fragment coded for a 36 kiloDalton (kD) protein and a 25kD protein . The DNA sequence was determined for a 654-base-pair open reading frame contained within a 1.2kb subcloned DNA fragment encoding albicidin resistance . The predicted molecular weight of the polypeptide translated from the open reading frame was 25.8kD . A putative Shine-Dalgarno sequence precedes the open reading frame but a potential promoter sequence was not detected . A possible rho-independent transcription termination signal was found directly following the stop codon . The functional protein for albicidin resistance was isolated and purified . Both the molecular weight and NH2-terminal amino acid sequence of this protein correspond with that predicted from the DNA sequence of the open reading frame . The cloned albicidin resistance gene had no effect on the tsx (nupA) nucleoside uptake gene associated with spontaneous albicidin resistance in E . coli; also, it did not complement any of a range of E . coli DNAts mutants at restrictive temperatures . The cloned resistance gene product remained intracellular in exponential cultures of K . oxytoca and E . coli . Cell-free extracts from E . coli containing the resistance gene protected a sensitive strain of E . coli from inhibition by albicidin, as did the purified albicidin resistance protein . The mechanism of this albicidin resistance protein involved binding to albicidin to form a complex without antibiotic activity, but without catalysing further chemical modification of the antibiotic. Ann Inst Pasteur Microbiol, 1988 Jul-Aug, 139(4), 435 - 51 Immunological comparison of constitutive beta-lactamases of gram-negative bacteria by neutralization in zymogram gels: properties of anti-TEM-1 and anti-TEM-2 sera; Paul G et al.; The zymogram technique was applied to a beta-lactamase neutralization assay with anti-TEM-1 and anti-TEM-2 sera . Both were shown to contain neutralizing antibodies directed towards various beta-lactamases of Gram-negative bacteria . The quantitative neutralization allowed classification into five groups of the 28 beta-lactamases used as standards and 61 from clinical isolates . In the first were enzymes such as TEM-1 and TEM-2 including TLE-1, SHV-1, SHV-2, penicillinases of Klebsiella pneumoniae and CTX-1 . Partial neutralization distinguished two groups containing the CARB group of enzymes, which are different from PSE-2 and PSE-3, and the MAL penicillinases of Levinea malonatica, which are different from L . amalonatica enzymes . Broad spectrum beta-lactamases of K . oxytoca constituted a unique group of partially neutralized enzymes . Among the beta-lactamases not neutralized by either serum were the plasmid-mediated OXA-enzymes, various species-specific beta-lactamases and cephalosporinases . The antigenic similarities of the enzymes appeared to correlate with the extent of similarities of their catalytic properties, namely those of penicillinases . Such comparisons between the beta-lactamase groups provide an indirect approach to the physiological and structural analysis of established and recently evolved beta-lactamases. Genetika, 1988 Jun, 24(6), 998 - 1007 {Expression of the nif and ntr genes of Klebsiella pneumoniae in Rhodobacter sphaeroides cells}; Zinchenko VV et al.; The genes glnA, ntr, nif or their promoters from Klebsiella pneumoniae cloned on the vectors, based on the plasmid RSF1010, were introduced into Rhodobacter sphaeroides cells . It was found that K . pneumoniae genes glnA, nifB, nifE, nifL and nifH are not expressed in R . sphaeroides . Neither was the glnA gene from cyanobacterium Anabaena 7120 expressed in R . sphaeroides . No functional activity of K . pneumoniae product of ntrA gene which is expressed from its own promoter, and the product of the gene nifA which is expressed from the constitutive promoter of the kanamycin resistance gene of the transposon Tn903, was detected . The implications of these findings are discussed. Clin Exp Immunol, 1988 Jun, 72(3), 406 - 9 Modulation of Klebsiella pneumoniae infection of mice by interferon-gamma; Hershman MJ et al.; The aim of this study was to determine the efficacy of interferon-gamma (IFN-gamma) as prophylaxis and therapy in a wound infection model with a 'surgical' pathogen . The bacterial challenge consisted of intramuscular injections of Klebsiella pneumoniae (10(3) organisms in 0.1 ml) . Groups of 12 CBA/J mice had either IFN-gamma or RPMI-1640 medium (controls) injected subcutaneously . Mice pretreated with IFN-gamma in a dose of 7,500 or 750 units per day, followed by infection and 2 days additional IFN-gamma treatment, survived significantly longer than controls or mice treated with 150 units of IFN-gamma per day . Significantly greater survival than controls was seen with only 5 or 3 days pretreatment with IFN-gamma but not with 1 day pretreatment . Administration of IFN-gamma to the opposite hind leg from the one receiving bacterial challenge was as effective as same leg treatment . When IFN-gamma therapy was commenced 1 h after bacterial challenge and continued for 7 days, 13 of 60 mice survived, which was significantly greater than four of 60 surviving controls . These effects may be secondary to IFN-gamma's immunoregulatory effects rather than by involving any antiviral properties. J Gen Microbiol, 1988 Jun, 134 ( Pt 6), 1635 - 44 Analysis of sucrose catabolism in Klebsiella pneumoniae and in Scr+ derivatives of Escherichia coli K12; Sprenger GA et al.; In contrast to a previous report, strains of Klebsiella pneumoniae were found to take up and phosphorylate the disaccharide sucrose via the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS) . In addition to the two soluble and general components enzymeI and HPr of the PTS, a sucrose-specific enzymeIIScr (gene scrA), together with the enzymeIII, coded for by the gene crr, were needed for the vectorial phosphorylation of sucrose to generate intracellular sucrose 6-phosphate . This sugar phosphate is hydrolysed by a hydrolase (invertase, gene scrB) to generate glucose 6-phosphate and free fructose . The latter is converted to fructose 6-phosphate by an ATP-dependent fructokinase (gene scrK), an enzyme which is part of the sucrose and not of the fructose catabolic pathway . Analysis of different mutants of K . pneumoniae strain 1033, and of Escherichia coli K12 derivatives carrying R'scr plasmids isolated from K . pneumoniae, showed that the genes scrA, B, and K, together with a gene scrR for a repressor, form a genetic unit located on the chromosome of K . pneumoniae . These genes and the corresponding sucrose metabolic pathway are very similar to a previously described scr system encoded on plasmid pUR400 and found in other enteric bacteria. J Appl Bacteriol, 1988 Jun, 64(6), 541 - 9 Starvation and nutrient resuscitation of Klebsiella pneumoniae isolated from oil well waters; Lappin-Scott HM et al.; Klebsiella pneumoniae isolated from oil well waters reduced in size in response to nutrient starvation . The cells remained viable during starvation and later were able to grow rapidly when stimulated by nutrients . The heterotrophic potential, culture absorbance and extracellular polysaccharide production decreased during cell starvation whereas an initial increase in colony-forming units was observed on agar plates . Transmission electron microscopy (TEM) after 24 d revealed that the cells had changed to small rods or cocci between 0.5 by 0.25 micron and 0.87 by 0.55 micron . When transferred to half-strength brain heart infusion medium, TEM showed cell division and rod-shaped cells after 45 min and full resuscitation within 4 h . Cell response was much slower in sodium citrate medium and resuscitation took 8 h. Clin Rheumatol, 1988 Jun, 7(2), 285 - 7 Acute rheumatoid factor positive (IgM) polyarthritis associated with a Klebsiella pneumonitis; Magaro' M et al.; The authors report a case of a patient suffering from acute polyarthritis with a high rheumatoid factor titre, associated with a Klebsiella pneumonitis . A polyclonal B lymphocyte activation or a possible cross reaction between rheumatoid factor and an antigen related to Klebsiella may explain the elevated production of rheumatoid factor observed. Biochem J, 1988 Jun 1, 252(2), 421 - 5 Iron K-edge X-ray absorption spectroscopy of the iron-molybdenum cofactor of nitrogenase from Klebsiella pneumoniae; Arber JM et al.; Iron K-edge X-ray absorption data for the iron-molybdenum cofactor ('FeMoco') from Klebsiella pneumoniae reported here provide the first evidence for long-range structural order in the cofactor {Fe...Fe(Mo) = 0.368 nm in addition to Fe...S = 0.22 nm and Fe...Fe(Mo) = 0.27 nm} and, in contrast with previously published data {Antonio, Teo, Orme-Johnson, Nelson, Groh, Lindahl, Kauzlarich & Averill (1982) J . Am . Chem . Soc . 104, 4703-4705}, indicate that most of the iron centres are not co-ordinated to light (oxygen, nitrogen) atoms . This demonstrates that presently available chemical models for FeMoco are inadequate. Br Med J (Clin Res Ed), 1988 May 21, 296(6634), 1432 - 4 Raised titres of anti-klebsiella IgA in ankylosing spondylitis, rheumatoid arthritis, and inflammatory bowel disease; Cooper R et al.; Serum titres of IgA are raised in ankylosing spondylitis and increased titres of antibodies to klebsiella have also been reported . The humoral response was investigated in ankylosing spondylitis and other inflammatory disorders . IgA antibodies to klebsiella pneumoniae K43 were measured in patients with ankylosing spondylitis, Crohn's disease, ulcerative colitis, and rheumatoid arthritis and in controls . Significantly raised median titres of anti-klebsiella IgA, measured as optical density at 405 nm with an enzyme linked immunosorbent assay (ELISA), were seen among the patients with ankylosing spondylitis (0.7), Crohn's disease (0.8), rheumatoid arthritis (0.6), and ulcerative colitis (0.8) compared with controls (0.4) . Activity of disease in ankylosing spondylitis and titres of anti-klebsiella IgA were not correlated . In contrast, titres of anti-klebsiella IgM were significantly lower in patients with ankylosing spondylitis and ulcerative colitis . The increase in the titres of anti-klebsiella IgA may be due to increased permeability of the gut to bacterial antigens, leading to an increased IgA response in the gut mucosa and permitting the release of IgA into the circulation . As the increased antibody titres were seen in Crohn's disease and rheumatoid arthritis as well as in ankylosing spondylitis the response may be nonspecific, occurring because of possible underlying inflammatory bowel disease in these conditions. Biochemistry, 1988 May 17, 27(10), 3647 - 52 Dinitrogenase with altered substrate specificity results from the use of homocitrate analogues for in vitro synthesis of the iron-molybdenum cofactor; Hoover TR et al.; The in vitro synthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase requires homocitrate (2-hydroxy-1,2,4-butanetricarboxylic acid) . Homocitrate is apparently synthesized by the nifV gene product . In the absence of homocitrate, no FeMo-co is formed in vitro, as determined from coupled C2H2 reduction assays and the lack of 99Mo label incorporation into apodinitrogenase . Several organic acids were tested for their ability to replace homocitrate in the FeMo-co synthesis system . With appropriate homocitrate analogues, aberrant forms of FeMo-co are synthesized that exhibit altered substrate specificity and inhibitor susceptibility . Homoisocitrate (1-hydroxy-1,2,4-butanetricarboxylic acid) and 2-oxoglutarate facilitated the incorporation of 99Mo into apodinitrogenase in the FeMo-co synthesis system, yielding a dinitrogenase that effectively catalyzed the reduction of protons but not C2H2 or N2 . Citrate also promoted the incorporation of 99Mo into apodinitrogenase, and the resulting holodinitrogenase reduced protons and C2H2 effectively but not N2 . In addition, proton reduction from this enzyme was inhibited by CO . The properties of the homodinitrogenase formed in the presence of citrate were reminiscent of those of the Klebsiella pneumoniae NifV- dinitrogenase . We also observed low rates of HD formation from NifV- dinitrogenase compared to those from the wild-type