J Med Microbiol, 1986 May, 21(3), 225 - 31 Adherence of multiresistant strains of Klebsiella pneumoniae to cerebrospinal fluid shunts: correlation with plasmid content; Denoya CD et al.; A nosocomial multiresistant Klebsiella pneumoniae strain (KMD01) isolated from a patient with an infected ventriculoperitoneal (V-P) shunt was found to contain three plasmids of mol . wts (10(6)) c . 85, 50 and 2.4 . A derivative isogenic strain (KMD11) carrying only the plasmids of mol . wts (10(6)) 50 and 2.4 was obtained spontaneously by plating the parent strain . The absence of the plasmid of mol . wt 85 X 10(6) in strain KMD11 correlated with an increased adherence to V-P catheters and glass surfaces, as well as autoagglutination in minimal medium . Bacterial cells containing the whole set of plasmids (strain KMD01) also showed the incorporation into the outer membrane of a new polypeptide (mol . wt, c . 41 X 10(3)), when grown in minimal medium . The presence of this polypeptide correlated with absence of autoagglutination, as shown by strain KMD01 under these cultural conditions . These data suggest that the cell-surface characteristics in K . pneumoniae may be affected by the plasmid content of the strain . Since nosocomial strains of K . pneumoniae usually contain one or more plasmids, and strains easily exchange these extrachromosomal elements, it seems reasonable to speculate that new variants with higher V-P shunt colonisation effectiveness, like the one described in this work, may also evolve in nature. J Bacteriol, 1986 May, 166(2), 545 - 51 Deletion analysis of the Klebsiella pneumoniae nitrogenase promoter: importance of spacing between conserved sequences around positions -12 and -24 for activation by the nifA and ntrC (glnG) products; Buck M; The nitrogen fixation promoters of Klebsiella pneumoniae are atypical procaryotic promoters lacking the usual -10 and -35 elements, requiring instead conserved sequences around -12 and -24 for transcriptional activation . By constructing a set of five deletions between the -12 and -24 elements in the nifH promoter, the spacing between the conserved GC and GG motifs at -12 and -24, respectively, has been reduced from the wild-type 10 bases to 9, 8, 6, 5, and 4 bases . The deletion of a single nonconserved nucleotide was sufficient to eliminate transcriptional activation by either nifA or ntrC (glnG) . All deletions relieved the multicopy inhibition of chromosomal nif expression normally shown by the nifH promoter . These results demonstrate a stringent requirement for the 10-base spacing found in ntr-activated promoters . In addition, specific sequences around the invariant GG at -24 were shown to be necessary for activation by either nifA or ntrC, with a minimal requirement for nucleotides through to position -27 for this activation. Ann Acad Med Singapore, 1986 Apr, 15(2), 176 - 81 Liver abscess--a clinical study; Teh LB et al.; Forty consecutive cases of liver abscess from the Department of Medicine III, Singapore General Hospital, from 1978 to July 1983 were reviewed . Nineteen (47.5%) were amoebic, 15 (37.5%) pyogenic and 6 (15%) of unknown aetiology . Of the first 20 cases from 1978 to 1980, amoebic abscesses (60%) predominated . An increased incidence of pyogenic abscess constituting 50% was seen in the next 20 cases . Though all racial groups were affected, a predilection among Indians was seen . Males outnumbered females (4:1), and peak incidence occurred in the 40 to 70 age group (62.5%) . Fifty percent presented early (less than one week of symptoms) to hospital . Common physical signs were fever (97.5%) and hepatomegaly (92.5%) . Investigations showed leucocytosis in excess of 10,000 WBCs/cmm (87.5%), an ESR of 80 mm/hr (80%) and an elevated alkaline phosphatase of at least twice normal (73.6%) . Single abscesses (72.5%) located in the right lobe were more likely to be amoebic . Where abscesses were multiple, they were more likely to be pyogenic (63.6%) . Two-thirds of the pyogenic abscesses were due to either Klebsiella species or E . coli . Medical treatment consisted of broad spectrum antibiotics, usually in combination with metronidazole . Aspiration or drainage (open or closed) was employed when indicated . These were carried out more often for pyogenic than amoebic abscesses . Amoebic abscesses responded faster to treatment compared to pyogenic abscesses . Mortality in the first 20 cases prior to 1981 was 30%, being mainly confined to pyogenic abscesses . However, after 1981, there has been no mortality in the ensuing 20 cases. J Gen Microbiol, 1986 Apr, 132 ( Pt 4), 1051 - 4 Purification and characterization of a bacteriocin from Klebsiella pneumoniae 158; Chhibber S et al.; Klebocin, a bacteriocin produced by Klebsiella pneumoniae 158, was purified to homogeneity by ammonium sulphate fractionation and sequential DEAE-Sephacel and Sephadex G-150 column chromatography . The purified preparation had an Mr of approximately 40 000 on SDS-PAGE . Chemical analysis of the purified preparation showed it to be a protein, and it was sensitive to digestion by various proteolytic enzymes. Ann Acad Med Singapore, 1986 Apr, 15(2), 172 - 5 Acute cholangitis in Singapore; Chew RK et al.; 115 cases of acute calculous cholangitis were analysed for their presentations, treatment and complications . There were slightly more Chinese males than females (1.4:1) and the mean age was 62 years old . Though pain was the commonest symptom (95.7%), only 35.7% of patients had complete Charcot's triad . 21.7% presented in septicaemic shock . E . Coli and Klebsiella were the commonest organisms isolated . All patients had cholecystectomy and exploration of common bile ducts . Twenty patients had additional choledochontrostomy performed . 50.4% of patients developed complications and the overall mortality was 16.5% . Patients over 60 presented in shock carried a significantly higher mortality (40%). J Clin Microbiol, 1986 Apr, 23(4), 687 - 90 Seroepidemiology of Klebsiella bacteremic isolates and implications for vaccine development; Cryz SJ Jr et al.; The frequencies of capsular serotypes among 703 Klebsiella strains isolated from the blood of hospitalized patients were determined . More than 90% of the isolates were typeable, with 69 of the 77 known serotypes being identified . Serotypes 2, 21, and 55, representing 8.9, 7.8, and 4.8% of all the isolates, respectively, were observed at a frequency significantly higher (P less than 0.05) than that for other capsular serotypes . Approximately 43% of the serotypes appeared at a frequency of less than 0.5% . Differences were found when the seroepidemiology of North American and European isolates was compared . The current findings indicate that a capsular polysaccharide-based vaccine against Klebsiella organisms is feasible and should be multivalent, eliciting antibodies directed against the 25 serotypes which make up approximately 70% of all the bacteremic isolates. J Bacteriol, 1986 Apr, 166(1), 353 - 6 Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae; Kong QT et al.; Oxygen sensitivity of the nifLA promoter of Klebsiella pneumoniae has been demonstrated . Studies on the oxygen regulation of nifB-lacZ and nifH-lacZ fusions in the presence of the nifLA operon, which contains either an intact or a deleted nifL gene, indicate that possibly both the nifL promoter and the nifL product are responsible for nif repression by oxygen. Eur J Biochem, 1986 Apr 1, 156(1), 157 - 62 Kinetic analysis of the reaction mechanism of oxaloacetate decarboxylase from Klebsiella aerogenes; Dimroth P et al.; The mechanism of oxaloacetate decarboxylase of Klebsiella aerogenes was investigated by enzyme kinetic methods . The activity of the decarboxylase was strictly dependent on the presence of Na+ or Li+ ions . For Li+ the Km was about 17 times higher and the Vmax about 4 times lower than for Na+ . No activity was detectable at Na+ concentrations less than 5 microM . The curve for initial velocity versus Na+ concentration was hyperbolic . Initial velocity patterns with oxaloacetate or Na+ as the varied substrate at various fixed concentrations of the cosubstrate produced a pattern of parallel lines which is characteristic for a ping-pong mechanism . Product inhibition by pyruvate was competitive