Klebsiella

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 Immun, 1987 Nov, 55(11), 2741 - 6
Role of lipopolysaccharide and complement in susceptibility of Klebsiella pneumoniae to nonimmune serum; Ciurana B et al.; The role of lipopolysaccharide (LPS) in the susceptibility of Klebsiella pneumoniae to serum and the mechanism of complement activation by serum-susceptible (SerS) strains were investigated . The classical and alternative complement pathways are involved in serum killing of susceptible K . pneumoniae strains . The LPS composition seems to play a very important role in the serum bactericidal reaction, while capsular polysaccharide from this bacterium does not play any role . High-molecular-weight LPS from serum-resistant (Serr) K . pneumoniae strains was able to inhibit completely the serum bactericidal activity . LPS from SerS K . pneumoniae strains was not able to inhibit completely the serum bactericidal activity; low-molecular-weight LPS from Serr K . pneumoniae strains could not either . All these findings suggested that LPS composition, especially the O-antigen polysaccharide chains, contributes to the susceptibility of K . pneumoniae strains to complement-mediated serum bactericidal activity.

Am J Vet Res, 1987 Nov, 48(11), 1617 - 25
Role of eicosanoids, histamine, and serotonin in the pathogenesis of Klebsiella pneumoniae-induced bovine mastitis; Zia S et al.; By inoculating Klebsiella pneumoniae into the teat canals of mammary glands, coliform mastitis was induced experimentally in 6 lactating cows . Release of eicosanoids, histamine, and serotonin in plasma and milk was studied in response to 2 doses of K pneumoniae . A low dose (mean, 5,000 organisms/ml) was inoculated into cows 1 through 4, and a high dose (mean, 200,000 organisms/ml) was inoculated into cows 5 and 6 . Milk and blood samples were collected before inoculation (0 hours), and hourly, from 3 to 24 hours after inoculation . Concentrations of prostaglandin F2 alpha (PGF2 alpha), prostaglandin E (PGE), thromboxane B2 (TxB2), histamine, and serotonin were measured in plasma and milk obtained from control (NaCl solution-inoculated) and infected quarters . Fluorometric analysis of milk from infected quarters revealed significantly increased histamine and serotonin concentrations regardless of the dose of K pneumoniae . The mean (+/- SEM) peak concentrations of histamine were significantly (P less than 0.01) increased from the preinoculation value of 44 (+/- 12) ng/ml to 312 (+/- 104) ng/ml in milk from infected quarters and 72 (+/- 24) ng/ml in milk from control quarters . The mean peak concentration of serotonin increased significantly from the preinoculation concentration of 436 (+/- 37) ng/ml to 1,754 (+/- 662) ng/ml and 4,867 (+/- 1,248) ng/ml in milk from control (P less than 0.02) and infected (P less than 0.001) quarters, respectively . However, serotonin concentration in milk from infected quarters was approximately 2.8 times greater than that in milk from control quarters . Concentrations of PGF2 alpha, PGE, and TxB2 in milk and plasma were evaluated by radioimmunoassay.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbohydr Res, 1987 Oct 15, 168(1), 89 - 102
Structure of the capsular polysaccharide of Escherichia coli O9:K32(A):H19; Annison G et al.; The capsular polysaccharide of the bacterium Escherichia coli O9:K32(A):H19 was analyzed using chemical methods (hydrolysis, sequential Smith degradation, methylation analysis) together with 1H- and 13C-n.m.r . spectroscopy . 13C-N.m.r . spectroscopy and chemical analyses indicated that the K32 polysaccharide is composed of equimolar proportions of glucose, galactose, rhamnose, and glucuronic acid, and carries O-acetyl groups . 1H-N.m.r . analysis of native K32 polysaccharide revealed five resonances in the anomeric region (delta 5.52, 5.16, 5.12, 5.02, and 4.73) and the presence of an acetyl group (delta 2.18) . O-Deacetylation of the polysaccharide resulted in the loss of the resonance at delta 2.18 and one of the resonances (delta 5.52) in the anomeric region . The "extra" anomeric resonance in the 1H-n.m.r . spectrum of the native K32 polymer was assigned to H-2 of rhamnose, which experiences a large downfield shift when the 2-position is O-acetylated . This was confirmed by a 2D-COSY n.m.r . experiment and studies of model compounds . The K32 capsular polysaccharide is of the "2 + 2" type, comprised of the following repeating unit: (sequence; see text) This structure is identical to that of Klebsiella K55 capsular polysaccharide.

Biochem J, 1987 Oct 15, 247(2), 287 - 91
Nucleotide sequence of the Klebsiella pneumoniae nifD gene and predicted amino acid sequence of the alpha-subunit of nitrogenase MoFe protein; Ioannidis I et al.; The nucleotide sequence of the Klebsiella pneumoniae nifD gene is presented and together with the accompanying paper {Holland, Zilberstein, Zamir & Sussman (1987) Biochem . J . 247, 277-285} completes the sequence of the nifHDK genes encoding the nitrogenase polypeptides . The K . pneumoniae nifD gene encodes the 483-amino acid-residue nitrogenase alpha-subunit polypeptide of Mr 54156 . The alpha-subunit has five strongly conserved cysteine residues at positions 63, 89, 155, 184 and 275, some occurring in a region showing both primary sequence and potential structural homology to the K . pneumoniae nitrogenase beta-subunit . A comparison with six other alpha-subunit amino acid sequences has been made, which indicates a number of potentially important domains within alpha-subunits.

Biochem J, 1987 Oct 15, 247(2), 277 - 85
A quantitative approach to sequence comparisons of nitrogenase MoFe protein alpha- and beta-subunits including the newly sequenced nifK gene from Klebsiella pneumoniae; Holland D et al.; The nucleotide sequence was determined for part of the Klebsiella pneumoniae nif gene cluster containing the 3' end of the nifD gene and the entire length of the nifK gene (encoding the alpha- and beta-subunits of the nitrogenase MoFe protein respectively), as well as the putative start of the nifY gene, a gene of as yet unknown function . A broad-based comparison of a number of MoFe protein alpha-subunits, beta-subunits and alpha-versus beta-subunits was carried out by the use of a computer program that simultaneously aligns three protein sequences according to the mutation data matrix of Dayhoff . A new kind of quantitative statistical measure of the similarity between the aligned sequences was obtained by calculating and plotting standardized similarity scores for overlapping segments along the aligned proteins . This calculation determines if a test sequence is similar to the consensus sequence of two other proteins that are known to be related to each other . The different beta-subunits compared were found to be significantly similar along most of their sequence, with the exception of two relatively short regions centred around residues 225 and 300, which contain insertions/deletions . The overall pattern of similarity between different alpha-subunits exhibits resemblance to the overall pattern of similarity between different beta-subunits, including regions of low similarity centred around residues 225 and 340 . Comparison of alpha-subunits with beta-subunits showed that a region of significant similarity between the two types of subunits was located approximately between residues 120 and 180 in both subunits, but other parts of the proteins were only marginally similar . These results provide insights into likely tertiary structural features of the MoFe protein subunits.

Nature, 1987 Oct 29-Nov 4, 329(6142), 855 - 7
Identification of the V factor needed for synthesis of the iron-molybdenum cofactor of nitrogenase as homocitrate; Hoover TR et al.; Nitrogenase catalyses the ATP-dependent reduction of N2 to NH3, and is composed of two proteins, dinitrogenase (MoFe protein or component I) and dinitrogenase reductase (Fe protein or component II) . Dinitrogenase contains a unique prosthetic group (iron-molybdenum cofactor, FeMoco) comprised of Fe, Mo and S, which has been proposed as the site of N2 reduction . Biochemical and genetic studies of Nif- (nitrogen fixation) mutants of Klebsiella pneumoniae which are defective in nitrogen fixation, have shown that the nifB, nifQ, nifN, nifE and nifV genes are required for the biosynthesis of FeMo-co . Recently, a system for in vitro synthesis of FeMoco was described . The assay requires at least the nifB, nifN and nifE gene products, and a low-molecular-weight factor (V factor) produced in the presence of the nifV gene product . We have used this system to study FeMoco biosynthesis . We report here the isolation of V factor and identify it as homocitric acid ({R}2-hydroxy-1,2,4-butanetricarboxylic acid).

Clin Exp Immunol, 1987 Oct, 70(1), 201 - 8
Polyclonal activation of murine B cells by a membrane proteoglycan of Klebsiella pneumoniae; Millet I et al.; The lymphocyte activating properties of a membrane proteoglycan (MPG) extracted from a mutant non-encapsulated strain of Klebsiella pneumoniae (Kp) (biotype a I-145) were investigated . Kp MPG induced a strong proliferative response of BALB/c spleen cells and Peyer's patches cells . Thymidine incorporation was dose-related (from 1 to 100 micrograms Kp MPG/ml) and reached a maximum at day 3 . It was not reduced by removal of most adherent cells, nor by depletion of Thy1-2 positive cells, but it was abrogated by removal of surface immunoglobulin bearing cells . Spleen cells from nude mice and those from C3H/Hej mice were strongly stimulated by Kp MPG . Conversely Kp MPG did not induce interleukin 2 production and did not trigger the proliferation of thymocytes but stimulated interleukin 1 production by adherent spleen cells . Finally, unfractionated or B-enriched spleen cells cultured with Kp MPG synthesized IgM and, to a lesser extent, IgG and IgA . It is concluded that Kp MPG is a T-independent polyclonal B cell activator and an inducer of interleukin 1 production.

J Biol Chem, 1987 Sep 25, 262(27), 12900 - 3
Isotopic hybrids of nitrogenase . Mössbauer study of MoFe protein with selective 57Fe enrichment of the P-cluster; McLean PA et al.; Previous Mossbauer and EPR studies of the MoFe protein (approximately 30 Fe and 2 Mo) of nitrogenase have revealed the presence of two unique clusters, namely, the P-clusters (presumably of the Fe4S4 type) and the molybdenum- and iron-containing cofactors (or M-clusters) . Mossbauer components D (approximately 10-12 Fe) and Fe2+ (approximately 4 Fe) represent subsites of the P-clusters while component S (approximately 2 Fe) appeared to belong to a separate, unidentified cluster . In order to refine the analyses of Mossbauer spectra, we have constructed an isotopic hybrid of the Klebsiella pneumoniae protein which contains 57Fe-enriched P-clusters and 56Fe-enriched M-clusters . The highly resolved 57Fe Mossbauer spectra of this hybrid show that component S behaves spectroscopically like the P-cluster sites D and Fe2+ in oxidized and reduced MoFe protein . This suggests that S is a subset of the P-clusters rather than a different cluster type . The present study shows, for the first time, that the Debye-Waller factors of different P-cluster subsites have a different temperature dependence . Thus, the Fe2+/D absorption ratio is 4.0:10.0 at 4.2 K and 4.0:11.6 at 173 K . We propose that the reduced MoFe protein contains two pairs of P-clusters: one pair containing one Fe2+ and three D-sites and the other one Fe2+, two D, and one S-site . We have argued previously that the oxidized P-clusters occur in pairs as well.

Carbohydr Res, 1987 Sep 15, 167, 279 - 90
Klebsiella serotype K39: structure of an unusual capsular antigen deduced by use of a viral endoglucosidase; Anderson AN et al.; Methylation analysis and graded acid hydrolysis showed that the capsular polysaccharide of Klebsiella K39 is of the "5 + 1" type and that the repeating unit contains two nonequivalent residues of D-glucopyranosyluronic acid, one of which constitutes the branch point . A decision between two possible structures, in favor of that shown below, was reached by examination of the hexasaccharide obtained by depolymerization using phage luminal diameter 39, which possessed an endo-beta-D-glucosidase . Anomeric configurations were assigned by analysis of 1H- and 13C-n.m.r . spectroscopic data for the polysaccharide and oligosaccharides derived therefrom . The following structure is proposed for the Klebsiella K39 antigen: -3(Glc beta 1-4)GlcA beta 1-2Man alpha 1-4GlcA beta 1-2Man alpha 1-3Glc beta 1- The polysaccharide is the first in the Klebsiella genus thus far reported to contain two uronic acid residues per repeating unit.

Carbohydr Res, 1987 Sep 15, 167, 257 - 67
Bacteriophage-associated lyase activity against Klebsiella serotype K64 capsular polysaccharide; Ravenscroft N et al.; Bacteriophage phi 64 possesses a lyase that depolymerises the capsular polysaccharide of Klebsiella K64 into a hexasaccharide having an unsaturated derivative of glucuronic acid at the non-reducing end (1) . The unsaturated hex-4-enuronic acid residue generated was characterised spectroscopically (u.v . and n.m.r.) and by g.l.c.-m.s . after hydrogenation of the double bond . Partial hydrolysis, Smith degradation, methylation analysis, and n.m.r . spectroscopy have been used to establish the structures of oligosaccharides produced from the polysaccharide . Evidence from 1H-n.m.r . spectroscopy indicates that the D-Man rho residue that undergoes fission is beta . (Formula: see text).

Infect Immun, 1987 Sep, 55(9), 2234 - 9
Direct activation of human monocyte-derived macrophages by a bacterial glycoprotein extract inhibits the intracellular multiplication of virulent Legionella pneumophila serogroup 1; Rajagopalan P et al.; Intracellular multiplication of virulent Legionella pneumophila serogroup 1 was inhibited by human monocyte-derived macrophages activated by a glycoprotein extract of Klebsiella pneumoniae, RU 41.740 . Macrophage cultures were infected with L . pneumophila in the presence of immune antibodies on day 7 of culture . Extracellular bacteria were removed an hour after infection, and various concentrations of RU 41.740 or an antibiotic, erythromycin, were added . Intracellular multiplication in the presence of RU 41.740 was significantly slowed down compared with that of cultures without RU 41.740 . The reduction was, however, significantly less than that effected by erythromycin, which was used as a positive control for inhibition of intracellular multiplication . Cultures incubated with RU 41.740 before infection also demonstrated a significant reduction in the intracellular multiplication of L . pneumophila . In addition, RU 41.740 increased superoxide anion production from human monocytes in suspension in the presence of L . pneumophila . These results show that direct nonspecific activation of macrophages by a bacterial glycoprotein inhibits the intracellular multiplication of L . pneumophila and may suggest a role for activated macrophages in host defense against intracellular pathogens.

Mol Microbiol, 1987 Sep, 1(2), 159 - 68
A gene fusion approach to the study of pullulanase export and secretion in Escherichia coli; d'Enfert C et al.; A series of fusions between the gene for the Klebsiella pneumoniae secreted lipoprotein pullulanase (pulA) and the genes for cytoplasmic beta-galactosidase (lacZ) or periplasmic alkaline phosphatase (phoA) were created by transposon mutagenesis using mini-MudII1681 or TnphoA, respectively . The hybrid genes were expressed in Escherichia coli K-12 with or without the K . pneumoniae genes that promote pullulanase secretion in E . coli . We characterized seven different pulA-lacZ gene fusions encoding hybrid polypeptides containing from 14 to c . 1060 residues of pro-pullulanase . All but the smallest hybrid were fatty acylated and were toxic to producing cells, causing the accumulation of precursors of other exported proteins . Four different pulA-phoA gene fusions encoded hybrids with alkaline phosphatase activity . All four hybrids were fatty acylated, but were not toxic . Although the hybrids were apparently membrane-associated, they were not secreted into the medium either by E . coli carrying pullulanase secretion genes or by K . pneumoniae . Immunofluorescence tests indicated that the pullulanase secretion genes promoted the localization of one of these hybrids to the outer face of the E . coli outer membrane, which may have important implications for the design of live vaccine strains and of immobilized enzymes.

Mol Microbiol, 1987 Sep, 1(2), 133 - 42
Analysis of the Klebsiella pneumoniae ntrB gene by site-directed in vitro mutagenesis; MacFarlane SA et al.; A number of in-frame insertion and deletion mutations have been constructed in vitro in the Klebsiella pneumoniae ntrB gene and the effects of each mutant NtrB protein on NtrC activity have been assessed after reintroduction of the ntrB mutation into the glnA ntrBC operon . These experiments suggest that the phosphorylation of NtrC catalysed by NtrB not only makes NtrC competent as a transcriptional activator but also improves the DNA-binding properties and hence the negative control functions of NtrC . The variety of NtrB phenotypes obtained suggest a structure/function model for the protein.

Biochem J, 1987 Sep 1, 246(2), 455 - 65
Nitrogenase of Klebsiella pneumoniae . Kinetic studies on the Fe protein involving reduction by sodium dithionite, the binding of MgADP and a conformation change that alters the reactivity of the 4Fe-4S centre; Ashby GA et al.; The kinetics of reduction of indigocarmine-dye-oxidized Fe protein of nitrogenase from Klebsiella pneumoniae (Kp2ox) by sodium dithionite in the presence and absence of MgADP were studied by stopped-flow spectrophotometry at 23 degrees C and at pH 7.4 . Highly co-operative binding of 2MgADP (composite K greater than 4 X 10(10) M-2) to Kp2ox induced a rapid conformation change which caused the redox-active 4Fe-4S centre to be reduced by SO2-.(formed by the predissociation of dithionite ion) with k = 3 X 10(6) M-1.s-1 . This rate constant is at least 30 times lower than that for the reduction of free Kp2ox (k greater than 10(8) M-1.s-1) . Two mechanisms have been considered and limits obtained for the rate constants for MgADP binding/dissociation and a protein conformation change . Both mechanisms give rate constants (e.g . MgADP binding 3 X 10(5) less than k less than 3 X 10(6) M-1.s-1 and protein conformation change 6 X 10(2) less than k less than 6 X 10(3) s-1) that are similar to those reported for creatine kinase (EC 2.7.3.2) . The kinetics also show that in the catalytic cycle of nitrogenase with sodium dithionite as reductant replacement of 2MgADP by 2MgATP occurs on reduced and not oxidized Kp2 . Although the Kp2ox was reduced stoichiometrically by SO2- . and bound two equivalents of MgADP with complete conversion into the less-reactive conformation, it was only 45% active with respect to its ability to effect MgATP-dependent electron transfer to the MoFe protein.

J Antimicrob Chemother, 1987 Sep, 20(3), 323 - 34
Transferable resistance to third-generation cephalosporins in clinical isolates of Klebsiella pneumoniae: identification of CTX-1, a novel beta-lactamase; Sirot D et al.; Approximately 10% (89 isolates) of Klebsiella pneumoniae isolated in 1985 from patients in intensive care units in Clermont-Ferrand exhibited a complex resistance phenotype towards antibiotics . They were resistant to amino-, carboxy- and ureidopenicillins, aminoglycosides (except gentamicin), chloramphenicol, sulphonamides, tetracyclines and, most importantly, to cephalosporins (except cefoxitin and latamoxef) and to aztreonam . The metabolic profile of fifty isolates was identical and seven were selected for further study . All the resistance characters in these isolates were transferable to Escherichia coli by conjugation and were lost en bloc after treatment with ethidium bromide . Agarose gel electrophoresis of crude lysates of the wild types and their transconjugants indicated that the multiple resistances were mediated by a 95kb plasmid, pCF04 . The seven isolates selected for study and their corresponding transconjugants, constitutively produced a plasmid-mediated beta-lactamase with a pI of 6.3 that was much more active against third-generation cephalosporins than against cephalothin . The substrate profile and the isoelectric-focusing behaviour of this enzyme differed from those of other known plasmid-mediated beta-lactamases, and the enzyme was designated CTX-1 . A chromosomally-encoded SHV-1 (PIT-2) penicillinase (pI 7.7) was also present in the seven K . pneumoniae isolates but did not transfer . Resistance to aminoglycosides in the K . pneumoniae isolates was due to synthesis of a 6'-aminoglycoside acetyltransferase type IV . Our data indicate an epidemic of antibiotic multiply-resistant strains of K . pneumoniae producing a new beta-lactamase.

Mol Gen Genet, 1987 Sep, 209(2), 352 - 9
Mapping of the sor genes for L-sorbose degradation in the chromosome of Klebsiella pneumoniae; Sprenger GA et al.; A series of mutants was isolated in Klebsiella pneumoniae strain 1033, among them mutants unable to grow on L-sorbose . Different R' plasmids carrying the sor genes and other surrounding chromosomal genes were also isolated . Each plasmid contained the structural genes sorA for an Enzyme II of the phosphoenolpyruvate-dependent carbohydrate:phosphotransferase system, sorD for a D-glucitol 6-phosphate dehydrogenase, sorE for an L-sorbose 1-phosphate reductase, and the corresponding regulator gene sorR . These structural genes are coordinately expressed and inducible by L-sorbose . Cis-dominant and pleiotropic mutations rendering the expression of the sor genes constitutive or eliminating it were isolated . Complementation of a series of mutations in Escherichia coli K12 and K . pneumoniae by various R' and F' plasmids and by P1 transduction in K . pneumoniae located the sor genes within the following gene sequence: rbs rha pfkA metB ppc argH ilv btuB rpoB metA ace sor pgi malB uvrA . The rbs-ilv gene loci tightly linked in E . coli K12 at 84 min, are separated in the map of K . pneumoniae 1033 and located at 86 and 89 min, respectively.