enzyme . No HD formation was observed with the dinitrogenase activated in vitro in the presence of citrate . We propose that in vivo NifV- mutants utilize citrate for FeMo-co synthesis. Nucleic Acids Res, 1988 May 11, 16(9), 4025 - 39 The effect on the function of the transcriptional activator NtrC from Klebsiella pneumoniae of mutations in the DNA-recognition helix; Contreras A et al.; We have constructed mutations in what we predict to be the DNA-recognition helix of Klebsiella pneumoniae NtrC, which regulates transcription from promoters under global nitrogen control . Mutations which disrupt the helix lead to complete loss of function . All point mutants tested were able to activate transcription from the sigma 54-dependent glnA promoter, but only those retaining some ability to recognise NtrC binding sites, as evidenced by their ability to repress the ntrB promoter and the upstream glnA promoter, were able to activate the nifL promoter . One mutant, which contained an amino acid substitution in the turn of the DNA-binding motif as well as in the recognition helix, suppressed mutations in the NtrC binding sites upstream from the nifL promoter, but only if both sites bore equivalent transitions . This confirms that the DNA-binding motif for this class of transcriptional activator has been correctly identified and suggests that binding of NtrC can be cooperative. J Biol Chem, 1988 May 5, 263(13), 6310 - 4 Purification and properties of a nitrilase specific for the herbicide bromoxynil and corresponding nucleotide sequence analysis of the bxn gene; Stalker DM et al.; A Klebsiella ozaenae nitrilase which converts the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) to 3,5-dibromo-4-hydroxybenzoic acid has been expressed at 5-10% of the total protein in Escherichia coli from a cloned K . ozaenae DNA segment and purified 10.3-fold to homogeneity . The purified polypeptide is molecular weight 37,000 in size, but the active form of the enzyme is composed of two identical subunits . The purified enzyme exhibits a pH optimum of 9.2 and a temperature optimum of 35 degrees C . The purified enzyme is also quite sensitive to thiol-specific reagents . The nitrilase is highly specific for bromoxynil as substrate with a Km of 0.31 mM and Vmax of 15 mumol of NH3 released/min/mg protein . Analysis of bromoxynil-related substrates indicates the enzyme exhibits preference for compounds containing two meta-positioned halogen atoms . Nucleotide sequence analysis of a 1,212-base pair PstI-HincII DNA segment containing the locus (bxn) encoding the bromoxynil-specific nitrilase reveals a single open reading frame encoding a polypeptide 349 amino acids in length . The predicted sequence of the purified enzyme was derived from the nucleotide sequence of the bxn gene. Zh Mikrobiol Epidemiol Immunobiol, 1988 May, (5), 49 - 52 {The intensity and dynamics of the development of immunity in the administration of experimental corpuscular ozena vaccines}; Molochko VA et al.; Experiments on the active protection of mice from ozenous infection in its two forms, generalized (acute sepsis) and local (plantar infiltration), have demonstrated that immunity, induced by experimental heat-killed ozena vaccine (Klebsiella ozaenae strain 2211, antigens 02B:K4) introduced in a single injection, is characterized by sufficiently high intensity (the degree of protection increases up to 10,000-fold) and duration (at least 30 days) . In both forms the development of immunity is characterized by a rapid rise of its intensity to the maximum level (achieved by the end of week 1), subsequent decrease by weeks 3-4 and disappearance by days 50-60 after immunization . Immunity becomes more intense with the increase of the number of injections if these injections are separated by sufficient intervals (up to 14 days) . The optimum schedule used in the study of postvaccinal immunity to experimental generalized and local ozenous infection consists of the subcutaneous injection of K . ozaenae strain 2211 in a dose of 250-500 million microbial bodies per mouse with the subsequent challenge with the virulent strain on week 2 from the date of immunization. Antimicrob Agents Chemother, 1988 May, 32(5), 626 - 30 Novel plasmid-mediated beta-lactamase in clinical isolates of Klebsiella pneumoniae more resistant to ceftazidime than to other broad-spectrum cephalosporins; Petit A et al.; Multiresistant Klebsiella pneumoniae strains isolated from three patients in the same intensive care unit were more resistant to ceftazidime than to cefotaxime and aztreonam but remained susceptible to moxalactam and imipenem . Resistance to beta-lactams, kanamycin, streptomycin, sulfonamides, and tetracyclines was transferable to Escherichia coli by conjugation and was lost en bloc after treatment with ethidium bromide . Agarose gel electrophoresis of wild types and transconjugants indicated that these resistances were mediated by a 150-kilobase plasmid, pCFF14 . The strains constitutively produced a beta-lactamase with isoelectric point close to 5.6 and which had a higher Vmax for ceftazidime and cephalothin than for cefotaxime . The substrate profile and isoelectric point of this enzyme thus differ from those of other known plasmid-mediated beta-lactamases, including the broad-spectrum enzyme CTX-1 . Hybridization studies support the derivation of the novel enzyme from a TEM-type beta-lactamase. J Med Microbiol, 1988 May, 26(1), 29 - 35 Penetration of immunoglobulins through the Klebsiella capsule and their effect on cell-surface hydrophobicity; Williams P et al.; The ability of antibodies to cell-surface components of Klebsiella to increase surface hydrophobicity and to gain access to antigens potentially masked by the capsule was investigated . Treatment of capsulate or non-capsulate strains with the respective autologous antiserum resulted in a marked increase in surface hydrophobicity . Antisera raised against a rough non-capsulate (K-O-) strain had little effect on the surface hydrophobicity of either of the capsulate strains K1+O1+ and K2+O1+, or of the non-capsulate K-O1+ strain . Whereas anti-K-O1+ sera or anti-K2+ sera increased the surface hydrophobicity of the K2+O1+ strain, only antisera containing anti-K1+ antibodies increased the hydrophobicity of the K1+O1+ strain . Immunoadsorption of anti-K-O1+ serum by whole capsulate cells revealed that neither the K1 nor the K2 capsular polysaccharide acted as a barrier to anti-O antibodies but that the K1 capsular polysaccharide masked the presence of the immunoglobulin at the cell surface . The Klebsiella capsular polysaccharide does not appear to present a permeability barrier to immunoglobulins although failure to detect outer-membrane proteins in the immune complexes of either of the capsulate strains or of the K-O1+ strain suggests that the O antigen may prevent access of antibodies to these antigens. J Clin Microbiol, 1988 May, 26(5), 1031 - 3 Alterations in the T-lymphocyte subpopulation in patients with rhinoscleroma; Berron P et al.; T-lymphocyte subpopulations were studied in a group of patients with rhinoscleroma due to Klebsiella rhinoscleromatis . The data demonstrated that these patients had a significantly greater number of T-suppressor/cytotoxic lymphocytes than did clinically healthy individuals . This finding correlated with a diminished response to the T-cell mitogen concanavalin A . The evidence indicated that the T-cell response of these patients was decreased and may reflect the host's response to the bacterial invader, thus explaining the chronicity of the disease. Biol Reprod, 1988 May, 38(4), 830 - 5 Bactericidal activity of testicular macrophages; Wei RQ et al.; The purpose of these studies was to determine if testicular macrophages are capable of bactericidal activity . Testicular macrophages were isolated from adult Wistar rats and studied in vitro . Studies were designed to determine if these cells could kill pathogenic gram-negative organisms and if these cells secreted lysozyme, an enzyme involved with the lysis of the cell wall of gram-positive bacteria . The regulation of lysozyme secretion by hormones and lipopolysaccharide was also studied . The secretion of this enzyme by testicular macrophages was also compared to enzyme secretion by macrophages isolated from other tissues . We also studied the secretion of superoxide anion, which is known to be involved in cytotoxic reactions . It was found that testicular macrophages were capable of killing up to approximately 38% of a virulent encapsulated strain of Klebsiella pneumoniae within 1 h . This process was in part dependent upon the presence of immune serum generated against these organisms but could not be mimicked by control serum or immune serum tested in the absence of macrophages . Testicular macrophages secreted lysozyme in culture for at least 8 days; however, macrophages from the peritoneal cavity and lung secreted significantly more lysozyme under the same conditions . Lipopolysaccharide suppressed lysozyme secretion in a dose-dependent manner, whereas neither follicle-stimulating hormone, testosterone, nor leuteininzing hormone had an effect on lysozyme secretion . Finally, testicular macrophages secreted superoxide anion in a manner similar to peritoneal macrophages . These studies indicate that testicular macrophages have the capability to mount an appropriate defense against pathogenic bacteria by opsonization-dependent phagocytosis, the secretion of lysozyme, and the production of super oxide anion. J Bacteriol, 1988 May, 170(5), 2240 - 6 Bidirectional promoter in the hut(P) region of the histidine utilization (hut) operons from Klebsiella aerogenes; Nieuwkoop AJ et al.; The hut(P) region (i.e., the region responsible for regulation of hutUH expression) of the Klebsiella aerogenes histidine utilization (hut) operons contains a bidirectional promoter . One transcript from this promoter encodes the hutUH operon; the role of the oppositely directed transcript is unknown, although it appears to be involved in regulating hutUH expression (A.J . Nieuwkoop, S.A . Boylan, and R.A . Bender, J . Bacteriol . 159:934-939, 1984) . A 247-base-pair (bp) fragment containing hut(P) carries two RNA-polymerase-binding sites agree with the start sites of the two transcripts produced from hut(P) DNA in vitro and in vivo . The binding sites share a 4-bp region, suggesting that occupancy of the regulatory site precludes occupancy of the hutUH promoter, and vice versa . In the absence of positive effectors, the binding to the site responsible for hutUH transcription is weaker than the binding to the site responsible for regulation . The nucleotide sequence of the 250-bp fragment containing hut(P) contains two possible matches to the consensus sequence for Escherichia coli promoters, a better and worse match, corresponding in position to the stronger and weaker RNA-polymerase-binding sites, respectively . The sequence also contains a region similar to the consensus sequence for binding of the catabolite gene activator protein of E . coli . A sequence similar to the consensus for Ntr-dependent promoters was also found, overlapping both RNA-polymerase-binding sites, but it is not a functional promoter . Finally, an initiation codon preceded by a Shine-Dalgarno consensus sequence and followed by an open reading frame identifies a probable start of the hutU gene coding sequences. Gene, 1988 Apr 29, 64(2), 231 - 40 Characterization of two genes encoding antigenically distinct type-1 fimbriae of Klebsiella pneumoniae; Gerlach GF et al.; A uropathogenic isolate of Klebsiella pneumoniae was shown to exhibit a mannose-sensitive hemagglutinating phenotype and to produce type-1 fimbriae consisting of subunits with a different electrophoretic mobility than those previously investigated . The gene cluster encoding expression of fimbriae was cloned and the genetic organization of the encoded polypeptides was determined . The gene encoding the major fimbrial subunit was localized and further examined by nucleotide sequence analysis . Comparison of two K . pneumoniae fimbrial genes revealed a nucleotide sequence agreement of 73%, and amino acid sequence agreement of 84% for the mature fimbrial subunits . Predictions