versus oxaloacetate and noncompetitive versus Na+ . Oxalate, a dead-end inhibitor, was competitive versus oxaloacetate and uncompetitive versus Na+ . The inhibition patterns are not consistent with a ping-pong mechanism comprising a single catalytic site but are analogous to kinetic patterns observed with the related biotin enzyme transcarboxylase, for which a catalytic mechanism at two different and independent sites has been demonstrated . The kinetic and other data support an oxaloacetate decarboxylase mechanism at two different sites of the enzyme with the intermediate formation of a carboxybiotin-enzyme complex . The first site is the carboxyltransferase which is localized on the alpha chain and the second site is the carboxybiotin-enzyme decarboxylase which is probably localized on the beta and/or gamma subunit . Binding studies with oxalate indicated that this is bound with high affinity to the alpha chain . The affinity was not affected by Na+ or by complex formation with the beta and gamma subunits . Oxalate protected the decarboxylase from heat inactivation but not from tryptic hydrolysis . The carboxybiotin-enzyme intermediate prepared from oxaloacetate decarboxylase with high specific activity was rapidly decarboxylated in the presence of Na+ ions alone . The effect of pyruvate on this reaction, noted previously, probably results from inhomogeneity of the enzyme preparation used which contained a considerable amount of free alpha subunits. Mol Gen Genet, 1986 Apr, 203(1), 101 - 9 Transcription termination within the Escherichia coli origin of DNA replication, oriC; Junker DE Jr et al.; Initiation of DNA replication from the Escherichia coli origin, oriC, is dependent on an RNA polymerase-mediated transcription event . The function of this RNA synthetic event in initiation, however, remains obscure . Since control of the synthesis of this RNA could serve a key role in the overall initiation process, transcription regulatory sites within and near oriC were identified using the galK fusion vector system . Our results confirm the existence of a transcription termination signal within oriC, first identified by Hansen et al . (1981), for the 16 kd transcript that is transcribed counterclockwise towards oriC . Termination is shown to be 92% efficient . A similar approach led to the detection of transcription termination within the chromosomal replication origin of Klebsiella pneumoniae . Approximately 50% of the E . coli 16 kd transcripts appear to terminate before reaching oriC between the XhoI (+416 bp) and the HindIII (+243 bp) sites . The predominant 3' ends of RNA that enter oriC, as determined by SI nuclease mapping, were located at positions +20 +/- 2, +23 +/- 2, +37, +39, +52, +66, +92, and +107 . These termination sites, which map cl to RNA . DNA junctions identified by Kohara et al . (1985), appear as triplets and quadruplets . The E . coli oriC Pori-L promoter described in in vitro transcription studies by Lother and Messer (1981) was not detected in this study in either wildtype cells or isogenic dnaA mutants at the nonpermissive temperature . A new promoter activity, Pori-R1, was identified within the E . coli origin in the clockwise direction. Biochemistry, 1986 Mar 11, 25(5), 1083 - 8 N2O as a substrate and as a competitive inhibitor of nitrogenase; Jensen BB et al.; We have investigated the inhibitory effect of N2O on NH3 formation by purified component proteins from Klebsiella pneumoniae and have confirmed that the inhibition is competitive with respect to N2 and that N2O is reduced to N2, which in turn is further reduced to NH3 . In addition, we have shown that N2O is unable to support HD formation from D2 and H2O . N2-supported HD formation from D2 and H2O was found to be inhibited by N2O . In contrast to N2, N2O was found to suppress nitrogenase-mediated H2 evolution completely at infinitely high pN2O . H2 was found to inhibit N2O-supported NH3 production but not N2O-supported N2 production . The steady-state kinetics of N2O reduction showed a good fit to Michaelis-Menten kinetics with a Km for N2O of 5 mM at 30 degrees C, corresponding to 24 kPa of N2O . A model is proposed that fits the observed results. Vaccine, 1986 Mar, 4(1), 15 - 20 Safety and immunogenicity of a polyvalent Klebsiella capsular polysaccharide vaccine in humans; Cryz SJ Jr et al.; A polyvalent Klebsiella vaccine composed of six serotypes of capsular polysaccharides (K2, K3, K10, K21, K30, and K55) was developed and its safety and immunogenicity evaluated in humans . Highly purified capsular antigens were treated in 0.1 N NaOH in 95% ethanol to detoxify trace amounts of contaminating lipopolysaccharide (LPS) . The vaccine was nontoxic and nonpyrogenic for animals . A total of 40 individuals received either 25 or 50 micrograms of each represented antigen subcutaneously . Reactions to vaccination, where noted, were transient and mild in nature . An immunizing dose of 50 micrograms of each antigen (300 micrograms total) elicited a fourfold or greater immunoglobulin G (IgG) response to all vaccine antigens in greater than 80% of vaccinees . Generally, the serospecificity of the antibody response was limited to those capsular antigens included in the vaccine . IgG isolated from the serum of vaccinees was found to be highly protective against fatal experimental Klebsiella K2 burn wound sepsis indicating that the functional antibody is elicited following vaccination. Biochem J, 1986 Mar 1, 234(2), 343 - 7 Structural and kinetic studies on beta-lactamase K1 from Klebsiella aerogenes; Emanuel EL et al.; beta-Lactamase K1 from Klebsiella aerogenes 1082E hydrolyses both penicillins and cephalosporins comparably and is inhibited by mercurials but not by cloxacillin . These properties distinguish it from those other beta-lactamases that have been allotted to classes on the basis of their amino sequences . beta-Lactamase K1 has been isolated by affinity chromatography; its composition shows resemblances to class A beta-lactamases . Moreover, the N-terminal sequence is similar to those of class A beta-lactamases: there is about 30% identity over the first 32 residues . Furthermore, a putative active-site octapeptide has been isolated and its sequence is similar to the region around the active-site serine residue in class A beta-lactamases . There is one thiol group in beta-lactamase K1; it is not essential for activity . The pH-dependence of kcat . and kcat./Km for the hydrolysis of benzylpenicillin by beta-lactamase K1 were closely similar, suggesting that the rate-determining step is cleavage of the beta-lactam ring. Prostaglandins Leukot Med, 1986 Mar, 21(3), 279 - 97 Regulation of prostaglandin E2 and plasminogen activator by various immunomodulators in human monocytes; Homo-Delarche F et al.; Mononuclear phagocyte populations and monocytes are able to produce, among numerous substances, a neutral protease, i.e . plasminogen activator (PA) and prostaglandins . Since it has been shown that prostaglandins (PGs) and particularly PGE2 could exert an inhibitory effect on PA production by macrophages, we have measured the in vitro production of PA and PGE2 by monocytes isolated from healthy donors . These monocytes were cultured either in the absence or the presence of various immunomodulators: lipopolysaccharide from E . coli, concanavalin A and RU 41740 or Biostim a broad spectrum immunostimulating agent isolated from Klebsiella pneumoniae (Cassenne Laboratories, France) . The production of PGE2 was proportional to the number of monocytes per incubation, and at a given cell concentration varied greatly from one subject to another . When considering PGE2 productions, the type of the response to the different immunomodulators varied from subject to subject and ranged from stimulation to no effect, or even inhibition . Moreover, a statistically significant, inverse relationship exists between the spontaneous production of PGE2 and the effect of each immunomodulator . For a given subject, all agents always acted in the same way and there was an inverse relationship between the effects of the immunomodulators on plasminogen activator and PGE2 production. J Med Microbiol, 1986 Mar, 21(2), 133 - 7 The role of K antigens as virulence factors in Klebsiella; Simoons-Smit AM