Clin Immunol Immunopathol, 1987 Sep, 44(3), 283 - 96
Ligand-binding and idiotypic cross-reactions between anti-DNA antibodies and antibodies to Klebsiella K30 polysaccharide in patients with systemic lupus erythematosus or rheumatoid arthritis; Harkiss GD et al.; Antibodies to DNA and K30p were purified by affinity procedures and tested for their ability to cross-react with K30p and DNA, respectively . Anti-ssDNA antibodies were shown to react with solid-phase K30p and be inhibited in a dose-dependent manner by soluble ssDNA and K30p . Conversely, anti-K30p antibodies were found to bind immobilized ssDNA and be inhibited with soluble inhibitor . These results show that certain subpopulations of anti-ssDNA and anti-K30p antibodies overlap in their ligand-binding specificities . Idiotypic (Id) analysis showed that anti-K30p antibodies did not react significantly with an anti-Id reagent directed against a common anti-DNA framework Id (AM Id), thus clearly separating K30p-binding anti-DNA antibodies into an AM Id-negative population . When anti-DNA antibodies were probed with an anti-Id reagent directed against a framework Id present on anti-K30p antibodies (SP Id), substantial reactivity was found . Thus the SP Id identifies a subpopulation of antibodies capable of binding both K30p and DNA . These results show that a subset of anti-ssDNA antibodies cross-react with K30p antigen and are idiotypically related to a subset of antibodies reactive with K30p.

J Bacteriol, 1987 Sep, 169(9), 4024 - 9
Comparative organization of nitrogen fixation-specific genes from Azotobacter vinelandii and Klebsiella pneumoniae: DNA sequence of the nifUSV genes; Beynon J et al.; In the facultative anaerobe Klebsiella pneumoniae 17 nitrogen fixation-specific genes (nif genes) have been identified . Homologs to 12 of these genes have now been isolated from the aerobic diazotroph Azotobacter vinelandii . Comparative studies have indicated that these diverse microorganisms share striking similarities in the genetic organization of their nif genes and in the primary structure of their individual nif gene products . In this study the complete nucleotide sequence of the nifUSV gene clusters from both K . pneumoniae and A . vinelandii were determined . These genes are identically organized on their respective genomes, and the individual genes and their products exhibit a high degree of interspecies sequence homology.

Immunology, 1987 Sep, 62(1), 61 - 7
Comparison of blocking effects of monoclonal antibodies anti-MO1-alpha and anti-LFA1-alpha on human neutrophil functions; Pham Huu T et al.; In order to analyse the role of LFA1 and MO1 on neutrophil functions, the blocking effects of two monoclonal antibodies (MAb), one (anti-MO1) recognizing an epitope of the MO1-alpha chain and the other (25.31) an epitope of the LFA1-alpha chain, were measured . Adherence of 51Cr-labelled control neutrophils was 66 + 8% (mean +/- 1 SD) on plastic nuclon plates; this figure decreased to 33 +/- 5% and 23 +/- 6% of control adherence when the neutrophils had been pretreated with anti-LFA1-alpha (anti-alpha L) and anti-MO1-alpha (anti-alpha M), respectively . On another support (plastic culture chambers), 84 +/- 6% of control neutrophils adhered and the adherence of neutrophils pretreated with anti-alpha L or anti-alpha M was 10% and 43% of the control figure, respectively . These results show that adherence of neutrophils is dependent upon the plastic used . Moreover, inhibition of adhesion by the two MAbs was also dependent upon the support used for the assay, suggesting that MO1 and LFA1 may be surface proteins with different specificities . Both antigens capped upon adhesion, while they were randomly distributed in resting neutrophils . Anti-alpha L inhibited (congruent to 50%) locomotion more than did anti-alpha M (congruent to 25%), without altering chemoattractant-induced shape changes . These results suggest that the two MAbs inhibit chemokinesis but not chemotaxis . Many other adherence-associated functions, such as ingestion of opsonized Klebsiella pneumoniae, and cytotoxicity towards K/562 cells were decreased more by anti-alpha L than by anti-alpha M . In contrast, chemiluminescence and iodination induced by opsonized zymosan were inhibited more by anti-alpha M than by anti-alpha L . Degranulation induced by zymosan or opsonized zymosan was altered by anti-alpha M only, and this alteration involved azurophilic and not specific granules . Chemiluminescence induced by phorbol myristate acetate was inhibited to a greater extent by anti-alpha M than by anti-alpha L, while degranulation induced by phorbol myristate acetate was not altered by either of the two Mabs.

Mol Microbiol, 1987 Sep, 1(2), 243 - 9
Transcriptional activation of the Klebsiella pneumoniae nitrogenase promoter may involve DNA loop formation; Buck M et al.; Transcriptional activation of nitrogen fixation genes by NifA in Klebsiella pneumoniae requires an upstream NifA binding site . We now report that the introduction of half turns of the DNA helix into the DNA separating the upstream NifA binding site from the downstream promoter element of the nifH promoter decreases NifA-mediated activation to a greater extent than does the introduction of full helical turns . Reducing the spacing between the upstream and downstream elements of the nifH promoter also results in a promoter down phenotype . Introduction of a tight protein-binding site, the lac operator, between the upstream and downstream promoter elements did not render activation of the nifH promoter sensitive to occupancy of this site by the lac repressor . These findings indicate that NifA-mediated activation of transcription requires that NifA is bound upstream, and to the correct face of the DNA helix, in order to interact with downstream transcription factors . This implies that the interaction is brought about by the formation of a DNA loop between upstream and downstream promoter elements rather than by NifA sliding downstream.

Ann Trop Paediatr, 1987 Sep, 7(3), 181 - 6
Acute illness in Nigerian children with sickle cell anaemia; Akinyanju O et al.; The pattern of illness in 60 consecutive children with homozygous sickle cell disease who attended the Paediatric Emergency Room of a busy Lagos hospital with acute illness was studied prospectively . Their ages ranged from 3 months to 13 years with a peak in the 2nd year . There were twice as many boys as girls . The commonest symptoms were fever, limb or abdominal pain and cough, and the commonest signs were pallor and hepatomegaly . Painful crises occurred in 27, anaemic crises in 11, and a combination of these in 12 children . Infection was detected in 76% of subjects in crises . Infection was found in 82% of all the children and was mainly bacterial . The commonest infections were pneumonia (35%), bacteraemia (32%), tonsillitis/pharyngitis (17%) and osteomyelitis (8%) . The predominant bacteria isolated were Klebsiella spp (38%), E . coli (23%), Staph . aureus (23%), Staph . albus (23%) and Pseudomonas spp (23%) . Some children had multiple isolates . Bacterial infection was a major cause of morbidity in very young children and merits appropriate control and preventive measures in this age group . The spectrum of bacteria isolated makes it unlikely that the specific anti-pneumococcal measures widely advocated in Europe and America for young children with SCA would be appropriate in Nigeria.

J Chromatogr, 1987 Aug 21, 403, 217 - 24
Use of chromatofocusing for separation of beta-lactamases . VIII . Analytical chromatofocusing of chromosomal cephalosporinases from four Klebsiella strains; Gal S et al.; Although still there are Klebsiella strains which do not harbour plasmids and produce constitutive chromosomal beta-lactamases, recently clinical isolates were found in ever increasing numbers carrying mainly TEM-, CARB- and OXA type R-factors . We selected four chromosomal cephalosporinase producing Klebsiella strains to study the pI values of the enzymes and their simultaneous separability from accompanying proteins by chromatofocusing techniques . We compared pI values of the pure and the crude preparations: K . pneumoniae K1 SC 10436: pIpure = 6.4, pIcrude = 6.42; K . aerogenes K1 1082 E: pIpure = 6.5, pIcrude = 6.5; K . oxytoca 1082 E: pIpure = 6.42, pIcrude = 6.4; K . oxytoca 20: pIpure = 7.62, pIcrude = 7.6 . Excellent agreement of the pI values among each other, but occasional differences with those obtained by analytical isoelectrofocusing are attributed to methodological diversities and to the presence of satellite enzymes, known to exist in Klebsiella.

Biochem Pharmacol, 1987 Aug 15, 36(16), 2609 - 15
Diclofenac binding to human polymorphonuclear neutrophils: effect on respiratory burst and N-formylated peptide binding; Perianin A et al.; The respiratory burst of human polymorphonuclear neutrophils (PMN) induced by particle or soluble stimuli was measured in the presence of the nonsteroidal anti-inflammatory drug, diclofenac sodium (Voltaren) . Diclofenac (25-100 micrograms/ml) inhibited the oxygen consumption of PMN stimulated by 5 X 10(-7) M of N-formyl-methionyl-leucyl-phenylalanine (FMLP) . The inhibition was linearly correlated to diclofenac concentration . By contrast, diclofenac did not affect the rate of heat-killed Klebsiella pneumoniae ingestion of PMN, or the PMN O2-uptake induced by (0.67 microgram/ml) serum-opsonized zymosan or (1 microgram/ml) phorbol myristate acetate (PMA) . The PMN production of superoxide anion induced by various FMLP concentrations (10(-7), 10(-6) and 10(-5) M) was also decreased by diclofenac . However, this inhibition declined when the formylated peptide concentration was raised suggesting that diclofenac could alter FMLP binding to the PMN membrane . Binding experiments of tritiated FMLP to intact PMN performed at 22 degrees and 4 degrees showed high- and low-affinity FMLP sites with dissociation constant (Kd) values of approximately 2 X 10(-8) M and 10(-5) M respectively . Diclofenac did not significantly alter the low-affinity component but induced modifications of the high-affinity component which were different at 22 degrees and 4 degrees . At 22 degrees only the dissociation constant value was enhanced by diclofenac (competitive inhibition) whereas at 4 degrees both binding parameters (i.e . dissociation constant and number of available binding sites) were modified (mixed inhibition) . Diclofenac was also shown to bind to PMN with a low affinity . This binding was not diminished at 4 degrees by various concentrations of FMLP which even increased the number of diclofenac binding sites on PMN at 22 degrees . These data suggest that diclofenac binding to PMN may decrease FMLP-induced PMN respiratory burst by interfering with the peptide recognition by specific FMLP receptors.

J Clin Microbiol, 1987 Aug, 25(8), 1553 - 4
Cerebral abscess caused by Klebsiella ozaenae; Strampfer MJ et al.; Klebsiella ozaenae is the putative cause of ozena or atrophic rhinitis . It has also been commonly isolated as a colonizing organism, but recent reports demonstrate its role as an invasive pathogen, especially in immunosuppressed hosts . We report the first known case of a cerebral abscess caused by K . ozaenae.

J Gen Microbiol, 1987 Aug, 133 ( Pt 8), 2053 - 7
A rapid and efficient method for plasmid transformation of Klebsiella pneumoniae and Escherichia coli; Merrick MJ et al.; A rapid and efficient method for plasmid transformation of Klebsiella pneumoniae M5a1 and Escherichia coli K12 has been developed . The method, which uses a freeze-thaw cycle in the presence of CaCl2 to facilitate DNA uptake, is substantially more efficient for K . pneumoniae M5a1 than the conventional transformation procedure for E . coli . The simplicity and speed of the method makes it very attractive for routine transformation of K . pneumoniae M5a1 and E . coli K12.

J Gen Microbiol, 1987 Aug, 133 ( Pt 8), 2253 - 62
Characterization of three group A klebicin plasmids: localization of their E colicin immunity genes; James R et al.; We have investigated the immunity to E colicins conferred by three group A klebicin plasmids . pP5a, which encodes klebicin A1-P5, like pClo-DF13, confers immunity to colicin E6 on Escherichia coli K12, whilst pP5b and pP3, which encode klebicins A2-P5 and A3-P3 respectively, both confer immunity to colicin E3 . We have determined the restriction endonuclease and functional maps of the three group A klebicin plasmids . By sub-cloning and transposon mutagenesis we have investigated the relationship between the klebicin immunity and the E colicin immunity conferred by these plasmids . The colicin E6 and the klebicin A1 immunity are encoded by a single gene present on pP5a . The colicin E3 and the klebicin A2 immunity are encoded by a single gene present on pP5b . The colicin E3 and the klebicin A3 immunity are encoded by separate genes present on pP3 . Recombinant pML8412, which is derived from the ColE6-CT14 plasmid and encodes colicin E6 immunity, confers klebicin A1-P5 immunity upon Klebsiella pneumoniae UNF5023 . Recombinant pKC23, which is derived from the ColE3-CA38 plasmid and confers colicin E3 immunity, confers immunity to klebicin A2-P5, but not to klebicin A3-P3.

Carbohydr Res, 1987 Jul 15, 165(1), 77 - 86
Structure of the capsular polysaccharide of Klebsiella serotype K40; Ray AK et al.; The capsular polysaccharide from Klebsiella Serotype K40 contains D-galactose, D-mannose, L-rhamnose, and D-glucuronic acid in the ratios of 4:1:1:1 . Methylation analysis of the native and carboxyl-reduced polysaccharide provided information about the glycosidic linkages in the repeating unit . Degradation of the permethylated polymer with base established the identity of the sugar unit preceding the glycosyluronic acid residue . The modes of linkages of different sugar residues were further confirmed by Smith degradation and partial hydrolysis of the K40 polysaccharide . The anomeric configurations of the different sugar residues were determined by oxidation of the peracetylated native and carboxyl-reduced polysaccharide with chromium trioxide . Based on all of these results, the heptasaccharide structure 1 was assigned to the repeating unit of the K40 polysaccharide . (Formula: see text)

Acta Chir Scand, 1987 Jul-Aug, 153(7-8), 447 - 52
Long-term effects on bacterial sensitivity patterns of preoperative antibiotic prophylaxis in colorectal surgery; Walder M et al.; Since 1973, when doxycycline was introduced as peroperative prophylaxis in elective colorectal surgery at Malmo General Hospital, Sweden, there has been an unchanged and low rate (8-12%) of septic complications in colonic surgery . For treating postoperative infections ampicillin, cefuroxime and piperacillin have been used since 1973, 1980 and 1982 respectively . The sensitivity pattern of E . coli and Klebsiella against these four antibiotics used for prophylaxis and treatment has been followed for the past five years and only minor changes have occurred through the period . However, a lower frequency of antibiotic resistance was recorded for bacteria isolated peroperatively than postoperatively after colorectal surgery or from infection sites from other patients presumably mostly due to selection caused by antibiotics used within the hospital . Due to the good clinical outcome and seemingly lack of development of antibiotic resistance in peroperative isolates, doxycycline still remains a choice for prophylaxis in bowel surgery.

Appl Environ Microbiol, 1987 Jul, 53(7), 1716 - 7
Enumeration of Klebsiella spp . in cold water by using MacConkey-inositol-potassium tellurite medium; Dutka BJ et al.; MacConkey-inositol-potassium tellurite agar was field tested for its ability to selectively enumerate Klebsiella species from the waters of the Saint John River Basin, which include fresh and marine waters . Water temperature varied from 1 to 6 degrees C during the survey period . Results of the study indicated that 77% of the typical colonies on MacConkey-inositol-potassium tellurite medium were Klebsiella species, but the total Klebsiella population enumerated was greatly underestimated.

J Child Neurol, 1987 Jul, 2(3), 201 - 4
Abnormal cerebral cortical neurons in a child with maternal PKU syndrome; Lacey DJ et al.; A child born to a phenylketonuric (PKU) woman not on dietary treatment was microcephalic and had congenital heart and other physical anomalies consistent with the maternal PKU syndrome . After a repeat cardiac catherization at 3 months of age, he suffered an embolic left cerebral infarct and seizures . He was found by his mother dead in bed at 4 months of age; general autopsy revealed Klebsiella sepsis and pneumonia . Neuropathologic studies of the right cerebral hemisphere showed neuronal loss of the third cortical layer; Golgi studies revealed persistently immature cortical pyramidal cell somata and dendritic spines . This is the first report of specific neuronal abnormalities in a child with the maternal PKU syndrome and may, in part, account for the clinical features of microcephaly, mental retardation, and seizures seen in affected children.

Mol Microbiol, 1987 Jul, 1(1), 92 - 100
Requirements for transcriptional activation in vitro of the nitrogen-regulated glnA and nifLA promoters from Klebsiella pneumoniae: dependence on activator concentration; Austin S et al.; Three proteins involved in nitrogen regulation in Klebsiella pneumoniae, NTRA, NTRB and NTRC, have been purified . In a defined in vitro system all three NTR proteins are required for initiation of transcription at the ntr activatable glnA and nifLA promoters . However, in crude S-30 extracts, transcription from the glnA promoter, but not the nifLA promoter, can be activated in the absence of NTRB . A higher concentration of NTRC is required for activation of nifLA transcription than for glnA transcription . Sequences located between -227 and -158 with respect to the nifL transcription start site are required for efficient activation of the nifLA promoter in vitro.

Cornell Vet, 1987 Jul, 77(3), 225 - 34
Studies on induced Klebsiella mastitis with relationships among N-acetyl-beta-D-glucosaminidase, bacterial and somatic cell counts; Kunkel JR et al.; Mastitis was induced by infusion of 3,000-7,000 colony-forming units of Klebsiella pneumoniae into the left rear quarter of four primiparous Holstein-Freisians . Greater than 100 organisms were recovered from a quarter milk sample within 2 hr and in 87.5% of the hourly cultures taken after 8 hr . Bacteria, California Mastitis Tests, somatic cell counts and N-acetyl-beta-D-glucosaminidase levels differed significantly from that of control quarters in infused cows after hr 6, 8, 9 and 9 respectively . High correlations were obtained between somatic cell count and N-acetyl-beta-D-glucosaminidase (r = .97) and N-acetyl-beta-D-glucosaminidase and natural log bacterial count (r . = .88) . N-acetyl-beta-D-glucosaminidase was related to natural log bacterial counts of samples taken 1, 2, 3, and 4 hr prior to the test (rN-acetyl-beta-D-glucosaminidase and natural log bacterial count (r = .91, .92, .93, and .95).

Biol Chem Hoppe Seyler, 1987 Jul, 368(7), 787 - 93
Electron microscopical investigation of citrate lyase single molecules; Ihle E et al.; Electron micrographs of citrate lyase from Rhodopseudomonas gelatinosa and Klebsiella aerogenes reveal two characteristic molecular forms . The "basket" form and the "star" form were subjected to two-dimensional image reconstruction using a technique involving averaging of superposed single molecular images after rotational correlation . A three-dimensional image reconstruction shows that the images of these forms can be interconverted by rotation and that they therefore represent different views of the same structure.

J Clin Microbiol, 1987 Jul, 25(7), 1228 - 32
New bacteriophage typing scheme for subdivision of the frequent capsular serotypes of Klebsiella spp; Gaston MA et al.; A bacteriophage typing scheme for hospital isolates of Klebsiella spp . was developed . The scheme was designed specifically as a secondary typing method to discriminate between strains of serotypes K2, K3, and K21 but proved to be an efficient general typing method for strains of most serotypes . The set of 15 phages gave 87.3% typeability on 236 strains of more than 70 different serotypes . Typeability within the K2, K3, and K21 strains was 93, 89, and 91%, respectively . There was a mean of 3.2 reactions strain-1 for all phage-typeable strains . Of the serologically nontypeable strains, 76.7% were susceptible to one or more phages . The most common pattern accounted for only 7% of the strains . The lytic patterns were reproducible if strains were typed on the same day, but differences were observed if strains were stored for 1 week or more before retyping . A total of 96.5% of the strains were typeable by a combination of capsular serology and phage typing.

Arch Biochem Biophys, 1987 Jul, 256(1), 212 - 22
Biosynthesis of o-succinylbenzoic acid . I: Cell free synthesis of o-succinylbenzoic acid from isochorismic acid in enzyme preparations from vitamin K producing bacteria; Weische A et al.; Escherichia coli K12 and a mutant of E . coli (viz., AN 154) as well as Aerobacter aerogenes 62-1 (i.e., Klebsiella pneumoniae) were used as sources of the enzyme catalyzing the formation of o-succinylbenzoic acid (OSB) from isochorismic acid and alpha-ketoglutaric acid in the presence of thiamine pyrophosphate . The product of the reaction (OSB) was identified by HPLC before and after derivatization to the methylester, dilactone, and coenzyme A ester . OSB synthase and alpha-ketoglutarate dehydrogenase are similar in that both decarboxylate alpha-ketoglutarate in the presence of thiamine pyrophosphate but the enzyme systems can be separated easily by several methods . Reexamination of mutants E . coli AN 154 and AN 191 showed that these mutants are leaky, rather than blocked, between chorismic acid and isochorismic acid . This finding, together with the observation that isochorismic acid rather than chorismic acid is the substrate of OSB synthase, invalidates previous assumptions on the reaction initiating vitamin K2 biosynthesis.

Ann Inst Pasteur Immunol, 1987 Jul-Aug, 138(4), 571 - 84
Immunological activities of RU-41740, a glycoproteic extract from Klebsiella pneumoniae . II.--Activation of macrophage cytotoxicity against tumour cells and production of a cytotoxic factor; Vacheron F et al.; RU-41740, a glycoprotein extract from Klebsiella pneumoniae, is an immunomodulating agent with a broad spectrum of activities . It enhances several macrophage functions, including interleukin-1 (IL-1) secretion . Present data show that RU-41740 was able to promote murine macrophage cytotoxicity against tumour cells . A soluble cytotoxic factor (CF) was found in supernatants from macrophage cultures stimulated with RU-41740 . These supernatants were shown to contain IL-1 . CF was detected on L-929 cells sensitized by actinomycin D . CF was analysed on the basis of MW and pHi: after gel filtration on "Ultrogel Aca54", a single peak of CF was found in the range of 50-60 Kd and was then distinct from IL-1, which was eluted at 15 Kd . After chromatofocusing, CF was found in a narrow peak of pH 4.8 . CF was detected in supernatants 2 h after macrophage stimulation by RU-41740, and its release was abolished by pretreatment of macrophages with cycloheximide (2 micrograms/ml) . CF described here shares several properties with tumour necrosis factor (TNF).