of putative antigenic sites were correlated with regions demonstrating amino acid variability . In agreement with these predictions, no serological cross-reactivity between both fimbrial proteins could be demonstrated using an enzyme-linked immunosorbent assay (ELISA). Carbohydr Res, 1988 Apr 1, 175(1), 103 - 9 Structural studies of the capsular polysaccharides from Klebsiella types 8 and 82, a reinvestigation; Jansson PE et al.; The structures of the capsular polysaccharides elaborated by Klebsiella types 8 (K8) and 82 (K82) have been reinvestigated . N.m.r . spectroscopy of the original and chemically modified polysaccharides was the principal method used . It is concluded that the polysaccharides are composed of repeating units having the following structures . (Formula: see text) . The presence of L-glutamic acid, linked as an amide to the carboxyl group of a uronic acid, has not been observed hitherto in bacterial polysaccharides. Carbohydr Res, 1988 Apr 1, 175(1), 93 - 102 Analysis by the reductive-cleavage method of linkage positions in a polysaccharide containing 4-linked D-glucopyranosyluronic residues; Vodonik SA et al.; The fate of 4-linked D-glucopyranosyluronic residues under reductive-cleavage conditions was investigated by using the Klebsiella aerogenes type 54 strain A3 capsular polysaccharide . Treatment of the fully methylated polysaccharide with triethylsilane and trimethylsilyl trifluoromethanesulfonate in dichloromethane, followed by in situ acetylation, yielded 1,5-anhydro-2,3,4,6-tetra-O-methyl-D-glucitol, 3,4-di-O-acetyl-1,5-anhydro-2,6-di-O-methyl-D-glucitol, and 3-O-acetyl-1,5-anhydro-2,4-di-O-methyl-L-fucitol, as expected, but the expected product of reductive cleavage of the 4-linked D-glucopyranosyluronic residue, namely, methyl 3-O-acetyl-2,6-anhydro-4,5-di-O-methyl-L-gulonate, was not observed . Instead, methyl 2-O-acetyl-3,6-anhydro-4,5-di-O-methyl-L-gulonate (6) was identified as the sole product of reductive cleavage of the 4-linked D-glucopyranosyluronic residue . That compound 6 arose as a result of rearrangement during reductive cleavage rather than by reductive cleavage of a 5-linked D-glucofuranosyluronic residue, was established by reductive cleavage of the fully methylated polysaccharide following reduction of its ester groups with either lithium aluminum hydride or lithium aluminum deuteride . The products of the latter reductive cleavage were the same as before, except for the absence of 6 and the presence of 4,6-di-O-acetyl-1,5-anhydro-2,3-di-O-methyl-D-glucitol, or its 6,6-dideuterio isomer . Although the reductive-cleavage technique is suitable for the direct analysis of polysaccharides containing 4-linked D-glucopyranosyluronic residues, it does not establish whether the uronic residue is a 4-linked pyranoside or a 5-linked furanoside . The expected product is, however, derived from the 4-linked D-glucopyranosyluronic residue after sequential methylation, reduction of its ester group and reductive cleavage. Carbohydr Res, 1988 Apr 1, 175(1), 77 - 83 The use of bacteriophage-mediated depolymerisation in the structural investigation of the capsular polysaccharide from Escherichia coli serotype K36; Parolis H et al.; The structure of the repeating unit of the capsular polysaccharide from Escherichia coli serotype K36 has been established from the results of spectroscopic and chemical analyses of (a) P1, the tetrasaccharide obtained on depolymerisation of the polysaccharide with a bacteriophage-borne endo-galactosidase, (b) P1-alditol, and (c) the original polysaccharide . The repeating unit, which is identical to that reported for Klebsiella K57, has the following structure . (Formula: see text). Klin Wochenschr, 1988 Apr 1, 66(7), 277 - 83 {Cold agglutinin disease}; Geissler RG et al.; Cold agglutinin disease is a normo- or macrocytic anemia due to antibodies, active under body temperature, mostly belonging to the immunoglobulin class M . Initially the agglutination of erythrocytes with acrocyanosis is reversible at body temperature . High antibody activity or long lasting period of coldness lead to intravascular or intrahepatic hemolysis, but high risk anemia is rare . Beside the idiopathic form, infection induced (especially infectious mononucleosis, cytomegalovirus, Mycoplasma pneumoniae, Klebsiella), and drug induced (especially quinidine, alpha methyldopa, penicillin, para-aminosalicylic acid, various analgetics, sulfonylurea), tumor associated (especially malignant lymphomas), and autoimmune disease associated (especially systemic lupus erythematosus) cold agglutinin anemias are discribed . "Cross reacting antigenity" to the hemolysing agent and the membrane of erythrocyte, exogenous induced changing of erythrocytic antigenity, and diversification concerning the production of antibodies are discussed as pathophysiological explanations. Eur J Clin Microbiol Infect Dis, 1988 Apr, 7(2), 279 - 84 Plasmid profiles and klebocin types in epidemiologic studies of infections by Klebsiella pneumoniae; Walia S et al.; The epidemiological methods of klebocin typing, antibiogram and plasmid DNA profile were evaluated using organisms isolated from a suspected epidemic of gentamicin-resistant Klebsiella pneumoniae and unrelated strains from different geographical areas as controls . The electrophoretic analysis of plasmid DNAs from Klebsiella pneumoniae showed the presence of at least one and up to as many as seven plasmids in each strain . The molecular weight of plasmid DNAs ranged from 1 to greater than 70 mega daltons . While none of the control Klebsiella pneumoniae strains showed identical plasmid profiles, 63% of the epidemic-related Klebsiella pneumoniae strains did . Klebocin typing and plasmid DNA profile gave different results for the same strains . Plasmid DNA profile was found to be a more valuable method than klebocin typing alone or klebocin typing in combination with antibiogram for differentiating epidemiologically related from unrelated isolates . Both plasmid DNA profile and klebocin typing methods were superior to antibiogram. J Bacteriol, 1988 Apr, 170(4), 1978 - 9 Homocitrate cures the NifV- phenotype in Klebsiella pneumoniae; Hoover TR et al.; Dinitrogenase was isolated from a culture of a Klebsiella pneumoniae NifV- strain derepressed for nitrogenase in the presence of homocitrate . The enzyme isolated from this culture was identical to the wild-type dinitrogenase . These data provide in vivo evidence that the absence of homocitrate is responsible for the NifV- phenotype. Infect Immun, 1988 Apr, 56(4), 966 - 71 Degradation of host defenses against respiratory tract infection by Klebsiella pneumoniae in aged mice; Yokota Y et al.; The host defense against respiratory tract infection with Klebsiella pneumoniae was much weaker in 60-week-old mice than in 4-week-old mice, but the resistance against systemic infection by intravenous and intraperitoneal challenge with K . pneumoniae in 60-week-old mice did not differ from that in 4-week-old mice . The number of alveolar macrophages at the resting stage in 60-week-old mice was the same as in 4-week-old mice, but the number of macrophages and polymorphonuclear leukocytes in the pulmonary cavity 4 h after challenge with formalinized K . pneumoniae in aerosol doubled in parallel with body weight . Phagocytosis and killing activities and superoxide anion production as measured by the Nitro Blue Tetrazolium reduction test of alveolar macrophages in 60-week-old mice were significantly weaker than in 4-week-old mice . The surfaces of the alveolar macrophages of the 60-week-old mice shrunk and a few adhered weekly to the glass plate, but the alveolar macrophages of the 4-week-old mice stretched to their full length and adhered firmly to the glass plate . These functions of alveolar macrophages clearly differed from those of peritoneal macrophages in 60-week-old mice, but those of the peritoneal phagocytes did not differ between 60-week-old and 4-week-old mice . The results suggest that the susceptibility to respiratory tract infection in 60-week-old mice is affected by a decline in the functions of alveolar macrophages rather than by the number of alveolar macrophages and exudated polymorphonuclear leukocytes in the lungs. Am Ind Hyg Assoc J, 1988 Mar, 49(3), 128 - 35 Nose-only versus whole-body aerosol exposure for induction of upper respiratory infections of laboratory mice; Stephenson EH et al.; The effectiveness of two aerosol delivery systems, nose-only and whole-body, were compared using Swiss-Webster mice and two pathogens, Klebsiella pneumoniae and Venezuelan equine encephalitis (VEE) virus . With K . pneumoniae the median lethal dose (LD50) and the mean time to death correlated with the inhaled dose . An LD50 value of 335 colony forming units (cfu) for nose-only exposure was significantly less than the LD50 value of 3741 cfu obtained for whole-body exposure . The LD50 values obtained with VEE virus for nose-only exposure {8 plaque forming units (pfu)} and whole-body exposure (11 pfu) were similar to each other . Following a 10-min nose-only exposure, concentrations of K . pneumoniae approximating 10(4)/g were present after 24 hr in the upper respiratory tract (URT) and lungs . The numbers of bacteria reached a peak at 72 hr, when resolution of the infection began . Detectable levels of bacteria in the blood and tissues were delayed in mice given whole-body exposure, plus there was a decreased concentration of bacteria per gram of tissue . Major pathological lesions induced by K . pneumoniae were mild suppurative rhinitis and minimal suppurative bronchopneumonia . Viremia was greatest at 96 hr following aerosol exposure to VEE . Virus concentrations in the URT, lungs, cerebrum, spleen and mesenteric lymph nodes reached maximum titers earlier for mice exposed by nose-only than for mice exposed to whole-body aerosols.(ABSTRACT TRUNCATED AT 250 WORDS) Antimicrob Agents Chemother, 1988 Mar, 32(3), 364 - 8 Influence of cephalosporins and iron on surface protein antigens of Klebsiella pneumoniae in vivo; Kadurugamuwa JL et al.; The outer membrane protein (OMP) profiles of Klebsiella pneumoniae grown in a rabbit peritonitis model in the presence or absence of cephalosporins were investigated . Six high-molecular-weight OMPs (Mr 69,000 to 83,000) were induced under iron-depleted conditions in vitro . Three of these proteins (the 69,000-Mr protein {69K protein} and the 70K and 78K proteins) and trace amounts of the 73K and 75K proteins were induced in the OM of bacteria infecting the peritoneal cavity of rabbits . Addition of iron either to the growth medium in vitro or to the peritoneum in vivo repressed the expression of these proteins . Cephaloridine had no significant effect on the OMP profiles . An additional 56,000-Mr protein was observed in the OM of bacteria cultivated in vivo in the presence of CGP 17520 and also to a lesser extent in vivo under conditions of iron excess . A difference in recognition of OM antigens between cells grown in vitro and in vivo was observed by immunoblotting techniques . The 26K, 27.5K, and 28.5K antigens present in the OM of cells grown in vitro (but not in vivo) were recognized by antibodies raised against bacteria cultivated in vitro under conditions of iron depletion, but were not recognized by antisera raised against bacteria harvested directly from infections . Antisera raised against a nonencapsulated K . pneumoniae strain caused no agglutination of encapsulated K . pneumoniae grown in vivo in the absence of cephalosporins . Rapid agglutination was observed with this antiserum when the same encapsulated strain was grown in vivo in the presence of either cephalosporin, indicating less occlusion of critical antigens by the capsule. J Antimicrob Chemother, 1988 Mar, 21(3), 301 - 7 Kinetic properties of two plasmid-mediated beta-lactamases from Klebsiella pneumoniae with strong activity against third-generation cephalosporins; Labia R et al.; We determined the kinetic constants for two plasmid-mediated beta-lactamases with strong activity against third-generation cephalosporins: CTX-1 and SHV-2 . The enzymes had many similar properties: their synthesis was constitutive and they were significantly active against