et al.; The importance of K antigen of Klebsiella as a virulence factor was studied in nine pairs of K+ and K- strains, each pair isogenic apart from the presence of K antigen . Loss of K antigen by nine K+ strains resulted in the reduced virulence of their K- variants in a mouse-skin model . This reduced virulence of K- strains for mice may be explained in all strains by a higher degree of phagocytosis as measured by chemiluminescence response of human polymorphonuclear leukocytes (PMNL) and in most strains by enhanced killing by either human PMNL or human serum or both . Although the protective role of the K antigen in serum-induced killing and killing by PMNL was generally evident, our results also suggested that other virulence factors were sometimes involved. Infect Immun, 1986 Mar, 51(3), 901 - 8 Mimicry of human histocompatibility HLA-B27 antigens by Klebsiella pneumoniae; Ogasawara M et al.; Anti-HLA-B27 monoclonal antibody M2, which was relatively specific for human histocompatibility antigen HLA-B27, was used to test several bacteria, some of which could potentially induce chronic arthritis in HLA-B27-positive individuals . Using the Western blot procedure, we observed positive reactions with 80,000- and 60,000-dalton antigens with one strain of Klebsiella pneumoniae . Reactivity was not observed with five other monoclonal antibodies which were not reactive with HLA-B27 antigens, nor was reactivity observed with seven other gram-negative bacteria, irrespective of their arthritis-causing potential . To test the validity of our observation, the 80,000-dalton Klebsiella cross-reactive antigen was isolated and used to generate an immune guinea pig serum . We found that the reactivity of this guinea pig serum with Klebsiella envelopes in an enzyme-linked immunosorbent assay was adversely affected by absorption with HLA-B27-positive cells . Our results support the existence of mimicry between HLA-B27 antigens and bacteria. Infect Immun, 1986 Mar, 51(3), 723 - 30 Effect of iron compounds on antibacterial function of human polymorphs and plasma; Ward CG et al.; Human plasma was bactericidal for small numbers of Klebsiella pneumoniae, but larger numbers grew slowly over a period of 24 h . Human polymorphs in a clot of autologous plasma had a bacteriostatic effect on relatively large numbers of bacteria for up to 24 h and were much more effective than plasma alone . The bactericidal effect of plasma could be abolished by saturating the plasma albumin and hemopexin with hematin, the haptoglobin with hemoglobin, and the transferrin with Fe3+ . Stimulation of bacterial growth in the presence of polymorphs depended on the degree of saturation of the plasma proteins which bind the Fe3+ or heme compounds . Hematin bound to albumin appeared to be readily available to K . pneumoniae . Free hemoglobin stimulated bacterial growth but the organism could not utilize the haptoglobin-hemoglobin complex . In the presence of polymorphs the addition of Fe3+ to give 60% saturation of the transferrin with iron led to rapid growth after a long delay (19 h) . Progressive increases in saturation above 60% gave correspondingly quicker growth, suggesting that the higher the saturation of transferrin, the easier it is for the bacteria to acquire iron . The antibacterial effect of the plasma appears to be critically dependent on the availability of iron for the bacteria, and this in turn affects the ability of the polymorphs to control bacterial growth. Ann Rheum Dis, 1986 Mar, 45(3), 190 - 7 Ankylosing spondylitis, HLA-B27, and Klebsiella: a study of lymphocyte reactivity of anti-Klebsiella sera; Singh B et al.; Twenty three anti-Klebsiella antisera were tested for their cytotoxic activity and four for their binding capacity for peripheral blood lymphocytes (PBL) from patients with HLA-B27 positive ankylosing spondylitis (AS+B27+) and from B27 positive (AS-B27+) and B27 negative (AS-B27-) healthy individuals . None of the antisera showed specific activity against PBL from any particular group . The antisera tested included two anti-Klebsiella K43 sera provided by an Australian group, who have reported them to be specifically cytotoxic for AS+B27+ PBL, four antisera raised against a Klebsiella K43 strain provided by this group, and an antiserum from another group, who have reported it as having increased binding capacity for AS+B27+ and AS-B27+ PBL compared with AS-B27- PBL . The results of other workers who have attempted to reproduce the results of either group are reviewed and the possible reasons for the repeated failure to confirm the reported findings are discussed. Proc Natl Acad Sci U S A, 1986 Mar, 83(6), 1636 - 40 In vitro synthesis of the iron-molybdenum cofactor of nitrogenase; Shah VK et al.; Molybdate- and ATP-dependent in vitro synthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase requires the protein products of at least the nifB, nifN, and nifE genes . Extracts of FeMo-co-negative mutants of Klebsiella pneumoniae and Azotobacter vinelandii with lesions in different genes can be complemented for FeMo-co synthesis . Both K . pneumoniae and A . vinelandii dinitrogenase (component I) deficient in FeMo-co can be activated by FeMo-co synthesized in vitro . Properties of the partially purified dinitrogenase activated by FeMo-co synthesized in vitro were comparable to those of dinitrogenase from the wild-type organism; e.g., ratios of acetylene- to nitrogen-reduction activities, as well as those of acetylene reduction activities to EPR spectrum peak height at g = 3.65, were very similar . A . vinelandii mutants UW45 and CA30 have mutations in a gene functionally equivalent to nifB of K . pneumoniae. Arthritis Rheum, 1986 Mar, 29(3), 358 - 62 Normal anti-Klebsiella lymphocytotoxicity in ankylosing spondylitis; Kinsella TD et al.; We compared in vitro lymphocytotoxicity (LCT) of peripheral blood lymphocytes (PBL), obtained from patients with ankylosing spondylitis (AS) and normal controls (NC) . Assays were performed with antibacterial antisera prepared from AS- and NC-derived Klebsiella and coliforms Escherichia coli . LCT assessed by eosin staining was not significantly different in PBL of 12 AS patients and 28 controls when reacted with 3 Klebsiella and 1 E coli antisera . LCT assessed by 51Cr release was not significantly different for PBL of 20 age- and sex-matched pairs of AS patients and NC when reacted with 3 Klebsiella and 1 E coli antisera . Similarly, LCT-51Cr of PBL of 15 matched AS and NC pairs was not significantly different for anti-K21, a serotype putatively implicated in Klebsiella-HLA-B27 antigenic cross-reactivity . Our results do not support the notion of molecular mimicry between Klebsiella and B27 in the pathogenesis of primary AS. J Med Microbiol, 1986 Mar, 21(2), 125 - 32 The influence of the O and K antigens of Klebsiella aerogenes on surface hydrophobicity and susceptibility to phagocytosis and antimicrobial agents; Williams P et al.; The surface hydrophobicity of Klebsiella aerogenes is influenced by the presence of capsular (K) and lipopolysaccharide (O) antigens . Loss of both K and O antigens (K-O-), but not the K antigen alone (K-O+), increased surface hydrophobicity and susceptibility to phagocytosis . Unheated serum (i.e., containing complement) increased the surface hydrophobicity and phagocytosis of the K-O+ and K-O- strains, but not of the K+O+ encapsulated parent strain . Despite the altered susceptibility to phagocytosis caused by the presence or absence of the K and O antigens, their loss did not influence sensitivity to a range of hydrophilic, hydrophobic or cationic antimicrobial agents. Antimicrob Agents Chemother, 1986 Feb, 29(2), 315 - 9 Plasmid-encoded amikacin resistance in multiresistant strains of Klebsiella pneumoniae isolated from neonates with meningitis; Woloj M et al.; Two multiresistant Klebsiella pneumoniae strains isolated from cerebrospinal fluid of human neonates were analyzed for their plasmid content . Two of the plasmids harbored by these strains, pJHCMW1 (11 kilobase pairs) and pJHCMW4 (75 kilobase pairs), carried genetic determinants for amikacin resistance . These plasmids also encoded resistance to kanamycin, tobramycin, and ampicillin which could be transferred to Escherichia coli by conjugation . Extracts from transconjugant derivatives carrying pJHCMW4 produced an acetyltransferase activity that acetylated all three aminoglycosides . Transconjugant derivatives carrying pJHCMW1 encoded both acetylating and phosphorylating activities . Southern blot hybridization analysis indicated considerable DNA homology between these two plasmids. Br J Ophthalmol, 1986 Feb, 70(2), 85 - 8 IgG and IgA immune response against klebsiella in HLA-B27-associated anterior uveitis; Kijlstra A et al.; Enteric infections with Gram-negative bacteria are thought to play an important part in HLA-B27-associated disease such as Reiter's syndrome and reactive arthritis . But the role of bacterial infections in HLA-B27-positive ankylosing spondylitis (AS) and acute anterior uveitis (AU) is still controversial . A special interest has recently been devoted to the role of klebsiella infection in HLA-B27-associated disease . We studied the humoral immune response against a 'cross-reactive' strain of Klebsiella pneumoniae in 62 patients with anterior uveitis and 33 healthy controls . The anterior uveitis patients were subdivided into 25 HLA-B27-negative patients without AS (B27- AU+ AS-), 17 HLA-B27-positive patients without ankylosing spondylitis (B27+ AU+ AS-), and 19 HLA-B27-positive patients with ankylosing spondylitis (B27+ AU+ AS+) . Total serum IgA was higher in patients than in controls in both the B27+ AU+ AS+ and B27+ AU+ AS- patients but not in the B27- AU+ AS- group . No abnormalities were observed in the total serum IgG levels . The level of both the IgG and IgA klebsiella antibodies did not differ in the various patient groups tested as compared with the controls . Comparisons between the patient groups showed that the IgG anti-klebsiella response was higher in B27-positive patients patients without AS than in those with AS . These results suggest that stimulation of mucosal surfaces may play a role in HLA-B27-associated anterior uveitis . Whether klebsiella organisms are involved in this stimulation remains unclear. Br J Exp Pathol, 1986 Feb, 67(1), 25 - 32 Passive immunization of mice against Klebsiella aerogenes; Roe EA et al.; Klebsiella vaccines were isolated by mild diafiltration techniques from culture filtrates of nine capsular types of K . aerogenes (K1, K2, K3, K15, K20, K35, K36, K44 & K63) . The bacteria were grown in a chemically defined medium in standardized conditions in a fermenter . The vaccines had a molecular weight of more than 20 000, a carbohydrate content of 40-89%, a protein content of between less than 1 and 16% and small amounts (0.6-1.2%) of lipopolysaccharide . Antisera raised in rabbits to the nine klebsiella vaccines were standardized by passive haemagglutination, immunoglobulin G enzyme-linked immunosorbent assay and by autologous passive protection tests . Each rabbit antiserum when passively transferred to mice showed a variable capacity to passively protect mice against lethal infections by a panel of ten capsular types of K . aerogenes (K1-K10) . Seven of the rabbit antisera protected mice against more than half of the challenge strains . A pool of six rabbit antisera (anti-K1, K2, K3, K20, K35 & K44) passively protected mice against lethal infections from strains of bacteria representing each of 77 capsular types of K . aerogenes. Eur J Clin Microbiol, 1986 Feb, 5(1), 13 - 7 Release of endotoxin from bacteria exposed to ciprofloxacin and its prevention with polymyxin B; Cohen J et al.; An in vitro system for studying the phenomenon of antibiotic-induced release of endotoxin from gram-negative bacteria is described . Endotoxin was measured by a quantitative limulus lysate microassay . Using a strain of Escherichia coli, ciprofloxacin was shown to cause an immediate and sustained increase in endotoxin release compared to control cultures whereas gentamicin did not, despite an equally rapid bactericidal effect . The potential value of the anti-endotoxin effect of polymyxin B was examined using a polymyxin B resistant strain of Klebsiella aerogenes . Ciprofloxacin alone caused endotoxin release, but when given with polymyxin B significantly less endotoxin was detected . These results indicate that in antibiotic-induced endotoxin release, the rate of cell death is less important than the site of antibiotic effect, and suggest that ciprofloxacin may act directly on the cell wall as well as on DNA gyrase . It is possible that polymyxin B could be used to reduce the extent of endotoxin release, and that this would be of clinical benefit. J Bacteriol, 1986 Feb, 165(2), 353 - 6 Influence of growth rate and iron limitation on the expression of outer membrane proteins and enterobactin by Klebsiella pneumoniae grown in continuous culture; Lodge JM et al.; The influence of the growth rate on outer membrane protein composition and enterobactin production was studied with Klebsiella pneumoniae grown under conditions of iron limitation in chemostats . More enterobactin was produced at fast (D = 0.4 h-1) and slow (D = 0.1 h-1) growth rates in continuous cultures than in either logarithmic- or stationary-phase batch cultures . When the growth rate was controlled under conditions of carbon limitation and the iron level was reduced to 0.5 microM, the iron-regulated outer membrane proteins and enterobactin were induced at the fast growth rate . At the slow growth rate, although the iron-regulated outer membrane proteins were barely visible, a significant level of enterobactin was still produced . These results suggest that under conditions of either carbon or iron limitation, the growth rate can influence the induction of the high-affinity iron uptake system of K . pneumoniae . Other outer membrane proteins, including a 39-kilodalton peptidoglycan-associated protein, were found to vary with the growth rate and nutrient limitation. EMBO J, 1986 Feb, 5(2), 399 - 407 Regulation of nitrogen metabolism in Azotobacter vinelandii: isolation of ntr and glnA genes and construction of ntr mutants; Toukdarian A et al.; The ntrA, ntrB and ntrC products are responsible for regulating the transcription of many genes involved in the assimilation of poor nitrogen sources in enteric bacteria . The presence of a similar system in the non-enteric bacterium Azotobacter vinelandii is reported here . Genes analogous to ntrA and ntrC were isolated from an A . vinelandii gene library by complementation of Escherichia coli mutants . The gene encoding glutamine synthetase, glnA, was also isolated and found to be adjacent to ntrC but distant from ntrA, as it is in enteric organisms . The cloned Azotobacter genes also complemented Klebsiella pneumoniae mutants and hybridized to K . pneumoniae ntrA, ntrC and glnA gene probes . The role of ntrA and ntrC in A . vinelandii was established by using Tn5 insertions in the cloned genes to construct mutants by marker exchange . These mutants show that both ntrA and ntrC are required for the utilization of nitrate as a nitrogen source . However, ntrC is not required for nitrogen fixation by A . vinelandii, in contrast with K . pneumoniae where both ntrA and ntrC are essential. J Biol Chem, 1986 Jan 15, 261(2), 772 - 8 Klebsiella pneumoniae nifM gene product is required for stabilization and activation of nitrogenase iron protein in Escherichia coli; Howard KS et al.; A series of plasmids encoding various Klebsiella pneumoniae nif (nitrogen fixation) genes were constructed to determine which were required to produce active iron (Fe) protein in Escherichia coli, a species which does not normally fix nitrogen . The greatest success was achieved with binary plasmid systems that produced nifA regulatory protein under the control of a tac promoter on one plasmid, which then induced synthesis of nifH and nifM proteins from their native promoter sites on a second plasmid . nifH protein, the monomeric subunit of Fe protein, produced in the presence of nifM constituted nearly 10% of the whole cell protein and exhibited the corresponding amount of C2H2-reducing activity in nitrogenase assays conducted in vitro . nifH protein formed in the absence of nifM constituted 4.7% of the whole cell protein and exhibited no detectable activity in assays of whole cell extracts . The plasmid-encoded Fe protein was purified to homogeneity and was found to be