Antimicrob Agents Chemother, 1987 Jul, 31(7), 982 - 5
Pharmacokinetics and therapeutic efficacy of gentamicin in an experimental pleural empyema rabbit model; Shohet I et al.; The pharmacokinetics and therapeutic efficacy of gentamicin were investigated in an experimental pleural empyema rabbit model . Pleural effusion was induced by the intrapleural administration of turpentine, and empyema was induced by direct inoculation of the effusion with Klebsiella pneumoniae . Pleural empyema compared with effusion was characterized by lower pH, oxygen tension (PaO2), and glucose levels and higher leukocyte count, lactic acid concentration, and PaCO2 . After a single administration, gentamicin was first detectable in the pleural fluid at 60 min, whereas peak levels in empyema were observed at 180 min . Gentamicin persisted in the empyema longer than in blood . Animals treated with gentamicin only had 60% bacterial cure on day 7; those treated with gentamicin in an oxygen chamber had 100% cure on day 5 (P = 0.004) . Low oxygen tension diminished the antibacterial efficacy of gentamicin in this model . An increase in oxygen tension improved the therapeutic results without alteration of the pharmacokinetics of gentamicin.

Mol Microbiol, 1987 Jul, 1(1), 37 - 44
Sequence of nifL from Klebsiella pneumoniae: mode of action and relationship to two families of regulatory proteins; Drummond MH et al.; We present the nucleotide sequence of K . pneumoniae nifL, which negatively regulates nif transcription in response to oxygen and fixed nitrogen . It shows partial sequence homology to the general nitrogen regulatory proteins NtrB of K . pneumoniae and Bradyrhizobium parasponiae . This homology is weaker than that shown between the NifA and NtrC activator components of the nif and general nitrogen control systems . The N-terminal section of the NifL protein includes a structural duplication sharing sequence homology with part of NtrB, and a region containing a cysteine pair which might be implicated in redox control Unlike NtrB, NifL appears to lack a DNA-binding motif, consistent with evidence that NifL represses by interacting directly with NifA . The C-terminal section of NifL shows clear homology to NtrB and to a family of proteins involved in transcriptional control or chemotaxis, each of which probably interacts with a member of the family of regulatory proteins showing homology to NtrC.

Mol Microbiol, 1987 Jul, 1(1), 107 - 16
Export and secretion of the lipoprotein pullulanase by Klebsiella pneumoniae; d'Enfert C et al.; Pullulanase, a secreted lipoprotein of Klebsiella pneumoniae, is initially localized to the outer face of the outer membrane, as shown by protease and substrate accessibility and by immunofluorescence tests . Freeze-thaw disruption of these cells released both membrane-associated and apparently soluble forms of pullulanase . Membrane-associated pullulanase co-fractionated with authentic outer membrane vesicles upon isopycnic sucrose-gradient centrifugation, whereas the quasi-soluble form had the same equilibrium density as inner membrane vesicles and extracellular pullulanase aggregates . The latter also contained outer membrane maltoporin, but were largely devoid of other membrane components including LPS and lipids . K . pneumoniae carrying multiple copies of the pullulanase structural gene (pulA) produced increased amounts of cell-associated and secreted pullulanase, but a large proportion of the enzyme was neither exposed on the cell surface nor released into the medium, even after prolonged incubation . This suggests that factors necessary for pullulanase secretion were saturated by the over-produced pullulanase . When pulA was expressed under lacZ promotor control, the pullulanase which was produced was not exposed on the cell surface at any time, suggesting that pullulanase secretion genes are not expressed constitutively, and raising the possibility that they, like pulA, may be part of the maltose regulon.

J Exp Med, 1987 Jul 1, 166(1), 173 - 81
Autoantibodies to HLA B27 in the sera of HLA B27 patients with ankylosing spondylitis and Reiter's syndrome . Molecular mimicry with Klebsiella pneumoniae as potential mechanism of autoimmune disease; Schwimmbeck PL et al.; Ankylosing spondylitis (AS) and Reiter's syndrome (RS) both show a strong correlation with the HLA B27 haplotype . We studied whether sharing of homologous amino acid sequences in the HLA B27 antigen with an invading microbe might occur, and if so, what is the biological significance of such homology . In a computer search of the Dayhoff data bank, we found a homology of six consecutive amino acids between HLA B27.1 antigen residues 72-77 and Klebsiella pneumoniae nitrogenase residues 188-193 . These shared sequences are hydrophilic, suggesting locations on molecules exposed to the cell surface . Immunochemical analysis showed that 18 of 34 sera from patients with RS (53%) and 7 of 24 sera from patients with AS (29%) contained antibodies that bound to a synthesized peptide sequence representing residues 69-84 of HLA B27.1 . In contrast, only 1 of 22 sera from healthy, B27+ controls (5%) contained antibodies to this peptide (p less than 0.01) . Sera from three HLA B27- patients with RS did not possess antibodies to the HLA B27 peptide . Additionally, greater than 40% of HLA B27 patients with AS or RS had antibodies to Klebsiella residues 184-193, while none of the normal nonarthritic HLA B27 haplotype subjects did . Our results suggest that an autoimmune response(s) directed against HLA B27.1 may be a pathogenic mechanism in a subset of patients with AS and RS . Further, this response may initially be induced against Klebsiella pneumoniae, a microorganism that shares sequence homology with HLA B27.

J Biol Chem, 1987 Jun 25, 262(18), 8814 - 20
Nitrogenase MoFe protein subunits from Klebsiella pneumoniae expressed in foreign hosts . Characteristics and interactions; Holland D et al.; The expression of selected nitrogen fixation (nif) genes from Klebsiella pneumoniae in foreign hosts provides an approach to determine the pathway, minimal genetic requirements, and host dependence of nitrogenase assembly . In this study, we investigated the assembly of the alpha 2 beta 2 MoFe protein, responsible for substrate binding and reduction, by introducing nifD and nifK (encoding respectively, the alpha and beta subunits) into Escherichia coli and the yeast Saccharomyces cerevisiae . In E . coli, both genes were expressed from the nifHDKY operon; in yeast, the genes, separately fused to the yeast ADH1 promoter, were introduced on two different plasmids . Denaturing immunoblot analyses demonstrated the presence of significant amounts of NifD and NifK in both hosts . In E . coli, the level or perhaps modification of NifD depended on the growth medium of the bacteria . Nondenaturing, anaerobic immunoblot assays revealed in E . coli, nif-specific antigens of lower electrophoretic mobility than Kp1, which may represent assembly intermediates . In yeast, no putative assembled products were evident, and the predominant antigens corresponded to the monomeric forms of the polypeptides . These results indicate that, unlike NifH, the Fe protein subunit (Berman, J., Gershoni, J . M., and Zamir, A . (1985) J . Biol . Chem . 260, 5240-5243), NifD and NifK are insufficient for the assembly of an electrophoretically Kp1-like structure . Homodimerization of nifK and probably of nifD primary gene products does not appear to occur spontaneously and hence is unlikely to represent the initial step in the assembly . The difference between the two hosts suggests that the cellular environment or mode of expression could affect the interaction between the two subunits.

J Biol Chem, 1987 Jun 25, 262(18), 8544 - 50
Activation and cleavage of the carbon-cobalt bond of adeninylethylcobalamin by diol dehydrase; Toraya T et al.; Adeninylethylcobalamin (AdeEtCbl) underwent cleavage of the C-Co bond by interaction with apoprotein of diol dehydrase from Klebsiella pneumoniae ATCC 8724, although this analog was quite inactive as coenzyme . Spectroscopic observation indicates that AdeEtCbl was converted to the enzyme-bound hydroxocobalamin without intermediates . The conversion was stoichiometric (1:1) and obeyed the second-order reaction kinetics (k = 0.027 min-1 microM-1 at 37 degrees C) depending upon concentrations of apoprotein and AdeEtCbl . This suggests that the complex formation is the rate-determining step and that AdeEtCbl undergoes rapid C-Co bond cleavage once it binds to the apoenzyme . Substrates and oxygen did apparently not affect the rate of the C-Co bond cleavage . The experiments using {adenine-U-14C}AdeEtCbl and {1(3)-3H}glycerol demonstrated that 9-ethyladenine was the only product formed from the adeninylethyl group of AdeEtCbl during the conversion and that an additional hydrogen atom in the 9-ethyladenine is not derived from the substrate . 1H NMR measurement of the 9-ethyladenine formed enzymatically from AdeEtCbl and DL-1,2-{1,1,2-2H3}propanediol also led to the same conclusion . All of these results indicate that the C-Co bond of AdeEtCbl is activated by diol dehydrase and undergoes heterolysis forming Co(III) and a carbanion or a carbanion-like species, in clear contrast to the homolysis of the C-Co bond of adenosylcobalamin in the normal catalytic process . 9-Ethyladenine formed remained tightly associated with the enzyme . Longer chain homologs, i.e . adeninylpropylcobalamin, adeninylbutylcobalamin, and adeninylpentylcobalamin did not undergo such cleavage of the C-Co bond by diol dehydrase.

J Chromatogr, 1987 Jun 19, 396, 281 - 6
Purification of the lipopolysaccharide fraction from Klebsiella pneumoniae O1 K2 by high-performance liquid chromatography; Kol O et al.; An high-performance liquid chromatography technique was applied to purify the lipopolysaccharide fraction from a lysate of Klebsiella pneumoniae O1 K2 (NCTC 5055) . The separation of the lipopolysaccharide fraction from the proteins was carried out with a reversed-phase column . By this method the lipopolysaccharide fraction was obtained in a pure state, devoid of proteins but possessing the same biological properties as the lipopolysaccharide fraction prepared by the classical phenol-water technique.

Immunol Lett, 1987 Jun, 15(2), 117 - 20
Serum immunoglobulin levels in patients with active pulmonary tuberculosis and patients with Klebsiella infection; Sela O et al.; Serum immunoglobulins levels were determined in 42 patients with active pulmonary tuberculosis as a prototype of chronic infection and 41 patients with Klebsiella infection representing acute infection, using a radial immunodiffusion technique . The mean serum concentration of IgG, IgA and IgM of the patients with pulmonary tuberculosis (1980 +/- 688, 314 +/- 152 and 222 +/- 123 mg/dl, respectively) were found to be significantly higher than normal control levels (P less than 0.005) . The mean serum IgG, IgA and IgM concentration of the patients with Klebsiella infection (1102 +/- 340, 287 +/- 133 and 168 +/- 105 mg/dl, respectively), were also higher than normal levels but only IgM level differed significantly from the normal control levels (P less than 0.05) . The difference in serum IgG and IgM between the patients with pulmonary tuberculosis and Klebsiella infection was statistically significant (P less than 0.05 and P less than 0.05, respectively) . These findings reflect the polyclonal hypergammaglobulinemia found in chronic infections, on the one hand, and the rise of IgM of the primary immune response in acute infection, on the other.

Antimicrob Agents Chemother, 1987 Jun, 31(6), 920 - 4
Enhancement of bronchoalveolar cell recovery and stimulation of alveolar macrophage chemiluminescence and resistance to influenza virus after treatment with RU 41821 aerosol; Rudent A et al.; Aerosol treatment with RU 41821, a glycoprotein extract from Klebsiella pneumoniae, was tested in mice for its effect on the kinetics of the induction of bronchoalveolar cells (i.e., alveolar macrophages, monocytes, lymphocytes, and polymorphonuclear leukocytes) . RU 41821 led to an increase in the total number of bronchoalveolar cells . The largest increase was observed for polymorphonuclear leukocytes, and more moderate increases occurred in the numbers of alveolar macrophages, monocytes, and lymphocytes . The alveolar macrophages recruited in response to RU 41821 were activated, as indicated by luminol-dependent chemiluminescence in response to stimulation by opsonized zymosan . The effects of five RU 41821 aerosol treatments and those of a single treatment were further examined in vivo by aerosol infection of mice inoculated with a mouse-pathogenic influenza virus . The maximum protective effect was obtained after five once-a-day treatments and was correlated with the largest increase in the total number of bronchoalveolar cells.

Br J Rheumatol, 1987 Jun, 26(3), 168 - 71
A study of antipolynucleotide antibodies, anti-Klebsiella (K30) antibodies and anti-DNA antibody idiotypes in ankylosing spondylitis; Isenberg DA et al.; Recent studies have indicated that both ankylosing spondylitis and the anti-DNA antibodies found in systemic lupus erythematosus may be related to Klebsiella surface antigens . In order to explore these possible relationships further, the sera of 24 patients with ankylosing spondylitis (AS), and 20 controls, have been examined for binding to a wide range of antipolynucleotide antibodies, antibodies binding to the Klebsiella pneumoniae polysaccharide K30 and two DNA antibody idiotypes designated 16/6 and 134 . We report that although 21% of the AS patients had IgG ssDNA antibodies it is evident that the aetiopathogenesis of this disease is not through the mechanism of autoantibodies or the common DNA antibody idiotypes tested.

Biochem J, 1987 Jun 1, 244(2), 485 - 8
Nitrogenase of Klebsiella pneumoniae . Rhodanese-catalysed restoration of activity of the inactive 2Fe species of the Fe protein; Pagani S et al.; The inactive 2Fe species of the Fe protein of the nitrogenase of Klebsiella pneumoniae was generated by treating oxidized Fe protein (Kp2) with MgATP and chelator . Incubation of the 2Fe species of Kp2 with the sulphurtransferase rhodanese in the presence of thiosulphate, ferric citrate and reduced lipoate reproducibly restored activity . The extent of restoration of activity depended on the molar ratio of 2Fe Kp2 to rhodanese and was time-dependent . Re-activation did not occur in the reaction mixture lacking rhodanese.

Pathol Biol (Paris), 1987 Jun, 35(5 Pt 2), 796 - 9
{Effects of beta-lactamases on the antibacterial activity of carumonam and aztreonam against Klebsiella sp . in comparison with 4 other beta-lactams}; Labia R et al.; Carumonam and aztreonam are two similar monocyclic beta lactam antibiotics . We have studied their antibacterial activities on Klebsiella, in comparison with cefotaxime, cefoxitin, cefotetan and imipenem . In the case of K . pneumoniae (53 strains) the antibacterial activities of the two monobactams are very similar . On the contrary, with K . oxytoca (22 strains) carumonam is significantly more active . This situation seems to be justified by an improved stability of carumonam to beta lactamases . In addition, the antibacterial activities of imipenem and of cephamycins remain quite constant within all the strains . Then the antibacterial activity of cefotaxime is fluctuant, which is in relation with a clear susceptibility to the Klebsiella beta lactamases.

J Antibiot (Tokyo), 1987 Jun, 40(6), 868 - 72
The mechanism of action of piperacillin-analogues in vitro; effect of the carbon number at the N-4 position of 2,3-dioxopiperazine on the outer membrane permeability, stability to beta-lactamase and binding affinity to penicillin-binding proteins; Mitsuyama J et al.; The relationship between the chemical structure and the mode of action of piperacillin-analogues (PIPC-analogues) against Escherichia coli and Klebsiella pneumoniae were investigated . The antibacterial activity of PIPC-analogues increased with an increase in the number of carbon atoms at the N-4 position of 2,3-dioxopiperazine . Their mode of action is discussed on the basis of the results of studies on outer membrane permeability, stability to beta-lactamase and binding affinity to penicillin-binding proteins (PBPs) . The outer membrane permeability and stability to beta-lactamase were hardly affected by the chain length of the alkyl group at the N-4 position . On the other hand, the affinity to PBPs, especially to PBP 3, became stronger with increase of the number of carbon atoms at N-4 position . These results suggest that increased affinity to PBPs is the main reason for the increased antibacterial activity of the PIPC-analogues reported here.

Tijdschr Diergeneeskd, 1987 Jun 1, 112(11), 653 - 9
{Klebsiella pneumoniae mastitis as a dairying problem}; Vecht U et al.; Quarter samples were taken from lactating and dry cows of a dairy herd in the Netherlands . Klebsiella pneumoniae was isolated from milk and dry udder secretum of cattle, and from the bedding of the cubicles . The animals concerned did not show any clinical symptoms . For some years Klebsiella mastitis caused severe problems on this farm . A strict hygienic program resulted in complete control of the disease . Diagnostics, therapy and prevention of Klebsiella mastitis are discussed.

J Bacteriol, 1987 Jun, 169(6), 2876 - 80
In vitro transcription of the nitrogen fixation regulatory operon nifLA of Klebsiella pneumoniae; Wong PK et al.; In vitro transcription from the promoter for the nitrogen fixation regulatory operon nifLA of K . pneumoniae requires four protein fractions: the core form of RNA polymerase; NTRA, an alternate sigma factor; NTRC, an auxiliary DNA-binding protein; and NTRB, a bifunctional enzyme that controls the activity of NTRC by covalent modification (A.J . Ninfa and B . Magasanik, Proc . Natl . Acad . Sci . USA 83:5909, 1986) . Two DNA-binding sites for NTRC lie approximately 150 base pairs upstream of the nifLA promoter.

FEBS Lett, 1987 May 25, 216(1), 133 - 9
The prephenate dehydrogenase component of the bifunctional T-protein in enteric bacteria can utilize L-arogenate; Ahmad S et al.; The prephenate dehydrogenase component of the bifunctional T-protein (chorismate mutase:prephenate dehydrogenase) has been shown to utilize L-arogenate, a common precursor of phenylalanine and tyrosine in nature, as a substrate . Partially purified T-protein from Klebsiella pneumoniae and from Escherichia coli strains K 12, B, C and W was used to demonstrate the utilization of L-arogenate as an alternative substrate for prephenate in the presence of nicotinamide adenine dinucleotide as cofactor . The formation of L-tyrosine from L-arogenate by the T-protein dehydrogenase was confirmed by high-performance liquid chromatography . As expected of a common catalytic site, dehydrogenase activity with either prephenate or L-arogenate was highly sensitive to inhibition by L-tyrosine.

Jpn J Antibiot, 1987 May, 40(5), 1041 - 6
{Clinical studies on cefuzonam in obstetrics and gynecological infections}; Matsuda S et al.; To evaluate the usefulness and the safety of cefuzonam (CZON, L-105) against obstetric and gynecological infections, we conducted clinical trials and obtained the results summarized as follows . 1 . Thirteen patients with obstetric and gynecological infections (intrauterine infection 2 cases, adnexitis 7, 1 case each of abscess of adnexa uteri, abscess of the vaginal wall, pyelonephritis, and mammitis) were treated with CZON . The CZON was administered by intravenous injection or intravenous drip infusion, 1 g twice daily for 4 to 7 days (8 g to 14 g in total) . The clinical effect was good in 12 and poor in 1 . 2 . Four cases, on which other antimicrobial agents were ineffective, responded well to CZON . 3 . The CZON displayed excellent effects on anaerobic bacteria (Peptostreptococcus anaerobius, Bacteroides fragilis) and Gram negative rods (Escherichia coli, Klebsiella pneumoniae) . 4 . No side effects or laboratory abnormalities were observed . 5 . From the above results, CZON appeared to be an effective and useful drug for obstetric and gynecological infections.

J Bacteriol, 1987 May, 169(5), 2050 - 4
Klebsiella pneumoniae 1,3-propanediol:NAD+ oxidoreductase; Johnson EA et al.; Fermentative utilization of glycerol, a more reduced carbohydrate than aldoses and ketoses, requires the disposal of the two extra hydrogen atoms . This is accomplished by sacrificing an equal quantity of glycerol via an auxiliary pathway initiated by glycerol dehydratase . The product, 3-hydroxypropionaldehyde, is then reduced by 1,3-propanediol NAD+:oxidoreductase (1,3-propanediol dehydrogenase; EC 1.1.1.202), resulting in the regeneration of NAD+ from NADH . The pathway for the assimilation of glycerol is initiated by an NAD-linked dehydrogenase . In Klebsiella pneumoniae the two pathways are encoded by the dha regulon which is inducible only anaerobically . In this study 1,3-propanediol:NAD+ oxidoreductase was purified from cells grown anaerobically on glycerol . The enzyme was immunochemically distinct from the NAD-linked glycerol dehydrogenase and was an octamer or hexamer of a polypeptide of 45,000 +/- 3,000 daltons . When tested as a dehydrogenase, only 1,3-propanediol served as a substrate; no activity was detected with ethanol, 1-propanol, 1,2-propanediol, glycerol, or 1,4-butanediol . The enzyme was inhibited by chelators of divalent cations . An enzyme preparation inhibited by alpha,alpha'-dipyridyl was reactivated by the addition of Fe2+ or Mn2+ after removal of the chelator by gel filtration . As for glycerol dehydrogenase, 1,3-propanediol oxidoreductase is apparently inactivated by oxidation during aerobic metabolism, under which condition the enzyme becomes superfluous.

Mol Gen Genet, 1987 May, 207(2-3), 492 - 8
Frameshifts close to the Klebsiella pneumoniae nifH promoter prevent multicopy inhibition by hybrid nifH plasmids; Buck M et al.; Certain multicopy plasmids bearing promoter sequences of nitrogen fixation (nif) genes inhibit expression of chromosomal genes in Nif+ Klebsiella pneumoniae, hence leading to a Nif- phenotype . This 'multicopy inhibition' has been attributed to the titration of the nif-specific activator protein NifA by the plasmid-borne promoter sequences . We now report that multicopy inhibition by nifH translational fusions is sensitive to frameshifts close to the nifH promoter . Transcriptional nifH fusion plasmids in which translation terminated near the nifH promoter were transcriptionally active and showed multicopy inhibition; introduction of a transcription terminator after the nifH coding sequence in these plasmids prevented their multicopy inhibition . Therefore it seems likely that premature termination of transcription prevents multicopy inhibition by the nifH promoter.