penicillins as well as cephalosporins . The two enzymes thus differed considerably from the chromosomal cephalosporinases, but bore some resemblance to the commonly-encountered plasmid-coded penicillinases, such as TEM beta-lactamases . Moreover, like the TEM enzymes, the plasmid-mediated CTX-1 and SHV-2 enzymes were highly sensitive to the action of the inhibitors clavulanic acid and sulbactam . These inhibitors protected cefotaxime from hydrolysis by these enzymes . Both CTX-1 and SHV-2 lacked activity against the cephamycins, cefoxitin, latamoxef (moxalactam) and cefotetan . The CTX-1 and SHV-2 enzymes had a low activity against oxacillin and were not sensitive to chloride ions . Thus, they were not related to the OXA type beta-lactamases . For the third-generation cephalosporins the rates of hydrolysis were high and thus bore no relation with those observed for the other presently-known beta-lactamases, with perhaps the exceptions of those produced by K . oxytoca . Imipenem was very resistant to the action of these CTX-1 and SHV-2 beta-lactamases. Eur J Epidemiol, 1988 Mar, 4(1), 115 - 8 Bacteriocin (klebocin) typing of clinical isolates of Klebsiella pneumoniae; Chhibber S et al.; Three-hundred-forty-two clinical isolates of Klebsiella pneumoniae were subjected to bacteriocin (klebocin) typing using six standard klebocin-producer strains (153-158) . The overall typability was 72.8 per cent . The predominant klebocin types found were 244 (14.3 per cent), 313 (13.7 per cent) and 113 (7.6 per cent) . Klebocin types 314 and 111 each contributed 5.2 per cent to the total number of isolates . No significant correlation was observed between the source of isolation and the klebocin type. Am J Otolaryngol, 1988 Mar-Apr, 9(2), 83 - 9 Experimental otitis media with effusion induced by lipopolysaccharide from Klebsiella pneumoniae: mucociliary pathology of the middle ear; Ohashi Y et al.; We inoculated 100 micrograms/ml of lipopolysaccharide (LPS) from Klebsiella pneumoniae into the tympanic cavity of guinea pigs and examined the mucociliary pathology in the middle ear . Serous effusion was observed in the tympanic cavity of every animal on the first, third, and seventh day following the procedure, but the volume of the effusion had decreased to 0.2 ml on day 7 . By that time, the ciliary activity in the opening to the eustachian tube within the middle ear had recovered to some extent, but in the middle ear distal to the opening no recovery was apparent . Our results show that cilia close to the eustachian tube play a more significant role in middle ear clearance than those in the middle ear distal to the tube . Compared with our previous study using 10 micron/ml of LPS, this study also demonstrates that inoculations with a higher concentration of LPS induces longer-term middle ear effusions. Plasmid, 1988 Mar, 19(2), 161 - 3 A demonstration that pCU1 tra gene products are not required in the killing of Klebsiella pneumoniae; Rotheim MB et al.; IncN group plasmids, including pCU1, are able to kill Klebsiella pneumoniae when conjugatively transferred from an Escherichia coli donor . Transposon mutagenesis and deletion analysis of the known tra complementation groups were used to demonstrate that the tra gene products inactivated are not required for the Kik phenotype. Biochim Biophys Acta, 1988 Feb 10, 952(3), 290 - 6 Cyanamide: a new substrate for nitrogenase; Miller RW et al.; (1) Cyanamide (N identical to C-NH2) has been shown to be a substrate for purified Mo-nitrogenases of Klebsiella pneumoniae and Azotobacter chroococcum, with apparent Km values near 0.8 mM . (2) Reduction products were CH4, CH3NH2 and NH3 formed by pathways requiring 6 or 8 electrons: N identical to CNH2 + 6e + 6H+----CH3NH2 + NH3; N identical to CNH2 + 8e + 8H+----CH4 + 2NH3 (3) Acetylene reduction and hydrogen evolution were inhibited more than 75% by cyanamide (10 mM) . Cyanamide also inhibited total electron flux at nitrogenase protein component ratios (Fe/MoFe) near 10 . (4) Cyanamide was also a substrate for the recently isolated Va-nitrogenase of A . chroococcum, but with an apparent Km of 2.6 mM showed weaker binding and an 8-fold lower Vmax than did either Mo-nitrogenase . (5) The component ratios of nitrogenase proteins favouring CH4 formation was 3.5 Fe/MoFe protein and 1 Fe/VaFe protein. Carbohydr Res, 1988 Feb 1, 172(2), 255 - 66 Analysis of linkage positions in a polysaccharide containing nonreducing, terminal alpha-D-glucopyranosyluronic groups by the reductive-cleavage method; Vodonik SA et al.; The fate of terminal (nonreducing) alpha-D-glucopyranosyluronic groups under reductive cleavage conditions was investigated by using the Klebsiella K2 (strain NCTC-418) capsular polysaccharide . Treatment of the fully methylated polysaccharide (1) with triethylsilane and a mixture of trimethylsilyl methanesulfonate (Me3SiOSO2CH3) and boron trifluoride etherate (BF3.Et2O) as the catalyst, resulted in complete cleavage of all glycosidic linkages to yield the expected products, namely 3-O-acetyl-1,5-anhydro-2,4,6-tri-O-methyl-D-glucitol (2), 3,4-di-O-acetyl-1,5-anhydro-2,6-di-O-methyl-D-mannitol (3), 4-O-acetyl-1,5-anhydro-2,3,6-tri-O-methyl-D-glucitol (4), and methyl 2,6-anhydro-3,4,5-tri-O-methyl-L-gulonate . Treatment of 1 with trimethylsilyl trifluoromethanesulfonate (Me3SiOSO2CF3) as the catalyst resulted in incomplete cleavage of the glycosidic linkage of the methylated D-glucopyranosyluronic group, to yield 4-O-acetyl-1,5-anhydro-2,6-di-O-methyl- 3-O-(methyl2,3,4-tri-O-methyl-alpha-D-glucopyranosyluronate )-D-mannitol (9) . Reductive cleavage of 1 in the presence of BF3.Et2O resulted in incomplete cleavage of all glycosidic linkages and gave rise to all four dimers (including 9) that could be formed from a tetrasaccharide repeating unit . The proposed structures of these dimers are based upon their composition, as established by chemical ionization mass spectrometry and by the reported structure of the polysaccharide . A small proportion of 1,5-anhydro-2,4,6-tri-O-methyl-3-O-(methyl 2,3,4-tri-O-methyl-alpha-D-glucopyranosyluronate)-D-mannitol (12) was also detected in the products of the BF3.Et2O-catalyzed reductive cleavage . The presence of 12 is chemical evidence for the phase of the tetrasaccharide repeating unit in the polysaccharide . The reductive cleavage of 1 was also accomplished after reduction of its ester groups with lithium aluminum hydride . Complete cleavage of all glycosidic linkages was observed when either Me3SiOSO2CF3 or Me3SiOSO2CH3-BF3.Et2O was used to catalyze reductive cleavage, and anhydroalditols 2, 3, 4, and 6-O-acetyl-1,5-anhydro-2,3,4-tri-O-methyl-D-glucitol were produced, as expected. Carbohydr Res, 1988 Feb 1, 172(2), 209 - 16 A structural investigation of the capsular polysaccharide of Klebsiella K69; Hackland PL et al.; The structure of the capsular polysaccharide isolated from Klebsiella serotype K69 has been investigated by a combination of chemical and spectroscopic methods . The repeating structure of the deacetylated polysaccharide is shown to be of the "3 + 1 + 1" type, and it carries a 1-carboxyethylidene acetal at positions 4 and 6 of a terminal galactosyl group . The location of acetyl groups in the polysaccharide has not been established . The repeating unit of the deacetylated polysaccharide has the following structure . (Formula: see text). Microb Pathog, 1988 Feb, 4(2), 165 - 8 Effects of interferon-gamma treatment on surgically simulated wound infection in mice; Hershman MJ et al.; Interferon-gamma (IFN-gamma) has been shown to have immunoregulatory properties and is able to modulate resistance to several microbial infections . This study was designed to determine the efficacy of IFN-gamma treatment in a mouse model of infection that simulates many clinical conditions associated with surgical wound infection: viz, a bacteria laden suture and tissue injury . The test bacteria were Klebsiella pneumoniae . Groups of CBA/J mice received either IFN-gamma or RPMI-1640 medium (controls) subcutaneously . IFN-gamma was administered daily at a dose of 7500 units for 5 days prior to bacterial challenge . Mice treated with IFN-gamma survived significantly longer than controls . Systemic bacterial recovery was significantly reduced in the IFN-gamma treated group but local bacterial recovery was unaffected. J Hosp Infect, 1988 Feb, 11(2), 144 - 9 Epidemiological characterization of Klebsiella isolates from patients in a renal department; Kolmos HJ; One-hundred-and-eighteen endemic patient isolates of Klebsiella pneumoniae, collected consecutively from clinical specimens of renal patients over a 3 1/2-year period, were studied using biotypes and antimicrobial sensitivity patterns as epidemiological markers . Minor temporal clusters were demonstrable among the more commonly occurring biotypes and resistance types, and comprised isolates acquired inside and outside the renal department . Among isolates acquired in the department two clusters could be demonstrated, in peritoneal dialysate and respiratory tract specimens . Both extended over the whole observation period and occurred independently of the temporal clusters described above . They may have represented common-source outbreaks, but further elucidation was not possible with the data obtained. J Gen Microbiol, 1988 Feb, 134 ( Pt 2), 425 - 32 Over-production and characterization of the nifA gene product of Klebsiella pneumoniae--the transcriptional activator of nif gene expression; Tuli R et al.; The nifA gene of Klebsiella pneumoniae, which encodes the transcriptional activator of nif gene expression, was cloned into a number of plasmid vectors to obtain high-level synthesis of nifA product (NifA) . When over-produced, NifA was very insoluble and it precipitated with the cell debris after cell lysis . Localization of beta-galactosidase activity from a nifA-lacZ translational fusion confirmed the insoluble nature of NifA . Analysis of two translational fusions in which the last six C-terminal amino acids of NifA were deleted suggests that these residues are required for activity. J Bacteriol, 1988 Feb, 170(2), 693 - 9 Identification and mapping of nitrogen fixation genes of Rhodobacter capsulatus: duplication of a nifA-nifB region; Klipp W et al.; Rhodobacter capsulatus mutants unable to fix nitrogen were isolated by random transposon Tn5 mutagenesis . The Tn5 insertion sites of 30 Nif- mutants were mapped within three unlinked chromosomal regions designated A, B, and C . The majority of Tn5 insertions (21 mutants) map within nif region A, characterized by two ClaI fragments of 2.5 and 25 kilobases (kb) . The 17-kb ClaI fragment of nif region B contains six nif::Tn5 insertions, and the three remaining mutations are located on a 32-kb ClaI fragment of nif region C . Hybridization experiments using all 17 Klebsiella pneumoniae nif genes individually as probes revealed homology to nifE, nifS, nifA, and nifB in nif region A . The nifHDK genes were localized in nif region B . About 2 kb away from this operon, a second copy of the DNA fragments homologous to nifA and nifB, originally found in nif region A, was identified. Biochim Biophys Acta, 1988 Jan 29, 952(2), 191 - 200 Roles of the beta-D-ribofuranose ring and the functional groups of the D-ribose moiety of adenosylcobalamin in the diol dehydratase reaction; Ichikawa M et al.; Four analogs of adenosylcobalamin (AdoCbl) modified in the D-ribose moiety of the Co beta ligand were synthesized, and their coenzymic properties were studied with diol dehydratase of Klebsiella pneumoniae ATCC 8724 . 2'-Deoxyadenosylcobalamin (2'-dAdoCbl) and 3'-deoxyadenosylcobalamin (3'-dAdoCbl) were active as coenzyme . 