indistinguishable from that isolated from derepressed wild type K . pneumoniae, having a similar specific activity, approximately 4 Fe/dimer of 68 kDa, and similar epr features . Although these experiments do not exclude the participation of other E . coli gene products in the maturation of nifH protein, they limit the nif-specific genes required for active Fe protein production to nifA, nifH, and nifM . Since nifA is thought to be the required activator protein involved in nif operon transcription, the simplest explanation for these observations is that nifH codes for the peptide of the Fe protein, while nifM acts to convert this nifH peptide to the functioning Fe protein of nitrogenase . In the absence of nifM, only an inactive nifH polypeptide is produced. Microbiol Immunol, 1986, 30(6), 553 - 9 Microbial adjuvant and autoimmunity . IV . The induction of thyroid lesions in syngeneic X-irradiated mice by the transfer of spleen cells from mice immunized with thyroid extract and Klebsiella O3 lipopolysaccharide; Yokochi T et al.; The role of humoral and cellular immune responses in the initiation and maintenance of autoimmune thyroiditis was investigated in mice immunized with syngeneic thyroid extract and Klebsiella O3 lipopolysaccharide (KO3 LPS) as an adjuvant . The transfer of spleen cells from hyperimmunized mice to 400R-irradiated syngeneic mice produced definite lesions in the thyroid glands, whereas the transfer of immune sera failed to do so . No lesions were induced in normal intact mice by the same transfer of sera and spleen cells from hyperimmunized mice . It was suggested that the induction of thyroiditis by immunization using KO3 LPS adjuvant is primarily due to cell-mediated immunity and that pretreatment of mice by X-irradiation is essential for production of the lesions . The role of X-irradiation in the induction of thyroiditis was discussed. Microbiol Immunol, 1986, 30(1), 25 - 33 Stability of the hexagonal lattice structure formed by an R-form lipopolysaccharide of Klebsiella: study of long-range stability; Kato N et al.; The R-form lipopolysaccharide (LPS) from Klebsiella strain LEN-111 (O3-:K1-) forms a hexagonal lattice structure with a lattice constant of 14 to 15 nm when it is precipitated by addition of two volumes of 10 mM MgCl2-ethanol . The stability of this hexagonal lattice structure in long-term incubation at 4 C was investigated . The hexagonal lattice structure was stable for at least 220 days when the LPS was suspended in distilled water, but it had been disintegrated into a rough mesh-like structure when the LPS was suspended in 50 mM tris(hydroxymethyl)aminomethane (Tris) buffer, pH 8.5, at 4 C for 60 days . Half of the Mg bound to the LPS was released when the LPS was suspended in Tris buffer for 60 days, whereas Mg was not released when it was suspended in distilled water even for 220 days . By contrast, it was stable for at least 220 days in Tris buffer containing 5 mM MgCl2 . The LPS suspended in Tris buffer for 60 days, at which time the structure had been disintegrated, could be restored to the original hexagonal lattice structure within 24 hr by addition of 5 mM MgCl2 . From these results it is concluded that the hexagonal lattice structure of the LPS retains long-range stability if Mg bound to the LPS is not released from the LPS. Microbiol Immunol, 1986, 30(1), 13 - 23 Stability of the hexagonal lattice structure formed by an R-form lipopolysaccharide of Klebsiella: decrease in the stability by electrodialysis and recovery by addition of the magnesium; Kato N et al.; The R-form lipopolysaccharide (LPS) from Klebsiella strain LEN-111 (O3-:K1-) forms a hexagonal lattice structure with a lattice constant of 14 to 15 nm when it is precipitated by addition of two volumes of 10 mM MgCl2-ethanol . When the LPS was suspended in various buffers (50 mM) at pH 2 to 12 for 24 hr at 4 C, at pH 2 and 3 pits of the hexagonal lattice structure markedly disappeared, at pH 4 to 8.5 the lattice structure was stable, and at pH 9 to 12 it tended to loosen somewhat . The LPS from which cations were removed by electrodialysis retained the ability of hexagonal assembly, although the lattice constant of the hexagonal lattice of the electrodialyzed LPS was large . The lattice structure of the electrodialyzed LPS was much more labile than that of the non-electrodialyzed LPS at alkaline pH levels and the former was completely disintegrated into ribbon-like structures when the LPS was suspended in 50 mM Tris buffer at pH 7.7 or higher . However, the electrodialyzed LPS formed a hexagonal lattice structure in Tris buffer at pH 8.5 containing 0.1 to 100 mM MgCl2 . The lattice constants of the hexagonal lattice formed by the electrodialyzed LPS at 10 or 100 mM MgCl2 were very similar to that of the lattice of the non-electrodialyzed LPS . From these results it is concluded that the lability of the hexagonal lattice structure of the electrodialyzed LPS at alkaline conditions is due to removal of Mg2+ by electrodialysis. Int J Immunopharmacol, 1986, 8(6), 589 - 92 Immunostimulation of blood monocyte function by RU 41.740 (Biostim) in patients with chronic bronchitis; Nielsen H et al.; RU 41.740 (Biostim), a glycoprotein extract from Klebsiella pneumoniae, which is effective in heightening resistance to experimental infections in animals, was examined with regard to influence on human blood monocyte function in vitro after administration to patients with chronic bronchitis in a randomized, double-blind clinical trial . Twelve patients were given Biostim orally (seven patients received 8 mg daily and five patients received 2 mg daily) while eight patients received placebo . In the Biostim group monocyte phagocytosis increased from 1.4 yeast cells per monocyte before therapy to 3.1 yeast cells per monocyte after 1 month (P less than 0.05) with no further increase after 3 months . Monocyte candidacidal activity increased in the Biostim group from 51% before therapy to 69% after 1 month (P less than 0.05) with a decrease to 57% after 3 months . As expected, placebo had no significant effect on monocyte functions . There was no difference between the improvement in the high dose vs the low dose group . These results suggest that Biostim exerts part of its immunostimulatory property by stimulating blood monocyte antimicrobial function. Int J Immunopharmacol, 1986, 8(7), 687 - 90 Enhancement of delayed cutaneous hypersensitivity by oral administration of RU 41740 (Biostim) in lymphoma patients--a randomized double blind multicentric trial; Lang JM et al.; RU 41740 (Biostim) is a glycoprotein extract from Klebsiella pneumoniae with immunostimulating properties in animal studies and in in vitro assays of human leukocyte functions . The present randomized double blind phase II trial showed that RU 41740 significantly (P less than 0.05, Mann & Whitney test) enhanced delayed cutaneous hypersensitivity to seven recall antigens (Multitest system) in lymphoma patients in unmaintained complete remission when given orally for 14 days at a daily dose of 8 mg, and compared to placebo . Daily doses of 2 and 32 mg were uneffective. Gene, 1986, 45(1), 59 - 65 Identification and cloning of Thiobacillus ferrooxidans structural nif genes in Escherichia coli; Pretorius IM et al.; The presence of nucleotide sequences which are homologous to the nifHDK genes of Klebsiella pneumoniae was demonstrated in total DNA preparations from five different iron-oxidizing Thiobacillus ferrooxidans strains . A non-iron-oxidizing Thiobacillus novellus strain and a heterotrophic Acidiphilium strain, which occurs in close association with T . ferrooxidans, did not contain DNA homologous to the nifHDK genes . The T . ferrooxidans ATCC33020 nifHDK genes were cloned and their arrangement was characterized on a 6.7-kb insert in pIMP16. Antimicrob Agents Chemother, 1986 Jan, 29(1), 165 - 7 Mechanism of resistance to silver ions in Klebsiella pneumoniae; Kaur P et al.; The uptake of silver by an experimentally derived silver-resistant Klebsiella pneumoniae strain was three to four times lower than the uptake by a susceptible strain . Spheroplasts of the two strains showed no difference in uptake . AgNO3 at a concentration of 40 micrograms/ml decreased the succinate dehydrogenase activity