Antimicrob Agents Chemother, 1987 May, 31(5), 758 - 62
Sub-MICs of cefuroxime and ciprofloxacin influence interaction of complement and immunoglobulins with Klebsiella pneumoniae; Williams P; Growth of encapsulated (K+) and nonencapsulated (K-) Klebsiella pneumoniae strains in media containing sub-MICs of either cefuroxime or ciprofloxacin resulted in cell elongation but had little effect on the outer membrane protein or lipopolysaccharide profiles . Exposure to serum complement increased the surface hydrophobicity of a K- strain but failed to interact or to increase the surface hydrophobicity of the K+ strains . However, after growth of the K+ strains in sub-MICs of the antibiotics, complement increased their surface hydrophobicity and complement C3 was detected bound to their surface . Antisera raised against a K-O- strain agglutinated the K+ strains grown in the presence but not in the absence of cefuroxime or ciprofloxacin . These findings suggest that the filamentous morphology induced by these antibiotics influences the distribution or amount of capsular polysaccharide such that cell envelope components previously masked by the capsule become accessible to complement and immunoglobulins.

J Bacteriol, 1987 May, 169(5), 2301 - 6
Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70; Katsuragi N et al.; We determined the entire nucleotide sequence of the Klebsiella aerogenes W70 pullulanase gene (pulA) contained on a 4.2-kilobase-pair fragment of plasmid pPB174 . The amino acid composition deduced from an open reading frame of 3,288 base pairs agreed closely with that determined for the intracellular pullalanase . The precursor enzyme consisted of 1,096 amino acid residues and contained a hydrophobic N-terminal signal peptide and the consensus sequence for the bacterial prelipoprotein signal peptide cleavage site.

J Gen Microbiol, 1987 May, 133 ( Pt 5), 1365 - 70
Identification of two chemical types of K21 capsular polysaccharide from klebsiellae; Allen PM et al.; Strains of Klebsiella pneumoniae of serotype K21 are frequently involved in outbreaks of nosocomial infections . The type strain of Klebsiella pneumoniae K21 (which we have renamed K21a) produces capsular polysaccharide that contains mannose, galactose and glucuronic acid in the ratio 2:2:1 . In contrast, all eight of the randomly chosen isolates of Klebsiella pneumoniae that were initially typed as K21 were shown by paper chromatography and NMR spectroscopy to produce a different capsular polysaccharide . We have designated this new polysaccharide K21b . The K21b capsular material appears to have a closely similar immunodominant side chain to K21a . However, K21b polysaccharide has two molecules of rhamnose in the polysaccharide backbone replacing the two molecules of mannose found in the K21a capsule . Our results suggest that the K21b Klebsiella serotype may be more frequently distributed than the classical K21a type.

Biochem J, 1987 Apr 15, 243(2), 561 - 7
The K1 beta-lactamase of Klebsiella pneumoniae; Joris B et al.; beta-Lactamase K1 was purified from Klebsiella pneumoniae SC10436 . It is very similar to the enzyme produced by Klebsiella aerogenes 1082E and described by Emanuel, Gagnon & Waley {Biochem . J . (1986) 234, 343-347} . An active-site peptide was isolated after labelling of the enzyme with tritiated beta-iodopenicillanate . A cysteine residue was found just before the active-site serine residue . This result could explain the properties of the enzyme after modification by thiol-blocking reagents . The sequence of the active-site peptide clearly established the enzyme as a class A beta-lactamase.

Biochemistry, 1987 Apr 7, 26(7), 1795 - 800
Characterization of the metal clusters in the nitrogenase molybdenum-iron and vanadium-iron proteins of Azotobacter vinelandii using magnetic circular dichroism spectroscopy; Morningstar JE et al.; Low-temperature magnetic circular dichroism (MCD) spectroscopy has been used to investigate the metal clusters in the conventional nitrogenase MoFe protein and alternative VFe protein from Azotobacter vinelandii . In the dithionite-reduced state, the MCD spectrum of the MoFe protein is extremely similar to that previously observed for the S = 3/2 spin state of the M clusters in the MoFe protein of Klebsiella pneumoniae . A paramagnetic cluster with an S = 3/2 ground state is also responsible for the temperature-dependent MCD transitions of dithionite-reduced VFe protein . However, the electronic and magnetic properties of this cluster are quite distinct from those of M centers in conventional nitrogenase . When these proteins are oxidized with thionine, the MoFe protein exhibits MCD spectra and magnetization characteristics identical with those observed for the P clusters in K . pneumoniae, while those of the VFe protein are only similar . However, the paramagnetism in the thionine-oxidized VFe protein, like the conventional enzyme, probably arises from an S = 5/2 spin system with near-axial symmetry and a negative zero-field splitting . Novel clusters with electronic, magnetic, and redox properties similar to those of conventional P clusters are, therefore, present in the VFe protein.

Eur J Biochem, 1987 Apr 1, 164(1), 237 - 41
Structural investigation of Klebsiella serotype K10 capsular polysaccharide; Chakrabarti A et al.; The primary structure of Klebsiella serotype K10 capsular polysaccharide has been investigated using mainly the techniques of methylation, partial hydrolysis, and 1H and 13C NMR spectroscopy . The polysaccharide was found to consist of hexasaccharide repeating units having the following structure: (formula; see text)

Arch Otolaryngol Head Neck Surg, 1987 Apr, 113(4), 374 - 6
Treatment of scleroma with ceforanide; Cone LA et al.; Three patients with rhinoscleroma who presented with nasal and pharyngeal symptoms are described . Treatment with ceforanide, a new second-generation cephalosporin with high intrinsic activity against Klebsiella, and possessing a long half-life, was used as outpatient treatment . A total of 120 g of ceforanide was administered over a period of two months . All three patients showed signs of clinical improvement, but only two of three became bacteriologically sterile at the end of two months; one patient relapsed bacteriologically but not clinically, once the drug was discontinued . Second- and third-generation cephalosporins appear to have excellent activity against the causative pathogens of rhinoscleroma . Because of the impracticality of administering these agents parenterally over prolonged periods of time, there is a need for the development of an oral cephalosporin with similar intrinsic activity and beta-lactamase stability . Perhaps the novel beta-lactam antibiotics such as the penems and monobactams, some of which can be administered orally, will answer that need.

Lab Anim Sci, 1987 Apr, 37(2), 159 - 66
The role of Klebsiella oxytoca in utero-ovarian infection of B6C3F1 mice; Davis JK et al.; A disease consisting of suppurative endometritis, salpingitis, perioophoritis and/or peritonitis has been an important problem in aging B6C3F1 mice on some chronic chemical carcinogenicity studies . Klebsiella oxytoca was identified as the most likely causative agent based on cultural isolations from lesions . A study was done to determine prevalence of K . oxytoca in the "normal" flora of mice from different breeding facilities . In a survey of 684 retired female breeder mice from 10 National Institutes of Environmental Health Sciences (NIEHS) and National Cancer Institute (NCI) production facilities, K . oxytoca was isolated from only 1% of nasopharynxes, vaginas and ceca in mice from 7 of 10 facilities . Epizootiology of the natural infection was investigated using the capsular and biochemical typing methods on 97 isolates of K . oxytoca from mice of 11 NIEHS and NCI production facilities and sentinel mice from three National Toxicology Program testing facilities . A few capsular types were associated with either lesions, nonlesion isolation sites, or certain facilities but the capsular typing method was not reproducible . No associations were found for any biotypes . A K . oxytoca isolate (capsular type 20, biotype A) from a typical case of perioophoritis was used in attempts to reproduce the natural disease in Klebsiella-free B6C3F1 female mice . Mice were inoculated at 6 months of age by the intravaginal, intrauterine or intraperitoneal route with one of four doses of K . oxytoca and killed at 4, 7 or 10 months post-infection . Some mice given high doses (10(6) or 10(8) colony forming units) of K . +oxytoca died of septicemia and a few developed mild inflammatory lesions in the uterus.(ABSTRACT TRUNCATED AT 250 WORDS)

J Bacteriol, 1987 Apr, 169(4), 1547 - 53
Products of the iron-molybdenum cofactor-specific biosynthetic genes, nifE and nifN, are structurally homologous to the products of the nitrogenase molybdenum-iron protein genes, nifD and nifK; Brigle KE et al.; The genes from Azotobacter vinelandii, which are homologous to the iron-molybdenum cofactor biosynthetic genes, nifE and nifN, from Klebsiella pneumoniae, have been cloned and sequenced . These genes comprise a single transcription unit and are located immediately downstream from the nitrogenase structural gene cluster (nifHDK) . DNA sequence analysis has revealed that the products of the nifE and nifN genes contain considerable homology when compared with the nifD (MoFe protein alpha subunit) and the nifK (MoFe protein beta subunit) gene products, respectively . These striking sequence homologies indicate a structural and functional relationship between a proposed nifEN product complex and the nitrogenase MoFe protein as well as imply an ancestral relationship between these gene clusters . The isolation and characterization of strains which contain deletions within the nifEN gene cluster demonstrate a role for these products in iron-molybdenum cofactor biosynthesis in A . vinelandii.

Lab Anim Sci, 1987 Apr, 37(2), 153 - 8
Utero-ovarian infection in aged B6C3F1 mice; Rao GN et al.; An unusually high number of ovarian masses and cysts with purulent material were observed in the B6C3F1 mice on 2 year chemical carcinogenicity studies sponsored by the National Cancer Institute-National Toxicology Program . To determine possible etiology, some of these lesions were cultured for bacteria and a majority yielded Klebsiella sp . Necropsy records of 14,029 female mice in 91 chronic studies necropsied from 1979 to 1983 at six toxicology testing laboratories were reviewed to determine the incidence of lesions and distribution of this disease . Animals for these studies were obtained from barrier production colonies of six suppliers . The incidence of this lesion was low in animals less than 14 months of age, increased with age and reached a peak in 24-26 month old mice . Most animals having this lesion either died or were sacrificed in moribund condition, indicating that this is a life shortening disease of aged B6C3F1 mice . The incidence of lesions ranged from less than 1% to 70% in different chronic studies . There was a marked difference in the incidence in mice from different suppliers and the incidence rate was 2.6 to 15% depending on the source of the animals . The incidence of this lesion in some testing laboratories was several-fold higher than in others and ranged from 0.9 to 20% . The proportion of mice with this lesion was low in some laboratories irrespective of the source of the animals, whereas in other laboratories the incidence was several-fold higher with animals from some, but not all suppliers, indicating testing laboratory-supplier interaction.(ABSTRACT TRUNCATED AT 250 WORDS)

J Antimicrob Chemother, 1987 Apr, 19(4), 487 - 91
Efficacy of different dosing schedules of tobramycin for treating a murine Klebsiella pneumoniae bronchopneumonia; Pechere M et al.; The efficacy of different dosing schedules of tobramycin for treating a murine Klebsiella bronchopneumonia was compared . Therapeutic efficacy depended upon dosing intervals . In mice treated for four days with a daily dose of 48 mg/kg, dosing intervals of 4 and 8 h allowed cure of 10/10 animals, whereas dosing intervals of 12 and 24 h yielded survival rates of only 6/10 (P less than 0.05) and 2/10 (P less than 0.01) respectively . Although observed in vitro, a post-antibiotic effect of tobramycin against K . pneumoniae was unlikely in vivo since residual concentrations of tobramycin were found in the lung tissue 12 h after dosing, when efficacy starts to decrease . Efficacy also depended to some extent upon the duration of therapy.

Immunol Lett, 1987 Apr, 14(4), 335 - 9
Dose-dependent efficacy of vaccination against K . pneumoniae in high and low antibody responder lines of mice; Mouton D et al.; Innate and acquired resistance to Klebsiella pneumoniae infection were investigated in high (HI) and low (LI) antibody responder lines of mice . The two lines were very susceptible to infection since even small inoculum doses of a virulent strain provoked a 100% mortality within a few days . However the mean survival time was significantly longer in LI than in HI . (HI X LI) F1 hybrids were more resistant than both parental lines . Immunization with heat killed K . pneumoniae was able to confer full protection on the mice in the two lines . However there was a large difference in the number of killed bacteria required to induce the protective effect in HI and in LI mice . The dose-effect relationship for protection correlated with that of antibody production . The protective role of antibodies was confirmed by the survival of HI and LI mice, when antibodies were passively given prior to lethal challenge . The results are in agreement with the fact already demonstrated, that the defect of LI mice in antibody responsiveness is a quantitative one . Therefore a satisfactory immune protection against K . pneumoniae could be obtained in LI mice by adapting the vaccination procedure.

Z Naturforsch {C}, 1987 Apr, 42(4), 353 - 9
Re-investigation of the protein structure of coenzyme B12-dependent diol dehydrase; Tanizawa K et al.; We have purified diol dehydrase, an adenosylcobalamin-dependent enzyme, from Klebsiella pneumoniae by two different procedures to re-investigate its protein structure; one including its extraction with detergent from the membrane fraction, and the other consisting of only chromatographic separations of the soluble fraction . The enzyme preparations obtained by these two methods were different in the subunit structure, but both are identical in molecular weight, and in-enzymological and immunochemical properties . In addition, the enzyme preparation obtained from the membrane fraction dissociated reversibly into two dissimilar protein components (F and S) in the absence of substrate, as did the preparation from the soluble fraction . Although the subunit multiplicity of component S might be partly due to proteolytic cleavage during the enzyme purification as revealed by limited digestion with trypsin, component F is not a product of proteolytic cleavage of component S, but a primordial and essential constituent of the enzyme.

Nucleic Acids Res, 1987 Mar 25, 15(6), 2757 - 70
The complete nucleotide sequence of the glnALG operon of Escherichia coli K12; Miranda-Rios J et al.; The nucleotide sequence of the E . coli glnALG operon has been determined . The glnL (ntrB) and glnG (ntrC) genes present a high homology, at the nucleotide and aminoacid levels, with the corresponding genes of Klebsiella pneumoniae . The predicted aminoacid sequence for glutamine synthetase allowed us to locate some of the enzyme domains . The structure of this operon is discussed.

Carbohydr Res, 1987 Mar 15, 161(1), 91 - 6
Studies of the primary structure of the capsular polysaccharide from Klebsiella serotype K15; Nath K et al.; The capsular polysaccharide of Klebsiella serotype K15 has been investigated mainly by methylation analysis, characterisation of the oligosaccharides obtained by partial acid hydrolysis, periodate oxidation, enzymic degradation, and 1H- and 13C-n.m.r . spectroscopy, and shown to have the hexasaccharide repeating-unit 1 . The glycan does not contain any pyruvic acetal or O-acetyl substituents . (formula; see text)

Arthritis Rheum, 1987 Mar, 30(3), 300 - 5
Failure of Klebsiella pneumoniae antibodies to cross-react with peripheral blood mononuclear cells from patients with ankylosing spondylitis; Cameron FH et al.; Cross-reactivity between antibodies to 2 strains of Klebsiella pneumoniae (K43 and F77) and the peripheral blood lymphocytes of patients with ankylosing spondylitis (AS) was examined in 3 separate antibody binding and cytotoxicity assays . Using K pneumoniae antisera in a chromium release cytotoxicity assay, we found no difference in the reactions of cells from AS patients and those from control subjects . This result contrasts with the results of previous studies . Similarly, using an enzyme-linked immunosorbent assay, we detected no significant increase in antibody binding to peripheral blood mononuclear cells (PBMC) in HLA-B27 positive patients with AS . Low levels of antibody binding were detected by a fluoresceinated antibody binding assay; however, normal rabbit serum, which was used as a control, was shown to have a binding affinity for PBMC that was significantly greater than that of specific K pneumoniae antisera . The results of our present study do not support the concept of a specific cross-reactivity between antibodies to K pneumoniae and the PBMC of patients with AS who are HLA-B27 positive.

J Med Chem, 1987 Mar, 30(3), 514 - 22
Synthesis and 3'-substituent effects of some 7 alpha-methoxy-1-oxacephems on antibacterial activity and alkaline hydrolysis rates; Narisada M et al.; Relationships between intrinsic antibacterial activity and beta-lactam chemical reactivity of 7 beta-(phenylacetamido)-7 alpha-methoxy-1-oxacephems with various 3'-substituents were studied in order to clarify the effect of the 3'-substituent on the antibacterial activity . The chemical reactivity of the beta-lactam ring estimated by pseudo-first-order rate constants log kobsdNMR of alkaline hydrolysis at pD 10.4 and 35.0 degrees C correlates well linearly with 13C NMR chemical shift differences (delta delta(4-3}, infrared stretching frequencies of the beta-lactam carbonyl (vC = O), and sigma I values . Values of log (1/CN), averaged for the MIC values for Escherichia coli, E . coli NIH JC-2, E . coli EC-14, and Klebsiella pneumoniae SRC-1, were taken as an estimation of the intrinsic antibacterial activity . The log (1/CN) values of the compounds without good leaving groups correlated fairly well with log kobsdNMR values . The comparatively high antibacterial activity of compounds with good leaving groups may be attributable to the different course of decomposition of these compounds.

Proc Natl Acad Sci U S A, 1987 Mar, 84(5), 1142 - 6
Promoter mapping and cell cycle regulation of flagellin gene transcription in Caulobacter crescentus; Minnich SA et al.; Caulobacter crescentus contains a 25- and a 27-kDa flagellin, which are assembled into the flagellar filament, and a 29-kDa flagellin, which is related in sequence but is of unknown function . We have used DNA sequence analysis and nuclease S1 assays to map the in vivo transcription start sites of the three flagellin genes and to study their regulation . These experiments lead to several conclusions . First, copies of the 29-, 25-, and 27-kDa flagellin genes are organized in a tandem array in the flaEY gene cluster of C . crescentus . Second, flagellin genes are under transcriptional control and each gene is expressed with a characteristic periodicity in the cell cycle . Third, flagellin gene promoters contain conserved nucleotide sequence elements at -13, -24, and -100 that are homologous to the fla genes in the hook gene cluster . The -13 and -24 sequences conform to a fla gene promoter consensus sequence (C/TTGGCC/GC-N5-TTGC) that is similar in sequence to the -12, -24 consensus sequence of the Klebsiella pneumonia nif gene promoters . Fourth, the sequence element at approximately -100 in the 25- and the 27-kDa flagellin genes is homologous to a 19-base-pair sequence {designated previously as II-1; see Chen, L.-S., Mullin, D . M . & Newton, A . (1986) Proc . Natl . Acad . Sci . USA 83, 2860-2864}at -101 in the promoter of transcription unit II of the hook gene cluster; the two flagellin genes, like the fla genes examined in the hook gene cluster that contain the -100 element, are under positive control by transcription unit III of the hook gene cluster . This result supports a model in which the timing of fla gene transcription in the C . crescentus cell cycle is determined in part by a cascade of trans-acting regulatory gene products.

Acta Pathol Jpn, 1987 Mar, 37(3), 475 - 86
Alveolar destruction in experimental Klebsiella pneumonia; Irifune K; Sequential histological changes of the lungs were studied in experimental Klebsiella pneumonia, using untreated control mice, cyclophosphamide-treated mice, and carrageenan-treated mice . Cyclophosphamide was used to deplete polymorphonuclear leukocytes and monocytes, and carrageenan was used to deplete mononuclear phagocytes selectively . At 72 hours, varying degree of alveolar necrosis could be seen in untreated control mice . However, the lung lesions of cyclophosphamide- or carrageenan-treated mice were significantly different from those of the control mice . The lung lesions of cyclophosphamide-treated mice indicated that destruction of the alveolar septa was not induced by K . pneumoniae itself but by inflammatory cells, because the alveolar walls were preserved very well in spite of considerable bacterial multiplication in alveolar lumina until infiltration of inflammatory cells occurred . The lung lesions of carrageenan-treated mice showed that alveolar spaces were packed with polymorphonuclear leukocytes, but the alveolar walls were preserved very well as far as the authors could tell after examining the lung lesions by silver impregnation staining . These results suggest that macrophages rather than polymorphonuclear leukocytes and organisms play an important role in alveolar injury in experimental Klebsiella pneumonia.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Mar, 263(4), 585 - 93
Haemagglutinins and adherence properties to HeLa and intestine 407 cells of Klebsiella pneumoniae and Klebsiella oxytoca isolates; Podschun R et al.; The occurrence of haemagglutination (HA) and adherence properties were examined in 50 strains of K . pneumoniae and 17 K . oxytoca strains isolated from humans . All isolates except three exhibited HA activity . Mannose-sensitive haemagglutinins (MSHA) were expressed by the majority of K . pneumoniae strains, but only by one K . oxytoca isolate . Mannose-resistant haemagglutination (MRHA) to human or guinea pig erythrocytes could not be detected; haemagglutinins of the MR/K type were found in both species with similar frequencies . Adhesive properties could be demonstrated in K . pneumoniae as often as in K . oxytoca: About half of the strains adhered to two human cell lines: HeLa and Intestine 407 . The incidence of HA activity was similar in adhering and nonadhering strains . A correlation between MSHA, MR/K-HA and adherence to tissue-cultured cells could not be detected.