2',3'-Secoadenosylcobalamin (2',3'-secoAdoCbl), an analog bearing the same functional groups as AdoCbl but nicked between the 2' and 3' positions in the ribose moiety, and its 2',3'-dialdehyde derivative (2',3'-secoAdoCbl dialdehyde) were totally inactive analogs of the coenzyme . It is therefore evident that the beta-D-ribofuranose ring itself, possibly its rigid structure, is essential and much more important than the functional groups of the ribose moiety for coenzymic function (relative importance: beta-D-ribofuranose ring much greater than 3'-OH greater than 2'-OH greater than ether group) . With 2'-dAdoCbl and 3'-dAdoCbl as coenzymes, an absorption peak at 478 nm appeared during enzymatic reaction, suggesting homolysis of the C-Co bond to form cob(II)alamin as intermediate . In the absence of substrate, the complexes of the enzyme with these active analogs underwent rapid inactivation by oxygen . This suggests that their C-Co bond is activated even in the absence of substrate by binding to the apoprotein . No significant spectral changes were observed with 2',3'-secoAdoCbl upon binding to the apoenzyme . In contrast, spectroscopic observation indicates that 2'3'-secoAdoCbl dialdehyde, another inactive analog, underwent gradual and irreversible cleavage of the C-Co bond by interaction with the apodiol dehydratase, forming the enzyme-bound cob(II)alamin without intermediates. Int J Immunopharmacol, 1988, 10(2), 121 - 33 In vitro and ex vivo effect of RU41740 on human polymorphonuclear leukocyte function; Capsoni F et al.; We investigated the effect of RU41740, a glycoprotein extracted from Klebsiella pneumoniae and possessing immunomodulating properties, on human neutrophil functions in vitro and ex vivo . Our in vitro results showed that RU41740 increased complement- and Fc receptor-dependent phagocytosis . Moreover, the drug enhanced the oxidative metabolism (assessed by chemiluminescence) both in resting and stimulated cells; in the latter case the RU41740-induced enhancement was observed when neutrophils were stimulated with opsonized particles of N-formyl-methionyl-leucyl-phenylalanine (FMLP) but not when phorbol myristate acetate was used . Using otherwise effective experimental conditions, RU41740 did not affect spontaneous or FMLP-induced neutrophil migration . For the ex vivo experience we tested neutrophils of ten elderly subjects with a previously demonstrated phagocytic defect . These subjects were treated orally with RU41740 at a daily dose of 2 mg for 1 week during the first month, and of 1 mg for 1 week in the second month . In this population, RU41740 was able to restore the impaired phagocytic activity and to induce a significant increase of spontaneous chemiluminescence (CL); stimulated CL was also positively influenced . These effects on neutrophils provide new explanatory bases for the immunostimulatory activity of RU41740. Invest Ophthalmol Vis Sci, 1988 Jan, 29(1), 108 - 11 Presence of Langerhans cells in the cornea of Klebsiella keratoconjunctivitis mice; Garcia-Olivares E et al.; Frozen sections of normal Balb/c corneas and corneas from Balb/c mice with Klebsiella keratoconjunctivitis were examined for the expression of class I, class II H-2 antigens and MAC-1 antigens using monoclonal antibodies in an immunoperoxidase technique . Class I antigens were readily detected, in both normal and diseased corneas, mainly in the epithelium . Class II (Ia) and MAC-1 antigens were not detected in the normal corneas . However, these two antigens were found mainly in the epithelium and to a lesser extent in the stroma of corneas from keratoconjunctivitis mice . Both normal and diseased corneas were furthermore shown to be peroxidase- . Since Langerhans cells (LC) are Ia+, MAC-1+, and peroxidase- cells, we conclude that although the normal mouse cornea is devoid of these cells, under bacterial infection LC infiltrate the corneal epithelium. Microbiol Immunol, 1988, 32(2), 151 - 60 In vitro hexagonal assembly of R-form lipopolysaccharides: effect of pH on the Mg+2-mediated hexagonal assembly; Kato N et al.; The R-form lipopolysaccharide (LPS) from Escherichia coli K-12, from which cationic material had been removed by electrodialysis and the pH of which had fallen to 3.6, formed a rough hexagonal lattice structure with the lattice constant of about 19 nm . The rough hexagonal structure was maintained in buffers at pH 5 or lower but disintegrated into the ribbon-like structures in buffers at pH 6 or higher . However, in the presence of 10 mM Mg2+, the hexagonal lattice structure was not disintegrated even at alkaline pH levels but conversely it became more dense . At pH 8.3 to 8.9, the hexagonal lattice structure with the shortest lattice constant (15 nm) was formed . The same optimal pH levels were obtained for formation of the dense hexagonal lattice structure (lattice constant, 14 to 15 nm) by the electrodialyzed LPS from Klebsiella pneumoniae strain LEN-111 (O3-:K1-) . The ability of Mg2+ to induce formation of the dense hexagonal lattice structure of the K-12 LPS depends upon the presence of buffers showing the optimal pH levels, since a very high concentration of Mg2+ such as 500 mM was required for the lattice formation in distilled water . The amount of the magnesium bound to the K-12 LPS did not significantly differ throughout the pH range of 3 to 9 . Therefore, the optimal pH range is another essential factor for formation of the dense hexagonal lattice structure of the LPS in addition to binding of the magnesium to the LPS. Infection, 1988, 16(1), 49 - 53 Comparative efficacy of ciprofloxacin, ceftazidime and gentamicin, given alone or in combination, in a model of experimental septicemia due to Klebsiella pneumoniae in neutropenic mice; Trautmann M et al.; The therapeutic efficacy of ciprofloxacin, used alone and in combination with either ceftazidime or gentamicin, was evaluated in a model of experimental Klebsiella pneumoniae septicemia in neutropenic mice . Therapeutic results were compared to those achieved by treatment with ceftazidime or gentamicin alone, as well as their combination . Infections were induced by i.v . injection of two strains of K . pneumoniae . Therapy with i.m . antibiotics was performed in 8 h intervals for a total of 13 doses, and survival rates were recorded at the end of treatment as well as seven days later . Ciprofloxacin alone proved to be significantly more effective than ceftazidime and gentamicin, producing survival rates similar to the combination of ceftazidime plus gentamicin . Bacterial counts of spleens and blood confirmed the superior bactericidal activity of ciprofloxacin . Except for the combination of ciprofloxacin with ceftazidime, there was no apparent advantage of combinations of ciprofloxacin with the other agents compared to ciprofloxacin alone . These data suggest a high in vivo activity of ciprofloxacin in systemic infection due to K . pneumoniae. Microbiologica, 1988 Jan, 11(1), 77 - 80 Unusual behaviour of Klebsiella rhinoscleromatis strains on API 20E strips; Berlutti F et al.; Seven Klebsiella rhinoscleromatis standard cultures and two wild isolates were examined for their responses to Api 20E (API System, S.A.) strips . Several strains yield incostant results for arabinose fermentation test in Api strips and, when positive, they were not identified . The arabinose positive test indeed, led to the numerical profiles 0004773 (not mentioned in the analytical catalogue), or 0004553 (two strains) corresponding to a Klebsiella ozaenae identification . The mathematical analysis of the biochemical results confirmed the identity of the strains as K . rhinoscleromatis. Am J Med Sci, 1988 Jan, 295(1), 55 - 9 Relapsing, bacteremic Klebsiella pneumoniae meningitis in an AIDS patient; Holder CD et al.; The acquired immunodeficiency syndrome (AIDS) is manifested by severe immunologic (predominantly T-lymphocyte) abnormalities and opportunistic infections . Central nervous system (CNS) infections are frequent . Pathogens causing CNS infections in AIDS patients include parasites, fungi, and viruses and are similar to those reported in other states of impaired cell mediated immunity (CMI) . A case of relapsing, bacteremic Klebsiella pneumoniae meningitis in an AIDS patient is presented. J Bacteriol, 1988 Jan, 170(1), 250 - 7 Regulation of nitrogenase synthesis in histidine auxotrophs of Klebsiella pneumoniae with altered levels of adenylate nucleotides; Stougaard J et al.; A histidine auxotrophic (hisA) mutant of Klebsiella pneumoniae is phenotypically Nif- when grown with 20 micrograms of histidine ml-1 but Nif+ when supplied with histidine at 100 micrograms ml-1 . Reversion to Nif+ at 20 micrograms of histidine ml-1 occurs phenotypically by the addition of 2-thiazolyl-DL-alanine or genetically by mutation in hisG; 2-thiazolyl-DL-alanine inhibits and hisG encodes phosphoribosyl phosphotransferase, the first enzyme of the histidine biosynthetic pathway which consumes ATP . Physiological studies of the hisA mutant JS85 showed that after removal of NH4+ from a culture of the mutant grown with 20 micrograms of histidine ml-1, synthesis of nitrogenase polypeptides occurred at a rate similar to that in the wild type for about 3 h and acetylene reduction activity reached about 10% of the fully derepressed wild-type level . Shortly thereafter the concentration of intracellular adenylates decreased; in particular, ATP fell to about 10% of normal levels . Also, nitrogenase proteins (nifHDK products) and the nifJ gene product stopped being synthesized . These effects were not due to impairment of growth or protein synthesis by histidine starvation . Inhibition of phosphoribosyl phosphotransferase with 2-thiazolyl-DL-alanine restored nitrogenase activity and synthesis, indicating that the effect of the hisA mutation on nif expression was probably a consequence of lowered energy resources that occurred during anaerobic N starvation . The loss of ATP was not associated with nitrogenase synthesis or activity, since hisA nifA and hisA nifH double mutants underwent a loss of ATP in derepressing conditions . Transcription from the nifL, nifN, and nifH promoters was examined in hisA strains with Mu d(Ap lac) fusions in these nif genes . Transcription was not significantly influenced under conditions where adenylates were decreased in concentration . Also nif mRNA apparently accumulated in cultures unable to synthesize nitrogenase, suggesting that translational control of nif gene product synthesis occurs under unfavorable energetic conditions. Infect Immun, 1988 Jan, 56(1), 45 - 50 Effect of recombinant human interleukin-2 on the course of experimental chronic respiratory tract infection caused by Klebsiella pneumoniae in mice; Iizawa Y et al.; The effect of recombinant human interleukin-2 (rIL-2) on the course of experimental chronic respiratory tract infection caused by Klebsiella pneumoniae in mice was examined . rIL-2 was administered subcutaneously once a day for 7 or 14 days, starting 2 weeks after the mice were infected . Administration of 2 or 20 micrograms of rIL-2 per mouse daily for 7 days reduced bacterial counts in the lungs dose dependently . At a dose of 0.2 microgram per day, proliferation of bacteria in the lungs was suppressed after 14 days of administration . Agglutinin titers in serum were not affected by rIL-2 treatment . Monocyte and lymphocyte counts in peripheral blood were increased by administration of 20 micrograms of rIL-2 daily for 14 days but not by treatment for 7 days . In addition, clearance of bacteria from the lungs after aerosol exposure was enhanced by treatment for 7 days before infection . Thus, rIL-2 acted therapeutically or prophylactically in the presence or absence, respectively, of a specific antigen . These effects were not abolished by anti-asialo GM1 antibody . This suggests that activation of natural killer cells does not play a critical role in the therapeutic and prophylactic effects of rIL-2. Clin Orthop, 1988 Jan, (226), 219 - 21 Emphysematous septic arthritis due to Klebsiella pneumoniae; Broom MJ et al.; A 60-year-old woman with rheumatoid arthritis developed acute emphysematous septic arthritis of the knee due to Klebsiella pneumoniae . She was brought to the hospital in septic shock with disseminated intravascular coagulation and had striking physical signs and roentgenograms showing distention of the knee with gas . She also had an infection of the hand with subcutaneous gas . After surgical drainage and institution of antibiotic therapy, she remained critically ill for several days but gradually improved . Two months later, she was ambulating independently . Emphysematous septic arthritis is rare . Four cases have previously been reported, but none were caused by Klebsiella. Cancer Detect Prev, 