in susceptible and resistant strains by 100 and 18%, respectively . More than one resistance mechanism may be involved. Int J Immunopharmacol, 1986, 8(2), 147 - 54 Modulation of murine hemopoiesis by repeated injections of a glycoprotein extract from Klebsiella pneumoniae; Andreux JP et al.; RU 41740, an immunomodulating agent extracted from the cell wall of Klebsiella pneumoniae, was tested for its ability to modulate hemopoiesis in mice treated intraperitoneally for ten days . In such conditions, a moderate anemia could be observed, which was rapidly reversible after the end of treatment . This anemia may partly result from a decrease of bone marrow erythroid colony forming units (CFU-E), this being incompletely compensated by the intrasplenic erythropoiesis . Moreover, the amount of granulocytes and their progenitors, i.e . granulocyte/macrophage colony forming cells (GM-CFC), increased both in the bone marrow and spleen . It is proposed that this effect of RU 41740 on granulopoiesis, which could explain the better resistance to infections in treated animals as observed elsewhere, is partly related to an increase of colony stimulating activity (CSA). Zentralbl Mikrobiol, 1986, 141(1), 3 - 9 {Occurrence of gentamicin-resistant coliforms in river water samples}; Stelzer W et al.; In river water samples 0.006...0.3% of total coliforms were detected as gentamicin resistant . Among gentamicin resistant coliforms most strains (86.5%) could be identified as Klebsiella . Most frequently a Gm-plasmid of about 150 Md was isolated harboring in Klebsiella oxytoca, K . pneumoniae and E . coli strains . All gentamicin resistant wild strains transferred the resistance to E . coli K-12 . Most Klebsiella strains harbored several plasmids, R-plasmids and others . There were not any differences in plasmid profiles between sensitive and resistant strains. J Bacteriol, 1986 Jan, 165(1), 175 - 80 Transcriptional analysis of promoter mutations in the Klebsiella pneumoniae nifHDKY operon; Lers A et al.; Previously isolated promoter mutations that allow expression of the Klebsiella pneumoniae nifHDKY operon in the absence of nifA (R . Bitoun, J . Berman, A . Zilberstein, D . Holland, J.B . Cohen, D . Givol, and A . Zamir, Proc . Natl . Acad . Sci . USA, 80:5812-5816, 1983) were further characterized . pRB1 and pRB5, containing, respectively, point and duplication mutations in the nifHDKY regulatory region, were transformed into Escherichia coli and K . pneumoniae hosts with different nifA and ntrA backgrounds . nif transcription start sites were determined by nuclease S1 mapping . The results indicated that nifA-independent expression from both mutants did not require ntrA . Transcription from pBR5 started 3 base pairs (bp) upstream of the start site of nif-regulated transcription and could stem from a canonical promoter sequence generated at the junction between the two copies of the duplicated sequence . In the presence of nifA-ntrA, transcription from pRB5 started predominantly at the site characteristic of the nif-regulated promoter . The site of constitutive transcription initiation in pRB1 was located 33 bp upstream of the point mutation and 40 bp upstream of the start of nifA-ntrA-activated transcription . Low-level transcription from the upstream site was also evident, in the absence of nifA or nifA or both, with the plasmid containing the wild-type nifHDKY regulatory region . However, when nifA and ntrA were present to activate transcription from the major nif promoter, no activity was evident from the upstream site in either pRB1 or the parental plasmid . Thus, the mutation enhanced the activity of a pre-existing constitutive promoter, the activity of which was repressed on nifA-ntrA activation of the major nif promoter. Gene, 1986, 45(3), 281 - 8 Deletion loop mutagenesis of the nifL promoter from Klebsiella pneumoniae: role of the -26 to -12 region in promoter function; Khan H et al.; Nine single C-to-T transitions were introduced into the -26 to -12 region of the Klebsiella pneumoniae nifL promoter by bisulphite mutagenesis of M13 heteroduplexes containing a 15 nucleotide single-stranded loop . Mutant promoter fragments were inserted into translational lac fusion vectors to utilise beta-galactosidase activity as a measure of promoter efficiency . Mutations in invariant nucleotides found in the consensus sequence for nif promoters gave a strong 'down' promoter phenotype with respect to transcriptional activation . Mutations in semi-conserved residues had a much weaker down phenotype, whereas a mutation which increased homology to the consensus sequence enhanced promoter strength . One mutant showed increased activation by ntrC and decreased activation by nifA. Med Pediatr Oncol, 1986, 14(5), 255 - 61 Infections in patients with non-small-cell lung cancer treated with intensive induction chemotherapy; Fuks JZ et al.; The records of 65 consecutive patients with non-small-cell lung cancer (NSCLC) treated with intensive induction chemotherapy were reviewed to study the infectious complications during therapy and to analyze the relationship of the frequency of infections to various predisposing factors . A total of 44 infectious episodes were observed among 30 of the 65 patients . Of the 44 infections, 18 were microbiologically documented and 19 clinically . Seven (16%) infections were without microbiological or clinical documentation and were categorized as "possible" infections . Among the 18 microbiologically documented infections, fifteen (83%) were caused by bacteria and three by fungi . The most frequent bacteria identified in 11 (61%) of the 18 infections were gram-negative organisms, Escherichia coli and Klebsiella pneumoniae being the most frequent . Eight of the 44 infections were associated with bacteremia and three with microbiologically documented pneumonias . There were three drug-related infectious deaths, two associated with bacteremia and one with possible infections . All 44 infectious episodes presented with WBC counts of less than 1,000/microliter, and 34 (79%) had WBC counts of less than 500/microliter . We observed that during therapy, patients with poor performance status (less than 80%) are at a much higher risk to develop infectious complications than those with good performance status (greater than 80%; p less than .001) . Although encouraging responses with intensive chemotherapy have been reported for NSCLC in several studies, a major impact of chemotherapy on the survival of patients with this disease has yet not been established . Thus, intensive chemotherapy for NSCLC should remain an experimental treatment modality and should be offered only to patients with good prognostic factors such as those defined by pretreatment performance status. Microbiol Immunol, 1986, 30(8), 761 - 76 Modification of susceptibility to Klebsiella pneumoniae during murine cytomegalovirus infection; Leung WC et al.; The effect of murine cytomegalovirus (MCMV) infection on susceptibility to bacterial infection was studied in mice by a combination of intraperitoneal (ip) inoculation of a sublethal dose of MCMV with subsequent ip challenge of 2 X 10(3) cfu of a strain of Klebsiella pneumoniae (KP) . When given alone, KP produced a mortality of 30-40% . Mortality was increased when KP was given 1 to 7 days after MCMV injection with the peak increase at the 4th to 5th day when 100% mortality occurred . Virus levels in various organs of mice infected with MCMV alone, or superinfected with KP did not differ . Bacterial counts on the other hand, showed that increased mortality in mixed MCMV and KP infected mice was due to an uncontrolled growth of bacteria at the site of primary lodgment, i.e., the peritoneum, and severe systemic infection . Neutrophil response to growth of KP during the first 3 days of bacterial infection was defective in MCMV infected mice . While the initial clearance of KP from the blood was more efficient in MCMV infected mice, probably due to activated reticuloendothelial function, it did not protect the mice against KP infection . Using the in vivo model, it was shown that poor neutrophil response and possibly other defective neutrophil functions were responsible for increased mortality to KP infection in MCMV infected mice. Infection, 1986, 