J Dairy Sci, 1987 Mar, 70(3), 696 - 704
Natural protective factors in bovine mammary secretions following different methods of milk cessation; Bushe T et al.; Bovine mammary secretions were obtained during late lactation and early involution to determine if different methods of milk cessation influenced milk yield, composition, and in vitro growth of coliform mastitis pathogens . Cows (n = 8/group) producing about 13 kg of milk prior to experimentation were dried off by abrupt or intermittent milk cessation . An additional group was dried off by intermittent milk cessation and fed only hay during the last week of lactation . Cows milked intermittently produced significantly less milk during the last week of lactation than cows dried off by abrupt milk cessation . Mammary secretions from cows milked intermittently and fed only hay contained higher concentrations of somatic cells, lactoferrin, immunoglobulin G, and bovine serum albumin, a lower citrate:lactoferrin molar ratio, and were more inhibitory to in vitro growth of Escherichia coli and Klebsiella pneumoniae throughout most of the experimental period than mammary secretions from cows dried off by intermittent or abrupt milk cessation . Few differences in mammary secretion composition or in vitro growth of mastitis pathogens were observed between cows dried off by intermittent or abrupt milk cessation . Data suggest that growth of mastitis pathogens in mammary secretions may be related to natural protective factors, which can be manipulated by different methods of milk cessation.

J Bacteriol, 1987 Mar, 169(3), 955 - 60
Cloning and expression in Escherichia coli of a Klebsiella ozaenae plasmid-borne gene encoding a nitrilase specific for the herbicide bromoxynil; Stalker DM et al.; An enzyme (nitrilase) that converts the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) to its metabolite 3,5-dibromo-4-hydroxybenzoic acid was shown to be plasmid encoded in the natural soil isolate Klebsiella ozaenae . The bromoxynil-specific nitrilase was expressed in Escherichia coli by direct transfer and stable maintenance in E . coli of a naturally occurring 82-kilobase K . ozaenae plasmid . Irreversible loss of the ability to metabolize bromoxynil both in E . coli and K . ozaenae was associated with the conversion of the 82-kilobase plasmid to a 68-kilobase species . In E . coli this conversion was the result of a host recA+-dependent recombinational event . A gene, designated bxn, encoding the bromoxynil-specific nitrilase was constitutively expressed in K . ozaenae and E . coli and subcloned on a 2.6-kilobase PstI DNA segment . The polarity and the location of the gene were determined by assaying hybrid constructs of the bromoxynil-specific nitrilase gene fused with the heterologous lac promoter.

J Hosp Infect, 1987 Mar, 9(2), 191 - 3
An outbreak of serious Klebsiella infections related to food blenders; Kiddy K et al.; An investigation, including environmental sampling, was undertaken after four leukaemic patients on the same hospital ward developed serious infections with Klebsiella aerogenes, capsular type K14 . The source of this organism, common to all four patients, was found to be a food blender used for preparing milk-based drinks on the ward.

Anal Biochem, 1987 Feb 1, 160(2), 480 - 2
Detection of pullulanase in polyacrylamide gels using pullulan-reactive red agar plates; Yang SS et al.; After electrophoresis, active pullulanase bands in acrylamide gels have been detected by overlaying and then incubating the gel on a replica gel containing 2.5% pullulan-reactive red conjugate and 2.1% agar . The enzyme activity is revealed as a clear band against a red background on the replica gel . The sensitivity of the replica plate is such that 0.0012 unit of Klebsiella aerogenes pullulanase can be detected easily . This procedure is effective in enzyme screening to distinguish pullulanase from other carbohydrases.

Sci Total Environ, 1987 Feb, 60, 105 - 19
Metal ion binding by biological surfaces: voltammetric assessment in the presence of bacteria; Goncalves ML et al.; Voltammetric techniques (differential pulse polarography (DPP) and differential pulse anodic stripping voltammetry (DPASV)) were evaluated for their capability to distinguish, without prior separation of the solid phase (e.g . filtration, centrifugation), between dissolved and particulate concentrations of Zn(II), Pb(II) and Cu(II), and to measure the extent of binding of these metals to the surface of a bacterium (Klebsiella pneumonia, formaldehyde treated) . From titration curves of bacterial cell suspensions with metals the specific adsorption of metals was determined and quantified in terms of average surface complex formation constants and differential equilibrium functions . The following stability sequence for surface complexes was found: Cu2+ greater than Pb2+ greater than Zn2+ much greater than Ca2+ . Simultaneous analytical determination permitted the measurement of both the binding of Cu(II) to the cell surface, and the binding to the solute exudate ligands . The affinity of the metal ions for the functional groups of the cell surface is strongly pH-dependent, and, at a given pH, decreases with increasing metal loading of the bacterial surfaces . This indicates that metal ions bind first to the highest affinity surface ligands and subsequently to those of lesser activity . Copper(II) appears to form stronger surface complexes with the high affinity ligands of the bacterial surface than with the functional groups of hydrous oxides.

J Bacteriol, 1987 Feb, 169(2), 849 - 55
Part of respiratory nitrate reductase of Klebsiella aerogenes is intimately associated with the peptidoglycan; Abraham PR et al.; Lysozyme digestion and sonication of sodium dodecyl sulfate (SDS)-purified Klebsiella aerogenes murein sacculi resulted in the quantitative release of both subunits of nitrate reductase, as well as a number of other cytoplasmic membrane polypeptides (5.2%, by weight, of the total membrane proteins) . Similar results were obtained after lysozyme digestion of SDS-prepared peptidoglycan fragments, which excluded the phenomenon of simple trapping of the polypeptides by the surrounding peptidoglycan matrix . About 28% of membrane-bound nitrate reductase appears to be tightly associated with the peptidoglycan . Additional evidence for this association was demonstrated by positive immunogold labeling of SDS-murein sacculi and thin sections of plasmolyzed bacteria . Qualitative amino acid analysis of trypsin-treated sacculi, a tryptic product of holo-nitrate reductase, and amino- and carboxypeptidase digests of both nitrate reductase subunits indicated the possible existence of a terminal anchoring peptide containing the following amino acids: (Gly)n, Trp, Ser, Pro, Ile, Leu, Phe, Cys, Tyr, Asp, and Lys.

J Gen Microbiol, 1987 Feb, 133 ( Pt 2), 445 - 51
Establishment of the steady state in glucose-limited chemostat cultures of Klebsiella pneumoniae; Rutgers M et al.; To investigate the relationship between growth rate and concentration of the nutrient that limits growth, 'Klebsiella aerogenes' NCTC 418 (K . pneumoniae) was grown in a glucose-limited chemostat . The actual time required to establish a steady-state glucose concentration exceeded that expected theoretically . Apparently, there is a long-term adaptation of the cells to nutrient limitation . As yet, it is not clear whether this has a phenotypic or genetic origin . In the final steady state, the dependence of the growth rate on glucose concentration could be mathematically described equally well by a hyperbolic and by a logarithmic function.

J Gen Microbiol, 1987 Feb, 133 ( Pt 2), 331 - 40
Isolation from Klebsiella and characterization of two rcs genes that activate colanic acid capsular biosynthesis in Escherichia coli; Allen P et al.; Two genes, designated rcsA (regulation of capsule synthesis) and rcsB, that had been cloned from the chromosome of Klebsiella aerogenes (K . pneumoniae) capsular serotype K21 were capable of activating expression of colanic acid capsular polysaccharide in Escherichia coli K12 . The Klebsiella rcsA gene encoded a polypeptide of 23 kDa that was required for the induction of a mucoid phenotype at less than or equal to 30 degrees C but not at greater than or equal to 37 C . The Klebsiella rcsB locus encoded no apparent polypeptides and was not capable by itself of causing the overproduction of colanic acid . However, when present in the same cell with rcsA, either in cis or in trans, rcsB caused expression of mucoidy in E . coli at all growth temperatures . These findings are best explained if the Klebsiella rcsA gene product acts as a positive regulator of colanic acid biosynthesis in E . coli and that activity of this protein is in turn subject to regulation by Lon protease . The Klebsiella rcsB locus may exert its effect by preferentially binding a negative regulator of capsular biosynthesis, possibly Lon itself . DNA sequences homologous to the Klebsiella K21b rcsA and rcsB genes were found in the genomes of all other capsular serotypes of klebsiellae examined, including K2, K12, K36 and K43 . However, there was no homology between such genes and the chromosome of E . coli . The ability of these rcs genes to induce a mucoid phenotype explains the apparent conjugative transfer from klebsiellae to E . coli of the ability to produce K21 or other Klebsiella capsular polysaccharides that are structurally and antigenically related to colanic acid.

Eur J Biochem, 1987 Jan 15, 162(2), 439 - 43
Structural studies on the capsular polysaccharide of Klebsiella serotype K40; Nath RK et al.; The capsular polysaccharide of Klebsiella serotype K40 contained D-mannose, D-glucuronic acid, D-galactose, and L-rhamnose in the approximate molar ratios 1:1:1:2 . The primary structure of the capsular polysaccharide has been investigated mainly by methylation analysis, periodate oxidation, characterization of oligosaccharides, base degradation reaction, and 1H and 13CNMR spectroscopy . The polysaccharide does not contain any pyruvic acetal or O-acetyl substitution . It has a pentasaccharide repeating unit of the following primary structure: alpha-D-Manp 1----4 ----4)-beta-D-GlcpA-(1----2)-alpha-L-Rhap-(1----3)-beta-D-Ga lp-(1----2)-alpha- L-Rhap-(1----.

J Biol Chem, 1987 Jan 5, 262(1), 254 - 9
Solid-state NMR studies of Klebsiella pneumoniae grown under nitrogen-fixing conditions; Jacob GS et al.; The carbon and nitrogen metabolism of Klebsiella pneumoniae M5a1 has been characterized using 13C and 15N labeling with detection by cross-polarization magic-angle spinning solid-state NMR . Cells grown on ammonium typically require some 20 h to derepress fully for nitrogenase when transferred to medium devoid of any source of fixed nitrogen . We have established that during this period some cellular proteins are catabolized with the liberated nitrogen being used for the synthesis of purines needed for formation of ribosomal RNA . The 20-h derepression period can be shortened to 6 h by the introduction of fixed nitrogen in certain specific forms . Serine is the most successful agent we have examined for shortening the derepression period and glycine among the least successful . We attribute this difference to the advantage of serine over glycine in providing both specific and nonspecific carbon and nitrogen sources for complete purine synthesis . These determinations were made by tracing the metabolism of 13C- and 15N-labeled chemical bonds from the 2 amino acids during derepression.

Am J Dis Child, 1987 Jan, 141(1), 111 - 2
Bacterial growth in refrigerated human milk; Sosa R et al.; Unprocessed breast milk is currently being used as a desirable feeding alternative for premature infants . To assess some of the potential risks of this practice, we studied the bacterial growth in 41 samples of unprocessed human milk for a period of five days . No bacteria were cultured in eight samples of milk; the bacteria that were cultured in the remaining 33 samples were similar to those found on the skin and nipple of the breast . With the exception of three samples, two with Klebsiella and one with Pseudomonas, all identified bacteria have been reported as normal skin flora . The initial concentration of bacteria found in milk was low, with a mean of 10,000 colonies (range, 1000 to 140,000 colonies) . Bacterial colony counts progressively decreased throughout the five-day refrigeration period.

Ann Biol Clin (Paris), 1987, 45(4), 397 - 401
{New epidemiologic markers of Klebsiella oxytoca strains isolated from nosocomial infections}; Richard C et al.; In addition to capsular antigen typing and biotyping (sorbose, d-tartrate, dulcitol), the epidemiology of Klebsiella oxytoca can be improved by new biochemical characterization . The authors propose to determine tetrathionate-reductase activity, D-melezitose fermentation and brown pigment production onto a mineral gluconate-ferric citrate medium . In order to assess the value of these new markers, several hospital epidemiological surveys were performed in France and Spain (112 strains studied).

Chemotherapy, 1987, 33(1), 68 - 76
Influence of dosage interval on the therapeutic response to gentamicin in mice infected with Klebsiella pneumoniae; Queiroz ML et al.; Without treatment all mice died after receiving 10(3) Klebsiella pneumoniae by intraperitoneal injection . Nevertheless, it was possible to delay treatment for 12 h and still observe a therapeutic response from im gentamicin (5 mg/kg) . This gave initial serum concentrations comparable to clinical levels, which fell rapidly (t 1/2 = 15 min) to reach the limit of detection by 90 min . Courses were given of 3 or 6 doses spaced at different intervals . Irrespective of dosage interval there was a marked fall in bacteraemia with each of the first two doses . Between doses separated by 8 or even 12 h there was no evidence of bacterial multiplication but this was obvious by 24 h . Both the bacteraemic responses and the lengths of survival were best with the 12-hour dosage interval . These results are consistent with other reports of the persistence of antibiotic effects despite undetectable serum concentrations and the compatibility of a substantial dosage interval with a successful therapeutic outcome.

Infect Immun, 1987 Jan, 55(1), 44 - 8
Production of an extracellular toxic complex by various strains of Klebsiella pneumoniae; Straus DC; Six isolates of Klebsiella pneumoniae (two serotype 1 isolates and a capsular variant of one of these, and two serotype 2 isolates and a capsular variant of one of these) possessing various degrees of virulence in rats and mice were examined for their in vitro production of an extracellular toxic complex (ETC) . The ETC has been shown to be lethal for and produce extensive lung pathology in mice . This compound has been shown to be composed of capsular polysaccharide, lipopolysaccharide, and a small amount of protein . All six isolates produced the ETC . Immunization of experimental animals with sublethal doses of the ETC was protective against both homologous and heterologous strains, and this protection was due to antibody production . An examination of the various phases of growth of K . pneumoniae showed that there was extracellular release of the component parts of the ETC occurring during all phases of growth . The presence of the ETC in the supernatant fluids was due to actual release of this material as opposed to cell lysis . Antibodies to the lipopolysaccharide portion (which has been shown to possess the observed toxicity) of the ETC were protective against the homologous bacterium.

Microbiol Immunol, 1987, 31(10), 1025 - 32
Characterization of autoantigens relevant to autoimmune ophthalmitis and thyroiditis in mice immunized with the syngeneic tissue extracts and Klebsiella O3 lipopolysaccharide; Yokochi T et al.; The histological localization and biochemical properties of the autoantigens relevant to experimental autoimmune ophthalmitis and thyroiditis were studied using sera from mice hyperimmunized with the corresponding tissue extract of syngeneic mice and Klebsiella O3 lipopolysaccharide (KO3 LPS) as a potent adjuvant . Specific antigens were detected in the lens of the eyeball by immunofluorescence test with sera from mice in which ophthalmitis had been induced and the antigens were lenticular proteins with molecular weights (MW) of 15,000 (15K) to 25K, and 45K . The lenticular proteins with MW of 15K to 25K correspond to the subunits of crystalline . These findings clearly demonstrated that our experimental model for autoimmune ophthalmitis was classified as the lens-induced uveitis . The colloids of the thyroid follicles and the follicular cells were markedly stained by sera from mice in which thyroiditis had been induced . One of the autoantigens detected in the thyroid gland was biochemically consistent with a thyroglobulin subunit . It was also shown that these autoantigens detected in the present study were organ-specific but not species-specific . The nature of autoantigens in the eye and the thyroid gland is discussed.

Anticancer Res, 1987 Jan-Feb, 7(1), 17 - 22
Augmentation of spontaneous cytotoxicity of human lymphocytes by RU 41.740, a glucoprotein extract of Klebsiella pneumoniae; Viland H et al.; RU 41.740 (Biostim) is an extract of Klebsiella pneumoniae with immunomodulating properties . This substance was observed to augment spontaneous cytotoxicity of human blood lymphocytes provided that monocytes were present during Biostim treatment . Maximal augmentation was observed after 2 h of incubation . The F1 fraction, a glucoprotein which comprises approximately 20% of Biostim, and LPS from Klebsiella pneumoniae were also observed to augment spontaneous cytotoxicity of lymphocytes . Biostim treatment of K562 cells, which were used as target cells for killing, did not change their sensitivity to the lytic action of spontaneously cytotoxic cells.

Bull Soc Pathol Exot Filiales, 1987, 80(5), 751 - 5
{Bactericidal activity of disinfectants against certain bacterial strains . Klebsiella pneumonia}; Economou-Stamatelopoulou C et al.; Twenty disinfectants were tested against a strain of K . pneumoniae . A suspension test with incremental addition of microorganisms was used . The influence of interfering substances (proteins and hard water) on the activity of disinfectants was also determined . Iodine compounds were influenced by proteins in distilled and hard water . The presence of mineral ions inactivated the polyvinylpyrrolidone-iodine 10% and chloroxylenol preparations against K . pneumoniae . The activity of some iodine products disappeared at the second and third incremental addition of microorganisms; the chloroxylenol and the phenolic compounds were inactivated by proteins at the third incremental addition . The sensibility of K . pneumoniae to the other disinfectants was not influenced by proteins, ions and incremental bacterial additions.

Immunopharmacol Immunotoxicol, 1987, 9(2-3), 235 - 41
Influence of heavy metals on the resistance of mice toward infection; Laschi-Loquerie A et al.; Previous studies have shown that heavy metals may exert marked immunomodulatory effects, at least in rodents, despite some discrepancies . However, the mechanism of their influence on the immune system is still unclear . As host resistance assays against experimental infections are generally considered as the most relevant criteria when predicting the immunotoxicity of drugs and chemicals, the effects of lead acetate, nickel chloride and sodium selenite on the resistance toward experimental Klebsiella pneumoniae infection was investigated in mice, with particular emphasis on the interference of the time of toxic exposure with the infectious challenge . Interestingly, one single intraperitoneal dose of 24 mg/kg lead or 4 mg/kg nickel enhanced the resistance of mice against Klebsiella pneumoniae when administered 24 hours before the infectious challenge, whereas host resistance proved to be impaired when the same dose was injected 5 hours after the infectious challenge . A 3-day pretreatment with 8 or 12 mg/kg lead also enhanced the resistance of mice but decreased it with 0.5 or 1 mg/kg nickel . In all cases, sodium selenite increased the resistance of mice toward infection . As lead, nickel and selenium appear to exert complex and possibly opposite effects on antibody response and phagocytosis, it remains to establish which immunotoxic consequences if any, an acute or chronic exposure to these heavy metals is likely to have in man.

Braz J Med Biol Res, 1987, 20(1), 35 - 41
Mathematical model of Klebsiella pneumoniae resistance to amikacin and gentamicin; Nicolelis MA et al.; 1 . The resistance of Klebsiella pneumoniae to amikacin and gentamicin was studied by a mathematical model to predict the rate of sensitivity decrease . The results accurately matched experimental data, showing that the model is a reliable predicting tool . 2 . The observations were carried out over six years and included 2677 cultures that were positive for K . pneumoniae . At the beginning of the observation period, 85.7% of the cultures were sensitive to amikacin and 40.8% were sensitive to gentamicin . Sensitivity to amikacin showed a surprisingly rapid decrease; at the end of the experimental period, amikacin and gentamicin sensitivities were 33.3% and 27.8%, respectively . 3 . We conclude that patterns of resistance of other bacteria could be investigated using this method.

Ital J Surg Sci, 1987, 17(1), 27 - 30
Postoperative acute acalculous cholecystitis; Jensen C et al.; A 5-year experience with postoperative acute acalculous cholecystitis is reported . The series concerns 9 male patients ranging in age from 28 to 69 years, with a mean age of 46 years . All underwent major surgical procedures and complications appeared in the postoperative course . Clinically, 89% of patients developed sepsis and 66% jaundice . Klebsiella pneumoniae was the microorganism most frequently isolated from the blood, intraabdominal and wound fluid collections . It is emphasized that the diagnosis of this form should be clinical and it should be immediately suspected whenever intraabdominal signs develop . A review of the international literature on the subject is presented . The etiology and pathogenetic mechanisms of postoperative acute acalculous cholecystitis are discussed.

Int J Clin Pharmacol Res, 1987, 7(1), 45 - 9
Serum bactericidal activity against Klebsiella pneumoniae in volunteers receiving increasing doses of tobramycin with or without cefamandole; Pascual-Lopez A et al.; Ten volunteers received intravenously in a randomly allocated order and on separate days the following regimens: cefamandole (15 mg/kg); tobramycin (1.5 mg/kg); tobramycin (3 mg/kg); cefamandole (15 mg/kg) + tobramycin (1.5 mg/kg); cefamandole (15 mg/kg) + tobramycin (3 mg-kg) . Tobramycin serum levels were 3.7 +/- 0.7 micrograms/ml one hour after infusion of the 1.5-mg/kg and 7.8 +/- 1.6 micrograms/ml after the higher dose . Cefamandole serum levels were 26.0 +/- 5.2 micrograms/ml at the same time . Serum bactericidal activity and killing-rate studies were performed on a collection of 18 strains of Klebsiella pneumoniae . Against cefamandole-sensitive strains, all regimens were satisfactory (median serum bactericidal activity 1:16 to 1:64) . Tobramycin (3 mg/kg) and both combinations were slightly more active on cefamandole-resistant strains (1:16 to 1:32) than cefamandole (1:2) and tobramycin low dose (1:8) . Except for cefamandole alone, all regimens were equivalent in killing studies.

Folia Microbiol (Praha), 1987, 32(2), 137 - 41
Effect of various inducing agents on bacteriocin (klebocin) production by Klebsiella pneumoniae; Chhibber S et al.; The influence of various inducing agents on growth, synthesis and release of klebocin by Klebsiella pneumoniae was studied . A significant level of klebocin was detected only after induction . The highest level of klebocin was achieved with mitomycin C followed by rifampicin and polymyxin B . Chloramphenicol and UV irradiation did not show any effect on klebocin production . Maximum klebocin release occurred after 8 h of induction with all the agents . Concentration of mitomycin C did not show any significant effect on klebocin production.