1988, 12(1-6), 231 - 6 The in vitro influence of rIL2 on the B-cell dysfunction in patients with persistent generalized lymph node enlargement (PGL) or AIDS; Kekow J et al.; In AIDS elevated serum Ig levels and autoimmune phenomena indicate that B cells are also involved . The human immunodeficiency virus (HIV) can be cultivated in B cells, and HIV can stimulate B cells . In order to characterize the B-cell dysfunction and conditions for modulating it, functional studies with highly purified B cells were done in four patients with PGL and HIV-positive sera . Data were compared with those from patients with AIDS and normal controls . The experiments consisted of an in vitro study of the differentiation response (IgM/G secretion into culture supernatants) to a T cell-independent polyclonal B-cell activator (PBA) {Klebsiella pneumoniae, KlebsM} . A weak increase in IgM/G levels under stimulatory conditions was characteristic . Addition of recombinant interleukin 2 (rIL2) failed to increase the spontaneous Ig levels . However, coculture experiments using KlebsM and rIL2 resulted in Ig levels like those known from healthy individuals . Patients with frank AIDS did not respond with increased IgG secretion . This indicates that the abnormal B-cell differentiation response to PBAs can be modulated by rIL2 in patients with PGL and partly in AIDS (only IgM). Int J Immunopharmacol, 1988, 10(8), 913 - 7 Increase in the number and the phagocytic function of guinea pig pulmonary and peritoneal macrophages following oral administration of RU 41740, a glycoprotein extract from Klebsiella pneumoniae; Radermecker M et al.; RU 41740 (Biostim) which is a purified glycoprotein extract from Klebsiella pneumoniae, is an orally active non-specific immunostimulant . In guinea pigs, 8 days after a 7 days oral administration of RU 41740 (10 or 100 mg/kg/day), an increase in the cell population of the pulmonary and peritoneal cavities was observed, especially in that of the macrophages . RU 41740 also enhanced the phagocytic activity of both the alveolar and peritoneal macrophages, when their chemotactic activity was not significantly modified . This increase in the number of pulmonary macrophages and the stimulation of their phagocytic function might explain the protective effect afforded by the oral administration of Biostim against respiratory infections in patients with chronic bronchitis. Cancer Detect Prev, 1988, 12(1-6), 211 - 6 Predominance of the IgG1 subclass in the hypergammaglobulinemia observed in pre-AIDS and AIDS; Kekow J et al.; In addition to the well-known T-cell dysfunctions in AIDS, hypergammaglobulinemia, mostly IgG, and autoimmune phenomena indicate that B cells are also involved . Reports of HIV-infected and activated B cells suggest T cell-independent B-cell abnormalities . In order to assess the IgG subclasses involved in hypergammaglobulinemia, we examined all four IgG subclasses in sera and in vitro with an enzyme-linked immunosorbent assay (ELISA) . The in vitro studies included 7-day cultures of mononuclear cells and highly purified B cells stimulated with a T cell-independent polyclonal B-cell activator (Klebsiella pneumoniae, KlebsM) . Cultures were done with cells from seven patients with AIDS, seven patients with persistent generalized lymph node enlargement and HIV antibodies, and normal controls . In vivo, hypergammaglobulinemia was found to be restricted to the IgG1 subclass . In vitro, high spontaneous levels of IgG were not elevated significantly under stimulatory conditions, as demonstrated by the measurement of all four IgG subclasses in the culture supernatants . In vitro, hypergammaglobulinemia also resulted from IgG1 . These results indicate that there are B-cell abnormalities in pre-AIDS and AIDS, in that the B-cell preactivation in vivo resulting in hypergammaglobulinemia is restricted to IgG1. Pediatr Hematol Oncol, 1988, 5(4), 293 - 7 Deferoxamine (Desferal)-induced ocular toxicity; Kaplinsky C et al.; A 4-year-old girl with juvenile chronic myeloid leukemia relapsed after an allogeneic bone marrow transplantation (BMT) and became refractory to conventional chemotherapy . Treatment with two courses of high-dose deferoxamine, an iron chelator (130-180 mg/kg/day), along with low-dose ARA-c (5 mg/kg/day) caused a remarkable decrease of the WBC and fetal Hb . Three days following the last dose of deferoxamine, the patient experienced an acute visual loss, confirmed by electroretinogram (ERG) and visual evoked response (VER) . Slight improvement occurred a few days later, but the patient developed severe pancytopenia and died of Klebsiella septic shock . The ocular manifestations were attributed to deferoxamine toxicity in light of the rapid onset after first exposure, the electrophysiological pattern of metabolic damage in the ERG and VER, and the long interval between the last chemotherapy and BMT . The pathogenesis of deferoxamine toxicity is discussed. Respiration, 1988, 54(3), 145 - 52 Double-blind trial RU 41740 vs . placebo: immunological and clinical effects in a group of patients with chronic bronchitis; Fietta A et al.; A double-blind trail was performed to investigate the effects of RU 41740, a glycoprotein extract from Klebsiella pneumoniae, on host defenses and its efficacy in reducing the number of exacerbation in 29 evaluable patients with chronic bronchitis, out of 36 patients who entered the study . The drug enhanced the phagocytosis indexes of both polymorphonuclear and mononuclear phagocytes . Increased candidacidal activity of monocytes was also observed . These effects, already detectable after one course of therapy and during the entire period of treatment, were no longer detectable when tested 6 months after the end of treatment . A significantly (p less than 0.05) larger number of patients in the treated group than in the placebo group had no exacerbations during drug administration (0-3 months) . Moreover, patients treated with RU 41740 had significantly fewer and shorter episodes of acute exacerbation . The positive decreases in these two parameters persisted throughout the follow-up. Microbiol Immunol, 1988, 32(9), 895 - 906 Experimental chronic pulmonary infection in mice caused by Klebsiella pneumoniae; Iizawa Y et al.; Examination of mouse strain differences in susceptibility to experimental respiratory tract infection with Klebsiella pneumoniae 27 revealed that a chronic pulmonary infection model could be established using CBA/J mice . After 6 X 10(5) colony-forming units of K . pneumoniae 27 were inoculated into the lung, the bacterial counts in the lungs changed with time showing four different phases: initial decrease, regrowth, steady-state, and final increase leading to death . Throughout the course of the infection, the challenge bacteria were isolated mainly from the respiratory organs . Pulmonary gross lesions appeared on day 2 after infection and persisted thereafter . Lobar consolidation was the primary lesion and occurred mainly in the anterior and middle lobes of the right lung, and the median lobe . Mice began to die from 4 weeks after aerosol exposure . This model may be useful for investigating the pathogenesis of chronic pulmonary infection by Klebsiella and its therapy. Microbiol Immunol, 1988, 32(5), 481 - 90 Formation of a hexagonal lattice structure by an R-form lipopolysaccharide of Klebsiella: effect of various divalent cations on the lattice formation; Kato N et al.; The R-form lipopolysaccharide from Klebsiella pneumoniae strain LEN-111 (O3-:K1-), from which cationic material had been removed by electrodialysis, was previously shown to form a hexagonal lattice structure with the lattice constant of 14 to 15 nm when suspended in 50 mM tris(hydroxymethyl)aminomethane buffer at pH 8.5 containing 10 mM Mg2+ . Under this experimental condition, effects of other divalent metal cations on the hexagonal assembly of the electrodialyzed LPS were compared with that of Mg2+ . The Zn2+, Hg2+, Cu2+, and Ni2+ could produce essentially the same hexagonal lattice structure with the lattice constant of 14.5 to 15.0 nm as that formed with Mg2+ . The Cd2+, Co2+, and Fe2+ produced the hexagonal lattice structure with the lattice constant of 15.5 to 16.0 nm, and Ba2+, Sr2+, and Ca2+ produced that with the lattice constant of 18 to 19 nm . In addition, the hexagonal lattice structures formed with the latter three cations were less orderly than those formed with the other cations . When the higher concentrations of Ba2+, Sr2+, and Ca2+ were used, the lattice constants were not shortened . The length of lattice constants of the hexagonal lattice structures formed with the divalent cations did not relate to the quantity of the cations bound to the LPS . Among the divalent cations tested, Hg2+ was bound to the LPS in the smallest amount (its atomic ratio to P, 0.07), and Zn2+ and Fe2+ were bound in very large amounts (their atomic ratios to P, 2.94 and 8.28, respectively). Autoimmunity, 1988, 1(1), 67 - 75 An analysis of autoimmunity through studies of DNA antibody idiotypes; Isenberg DA et al.; The existence of idiotypic networks, first postulated over 12 years ago, is now widely recognised . Idiotypic analyses of autoantibodies have been reported among both hybridoma-derived and naturally occurring immunoglobulins . In this review the many studies of idiotypes detected on anti-DNA antibodies, notably one designated 16/6, are analysed to see what clues they offer to our understanding of autoimmunity . The links between infection and autoimmunity are emphasised by this analysis . It is also obvious that idiotypes first identified an autoantibodies are not confined to these immunoglobulins . Thus, the 16/6 idiotype originally described on a hybridoma-derived monoclonal anti-DNA antibody has also been identified on naturally occurring antibodies binding the Klebsiella polysaccharide K30. FEMS Microbiol Immunol, 1988 Jan, 1(1), 19 - 25 Plasmid-mediated conjugative transfer of Klebsiella sp . rcs genesable to induce colanic acid capsular polysaccharide biosynthesis in Escherichia coli; Allen PM et al.; Regulation of capsular biosynthesis (rcs) genes, encoding the ability to induce the production of a colanic acid polysaccharide capsule, were transferred to Escherichia coli by conjugation with Klebsiella pneumoniae (aerogenes) of capsular serotype K36 . Transfer was mediated by a 58.4-MDa conjugative plasmid of incompatibility group IncM, which carried a copy of Tn7 (specifying resistance to trimethoprim and streptomycin) together with determinants for several further resistances . This plasmid did not carry the rcs genes itself, but mediated the conjugative recA-dependent transfer of part of the Klebsiella chromosome to E . coli . Once resident in E . coli, the rcs gene(s) could not be mobilised to other strains of E . coli, and the mobilising plasmid could be cured from capsulate transconjugants without loss of the ability to produce colanic acid . All such cured transconjugants contained an insertion of Tn7 in the chromosome, suggesting that the transposon might be involved in mobilisation of the rcs genes from Klebsiella sp . to E . coli . These findings explain previous observations that the ability to manufacture capsular polysaccharide could be transferred by plasmids between Klebsiella sp . and E . coli. Immunol Cell Biol, 1988 Jan, 66 ( Pt 3), 247 - 9 Production of monoclonal antibodies against surface antigenic determinants of Klebsiella pneumoniae; Uchiyama J et al.; The production of four murine monoclonal antibodies (Kp26, Kp53, Kp62 and Kp71) to Klebsiella pneumoniae surface antigen(s) is described . The binding of all four monoclonal antibodies to K . pneumoniae was inhibited by F(ab')2 fragments of normal human serum IgG, suggesting that the antigenic determinants of K . pneumoniae detected by the four monoclonal antibodies may be similar to those recognized by human serum IgG . The antigen identified by Kp62 was purified from a deoxycholate-solubilized bacterial fraction using immunoaffinity chromatography . The molecular weight of the antigen was determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis to be 50,000-70,000. Int J Immunopharmacol, 1988, 10(7), 879 - 87 Human monocytes exposed