14 Suppl 1, S36 - 9 Chemotherapeutic effects of ofloxacin (HOE 280) and other quinolone-carboxylic derivates in the treatment of experimental lung infections due to Klebsiella pneumoniae DT-S in mice; Klesel N et al.; The chemotherapeutic activity of ofloxacin (HOE 280), a new pyridone-carboxylic acid derivative, was compared with that of other drugs in the same group, including norfloxacin, ciprofloxacin, enoxacin and in some cases pipemidic acid and nalidixic acid . The test model used was experimental pneumonia caused by Klebsiella pneumoniae DT-S in mice . In the treatment of pneumonic mice, ofloxacin was 4 to 18 times more effective than norfloxacin and enoxacin and in most cases slightly more effective than ciprofloxacin . Pipemidic acid and nalidixic acid showed only low activity or proved to be inactive . In studies on the bactericidal activity of the compounds in vivo, ofloxacin produced a more pronounced effect than ciprofloxacin . With norfloxacin and enoxacin, a bactericidal effect in the infected tissue of the animals was only observed during the first hours after treatment. Tokai J Exp Clin Med, 1986, 11 Suppl, 59 - 63 Experimental analysis of concurrent viral and bacterial infection in the mouse; Hashimoto K; The roles of both microorganisms in combined murine cytomegalovirus (MCMV) and Klebsiella pneumoniae (KP) infection in the mouse, and host response to the infection were investigated . Increased mortality in mixed MCMV and KP infected mice was observed, and this was due to uncontrolled growth of bacteria in the peritoneal cavity, and severe generalized infection . Neutrophil response to growth of KP was defective in MCMV infected mice . While the initial clearance of KP from the blood was more efficient in MCMV-infected mice, it did not protect the mice against KP infection . In the in vitro experiments, the neutrophil chemotaxis of MCMV-infected mice to KP was found to be lowered . Tissue extract and serum of MCMV-infected mice exhibited chemotactic activity for neutrophils, and, at the same time contained some factor(s) which reduced chemotaxis of normal neutrophils to KP. Carbohydr Res, 1985 Dec 15, 145(1), 67 - 80 Structural investigation of the capsular polysaccharide of Klebsiella serotype K35; Dutton GG et al.; The structure of the capsular polysaccharide (K antigen) of Klebsiella K35 has been established as having the pentasaccharide repeating unit shown ("four plus one" type) . The structural investigation utilized the techniques of methylation, beta-elimination, Smith degradation, and partial hydrolysis . N.m.r . spectroscopy (1H and 13C) was used extensively to establish the configurations of the anomeric linkages and to delineate the sequence of the sugars in the structure of the polysaccharide . (Formula: see text). Carbohydr Res, 1985 Dec 15, 145(1), 59 - 66 Structural studies of the O-antigen polysaccharides of Klebsiella O5 and Escherichia coli O8; Jansson PE et al.; The O-antigen polysaccharides of Klebsiella serotype O5 and Escherichia coli serotype O8 are serologically very similar or identical . The structures of these two polysaccharides have now been re-investigated . N.m.r . spectroscopy, chromium trioxide oxidation, hydrolysis with a specific phage enzyme, and f.a.b . mass spectrometry were the principal methods used . It is concluded that the O-antigen has the following structure, in which D-Man3Me is 3-O-methyl-D-mannose and n is approximately 10 . (Formula: see text) Biosynthetic studies indicate that these antigens are synthesised by addition of D-mannopyranosyl groups to the "non-reducing" end of the mannan chain, and it seems possible that addition of a 3-O-methyl-D-mannopyranosyl group involves termination. Biochim Biophys Acta, 1985 Dec 13, 843(3), 214 - 29 Regulation of glutamine synthetase, aspartokinase, and total protein turnover in Klebsiella aerogenes; Fulks RM et al.; When suspensions of Klebsiella aerogenes are incubated in a nitrogen-free medium there is a gradual decrease in the levels of acid-precipitable protein and of aspartokinase III (lysine-sensitive) and aspartokinase I (threonine-sensitive) activities . In contrast, the level of glutamine synthetase increases slightly and then remains constant . Under these conditions, the glutamine synthetase and other proteins continue to be synthesized as judged by the incorporation of {14C}leucine into the acid-precipitable protein fraction and into protein precipitated by anti-glutamine synthetase antibodies, by the fact that growth-inhibiting concentrations of chloramphenicol also inhibit the incorporation of {14C}leucine into protein and into protein precipitated by anti-glutamine synthetase antibody, and by the fact that chloramphenicol leads to acceleration in the loss of aspartokinases I and III and promotes a net decrease in the level of glutamine synthetase and its cross-reactive protein . The loss of aspartokinases I and III in cell suspensions is stimulated by glucose and is inhibited by 2,4-dinitrophenol . Glucose also stimulates the loss of aspartokinases and glutamine synthetase in the presence of chloramphenicol . Cell-free extracts of K . aerogenes catalyze rapid inactivation of endogenous glutamine synthetase as well as exogenously added pure glutamine synthetase . This loss of glutamine synthetase is not associated with a loss of protein that cross-reacts with anti-glutamine synthetase antibodies . The inactivation of glutamine synthetase in extracts is not due to adenylylation . It is partially prevented by sulfhydryl reagents, Mn2+, antimycin A, 2,4-dinitrophenol, EDTA, anaerobiosis and by dialysis . Following 18 h dialysis, the capacity of extracts to catalyze inactivation of glutamine synthetase is lost but can be restored by the addition of Fe2+ (or Ni2+) together with ATP (or other nucleoside di- and triphosphates . After 40-60 h dialysis Fe3+ together with NADH (but not ATP) are required for glutamine synthetase inactivation . The results suggest that accelerated protein degradation in cells exposed to nitrogen-limited conditions reflects the differential destruction of some proteins, including aspartokinases I and III, in order to sustain the biosynthesis of others such as glutamine synthetase . The loss of glutamine synthetase activity in cell-free extracts is likely mediated in part by mixed-function oxidation systems and could represent a 'marking' step in protein turnover. S Afr Med J, 1985 Dec 7, 68(12), 858 - 63 Factors associated with fatal cases of measles . A retrospective autopsy study; Beckford AP et al.; Clinical and laboratory records at Red Cross War Memorial Children's Hospital, Cape Town, for the period 1976-1982 were reviewed to determine factors associated with fatal cases of measles . Pneumonia was found to be the commonest lethal complication of measles . Supervening infections by both viral (especially adenovirus and herpesvirus) and bacterial (especially Klebsiella species and Pseudomonas) agents played a prominent role in causing the deaths of children who had recently been infected with measles . Severe malnutrition was present with almost equal frequency among those dying of measles and those dying from other causes, but was significantly (P less than 0,02) less common in measles patients who survived . Peripheral lymphopenia and depletion of T-cell zones in the lymph nodes and spleen were more common in those who died from measles than in others. J Antimicrob Chemother, 1985 Dec, 16(6), 727 - 34 The therapeutic efficacy of AC-1370 against experimental infections in mice; Obana Y et al.; The therapeutic efficacy of AC-1370, a new cephalosporin, was compared with that of cefoperazone in experimental infections in mice . For intraperitoneal infections caused by Escherichia coli and Klebsiella pneumoniae, the efficacy of AC-1370 was inferior to that of cefoperazone . On the other hand, AC-1370 was more potent than cefoperazone against pseudomonal infections . The MICs of AC-1370 against the test strains, however, were inferior or equivalent to those of cefoperazone . Serum and peritoneal exudate levels of AC-1370 in infected mice were more durable than those of cefoperazone . The relationship between AC-1370 and neutrophils was investigated . The efficacy of AC-1370 was more dependent on neutrophils