Anticancer Res, 1987 Jan-Feb, 7(1), 23 - 8
Augmentation of spontaneous cytotoxicity of human lymphocytes by RU 41.740 is due to monocyte-derived factors distinct from interleukin 2, interferon alpha and gamma; Viland H et al.; In vitro exposure of human blood lymphoid cells to RU 41.740 (Biostim), a glucoprotein extract of Klebsiella pneumoniae, augments spontaneous cytotoxicity of the lymphocytes . It was observed that the augmentation could be blocked by inhibitors of RNA- and protein-synthesis, but not by an inhibitor of DNA-synthesis . The increased cytotoxicity of Biostim-treated cells could not be explained by an increased proportion of lymphocytes binding to the target cells (K562) . This indicates that Biostim acts by increasing the lytic activity of lymphocytes rather than by increasing the expression of target cell recognition structures . Further, it was shown that culture supernatants of Biostim-exposed monocytes, but not purified lymphocytes, contain factors that augment cytotoxicity of purified lymphocytes . This finding is in line with the previously reported monocyte-dependence of Biostim-induced augmentation of lymphocyte cytotoxicity . Interleukin-2 (IL-2) activity was not observed in supernatants of Biostim-exposed lymphoid cells and augmentation of lymphocyte cytotoxicity could not be inhibited by antibodies directed against interferon (IFN) alpha or gamma . This indicates that Biostim stimulates monocytes to liberate factors other than IL-2 or IFN which augment spontaneous cytotoxicity of lymphocytes.

Int J Immunopharmacol, 1987, 9(4), 417 - 24
Effects exerted by RU 41740 on oxidative metabolism and migration of rat polymorphonuclear leukocytes collected after induction of one acute non specific inflammatory reaction; Roch-Arveiller M et al.; The activity of RU 41740, a glycoprotein extract from Klebsiella pneumoniae, endowed with immuno-modulating properties, has been investigated on polymorphonuclear (PMN) leukocyte functions . This report deals with the effect of RU 41740 on oxidative metabolism (assessed by chemiluminescence, 02 consumption and O2- production) and on chemotaxis and random migration (using agarose and Boyden chamber techniques) . PMNs were collected from the rat pleural cavity after induction of one acute inflammatory reaction (pleurisy induced by injection of a suspension of calcium pyrophosphate crystals) . Experiments were performed in parallel after in vivo treatment or incubation in vitro . RU 41740 enhanced PMN oxidative metabolism and inhibited PMN chemotaxis while random migration was only affected using agarose assay at high concentration . This effect on PMN migration was observed with at least two attractants . These observations have been obtained either after incubation in vitro or administration in vivo . The minimal effective dose was 1 mg/kg in vivo and 0.1 microgram/ml in vitro . These data suggest that RU 41740 acts directly on PMN membrane receptors.

Infect Control, 1987 Jan, 8(1), 36 - 40
Beta-lactamase inhibitors: another approach to overcoming antimicrobial resistance; Parker RH et al.; Beta-lactamase inhibitors in combination with a beta-lactam antimicrobial agent provide another approach to the treatment of infections caused by many bacteria resistant to commonly used agents . Currently two fixed combinations are commercially available . One is the oral preparation of amoxicillin and potassium clavulanate (Augmentin) . The other is for intravenous therapy and combines ticarcillin and potassium clavulanate (Timentin) . In vitro studies confirm that the combination of a beta-lactamase inhibitor and a beta-lactam can increase the activity of the beta-lactam to such important pathogens as S aureus, Klebsiella, H influenzae, Bacteroides spp and other bacteria which produce plasmid-mediated beta-lactamase . Clinical efficacy studies have demonstrated potential usefulness of these combinations particularly for therapy of suspected or proven infections caused by mixed aerobic and anaerobic bacteria . The combination of amoxicillin or ticarcillin with clavulanic acid can be used to improve cost-effectiveness of antimicrobial therapy if they replace regimens that use multiple drugs or drugs with a relatively high incidence of adverse reactions.

J Bacteriol, 1987 Jan, 169(1), 260 - 71
Complementation of nitrogen-regulatory (ntr-like) mutations in Rhodobacter capsulatus by an Escherichia coli gene: cloning and sequencing of the gene and characterization of the gene product; Allibert P et al.; In vivo genetic engineering by R' plasmid formation was used to isolate an Escherichia coli gene that restored the Ntr+ phenotype to Ntr- mutants of the photosynthetic bacterium Rhodobacter capsulatus (formerly Rhodopseudomonas capsulata; J . F . Imhoff, H . G . Truper, and N . Pfenning, Int . J . Syst . Bacteriol . 34:340-343, 1984) . Nucleotide sequencing of the gene revealed no homology to the ntr genes of Klebsiella pneumoniae . Furthermore, hybridization experiments between the cloned gene and different F' plasmids indicated that the gene is located between 34 and 39 min on the E . coli genetic map and is therefore unlinked to the known ntr genes . The molecular weight of the gene product, deduced from the nucleotide sequence, was 30,563 . After the gene was cloned in an expression vector, the gene product was purified . It was shown to have a pI of 5.8 and to behave as a dimer during gel filtration and on sucrose density gradients . Antibodies raised against the purified protein revealed the presence of this protein in R . capsulatus strains containing the E . coli gene, but not in other strains . Moreover, elimination of the plasmid carrying the E . coli gene from complemented strains resulted in the loss of the Ntr+ phenotype . Complementation of the R . capsulatus mutations by the E . coli gene therefore occurs in trans and results from the synthesis of a functional gene product.

Arch Immunol Ther Exp (Warsz), 1987, 35(3), 283 - 7
Cell surface hydrophobicity of Klebsiella strains; Przondo-Hessek A et al.; On 14 encapsulated Klebsiella strains with different K-antigens and their non-encapsulated mutants the type of fimbriae present and the grade of hydrophobicity of their cell surface (expressed as SAT-value) were investigated . It could be demonstrated that clear correlations exist between the fimbriation of Klebsiella bacteria and their cell surface hydrophobicity . On the other hand, the presence of capsules and the type of K-antigen showed no influence on the degree of hydrophobicity.

Braz J Med Biol Res, 1987, 20(3-4), 321 - 30
Construction of a gene library from Azospirillum brasilense and characterization of a recombinant containing the nif structural genes; Schrank IS et al.; 1 . We have constructed a gene library, from Azospirillum brasilense using the vector EMBL4 . 2 . A recombinant containing the nif structural genes from A . brasilense was isolated and characterized . This recombinant contains a DNA insert of about 15 kilobases (kb) which gives rise to five fragments after cleavage with EcoRI . Only one of the DNA fragments (6.5 kb) hybridized to the nifHDK genes of Klebsiella pneumoniae . 3 . The organization of the nif genes in this DNA fragment was determined using different DNA segments containing the nifH, nifK or nifD genes of K . pneumoniae as probes.

Int J Immunopharmacol, 1987, 9(7), 775 - 81
Effects of RU 41740 aerosol treatment on mouse bronchoalveolar cells, and protection afforded against influenza virus infection; Michel F et al.; RU 41740, an immunomodulating compound extracted from Klebsiella pneumoniae, was previously shown to enhance mice resistance to bacterial and viral lung infections . To explore lung defense mechanisms, we studied the influence of RU 41740 aerosol treatment on the bronchoalveolar cell populations . Five successive daily RU 41740 aerosol treatments induced a large accumulation of leukocytes in the lungs 4h after the last treatment . Polymorphonuclear leukocytes predominated . The numbers of lymphocytes and monocytes rose significantly . A single RU 41740 aerosol treatment significantly raised the number of polymorphonuclears only . A luminol-dependent chemiluminescence assay was used to test the effect of RU 41740 on the opsonized zymosan induced response of alveolar macrophages . In vitro, addition of RU 41740 enhanced this chemiluminescence . After a single RU 41740 aerosol treatment of mice, the chemiluminescence of purified alveolar macrophages from these mice increased significantly . The protective effect of five daily RU 41740 aerosol treatments against influenza virus infection was believed to be due to the great intensity of the cellular response and the polymorphonuclear influx . The alveolar macrophage activation observed might also explain the enhanced resistance of mice to influenza virus infection.

Nat Immun Cell Growth Regul, 1987, 6(2), 65 - 76
Modulation of natural killer activity by muramyl peptides: relationship with adjuvant and anti-infectious properties; Le Garrec Y et al.; Natural killer (NK) activity of spleen cells was studied in DBA/2 mice, 24 and 72 h after intravenous injection of various muramyl peptides: muramyl dipeptide (MDP) and derivatives which are both adjuvant-active and able to increase resistance against Klebsiella pneumoniae; derivatives which are adjuvant-active but devoid of anti-infectious properties; derivatives which are anti-infectious but devoid of adjuvant activity, and derivatives which are devoid of both activities such as the stereoisomer MDP{D-Ala}1 . An early increase in NK activity was observed 24 h after injection of all nonadjuvant derivatives, whatever their effect on infection . A stimulation of natural cytotoxicity was always induced 72 h after injection of MDP and derivatives able to protect mice against Klebsiella pneumoniae infection . So, even if the reverse was not true, there seems to exist some correlation between the anti-infectious effect of muramyl peptides and the late increase in NK activity . The modulation of NK activity by muramyl peptides appeared to be independent of interferon production . Moreover, inhibition of the stimulatory effect by a cell cycle-specific drug, hydroxyurea, observed 72 h after MDP suggests a requirement for proliferation.

Carbohydr Res, 1986 Dec 1, 157, 27 - 51
Depolymerization of the capsular polysaccharide from Klebsiella K19 by the glycanase associated with particles of Klebsiella bacteriophage luminal diameter 19; Beurret M et al.; The site of cleavage of the capsular polysaccharide from Klebsiella K19 by the endoglycanase associated with particles of Klebsiella bacteriophage luminal diameter 19 was determined . The specific cleavage of the bond Rhap-(1----2)-Rhap provided a series of oligosaccharides having rhamnose at the reducing end . The enzyme is thus an alpha-rhamnosidase . Structural studies on the oligomers confirmed the sequence of the repeating unit of the polysaccharide from K19 . The 1H- and 13C-n.m.r . spectra of the homologous series of oligosaccharides corresponding to one, two, three, and four repeat-units exhibit important differences that denot variation of conformation with chain length . The bacteriophage acted on modified forms of K19 polysaccharide to provide a series of linear oligomers, and emphasized the essential role of the negative charge on the uronic acid in the action of the glycanase.

Carbohydr Res, 1986 Dec 1, 157, 13 - 25
Structural investigation of the capsular polysaccharide from Klebsiella K19 by chemical and N.M.R . analyses; Beurret M et al.; Sugar analysis of the capsular antigen K19 from Klebsiella and of the carboxyl-reduced derivative confirmed its classification into the chemotype containing rhamnose, galactose, glucose, and glucuronic acid residues . Partial acid hydrolysis and phage depolymerization of K19 provided respectively a modified, linear form of the polysaccharide and oligosaccharides of the repeating unit, these were used for the structural elucidation of the original polymer . Methylation analysis, Smith degradation, and 1H- and 13C-n.m.r . spectroscopy of the polysaccharide and derivatives permitted formulation of the following structure for K19: (formula; see text)

Compr Ther, 1986 Dec, 12(12), 38 - 42
Acute pyelonephritis in pregnancy; Gilstrap LG 3rd et al.; Acute pyelonephritis is one of the most common serious medical complications of pregnancy and may result in significant maternal morbidity . E . coli and Klebsiella are the two most common causative organisms . Pregnant women with acute pyelonephritis should be hospitalized and treated with parenteral antibiotics . Clinical response is usually dramatic, with the majority of patients being asymptomatic within 48 to 72 hours of therapy . Recurrent infection is common, and these women must be followed closely with frequent surveillance cultures . Detection and eradication of bacteriuria at the initial prenatal visit is important in preventing this complication.

Antimicrob Agents Chemother, 1986 Dec, 30(6), 964 - 5
Association between colistin resistance and broad-spectrum recipient deficiency in Klebsiella pneumoniae; Lamousin-White M et al.; A colistin-resistant mutant of Klebsiella pneumoniae served well as a donor but not as a recipient in conjugation . A nearly 1,000-fold difference between colistin-susceptible and colistin-resistant forms of this strain was observed by using donors of plasmids of four incompatibility groups . Recipient efficiency was not restored by filter matings.

Ann Gastroenterol Hepatol (Paris), 1986 Dec, 22(7), 395 - 6
{Post-jejunoscopy bacteremia caused by Klebsiella pneumoniae}; Jemni L et al.; We report a case of symptomatic Klebsiella pneumoniae bacteremia following a jejunoscopy . Occurrence of bacteremia in patients undergoing gastro-intestinal endoscopy is reviewed.

Environ Res, 1986 Dec, 41(2), 497 - 504
Impaired acquired resistance of mice to Klebsiella pneumoniae infection induced by acute NO2 exposure; Bouley G et al.; The natural resistance of nonimmunized C57Bl/6 mice to an intraperitoneal Klebsiella pneumoniae challenge was not significantly affected by prior continuous exposure to 20 ppm NO2 for 4 days . In contrast, the acquired resistance of mice immunized just before and infected just after NO2 exposure was seriously impaired . This could not be explained by the loss of appetite (about 30%) observed in NO2 treated mice, for neither the natural nor acquired resistance of control air exposed mice given approximately 70% ad libitum food and water were significantly modified.

J Bacteriol, 1986 Dec, 168(3), 1220 - 7
Comparison of the malA regions of Escherichia coli and Klebsiella pneumoniae; Bloch MA et al.; Using the mini-Mu-duction technique, we cloned the malA regions from Escherichia coli K-12 and Klebsiella pneumoniae . A comparison of the structures of the cloned DNAs indicated that the malT, malP, and malQ genes, encoding the transcriptional activator of the maltose regulon, maltodextrin phosphorylase, and amylomaltase, respectively, are similarly organized in both species; malP and malQ constitute an operon divergent from the malT gene . We sequenced 1,200 nucleotides encompassing the beginnings of the malT and malP genes, their promoters, and the intergenic region . The DNA sequences from the two species were very different; the levels of homology ranged from 28 to 80%, depending on the region . The sequences of the coding regions and of elements known to be important for the functions of these two promoters in E . coli were well conserved between the two bacteria, whereas the sequence of the malT-malP intergenic region had totally diverged.

Infect Immun, 1986 Dec, 54(3), 603 - 8
Correlation of the virulence of Klebsiella pneumoniae K1 and K2 with the presence of a plasmid encoding aerobactin; Nassif X et al.; Nine isolates of Klebsiella pneumoniae belonging to capsular serotypes K1 and K2 were assayed for virulence in mice . Virulent isolates (50% lethal dose of less than 10(3) microorganisms) and avirulent isolates (50% lethal dose of over 10(6) microorganisms) were selected . Supplementation of a defined minimal medium with transferrin markedly reduced the growth of avirulent strains but had no significant effect on the growth of virulent strains . All isolates produced enterochelin, but only production of aerobactin could be correlated with virulence . The genes encoding aerobactin and its receptor protein were located on a 180-kilobase plasmid . They were cloned into the mobilizable vector pSUP202 . Homology was demonstrated with the aerobactin operon of the Escherichia coli plasmid pColV-K30 . Transfer of the recombinant plasmid pKP4 into an avirulent recipient enhanced virulence by 100-fold . These experiments demonstrated that aerobactin is an essential factor of pathogenicity in K . pneumoniae.

Can J Microbiol, 1986 Nov, 32(11), 884 - 8
Molecular weight determination and partial characterization of Klebsiella pneumoniae hemolysins; Barberis LI et al.; Two thiol-activated Klebsiella pneumoniae hemolysins were purified from growth media by means of salt precipitation, gel filtration, ion-exchange chromatography, and polyacrylamide gel electrophoresis . The hemolysins peaks coincided with the protein and glycoprotein peaks as determined by chromatography and electrophoresis . The molecular weights, estimated by gel filtration, were 8400 and 19,000; by sodium dodecyl sulfate--polyacrylamide gel electrophoresis, the values were calculated as 15,500 and 27,000 . The electrophoretic bands were best detected by the periodic acid--Schiff method . Reduction of the disulfide linkages did not cause the originally larger molecule to break into 8400 and 19,000 hemolysins . However, trypsin treatment cleaved the 19,000 hemolysin into an active moiety, with an electrophoretic migration similar to the 8400 hemolysin . A naturally occurring proteolytic activity was investigated using pepstatin and antipain . When the trypsin inhibitor was added to the system, the hemolytic activity was detected only in the 19,000 hemolysin and the smaller hemolysin was absent.

J Antimicrob Chemother, 1986 Nov, 18(5), 599 - 608
Evaluation of ceftazidime in experimental Klebsiella pneumoniae pneumonia: comparison with other antibiotics and measurement of its penetration into respiratory tissues and secretions; McColm AA et al.; The activity of ceftazidime was examined in a murine model of Klebsiella pneumoniae pneumonia in which the antibiotic was administered subcutaneously 6 h after intranasal infection and then twice daily for the next three days (i.e . seven doses) . In a series of experiments using this test, the dose of ceftazidime giving 50% survival relative to controls (SD50) ranged from 1.0-9.0 mg/kg/dose while the dose required to reduce the log10 cfu/lung by 50% (CD50) ranged from 24-64 mg/kg/dose . Ceftazidime was considerably more effective than cefotiam, amoxycillin-clavulanic acid or kanamycin in the test . Pharmacokinetic studies with ceftazidime showed that no differences in respiratory tract penetration existed between uninfected mice and mice infected for 48 h with K . pneumoniae . The percentage penetration of ceftazidime from serum was 73% for pleural fluid, 44% for tracheal fluid, 27% for tracheal wall tissue and 17% for whole lung tissue after a subcutaneous injection of 100 mg/kg . At this dose, ceftazidime remained at supra-MIC concentrations for 2-3 h in all compartments examined.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Nov, 262(4), 512 - 21
{In vitro model for the study of the concentration kinetics of antibiotic combinations and their antibacterial activity}; Patsch R et al.; The concentration kinetics of combinations of antibiotics similar to those taking place after i.v . administration was simulated in an in vitro model taking into account the different half-life periods of the antibiotics . A hose pump was used for continuous supply of sterile nutrient to a liquid bacterial culture containing both antibiotics in a preselected concentration and for removal of fluid from the system at an identical flow rate . To account for the different half-life periods of the antibiotics, a solution containing the antibiotic having a longer half-life in a corresponding concentration was continuously added by pumping . Samples were taken at fixed intervals to determine bacterial counts and check the expected concentrations of active substance did not contain antibiotics . This model has been developed to enable a determination of the dynamic relationship between the course of concentrations of antibiotics and their antibacterial activity . To permit a statement on the synergistic, indifferent or antagonistic activity of the combination, in analogy to the FIC indices in the checkerboard technique, the relative reduction of the bacterial count by the combination as compared to the individual components at time t was calculated . The model has been demonstrated by the example of the combined action of azlocillin and gentamicin against a strain of Klebsiella pneumoniae . In a simple way, the model permits any increase in the number of combination partners having different half-life periods . The mathematical development of the model is explained in detail.

Hum Immunol, 1986 Nov, 17(3), 224 - 38
Biochemical studies on a factor isolated from Klebsiella K43-BTS1 that cross-reacts with cells from HLA-B27 positive patients with ankylosing spondylitis; Upfold LI et al.; A component of the cell walls of certain enteric bacteria has been identified that cross-reacts with an HLA-B27-associated cell-surface structure on lymphocytes and other cell types from patients with ankylosing spondylitis . This component, or "modifying factor," from one particular organism, Klebsiella K43-BTS1, has been studied in detail . A purification scheme has been developed using preparative electrofocusing and gel-permeation high performance liquid chromatography techniques and the purified material used in various characterization studies . A previous study demonstrated that the modifying factor has an approximate molecular weight of 30,000 and an isoelectric point of 5.4-5.5 . In this study two-dimensional gel electrophoresis experiments demonstrated that the modifying factor is associated with a single protein component of the cell wall of this organism . Pronase and papain destroyed the modifying factor activity whereas trypsin and alpha-chymotrypsin degraded the factor into smaller fragments without destroying its ability to modify B27+ AS- lymphocytes . Neuraminidase did not affect the modifying factor itself but did affect B27+ AS- lymphocytes such that they became unresponsive to modification . Sugar inhibition studies suggested that sugar groups are probably not involved in the function of the modifying factor . The availability of purified modifying factor should permit more detailed chemical analyses as well as functional studies to determine the significance of this molecule to the pathogenesis of ankylosing spondylitis.

Infect Immun, 1986 Nov, 54(2), 403 - 7
Immunization against fatal experimental Klebsiella pneumoniae pneumonia; Cryz SJ Jr et al.; The ability of serospecific anti-capsular polysaccharide (CPS) antibody to prevent fatal Klebsiella pneumoniae pneumonia was evaluated in a rat lung model . Rats were immunized intramuscularly with 100 micrograms of purified serotype 2 CPS and challenged intrabronchially 14 days later with a serotype 2 strain of K . pneumoniae . Vaccination engendered high levels of serum anti-CPS antibody which afforded significant protection (P less than 0.01) against fatal pneumonia . Immunization promoted clearance of the challenge bacteria from the lungs and prevented bacteremia . Histological examination of lung tissue from infected control animals showed pronounced inflammatory cellular infiltrate in the alveolar spaces, intra- and peribronchial inflammation, and tissue necrosis . In contrast, pathological changes noted in lungs from immunized animals were restricted to infrequent intra- and peribronchial involvement.