to Biostim (RU 41740) alter lymphocyte mitogenesis: mechanisms of action; Viland H et al.; The immunomodulatory agent RU 41740 (Biostim), which is derived from Klebsiella pneumoniae, may augment mitogenic responses of purified human blood lymphocytes . In non-purified preparations, however, responses may be sharply reduced due to the fact that Biostim induces monocytes to secrete immunosuppressive factors . This investigation has shown that both these biological activities can be exerted by a single, major glucoprotein fraction of Biostim termed F1 . The Biostim-induced suppression of mitogen responses was not blocked by antibodies directed against IFN alpha or IFN gamma, thus speaking against IFN as being a mediator of suppression . Reduced suppression, however, was observed in the presence of drugs which inhibit arachidonic acid transformation . The cyclo-oxygenase inhibitors meclofenamic acid and indomethacin, which diminish biosynthesis of prostaglandins, could partially block the Biostim-induced suppression . Such an effect was not observed with 5,8,11-eicosatrynoic acid (ETI) which is an inhibitor of 12-lipoxygenase and leukotriene biosynthesis . Combinations of ETI and meclofenamic acid, however, were more potent than the latter tested separately . Another drug termed diclofenac Na, which apart from being an inhibitor of cyclo-oxygenase, rapidly clears cells of free arachidonic acid by binding to triglycerides, was found to be the most potent in preventing Biostim-induced suppression of mitogen responses . It is concluded that Biostim-exposed monocytes liberate increased amounts of immunosuppressive eicosanoids such as prostaglandins. Eur J Biochem, 1987 Dec 30, 170(1-2), 259 - 65 The nucleotide sequence of the nifM gene of Klebsiella pneumoniae and identification of a new nif gene: nifZ; Paul W et al.; A 1.4-kb PstI-HpaI DNA fragment carrying the Klebsiella pneumoniae nifM gene has been sequenced; nifM has been shown to encode a 30.6-kDa polypeptide . Two other open-reading frames were identified upstream of nifM . The one immediately upstream of nifM encodes a 16.6-kDa polypeptide which has been identified by in vitro transcription/translation in an Escherichia coli 30,000 x g supernatant system; we propose to designate this gene nifZ . The sequence of the second open reading frame is incomplete but it does not correspond to nifV, the gene previously thought to be immediately upstream of nifM, and may therefore identify another new nif gene . Both nifM and nifZ have functional nif promoters with the characteristic-24, -12 consensus sequence, we find no evidence for a nifM upstream activator sequence . The role of nifZ in nitrogenase biosynthesis is unknown but its identification calls into question previous assertions that only nifM and nifH are required for the synthesis of nitrogenase Fe protein. Eur J Biochem, 1987 Dec 15, 169(3), 457 - 65 Quantitative EPR of an S = 7/2 system in thionine-oxidized MoFe proteins of nitrogenase . A redefinition of the P-cluster concept; Hagen WR et al.; Thionine-oxidized nitrogenase MoFe proteins from Azotobacter vinelandii . Azotobacter chroococcum and Klebsiella pneumoniae exhibit excited-state EPR signals with g = 10.4, 5.8 and 5.5 with a maximal amplitude in the temperature range of 20-50 K . The magnitude of these effective g values, combined with the temperature dependence of the peak area at g = 10.4 from 12 K to 86 K, are consistent with an S = 7/2 system with spin Hamiltonian parameters D = -3.7 +/- 0.7 cm-1, {E} = 0.16 +/- 0.01 cm-1 and g = 2.00 . This interpretation predicts nine additional effective g values some of which have been detected as broad features of low intensity at g approximately 10, approximately 2.5 and approximately 1.8 . The S = 7/2 EPR is ascribed to the multi-iron exchange-coupled entities known as the P clusters . Quantification relative to the S = 3/2 EPR signal from dithionite-reduced MoFe protein indicates a stoichiometry of one P cluster per FeMo cofactor . Two possible interpretations for these observations, together with data from the literature, are proposed . In the first model there are two P clusters per tetrameric MoFe protein . Each P cluster encompasses approximately 8Fe ions and releases a total of three electrons on oxidation with excess thionine . In the second model the conventional view of four P clusters, each containing approximately 4Fe, is retained . This alternative requires that following one-electron oxidation, the P clusters factorize into two populations, Pa and Pb, only one of which is further oxidized with thionine resulting in the S = 7/2 system . Both models require eight-electron oxidation of tetrameric MoFe protein to reach the S = 7/2 state. Pharm Weekbl Sci, 1987 Dec 11, 9 Suppl, S33 - 40 Influence of dose frequency on the therapeutic efficacies of ciprofloxacin and ceftazidime in experimental Klebsiella pneumoniae pneumonia and septicemia in relation to their bactericidal activities in vitro; Roosendaal R et al.; The antibacterial activities of ciprofloxacin versus ceftazidime against Klebsiella pneumoniae in vitro and in vivo were compared . Although there was only a minor difference in MBC values between both drugs ciprofloxacin demonstrated a high and dose-dependent bacterial killing rate in vitro and in lungs of leukopenic rats in contrast to the more time-dependent bactericidal activity of ceftazidime . After treatment of a K.pneumoniae pneumonia and septicemia the efficacy of ciprofloxacin was only slightly influenced by the mode of administration, either at 6-h intervals or continuously, whereas ceftazidime was far more effective after continuous administration . This resulted in a superior efficacy of ciprofloxacin after intermittent treatment as compared to ceftazidime, whereas ceftazidime was more effective after continuous administration as compared to ciprofloxacin . Also ciprofloxacin proved to be bactericidal against bacteria that were not actively growing, both in vitro and in vivo, whereas ceftazidime was not. Nucleic Acids Res, 1987 Dec 10, 15(23), 9945 - 56 Mutational analysis of upstream sequences required for transcriptional activation of the Klebsiella pneumoniae nifH promoter; Buck M et al.; Upstream sequences of the Klebsiella pneumoniae nifH promoter were mutagenised and activation of the mutated promoters by the nif-specific transcriptional activator protein NifA examined in vivo . Of the sixteen mutations analysed, only those within the nifH upstream activator sequence (UAS), characterised by a TGT-N10-ACA motif, influenced nifH promoter activity . Mutations altering the two-fold rotational symmetry of the UAS or the spacing between the TGT and ACA motifs reduced promoter activity, consistent with the UAS functioning as a NifA binding site . The bases flanking the TGT-ACA motif of the UAS also appear to influence activation by NifA . Substituting the nifH UAS with a binding site for the transcriptional activator NtrC resulted in improved NtrC-dependent activation of the nifH promoter demonstrating that the activator specificity of the nifH promoter is dependent upon the presence of the appropriate upstream sequences to which the activator binds. Epidemiol Infect, 1987 Dec, 99(3), 625 - 34 Comparison of biochemical and serological typing results and antimicrobial susceptibility patterns in the epidemiological investigation of Klebsiella spp; Simoons-Smit AM et al.; An analysis of the serological and biochemical typing results of 925 clinical isolates of klebsiella revealed that biotyping and serotyping of klebsiella could replace each other for epidemiological purposes . The combination of both typing methods provided even more epidemiological information in analysing clusters of particular serotypes and biotypes in time . Clustering serotypes, mainly of neonatal origin, were nearly uniformly more resistant to the antibiotics in common use than other serotypes . Biotyping as well as serotyping of klebsiella isolates recovered from environmental surveys in the neonatal ward showed that epidemic and non-epidemic klebsiella isolates could occasionally be cultured from the environment and from the staff. Biochem J, 1987 Dec 1, 248(2), 351 - 8 Metabolism of 1-aminoethylphosphinate generates acetylphosphinate, a potent inhibitor of pyruvate dehydrogenase; Laber B et al.; The alanine analogue 1-aminoethylphosphinate {H3C-CH(NH2)-PO2H2} effectively inhibited anthocyanin synthesis in buckwheat hypocotyls and caused an increase in the concentrations of alanine and alanine-derived metabolites . Aminotransferase inhibitors partially alleviated the effects of the analogue . 1-Aminoethylphosphinate did not affect the growth of Klebsiella pneumoniae under anaerobic conditions, but under aerobic conditions it inhibited growth and caused the massive excretion of pyruvate . The analogue inhibited the pyruvate dehydrogenase complex in vitro in the presence of an aminotransferase activity . The transamination product of 1-aminoethylphosphinate, acetylphosphinate (H3C-CO-PO2H2), was found to inhibit the pyruvate dehydrogenase complex in a time-dependent reaction that followed first-order and saturation kinetics and required the presence of thiamin pyrophosphate. Am J Vet Res, 1987 Dec, 48(12), 1669 - 73 Growth inhibition of Escherichia coli and Klebsiella pneumoniae during involution of the bovine mammary gland: relation to secretion composition; Oliver SP et al.; Mammary secretions from 12 Holstein dairy cows were collected to evaluate growth inhibition of Escherichia coli and Klebsiella pneumoniae during involution and during physiologic transitions of the mammary gland . Mammary secretions obtained during late lactation poorly inhibited growth of E coli and K pneumoniae . However, as involution progressed, mammary secretions increasingly inhibited growth of both coliform mastitis pathogens . Greatest inhibition of E coli and K pneumoniae growth was observed when mammary glands were fully involuted . Growth inhibition remained high until 7 days before parturition, and then it decreased significantly (P less than 0.05) to that observed during late lactation . Inhibition of coliform mastitis pathogen growth was associated with high concentrations of lactoferrin and immunoglobulin G, decreased citrate concentration, and a low citrate to lactoferrin molar ratio . These data suggested that differences in susceptibility or resistance to new intramammary infection with coliform mastitis pathogens during the nonlactating period may be attributable, in part, to marked changes in mammary secretion composition that develop during physiologic transitions of the mammary gland . Resistance of the fully involuted mammary gland to coliform infection may be associated with high concentrations of natural protective factors. J Med Microbiol, 1987 Dec, 24(4), 363 - 70 The role of capsular polysaccharide K21b of Klebsiella and of the structurally related colanic-acid polysaccharide of Escherichia coli in resistance to phagocytosis and serum killing; Allen PM et al.; The behaviour of strains of Klebsiella aerogenes of capsular serotype K21 and strains of Escherichia coli producing a structurally related polysaccharide (colanic acid) was analysed by phagocytic and serum-killing assays . The cell-surface characteristics of these strains and of non-capsulate strains derived from them were also investigated by partitioning experiments in aqueous two-polymer phase systems . The possession of K21-type capsule by K . aerogenes or colanic-acid polysaccharide by E . coli conferred a strong negative charge on capsulate bacteria . Negatively charged bacteria of E . coli producing colanic-acid capsules, however, like non-capsulate K . aerogenes, were susceptible to uptake by polymorphonuclear leukocytes . In contrast, K21 polysaccharide conferred on klebsiellae considerable resistance to phagocytic uptake . The finding that ingested non-capsulate derivative strains of K . aerogenes were less rapidly degraded by phagocytes than E . coli strains suggested that other components of the cell surface of Klebsiella, notably lipopolysaccharide, may be involved in protection against phagocytic killing . The presence of colanic-acid capsules on E . coli conferred little resistance