Infect Immun, 1986 Nov, 54(2), 365 - 70
Immunogenic properties of Klebsiella pneumoniae type 2 capsular polysaccharide; Robert A et al.; The immunoprotective activity of Klebsiella pneumoniae K2 cell surface preparations and purified capsular polysaccharide was tested in mice . The 50% protective dose (PD50), expressed as capsular polysaccharide content, was 2 ng for cell surface preparations and 50 ng for purified capsular polysaccharide . Both preparations lost their immunoprotective activity after alkali treatment . Immune sera were raised in rabbits immunized with cell surface preparations . The precipitating and hemagglutinating capacity of these antisera was tested against either purified capsular polysaccharide or alkali-treated capsular polysaccharide . No difference was observed between the reactivity of the antisera against each antigen . The protective activity of these sera was tested on mice in passive transfer experiments, before and after absorption with either purified capsular polysaccharide or alkali-treated capsular polysaccharide . The sera lost their protective activity after absorption with purified capsular polysaccharide and after absorption with alkali-treated capsular polysaccharide . These experiments show that the difference in immunoprotective activity of cell surface preparations, purified capsular polysaccharide, and alkali-treated capsular polysaccharide is not due to a difference in their antigenic determinants . Cell surface preparations and purified capsular polysaccharide were fractionated by gel filtration on Sepharose 4B and by ultracentrifugation on cesium chloride density gradients . Three forms of capsular polysaccharide have been characterized . (i) A form of capsular polysaccharide with a very high protective activity (PD50 = 2 ng) that copurified with protein and lipopolysaccharide and was characterized by a low coefficient of distribution (Kd = 0.20) and a low density (1.5 to 1.6 g/cm3) . (ii) A form of capsular polysaccharide with an intermediate protective activity (PD50 = 50 ng), contamined by less than 3% protein and 1% lipopolysaccharide, with a Kd of 0.35, and a density of 1.7 to 1.8 g/cm3 . (iii) A nonimmunoprotective capsular polysaccharide obtained after alkali treatment of either cell surface preparations or purified capsular polysaccharide . The Kd of these fractions varied from 0.20 to 0.90 and their density from 1.7 to 1.8 g/cm3.

Mol Gen Genet, 1986 Nov, 205(2), 253 - 9
Nucleotide sequence of the nifLA operon of Klebsiella oxytoca NG13 and characterization of the gene products; Kim YM et al.; The complete nucleotide sequence of the regulatory operon nifLA of a nitrogen fixer Klebsiella oxytoca NG13 was determined, and the transcriptional start point was assigned by S1 mapping . The nifL protein (a repressor) was coded by an open reading frame of 1,485 bases, corresponding to a protein of 495 amino acids with a calculated molecular weight of 55,242 . The open reading frame (1,572 bases) of the nifA protein (an activator), corresponding to a molecular weight of 58,649, was confirmed by in vitro transcription-translation experiments, using the wild type and artificially deleted nifA genes . The initiation codon (ATG) of nifA overlapped the termination codon(TGA) of nifL, sharing the two bases T and G . A conserved DNA contact point {Gln-(X)3-Ala-(X)3-Gly-(X)5-Val} common in many DNA binding proteins was found in the C-terminal region of the nifA sequence . The promoter sequences of nifLA, nifB and nifF in K . oxytoca coincided exactly with those of K . pneumoniae in the consensus regions at -12 and -26, although the overall homology in the promoter regions was 96% . Changes of four amino acids were found between the nifA coding sequences of K . oxytoca and K . pneumoniae.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Nov, 262(4), 522 - 30
Opsonin independent interaction of Klebsiella strains with human polymorphonuclear leukocytes; Przondo-Hessek A et al.; Nine encapsulated Klebsiella strains with different types of fimbriation and their nonencapsulated mutants were tested for their stimulatory potency for human polymorphonuclear leukocytes in the absence of opsonins . The luminol chemiluminescence assay was used for these experiments . It could be shown that the interaction between Klebsiella bacteria and human leukocytes is rather complex depending not only on the presence of capsules but also on the hydrophobicity of Klebsiella surface and on the type of fimbriation existing.

FEBS Lett, 1986 Oct 20, 207(1), 69 - 74
Close evolutionary relationship between the chromosomally encoded beta-lactamase gene of Klebsiella pneumoniae and the TEM beta-lactamase gene mediated by R plasmids; Arakawa Y et al.; Sixty-three percent homology of nucleotide sequence and 67% homology of deduced amino acid sequence were found between the chromosomally encoded beta-lactamase gene of Klebsiella pneumoniae and the TEM beta-lactamase of transposon Tn3 . Moreover, 22 out of 24 amino acid residues are identical around the predicted active site . It is therefore suggested that these two kinds of beta-lactamases share a common evolutionary origin . The 0.5 kb DNA fragment of the cloned gene hybridized specifically with the chromosomal DNA of all the K . pneumoniae strains tested which had been isolated in Japan, USA and Europe.

Eur J Biochem, 1986 Oct 15, 160(2), 371 - 7
The nifH gene product is required for the synthesis or stability of the iron-molybdenum cofactor of nitrogenase from Klebsiella pneumoniae; Filler WA et al.; The MoFe protein of nitrogenase from Klebsiella pneumoniae contains an iron-molybdenum cofactor, FeMoco, the synthesis or processing of which involves the products of at least five genes, nifQ, nifB, nifN, nifE and nifV . We have detected FeMoco activity in extracts of strains which synthesise neither of the MoFe protein subunits, indicating that FeMoco can be synthesised prior to combination with the MoFe protein polypeptides . Expression of the nifH gene (or a large part of it), was essential for FeMoco activity to be observed either in the presence or in the absence of the MoFe protein subunits . The nifH gene product was not involved in the control of the transcription of other nif gene products known to be involved in FeMoco synthesis or processing, nor was it essential for the stability of performed FeMoco before its combination with the MoFe protein polypeptides.

J Antimicrob Chemother, 1986 Oct, 18 Suppl C, 223 - 33
The epidemiology of antibiotic resistance in hospitals; Mayer KH; Changes in antibiotic susceptibility among different species are a function of the interaction of bacterial genomes (chromosomes, plasmids, transposons) with specific host and environmental factors . Computer-assisted surveillance of resistance patterns and plasmid fingerprinting with restriction endonucleases have facilitated resistance gene tracking . Certain nosocomial plasmids have persisted for many years within specific centres, others are widely distributed by species and location, and others may have a narrow host range or undergo rapid molecular evolution . Klebsiella pneumoniae is most often the index species for the dissemination of nosocomial plasmids, which are frequently multiresistant . Acute increases in antibiotic resistance may be due to antibiotic selection pressures, introduction of new, well-adapted species, over-crowding and other environmental changes . The most resistant organisms are found in units where patients are immunocompromised, are subject to invasive procedures or remain for long periods . Although large-scale studies have not shown uniform increases in the prevalence of resistant bacteria, acute outbreaks in specific niches have resulted in increased patient morbidity, necessitating rapid changes in antibiotic usage . Further molecular epidemiological studies are required in order to understand better how to anticipate the spread of antibiotic resistance.

Immunobiology, 1986 Oct, 173(1), 63 - 71
Augmentation of antibody responses of mice to inhaled protein antigens by simultaneously inhaled bacterial lipopolysaccharides; Mizoguchi K et al.; Serum antibody responses of mice to repeatedly inhaled protein antigens such as bovine serum albumin and ovalbumin, plus or minus bacterial lipopolysaccharide (LPS) in the form of an aerosol were studied . Results showed that the levels of responses to inhaled protein antigens varied, depending on the mouse strain-antigen combination and that LPS inhaled simultaneously with the antigens definitely augmented the responses which were not otherwise very high . LPS extracted from Klebsiella O3 (LPS-K) but not LPS from Escherichia coli O55 (LPS-E), which was inhaled at the time of initial inhalation of antigen, significantly intensified the priming for the secondary antibody response to the antigen subsequently inhaled . Both LPS-K and LPS-E, however, definitely acted to augment the response when they were inhaled repeatedly together with the antigen . Oral administration of antigen or antigen plus LPS-K did not induce any detectable antibody response in our experiment, ruling out the possibility that the antigen and LPS stimulated the immune system via alimentary canal rather than via lung . Tissue distribution of the radioactivity soon after inhalation of 131I-labeled antigen and decay speed of the radioactivity were not significantly changed by LPS-K inhaled simultaneously . This suggested that the augmentation of responses was not mediated by the action of LPS to modulate the air-blood barrier against the entry of antigen via lung . All the results prove for the first time that inhaled LPS displays a definite adjuvant action on antibody responses to inhaled antigens.

Biochem J, 1986 Oct 1, 239(1), 69 - 75
Electron transfer to nitrogenase . Characterization of flavodoxin from Azotobacter chroococcum and comparison of its redox potentials with those of flavodoxins from Azotobacter vinelandii and Klebsiella pneumoniae (nifF-gene product); Deistung J et al.; Flavodoxin in the hydroquinone state acts as an electron donor to nitrogenase in several nitrogen-fixing organisms . The mid-point potentials for the oxidized-semiquinone and semiquinone-hydroquinone couples of flavodoxins isolated from facultative anaerobe Klebsiella pneumoniae (nifF-gene product, KpFld) and the obligate aerobe Azotobacter chroococcum (AcFld) were determined as a function of pH . The mid-point potentials of the semiquinone-hydroquinone couples of KpFld and AcFld are essentially independent of pH over the range pH 7-9, being -422 mV and -522 mV (normal hydrogen electrode) at pH 7.5 respectively . The mid-point potentials of the quinone-semiquinone couples at pH 7.5 are -200 mV (KpFld) and -133 mV (AcFld) with delta Em/pH of -65 +/- 4 mV (KpFld) and -55 +/- 2 mV (AcFld) over the range pH 7.0-9.5 . This indicates that reduction of the quinone is coupled to protonation to yield a neutral semiquinone . The significance of these values with respect to electron transport to nitrogenase is discussed . The amino acid compositions, the N- and C-terminal amino acid sequences and the u.v.-visible spectra of KpFld and AcFld were determined and are compared with published data for flavodoxins isolated from Azotobacter vinelandii.

Arch Intern Med, 1986 Oct, 146(10), 1913 - 6
Klebsiella pneumoniae liver abscess associated with septic endophthalmitis; Liu YC et al.; Metastatic septic bacterial endophthalmitis is a rare, but devastating disease . We encountered seven cases of pyogenic liver abscess associated with septic endophthalmitis during a recent four-year period . The causative organism was a pure culture of Klebsiella pneumoniae . The diagnosis was made by results of a blood culture in seven of the cases, liver aspirate culture in four, and eye contents or conjunctival culture in four . Chest roentgenographic examination showed pulmonary embolization in four patients, purulent meningitis in one patient, and suspicious prostatic abscess in one patient . Despite diligent antibiotic therapy, six patients lost their vision and one had impaired vision . This poor outcome for septic endophthalmitis seems to result from delayed diagnosis and lack of scheduled periocular injections of antibiotics . A combination course of treatment by the internist and ophthalmologist acting aggressively, both diagnostically and therapeutically, is needed in the future . To have seven cases of K pneumoniae liver abscess complicated by septic endophthalmitis during a period of four years in one hospital is very unusual . To our knowledge, it has never been reported in the literature.

Infect Immun, 1986 Oct, 54(1), 85 - 9
Role of capsule and O antigen in resistance of Klebsiella pneumoniae to serum bactericidal activity; Tomas JM et al.; The ability of Klebsiella pneumoniae strains to resist the bactericidal activity of serum was quantitated . The K . pneumoniae strains tested included mutants lacking the capsular polysaccharide and mutants having a modified lipopolysaccharide structure . The last mutants were obtained as phage-resistant mutants, and their lipopolysaccharide was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and chemical analysis . Serum-resistant mutants derived from phage-resistant mutants (lipopolysaccharide mutants) were also characterized . Resistance to the bactericidal activity of complement was mediated by the lipopolysaccharide, especially by the O-antigen polysaccharide chains . The capsular polysaccharide seemed not to play any important role in resistance to serum bactericidal activity in this bacterium.

Eur J Clin Invest, 1986 Oct, 16(5), 415 - 22
Ciclosporin and prednisone v . prednisone in treatment of Graves' ophthalmopathy: a controlled, randomized and prospective study; Kahaly G et al.; Forty patients with Graves' ophthalmopathy stages III-V were divided into two groups in a random manner according to their year of birth . Group I received prednisone in decreasing dosage . Group II received prednisone at a comparable dosage and ciclosporin . Steroids were discontinued after 10 weeks in the two groups . In the patients of group II, ciclosporin was continued over 12 months . The therapeutic effect was assessed by an activity score based on subjective and objective symptoms (computerized tomography and sonography of the orbit, Hertel values, clinical findings) . All signs of endocrine ophthalmopathy improved significantly in both groups (P less than 0.01 in group I; P less than 0.001 in group II) . The improvement was significantly greater in group II (P less than 0.05) according to the predefined score . After corticosteroids were discontinued, inflammatory signs recurred in nine patients in group I and in one of group II . During the observation period of 12 months, relapses occurred in eight out of twenty patients in group I and in only one out of twenty in group II . Muscle thickness decreased in nine patients in group II, 6 months after beginning therapy . At this time, the results were not influenced in any of the twenty patients in group I . Microsomal antibodies decreased significantly (P less than 0.001) in the ciclosporin group, whereas no change was seen in the other group . Renal values rose within the normal range in group II . In this group, an infection with Klebsiella pneumoniae occurred in one patient after 4 months of therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Clin Invest, 1986 Oct, 16(5), 338 - 46
Lymphocyte response to Klebsiella in ankylosing spondylitis; Gross WL et al.; The lymphocyte response of mononuclear cells (MNC) to Klebsiella (Klebs) was studied in ankylosing spondylitis (ASP) and, for comparison, in patients convalescing from Klebs infection and in HLA-B27+ and B27- healthy blood donors . When large doses of various Klebs cell envelope structures were used, MNC from all healthy blood donors were stimulated to produce LIF but not to synthesize significant amounts of DNA . In contrast, MNC from convalescent patients showed LIF synthesis (but no significant proliferation) even when small doses of the Klebs biostructures were used . This heightened LIF response to Klebs was long-standing and could be demonstrated even after serum antibodies had vanished . Very similarly, MNC from ASP patients did not proliferate significantly, but could be activated to produce LIF even with small doses of Klebs . The heightened LIF response to Klebs in ASP appeared to be correlated with the clinical activity and/or duration of the disease . In contrast, there was no correlation between the presence of HLA-B27 in ASP patients or healthy controls and the LIF or proliferative response to Klebs.

J Gen Microbiol, 1986 Oct, 132 ( Pt 10), 2827 - 35
Expression and purification of glutamine synthetase cloned from Bacteroides fragilis; Southern JA et al.; A glutamine synthetase (GS) gene, glnA, from Bacteroides fragilis was cloned on a recombinant plasmid pJS139 which enabled Escherichia coli glnA deletion mutants to utilize (NH4)2SO4 as a sole source of nitrogen . DNA homology was not detected between the B . fragilis glnA gene and the E . coli glnA gene . The cloned B fragilis glnA gene was expressed from its own promoter and was subject to nitrogen repression in E . coli, but it was not able to activate histidase activity in an E . coli glnA ntrB ntrC deletion mutant containing the Klebsiella aerogenes hut operon . The GS produced by pJS139 in E . coli was purified; it had an apparent subunit Mr of approximately 75,000, which is larger than that of any other known bacterial GS . There was very slight antigenic cross-reactivity between antibodies to the purified cloned B . fragilis GS and the GS subunit of wild-type E . coli.

J Bacteriol, 1986 Oct, 168(1), 173 - 8
Posttranscriptional control of Klebsiella pneumoniae nif mRNA stability by the nifL product; Collins JJ et al.; Posttranscriptional control of nif mRNA stability was demonstrated by functional and chemical analyses, using specific probes for four nif transcripts . In the wild type, nif transcripts (except nifLA) were stable during derepression, with half-lives of approximately 30 min . They were dramatically destabilized by O2 or elevated temperature (41 degrees C) and to a lesser extent by NH4+ . In contrast, the nifLA message was not particularly stable, and posttranscriptional control was not evident . In NifL- strains, both forms of analysis indicated that the nifL product was involved in nif mRNA destabilization in the presence of O2 and NH4+.

Eur J Biochem, 1986 Sep 15, 159(3), 549 - 53
Structure predictions and surface charge of nitrogenase flavodoxins from Klebsiella pneumoniae and Azotobacter vinelandii; Drummond MH; A first approximation to the tertiary structure of the nitrogenase flavodoxins of Klebsiella pneumoniae and Azotobacter vinelandii can be obtained by superimposing their amino acid sequences upon the crystallographically determined structure of the long-chain flavodoxin from Anacystis nidulans . This procedure is validated by secondary structure predictions based on the sequence alone and by the distribution of polar and hydrophobic residues . It reveals, among other things, a distinctive distribution of surface charge peculiar to the nitrogenase flavodoxins, which is probably important in determining the kinetics of electron transfer with their physiological redox partners . The most likely positions of the phosphodiester bridge which has been described in the A . vinelandii molecule can also be assessed.

Carbohydr Res, 1986 Sep 1, 152, 249 - 59
The structural elucidation of the capsular polysaccharide of Klebsiella K68; Dutton GG et al.; The structure of the capsular polysaccharide isolated from Klebsiella K68 bacteria has been elucidated by both chemical and spectroscopic methods . The structure is of the "3 + 1" type, similar in pattern to the structures of the polysaccharides of Klebsiella K11 and K57, having a single branch point on the uronic acid . The polysaccharide is shown to consist of the following repeating unit: (Formula: see text).

Carbohydr Res, 1986 Sep 1, 152, 205 - 15
Structure of the Klebsiella type 10 capsular polysaccharide; Sarkar AK et al.; The capsular polysaccharide from Klebsiella Type 10 was found to contain D-galactose, D-glucose, D-mannose, and D-glucuronic acid in the ratios 3:1:1:1 . Acid hydrolysis of the polysaccharide gave one aldobiouronic acid, one aldotriouronic acid, one aldotetraouronic acid, and two neutral disaccharides the structures of which were established . The native and carboxyl-reduced polysaccharide have been subjected as appropriate to methylation analysis and Smith degradation . Degradation of the methylated polysaccharide with base established the identity of the sugar unit preceding the glucosyluronic acid residue . The anomeric configurations of the sugar residues were determined by oxidation of the acetylated native and carboxyl-reduced polysaccharides with chromium trioxide . Based on these studies, the hexasaccharide structure 1 has been assigned to the repeating unit of the K-10 polysaccharide.

J Lab Clin Med, 1986 Sep, 108(3), 182 - 9
Activity of intravenous immune globulins against Klebsiella; Cryz SJ Jr et al.; Seven commercial intravenous immune globulin (IVIG) preparations and an experimental hyperimmune globulin (Klebsiella immune-IVIG) were analyzed for antibody levels to Klebsiella capsular polysaccharides, opsonic activity, and protective capacity against Klebsiella . With one exception, all IVIGs reacted immunologically with the various capsular antigens, although titers between the preparations varied considerably . Klebsiella immune-IVIG possessed substantially higher titers (32-fold to 128-fold) than the commercial preparations . Only the hyperimmune globulin was capable of promoting the phagocytosis and killing of six Klebsiella test strains . When prophylactically administered, a commercial IVIG afforded significant protection (P less than 0.01) against fatal experimental Klebsiella sepsis only at a dose of 500 mg/kg . Klebsiella immune-IVIG provided a similar level of protection at a dose of 5 mg/kg . Whereas the commercial IVIG was ineffective when used therapeutically, the hyperimmune globulin was highly protective . Combined antibiotic-immunoglobulin therapy was most effective at reducing mortality.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Sep, 262(3), 335 - 45
Comparative investigations of Klebsiella species of clinical origin: plasmid patterns, biochemical reactions, antibiotic resistances and serotypes; Podschun R et al.; A total of 124 K . pneumoniae and 52 K . oxytoca isolates obtained from clinical specimens was investigated for plasmid patterns, biochemical reactions, antibiotic resistances and serotypes regarding to the distribution and relationships of these characters . A great diversity of plasmid patterns, bio/serotypes and resistance patterns was revealed . About 90% of strains contained plasmid DNA and up to seven plasmid bands per isolate could be shown . For K . pneumoniae, serotype 7 and for K . oxytoca, type 55 were most common . In general, little difference between both species was found and characters were similarly distributed . With respect to the site of isolation, serotype 7 was predominating in K . pneumoniae strains from the respiratory tract . Highly multiple-resistant organism were found in the largest number in specimens from the urogenital tract, in the lowest in specimens from wounds . Extensive statistical analyses did not detect any relationship among the characters investigated.

Antimicrob Agents Chemother, 1986 Sep, 30(3), 403 - 8
Continuous versus intermittent administration of ceftazidime in experimental Klebsiella pneumoniae pneumonia in normal and leukopenic rats; Roosendaal R et al.; Experimental Klebsiella pneumoniae pneumonia was used to study the influence of cyclophosphamide-induced leukopenia on the relative therapeutic efficacy of continuous and intermittent (6-h intervals) administration of ceftazidime . The antimicrobial response was evaluated with respect to the calculated daily dose that protected 50% of the animals from death (PD50) until 16 days after the termination of a 4-day treatment . When ceftazidime was administered intermittently to leukopenic rats, the PD50 was 24.37 mg/kg per day, 70 times (P less than 0.001) the PD50 of 0.35 mg/kg per day for normal rats . Continuous administration of ceftazidime to leukopenic rats resulted in a PD50 of 1.52 mg/kg per day, four times (P less than 0.001) the PD50 of 0.36 mg/kg per day for normal rats . Continuous administration of ceftazidime in daily doses that protected 100% of normal and leukopenic rats from death resulted in serum levels of 0.06 and 0.38 micrograms/ml, respectively, whereas the MIC for the infecting K . pneumoniae strain was 0.2 micrograms of ceftazidime per ml . The effect of the duration of ceftazidime treatment by continuous infusion on the therapeutic efficacy in relation to the persistence of leukopenia was then investigated in leukopenic rats . The administration of 3.75 mg of ceftazidime/kg per day for 4 days protected all leukopenic rats from death, provided the circulating leukocytes returned at the end of antibiotic treatment . When leukopenia persisted for 8 days this ceftazidime treatment schedule resulted in the mortality of rats (P less than 0.05) . However, when ceftazidime treatment was continued for 8 days, until the return of the leukocytes, there was no significant mortality (P greater than 0.05).