to the bactericidal activity of human serum or phagocytic uptake and did not protect against intracellular killing by polymorphonuclear leukocytes. Antimicrob Agents Chemother, 1987 Dec, 31(12), 1955 - 60 Tn1331, a novel multiresistance transposon encoding resistance to amikacin and ampicillin in Klebsiella pneumoniae; Tolmasky ME et al.; A 7.5-kilobase-pair multiresistance transposon, Tn1331, harboring amikacin resistance was identified as part of Klebsiella pneumoniae plasmid pJHCMW1 . Restriction mapping, hybridization, and transposition complementation experiments demonstrated that Tn1331 belongs to the Tn3 family . Its structure is similar to that of Tn3 with the insertion of a DNA fragment encoding resistance to amikacin, kanamycin, and tobramycin. J Mol Biol, 1987 Nov 20, 198(2), 211 - 22 Nucleotide sequence of korB, a replication control gene of broad host-range plasmid RK2; Kornacki JA et al.; The korB gene is a major regulatory element in the replication and maintenance of broad host-range plasmid RK2 . It negatively controls the replication gene trfA, the host-lethal determinants kilA and kilB, and the korA-korB operon . Here, we present the nucleotide sequence of an 1167 base-pair region that encodes korB . Using sequence data from korB mutants, we identified the korB structural gene . The predicted polypeptide product is negatively charged and has a molecular weight of 39,015, which is considerably less than that estimated by its electrophoretic mobility in SDS/polyacrylamide gels . Secondary-structure predictions of korB polypeptide revealed three closely spaced helix-turn-helix regions with significant homology to similar structures in known DNA-binding proteins . The korB gene, like all other sequenced RK2 genes, shows a strong preference for codons ending in a G or C residue . This is similar to codon usage by genes of Klebsiella and Pseudomonas, the original hosts for RK2 and some closely related plasmids . We also sequenced the site of transposon Tn76 insertion in the host-range mutant pRP761 and found it to be located immediately upstream from korB in the incC gene . Finally, we report the presence of sequences resembling a replication origin within the korB structural gene: a cluster of four 19 base-pair direct repeats and a nearby potential binding site for Escherichia coli dna A replication protein. Jpn J Antibiot, 1987 Nov, 40(11), 1891 - 4 {The clinical effect of fosfomycin on cystitis}; Fujita K et al.; Fosfomycin (FOM) is an antibiotic which inhibits phospho(enol)pyruvic acid transferase . Fifty-five patients with cystitis were treated with the drug for 7 days, at oral doses of 1 g for 3 times a day . Adult females under 70 years of age, visiting the clinic within 2 weeks after the onset, were classified as cases with typical simple cystitis with confirming the bladder irritability, pyuria and bacteriuria . The treatment was effective for 95.0% of these cases . Two cases with atypical cystitis caused by Klebsiella pneumoniae poorly responded to the drug. Am J Otolaryngol, 1987 Nov-Dec, 8(6), 387 - 90 Atrophic rhinitis: antibiotic treatment; Dudley JP; Atrophic rhinitis is a term used to describe a rare nasal infection . Although it does not have a fatal outcome, cause osteomyelitis, or produce pain, it does induce bilateral nasal obstruction and a persistent foul odor of which the subject and others are painfully aware . The organism most often associated with atrophic rhinitis is Klebsiella ozenae . Antibiotic susceptibility patterns of this microorganism have made treatment with orally administered antibiotics difficult . K ozenae was cultured from the nasal cavity of three patients . Two patients were treated for two weeks with tobramycin (MIC, 4 micrograms/ml; 4 mg/kg/day) . Odor decreased in one patient, but K ozenae failed to clear . In the second patient both odor and K ozenae disappeared . The third patient was treated for 1 week with tobramycin (MIC, 4 micrograms/ml; 4 mg/kg/day); odor decreased, but K ozenae could still be cultured . She was treated for an additional 2 weeks with topical gentamicin (MIC, 0.5 micrograms/ml) with disappearance of both odor and K ozenae . Intravenous aminoglycoside may be helpful in treating atrophic rhinitis, but topical aminoglycoside may provide an effective and cheaper form of treatment. J Burn Care Rehabil, 1987 Nov-Dec, 8(6), 487 - 91 Iron and immunologic function; Ward CG; This short review focuses on the immunological functions of iron-containing compounds . Five such compounds--transferrin, haptoglobin, hemopexin, albumin, and lactoferrin--are thought to contribute to normal infection resistance . Experimental results that demonstrate the ability of these compounds to inhibit the growth of an important pathogen, Klebsiella pneumoniae, are discussed. Antimicrob Agents Chemother, 1987 Nov, 31(11), 1809 - 15 Comparative activities of ciprofloxacin and ceftazidime against Klebsiella pneumoniae in vitro and in experimental pneumonia in leukopenic rats; Roosendaal R et al.; The antibacterial activities of ciprofloxacin and ceftazidime against Klebsiella pneumoniae in vitro and in vivo were compared . Although there was only a minor difference between the MBCs of both drugs, the bacterial killing rate of ciprofloxacin in vitro was very fast in comparison with that of ceftazidime . Similarly, the intravenous administration of ciprofloxacin at 1 h after bacterial inoculation resulted in effective bacterial killing in the lungs of leukopenic rats . This killing was dose dependent, in contrast to the dose-independent bactericidal effect of ceftazidime . The high antibacterial activity of ciprofloxacin in the lungs as compared with that of ceftazidime was also reflected in its therapeutic efficacy in K . pneumoniae pneumonia and septicemia in leukopenic rats when these infections were treated at 6-h intervals over 4 days, starting at 5 h after bacterial inoculation . Concentrations of ciprofloxacin and ceftazidime in lung tissue were not significantly different . Regarding the antibacterial activity of both drugs against K . pneumoniae in relation to the bacterial growth rate in vitro and in the lungs of leukopenic rats, ciprofloxacin killed K . pneumoniae organisms that were not actively growing, whereas ceftazidime did not . In addition, it was demonstrated that when the intravenous administration of antibiotic was delayed from 1 h up to 24 h after bacterial inoculation, ceftazidime lost its antibacterial activity in the lungs and blood of leukopenic rats, whereas ciprofloxacin was still very effective . These data suggest that the capacity of an antibiotic to kill bacteria at a slow growth rate may be relevant for its therapeutic effect in established infections, in which slowly growing bacteria form a substantial part of the total bacterial population. Vet Microbiol, 1987 Nov, 15(3), 219 - 28 Capsule types of Klebsiella pneumoniae isolated from the genital tract of mares with metritis, extra-genital sites of healthy mares and the genital tract of stallions; Kikuchi N et al.; A survey of K . pneumoniae was performed on cervical swabs, feces and nasal swabs of mares and on samples from the genital tract of stallions from 1980 to 1986 in south-western Hokkaido, Japan . K1 was the predominant type (79 of 88, 89.8%) in the metritis cases due to K . pneumoniae in mares of racing breeds . The same type was isolated from semen and swabs of the fossa glandis of 6 of 20 (30.0%) of the stallions of racing breeds . Heavily encapsulated and less heavily encapsulated K1 strains were isolated from the stallions . Mares bred to stallions carrying heavily encapsulated strains developed metritis, while those bred to stallions carrying less heavily encapsulated strains did not . K39 was isolated from cervical swabs solely from metritis-infected mares of draft breeds and not from any mares of the racing breeds examined . Untypable strains were isolated from cervical swabs in 7 of 88 (8.0%) metritis cases of mares of racing breeds and from semen in 7 of 19 (36.8%) stallions of racing breeds and they were predominant in feces (19 of 21, 90.5%) and nasal swabs (3 of 4, 75.0%) of healthy mares of racing breeds. Mol Gen Genet, 1987 Nov, 210(1), 140 - 4 Positional requirements for the function of nif-specific upstream activator sequences; Buck M et al.; The upstream activator sequence (UAS) found in Klebsiella pneumoniae nif promoters and required for the activation of transcription by nifA, is absent from the nifF-nifL intergenic region, but is present downstream from the nifLA transcription start at +59 . To determine whether nif upstream activator sequences can function in a 3' position, the nifH UAS was cloned downstream from the NifH transcription start, but no activation of transcription by nifA dependent upon the UAS in its 3' location could be detected . A mild repressive effect was detectable when the nifH UAS was placed downstream of the nifH promoter, but not when the cat promoter was substituted for the nifLA promoter upstream from the motif at +59 described above . However, deletion analysis showed that the UAS motif located downstream of the nifLA promoter has a role in transcription from the nifF promoter, although it is situated at position -263 with respect to the nifF transcription start, about 100 bp further upstream than previously described occurrences of the activator sequence. EMBO J, 1987 Nov, 6(11), 3531 - 8 Cloning and expression in Escherichia coli of the Klebsiella pneumoniae genes for production, surface localization and secretion of the lipoprotein pullulanase; d'Enfert C et al.; This article describes the reconstitution in Escherichia coli of a heterologous protein secretion system comprising a gene for an extracellular protein together with its cognate secretion genes . The protein concerned, pullulanase, is a secreted lipoprotein of the Gram-negative bacterium Klebsiella pneumoniae . It is initially localized to the cell surface before being specifically released into the medium . E . coli carrying the cloned pullulanase structural gene (pulA) produces pullulanase but does not expose or secrete it . Secretion genes were cloned together with pulA in an 18.8 kbp fragment of K . pneumoniae chromosomal DNA . E . coli carrying this fragment exhibited maltose-inducible production, exposition and specific secretion of pullulanase . Transposon mutagenesis showed that the secretion genes are located on both sides of pulA . Secretion genes located 5' to pulA were transcribed in the opposite orientation to pulA under the control of the previously identified, malT-regulated malX promoter . Thus these secretion genes are part of the maltose regulon and are therefore co-expressed with pulA . Transposon mutagenesis suggested that secretion genes located 3' of pulA are not co-transcribed with pulA, raising the possibility that some secretion functions are not maltose regulated. Biochem J, 1987 Nov 1, 247(3), 547 - 54 Klebsiella pneumoniae nitrogenase . Inhibition of hydrogen evolution by ethylene and the reduction of ethylene to ethane; Ashby GA et al.; Ethylene (C2H4) inhibited H2 evolution by the Mo-containing nitrogenase of Klebsiella pneumoniae . The extent of inhibition depended on the electron flux determined by the ratio of Fe protein (Kp2) to MoFe protein (Kp1) with KiC2H4 = 409 kPa ({Kp2}/{Kp1} = 22:1) and KC2H4i = 88 kPa ({Kp1}/{Kp2} = 21:1) at 23 degrees C at pH 7.4 . At {Kp2}/{Kp1} = 1:1, inhibition was minimal with C2H4 (101 kPa) . Extrapolation of data obtained when C2H4 was varied from 60 to 290 kPa indicates that at infinite pressure of C2H4 total inhibition of H2 evolution should occur . C2H4 inhibited concomitant S2O4(2-) oxidation to the same extent that it inhibited H2 evolution . Although other inhibitors of total electron flux such as CN- and CH3NC uncouple MgATP hydrolysis from electron transfer, C2H4 did not affect the ATP/2e ratio . Inhibition of H2 evolution by C2H4 was not relieved by CO . C2H4 was reduced to C2H6 at {Kp2}/{Kp1} ratios greater than or equal to 5:1 in a reaction that accounted for no more than 1% of the total electron flux . These data are discussed in terms of the chemistry of alkyne and alkene reduction on transition-metal centres. Infect Immu