J Antimicrob Chemother, 1986 Sep, 18(3), 387 - 91
Comparative efficacy of different beta-lactam antibiotics and gentamicin in Klebsiella pneumoniae septicaemia in neutropenic mice; Trautmann M et al.; The in-vivo activity of ceftazidime, cefotetan, imipenem/cilastatin, piperacillin and gentamicin against two strains of Klebsiella pneumoniae was evaluated in a model of experimental septicaemia in neuropenic mice . Single agent therapy with the aminoglycoside was highly effective against both strains . Among the beta-lactams, ceftazidime and cefotetan were nearly as active as gentamicin, whereas imipenem/cilastatin was slightly less effective, and the results achieved with piperacillin were markedly inferior.

J Bacteriol, 1986 Sep, 167(3), 999 - 1003
nifV-dependent, low-molecular-weight factor required for in vitro synthesis of iron-molybdenum cofactor of nitrogenase; Hoover TR et al.; The molybdate- and ATP-dependent in vitro synthesis of the iron-molybdenum cofactor of nitrogenase requires a low-molecular-weight factor . The factor is present in extracts of nitrogen fixation-derepressed cultures of Klebsiella pneumoniae and Azotobacter vinelandii, but not in extracts of repressed cultures of these bacteria . Analysis of K . pneumoniae Nif mutants has indicated that the nifV gene product is the only nif protein (besides nifA) necessary for the synthesis and accumulation of the factor . The factor is stable to oxygen, temperatures below 120 degrees C, and extremely acidic and basic conditions . The activity of the factor was completely destroyed by dry ashing or digestion with sulfuric acid . The factor has been partially purified by filtration through an Amicon PM-10 DIAFLO membrane and chromatography on DEAE-cellulose, hydroxylapatite, silica gel, and Sephadex G-25.

Appl Environ Microbiol, 1986 Aug, 52(2), 325 - 30
Metabolism of the herbicide bromoxynil by Klebsiella pneumoniae subsp . ozaenae; McBride KE et al.; Enrichment of soil samples for organisms able to utilize the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) as a nitrogen source yielded bacterial isolates capable of rapidly metabolizing this compound . One isolate, identified as Klebsiella pneumoniae subsp . ozaenae, could completely convert 0.05% bromoxynil to 3,5-dibromo-4-hydroxybenzoic acid and use the liberated ammonia as a sole nitrogen source . Assays of cell extracts of this organism for the ability to produce ammonia from bromoxynil revealed the presence of a nitrilase (EC 3.5.51) activity . The enzyme could not utilize 3,5-dibromo-4-hydroxybenzamide as a substrate, and no 3,5-dibromo-4-hydroxybenzamide could be detected as a product of bromoxynil transformation . Comparison of related aromatic nitriles as substrates demonstrated that the Klebsiella enzyme is highly specific for bromoxynil.

Antimicrob Agents Chemother, 1986 Aug, 30(2), 315 - 20
Molecular cloning of amikacin resistance determinants from a Klebsiella pneumoniae plasmid; Tolmasky ME et al.; A multiresistant Klebsiella pneumoniae strain, JHCK1, harbored several plasmids . One of these, plasmid pJHCMW1, carried determinants for resistance to amikacin in addition to kanamycin, tobramycin, and ampicillin . The amikacin resistance determinant(s) was cloned and studied by restriction mapping, insertion, and deletion analysis . The amikacin resistance gene(s) was localized in a 1.5-kilobase DNA fragment . This pJHCMW1 DNA region was responsible for not only the resistance to amikacin but also the resistance to kanamycin and tobramycin . The cloned DNA fragment specified both an acetyltransferase activity and a low level of phosphotransferase activity . The two activities were absent from mutants that did not confer resistance to amikacin, kanamycin, and tobramycin.

Biol Chem Hoppe Seyler, 1986 Aug, 367(8), 813 - 23
Citrate transport in Klebsiella pneumoniae; Dimroth P et al.; Sodium ions were specifically required for citrate degradation by suspensions of K . pneumoniae cells which had been grown anaerobically on citrate . The rate of citrate degradation was considerably lower than the activities of the citrate fermentation enzymes citrate lyase and oxaloacetate decarboxylase, indicating that citrate transport is rate limiting . Uptake of citrate into cells was also Na+ -dependent and was accompanied by its rapid metabolism so that the tricarboxylic acid was not accumulated in the cells to significant levels . The transport could be stimulated less efficiently by LiCl . Li+ ions were cotransported with citrate into the cells . Transport and degradation of citrate were abolished with the uncoupler {4-(trifluoromethoxy)phenylhydrazono}propanedinitrile (CCFP) . After releasing outer membrane components and periplasmic binding proteins by cold osmotic shock treatment, citrate degradation became also sensitive towards monensin and valinomycin . The shock procedure had no effect on the rate of citrate degradation indicating that the transport is not dependent on a binding protein . Citrate degradation and transport were independent of Na+ ions in K . pneumoniae grown aerobically on citrate and in E . coli grown anaerobically on citrate plus glucose . An E . coli cit+ clone obtained by transformation of K . pneumoniae genes coding for citrate transport required Na specifically for aerobic growth on citrate indicating that the Na-dependent citrate transport system is operating . Na+ and Li+ were equally effective in stimulating citrate degradation by cell suspensions of E . coli cit+ . Citrate transport in membrane vesicles of E . coli cit+ was also Na+ dependent and was energized by the proton motive force (delta micro H+) . Dissipation of delta micro H+ or its components delta pH or delta psi by ionophores either totally abolished or greatly inhibited citrate uptake . It is suggested that the systems energizing citrate transport under anaerobic conditions are provided by the outwardly directed cotransport of metabolic endproducts with protons yielding delta pH and by the decarboxylation of oxaloacetate yielding delta pNa+ and delta psi . In citrate-fermenting K . pneumoniae an ATPase which is activated by Na+ was not found . The cells contain however a proton translocating ATPase and a Na+/H+ antiporter in their membrane.

J Biol Chem, 1986 Jul 15, 261(20), 9289 - 93
The synthesis of adenine-modified analogs of adenosylcobalamin and their coenzymic function in the reaction catalyzed by diol dehydrase; Toraya T et al.; Five analogs of adenosylcobalamin modified in the adenine moiety of the Co beta ligand were synthesized and tested for coenzymic function with diol dehydrase of Klebsiella pneumoniae ATCC 8724 . 1-Deaza and 3-deaza analogs of adenosylcobalamin were active as coenzyme, whereas 7-deaza and N6,N6-dimethyl derivatives and guanosylcobalamin did not show detectable coenzymic activity . 7-Deaza and N6,N6-dimethyl analogs acted as strong competitive inhibitors with respect to adenosylcobalamin . The formation of cob(II)alamin as intermediate in the catalytic reaction was spectroscopically observed with catalytically active complexes of the enzyme with 1-deaza and 3-deaza analogs in the presence of 1,2-propanediol, but not with complexes with the inactive analogs . Oxygen sensitivity of the enzyme-analog complexes suggests that the carbon-cobalt bond of 1-deaza and 3-deaza analogs becomes activated by the enzyme even in the absence of substrate . These results indicate that the importance of the nitrogen atoms in the adenine moiety of the coenzyme for manifestation of catalytic function and for activation of the carbon-cobalt bond decreases in the following order: N-7 greater than 6-NH2 greater than N-3 greater than N-1 . The dissociation constant for 5'-deoxyadenosine determined by equilibrium dialysis at 37 degrees C was about 23 microM.

Carbohydr Res, 1986 Jul 1, 149(2), 411 - 23
The use of bacteriophage depolymerization in the structural investigation of the capsular polysaccharide from Klebsiella serotype K3; Dutton GG et al.; The structure of the repeating unit of the capsular polysaccharide from Klebsiella serotype K3 has been established from the results of n.m.r . (1H and 13C) spectroscopy and methylation analysis of P1, the pyruvic acetal-bearing pentasaccharide obtained on depolymerization of the polysaccharide with a bacteriophage-borne endogalactosidase, reduced deacetalated P1, and the native polysaccharide . The data permit the assignment of the following structure to the repeating unit: (formula see text)

J Antibiot (Tokyo), 1986 Jul, 39(7), 971 - 7
Comparative effects of cefpirome (HR 810) and other cephalosporins on experimentally induced pneumonia in mice; Klesel N et al.; The chemotherapeutic efficacy of cefpirome (HR 810), a new polar aminothiazolylcephalosporin and that of ceftazidime, cefotaxime, cefoperazone, latamoxef and cefodizime were examined against experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice . When compared in terms of MIC values against the infecting organism and the pharmacokinetic pattern, cefpirome showed equal activity and a similar pharmacokinetic behavior to ceftazidime and cefotaxime in mice . Trials to assess the bactericidal activity in vivo, however, showed that cefpirome displayed a more marked bactericidal effect in pneumonic mice than the other cephalosporins tested . Only cefodizime, a cephalosporin with extremely high and prolonged blood and tissue levels in experimental animals exerted chemotherapeutic effects similar to cefpirome . After cefpirome or cefodizime medication (50 mg/kg), the viable counts in the lungs of experimental animals fell steadily to 1/10,000 of the pretreatment level and, in contrast to the reference compounds, no regrowth of the challenge organisms could be observed with both drugs . Moreover, with ED50s ranging from 1.1 to 59.1 mg/kg in treatment studies, cefpirome as well as cefodizime were two to ten times more effective than ceftazidime and cefotaxime, whereas cefoperazone and latamoxef were considerably less effective.

Ann Inst Pasteur Microbiol, 1986 Jul-Aug, 137B(1), 29 - 36
Klebsiella pneumoniae C3 lipopolysaccharide mutants obtained by resistance to bacteriophage FC3-9; Benedi VJ et al.; Bacteriophage FC3-9 is one of the several bacteriophages of Klebsiella pneumoniae C3 isolated in our laboratory . Mutants resistant to this bacteriophage were isolated and found to be devoid of lipopolysaccharide O antigen, modified in outer-membrane protein composition and sensitive to complement killing (serum-sensitive), unlike the wild-type strain . Serum-resistant mutants were isolated from these strains . They regained the lipopolysaccharide O antigen and the wild-type outer membrane protein composition and became sensitive to bacteriophage FC3-9.

Toxicol Lett, 1986 Jul-Aug, 32(1-2), 41 - 9
Effects of acrolein on macrophage functions in rats; Sherwood RL et al.; Male Sprague-Dawley rats were exposed to 0.1, 1.0 or 3.0 ppm acrolein or filtered air 6 h/day, 5 days/week for 3 weeks . Rats were tested one day following the last exposure and exhibited no change in pulmonary clearance of inhaled 35S-labeled Klebsiella pneumoniae at any acrolein concentration . Decreased numbers of peritoneal cells were obtained from exposed rats while the number of cells lavaged from the lungs was unchanged . Macrophages of acrolein-exposed rats had altered phagocytic and enzymatic patterns as compared to macrophages from animals breathing filtered air . However, these changes had no apparent effect on macrophage killing of inhaled bacteria and were therefore probably not indicative of extreme chemical toxicity.

Ann Inst Pasteur Microbiol, 1986 Jul-Aug, 137B(1), 3 - 18
Cloning and characterization of the glnA gene of Azospirillum brasilense Sp7; Bozouklian H et al.; A plasmid which, by complementation, restored a Gln+Nif+ phenotype to the Gln-Nif- Azospirillum brasilense mutant 7029, was isolated from a gene bank of total DNA of A . brasilense Sp7 (ATCC 29145) constructed in the broad host range vector pVK100 . This plasmid contained the structural gene (glnA) for glutamine synthetase . The glnA gene was mapped by Tn5 insertion and DNA hybridization with a Klebsiella pneumoniae glnA probe . The direction of transcription of glnA was determined . The glnA product was identified as a 50-Kd polypeptide which could be adenylylated in Escherichia coli, and glutamine synthetase activity was characterized in E . coli . Plasmids containing the glnA gene restored glutamine-independent growth and a Nif+ phenotype to Gln-Nif- and Gln-Nifc mutants of Azospirillum.

J Hosp Infect, 1986 Jul, 8(1), 39 - 46
The effects of sub-lethal concentrations of chlorhexidine on bacterial pathogenicity; Holloway PM et al.; Suspensions of Escherichia coli and Klebsiella aerogenes were exposed to low levels of chlorhexidine, which had little effect on their in vitro viability . A substantial reduction in their in vivo infectivity measured by intraperitoneal inoculation of mice was found . This selective reduction in bacterial infectivity by chlorhexidine should be taken into account when conventional in vitro tests are used to assess the clinical effectiveness of chlorhexidine-containing antiseptics.

Tohoku J Exp Med, 1986 Jul, 149(3), 341 - 2
Rapid identification of Klebsiella pneumoniae in the lung tissue by fluorescence microscopy; Senba M et al.; Klebsiella pneumoniae is stained weekly with hematoxylin and eosin, moderately with periodic acid Schiff (PAS), and strongly by the Warthin-Starry method . Staining by the Warthin-Starry method, however, takes more than one hour . Therefore, we developed a new procedure for rapid identification of K . pneumoniae in paraffin sections of the lung under a fluorescence microscope.

Appl Environ Microbiol, 1986 Jun, 51(6), 1361 - 3
New, simple medium for selective, differential recovery of Klebsiella spp; Tomas JM et al.; A highly selective, differential medium for the enumeration and isolation of Klebsiella spp . was developed . With pure cultures, 100% recovery of Klebsiella spp . was observed . Recovery of Klebsiella spp . on MacConkey-inositol-potassium tellurite (MCIK) agar was as good as or better than on MacConkey-inositol-carbenicillin agar either with pure cultures or environmental samples . Recovery and percent colony confirmation with MCIK agar were greater and easier to obtain than for other proposed Klebsiella selective media.

Pathol Biol (Paris), 1986 Jun, 34(5 Pt 2), 611 - 5
{Klebsiella oxytoca beta-lactamases: study of their action on 3d-generation cephalosporins}; Labia R et al.; Indole-positive Klebsiella pneumoniae or K . oxytoca are usually resistant to penicillins as a result of the production of a chromosomally-mediated beta-lactamase with a low level of synthesis (specific activity approximately 50 to 100 mU/mg) . Although most strains are susceptible to the majority of cephalosporins, some strains exhibit resistance to cephalosporins including third-generation drugs . These resistant strains produce a chromosomally-mediated beta-lactamase with a high level of synthesis (specific activity approximately 5,000 mU/mg or higher) . Four beta-lactamases have been identified on the basis of their isoelectric points: pI = 5.5, 5.7, 6.0 and 6.3; nevertheless they have similar kinetic constants, and are inhibited by clavulanic acid . These enzymes hydrolyze most third-generation cephalosporins, in the following order of decreasing velocities: cefoperazone, ceftriaxone, cefotaxime, cefodizime, cefpirome; ceftazidime, and cefoxitin, cefotetan, latamoxef, cephamycins which are totally resistant to these enzymes.

J Bacteriol, 1986 Jun, 166(3), 1083 - 8
Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein; Pugsley AP et al.; Pullulanase is a starch-debranching enzyme produced by the gram-negative bacterium Klebsiella pneumoniae . In this organism, the enzyme is first exported to the outer membrane and is subsequently released into the growth medium . Evidence reported here indicates that pullulanase is a lipoprotein . It is apparently synthesized as a precursor with a 19-residue-long signal sequence and modified by the covalent attachment of palmitate to the cysteine residue which becomes the amino terminus after cleavage of the signal sequence . In this respect, pullulanase is similar to some penicillinases produced by gram-positive bacteria which are initially exported to the cell surface and subsequently released into the medium . However, pullulanase and the penicillinases differ in one important aspect, namely, that the extracellular pullulanase still carries the covalently attached fatty acyls, whereas extracellular penicillinases lack the modified amino-terminal cysteine together with a limited number of other residues from the amino terminus.

Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 403 - 12
The immune response of a marine teleost, Pseudopleuronectes americanus, (winter flounder) to the protozoan parasite Glugea stephani; Laudan R et al.; G . stephani is an intracellular cyst-forming microsporidan parasite that is found in the intestine of winter flounder (WF) Pseudopleuronectes americanus . No detectable humoral response was seen in parasitized fish or in fish injected with either spores or spore homogenate from this parasite . Quantification of total immunoglobulin (Ig) levels showed a decrease in Ig levels rather than enhancement, 21 days after intramuscular (IM) injections of spores (3 X 10(6)/ml) . When a second injection of spores was administered on day 21 and tested 3 weeks later, a further decrease in total serum Ig's occurred . A decrease in total IgM levels also occurred in WF that were simultaneously injected with G . stephani and the antigens, horse red blood cells (HRBC) or formalin-killed Klebsiella pneumonia (KP) . The total Ig levels of fish injected with an antigen plus spores was not as low as those injected with the parasite alone . The Ig levels, as well as antibody titers to HRBC and KP were however, lower when compared to fish injected only with the HRBC or bacteria . Disrupted spore homogenate injected into winter flounder, showed a less marked decrease in Ig levels when compared with whole spores . When a single IM injection of spores was given, followed by two weekly injections of indomethacin (a drug that inhibits prostaglandin activity), no decrease in Ig levels occurred and levels were comparable to control (saline injected) fish.

Ann Trop Paediatr, 1986 Jun, 6(2), 123 - 8
Nosocomial Klebsiella pneumoniae colonization in a neonatal special care unit; Denny F et al.; A clinical study is reported of nosocomial Klebsiella pneumoniae colonization and infection in a neonatal special care unit in Barbados . Of the 14 infants who were affected nine were symptomatic and two died (15%) during a 9-day period . This outbreak appeared to have been an indirect result of an acute shortage of nursing personnel coupled with a breakdown in adequate handwashing techniques . We feel that periodic review and continuous reinforcement of infection control policies in areas with patients at high risk of nosocomial infection, such as the neonatal unit, are of paramount importance.

Zh Mikrobiol Epidemiol Immunobiol, 1986 May, (5), 32 - 4
{Characteristics of Klebsiella strains isolated from patients with acute intestinal diseases}; Gaziev GM et al.; A total of 203 Klebsiella strains isolated from patients with acute intestinal diseases were studied . The biochemical variants were determined by taking into account the complex of such signs as the capacity for producing indole, hydrolyzing urea, utilizing sodium malonate, fermenting inositol, dulcitol, sorbose, adonitol, synthesizing acetylmethylcarbinol, reacting with methyl red . The strains under study were found to belong to 36 K-types . Klebsiella strains with K-antigens 20, 2, 62, 60, 21, 40 showed the highest isolation rate.

J Clin Microbiol, 1986 May, 23(5), 948 - 50
Nosocomial infection and colonization by Klebsiella trevisanii; Freney J et al.; During an 18-month period we identified two cases of septicemia and 24 examples of colonization of humans by Klebsiella trevisanii . Organisms were identified using the API 20EC and API 147 assimilation galleries . Of 147 clinical isolates initially identified as K . oxytoca, 18% were found to be K . trevisanii . Tracheal aspirate was the most common source of the organism . An extensive environmental sampling survey in the rooms of 12 colonized patients revealed a possible reservoir of the organism only once (a face cloth).

J Antimicrob Chemother, 1986 May, 17 Suppl C, 141 - 8
The efficacy of the combination of Timentin and tobramycin in the treatment of patients with bacteraemia; Degener JE et al.; Twenty-eight patients with bacteraemia were treated with Timentin, a combination of ticarcillin and clavulanic acid, and tobramycin . According to a simple physiological classification scheme 20 patients were moderately and eight severely ill . Clinical cure was achieved in 15 of 20 moderately ill and in four of eight severely ill patients . The nine therapeutic failures were due to non-removable foreign material (2 cases), abscesses that were not effectively evacuated (5 cases), urolithiasis (1 case), and decubitus ulcer (1 case) . One of these cases was complicated by infection with a Klebsiella strain with Timentin and tobramycin MICs of 64 mg/l . Thirteen of 31 bacterial isolates from blood were resistant to greater than or equal to 128 mg/l of ticarcillin of which 11 were susceptible to less than or equal to 64 mg/l of Timentin . The addition of Timentin to tobramycin offered a better coverage in vitro than ticarcillin combined with tobramycin against the strains isolated from the blood of the patients included in this study.

J Appl Bacteriol, 1986 May, 60(5), 375 - 9
Submicroscopical changes in Klebsiella pneumoniae cells treated with concentrated sucrose and polyethylene glycol 400 solutions; Bozzini JP et al.; This study reports the extent and character of plasmolysis and other morphological changes as shown by electron microscopy in a strain of Klebsiella pneumoniae and with sucrose or polyethylene glycol 400 (PEG-400) as the plasmolysing agent at a water activity of 0.935 . Both solutes produced severe plasmolysis in K . pneumoniae cells; PEG-400 also caused some cell wall collapse and finger like extrusions to emerge from the bacterial cell.

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