than that of cefoperazone . AC-1370 did not enhance the neutrophil functions of migration and phagocytosis to any significant extent. Carbohydr Res, 1985 Dec 1, 144(2), 263 - 76 Structure of the capsular polysaccharide of Klebsiella serotype K79; Dutton GG et al.; The structure of the capsular polysaccharide from Klebsiella K79 was determined by the techniques of methylation, periodate oxidation, beta-elimination, chromic acid oxidation, and partial hydrolysis . N.m.r . spectroscopy (1H and 13C) was used extensively to establish the nature of the anomeric linkages of the polysaccharide and of oligosaccharides derived through degradative procedures . The polysaccharide was found to have the heptasaccharide, "5 + 2" repeating unit: (Formula: see text). Carbohydr Res, 1985 Dec 1, 144(2), 251 - 61 Novel oligosaccharides obtained by bacteriophage degradation of the polysaccharide from Klebsiella serotype K26; Di Fabio JL et al.; Bacteriophage phi 26 was used to depolymerize the polysaccharide of Klebsiella K26, yielding three oligosaccharides . The major product was the heptasaccharide repeating unit, with one of the minor products being the fourteen-sugar oligosaccharide corresponding to two repeating units . The other minor product was unusual since it was a hexasaccharide devoid of the terminal, pyruvate-containing galactose unit present in the side chain of the normal repeating unit . Phage phi 26 was shown to act as a beta-galactosidase, and hence it may have the ability to remove the terminal beta-galactose residue in the side chain. Acta Pathol Microbiol Immunol Scand {C}, 1985 Dec, 93(6), 233 - 43 NK-cell stimulating properties of a membrane proteoglycane from non-capsulated Klebsiella pneumoniae biotype a; Normier G et al.; Upon testing the individual fractions of a composite bacterial vaccine for biological activities, a potent immuno-stimulatory capacity could be demonstrated within a crude membrane proteoglycan preparation from Klebsiella pneumoniae . One of its characteristic features was the capacity to induce alpha-type interferon and increased NK activity in vivo in mice, following intraperitoneal or oral administration . A highly purified fraction from the crude preparation was obtained using alkaline hydrolysis, delipidation and size fractionation . This fraction was shown to be a very potent inducer of NK cells in vivo or in vitro, where in the latter systems concentrations as low as 0.1 microgramme per ml were highly efficient. Appl Environ Microbiol, 1985 Dec, 50(6), 1444 - 50 Optimizing aerobic conversion of glycerol to 3-hydroxypropionaldehyde; Slininger PJ et al.; When cells of Klebsiella pneumoniae NRRL B-199 (ATCC 8724) were grown aerobically on a rich glycerol medium and then suspended in buffer supplemented with semicarbazide and glycerol, aerobic conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA) ensued . Depending on conditions, 0.38 to 0.67 g of 3-HPA were formed per gram of glycerol consumed . This means that up to 83.8% of the carbon invested as glycerol could potentially be recovered as the target product, 3-HPA . Production of 3-HPA was sensitive to the age of cells harvested for resuspension and was nonexistent if cells were cultivated on glucose instead of glycerol as the sole carbon source . Compared with 24- and 72-h cells, 48-h cells produced 3-HPA at the highest rate and with the greatest yield . The cell biomass concentration present during the fermentation was never particularly critical to the 3-HPA yield, but initial fermentation rates and 3-HPA accumulation displayed a linear dependence on biomass concentration that faded when biomass exceeded 3 g/liter . Fermentation performance was a function of temperature, and an optimum initial specific 3-HPA productivity occurred at 32 degrees C, although the overall 3-HPA yield increased continuously within the 25 to 37 degrees C range studied . The pH optimum based on fermentation rate was different from that based on overall yield; 8 versus 7, respectively . Initial glycerol concentrations in the 20 to 50 g/liter range optimized initial 3-HPA productivity and yield. J Bacteriol, 1985 Dec, 164(3), 1081 - 7 Biosynthesis of the iron-molybdenum cofactor and the molybdenum cofactor in Klebsiella pneumoniae: effect of sulfur source; Ugalde RA et al.; NifQ- and Mol- mutants of Klebsiella pneumoniae show an elevated molybdenum requirement for nitrogen fixation . Substitution of cystine for sulfate as the sulfur source in the medium reduced the molybdenum requirement of these mutants to levels required by the wild type . Cystine also increased the intracellular molybdenum accumulation of NifQ- and Mol- mutants . Cystine did not affect the molybdenum requirement or accumulation in wild-type K . pneumoniae . Sulfate transport and metabolism in K . pneumoniae were repressed by cystine . However, the effect of cystine on the molybdenum requirement could not be explained by an interaction between sulfate and molybdate at the transport level . Cystine increased the molybdenum requirement of Mol- mutants for nitrate reductase activity by at least 100-fold . Cystine had the same effect on the molybdenum requirement for nitrate reductase activity in Escherichia coli ChlD- mutants . This shows that cystine does not have a generalized effect on molybdenum metabolism . Millimolar concentrations of molybdate inhibited nitrogenase and nitrate reductase derepression with sulfate as the sulfur source, but not with cystine . The inhibition was the result of a specific antagonism of sulfate metabolism by molybdate . The effects of nifQ and mol mutations on nitrogenase could be suppressed either by the addition of cystine or by high concentrations of molybdate . This suggests that a sulfur donor and molybdenum interact at an early step in the biosynthesis of the iron-molybdenum cofactor . This interaction might occur nonenzymatically when the levels of the reactants are high. Infect Immun, 1985 Dec, 50(3), 787 - 95 Importance of a lipopolysaccharide-containing extracellular toxic complex in infections produced by Klebsiella pneumoniae; Straus DC et al.; A Klebsiella pneumoniae serotype 2 strain was examined for its ability to produce extracellular toxic material . The organism was grown to the stationary phase in a defined medium, and the toxic material was isolated by ultrafiltration-ion-exchange chromatography on DEAE-Sephacel and gel filtration chromatography on Sepharose 4B or 2B . It was found to be comprised of 63% capsular polysaccharide, 30% lipopolysaccharide, and 7% protein and possessed a 50% lethal dose (when injected intraperitoneally into mice) of 393 +/- 45 micrograms . The toxicity appeared to be associated with the endotoxin portion of the compound, because boiling for 15 min and exposure to proteolytic enzymes had no effect on the toxicity . However, saponification destroyed the toxicity of the compound . Studies employing radial immunodiffusion examining the sera of mice infected with this organism demonstrated in vivo production of the complex at levels sufficiently high to produce death . When sublethal amounts of this complex were placed in the lungs of specific-pathogen-free mice, the lung pathology observed after 24, 48, and 72 h was similar to the damage caused by an active K . pneumoniae lobar pneumonia . These data indicate that this extracellular toxic compound produced by K . pneumoniae may be responsible for the lethality and lung tissue destruction normally associated with an active lobar pneumonia caused by this organism. Clin Exp Immunol, 1985 Dec, 62(3), 672 - 7 The modification of HLA-B27-positive lymphocytes by the culture filtrate of Klebsiella K43 BTS 1 is a metabolically active process; Sullivan JS et al.; The culture filtrate of some bacteria contains a modifying factor which specifically interacts with the cells of HLA-B27-positive normal individuals (BS+ AS-); this modification results in a serologically detectable change similar to that seen on the cells of patients with ankylosing spondylitis (B27+ AS+) . Puromycin, 5-azacytidine, actinomycin D, cyclosporin A, dexamethasone, indomethacin, colchicine, quinacrine, chlorpromazine but not tunicamycin can effectively inhibit the in vitro modification of HLA-B27-positive lymphocytes from normal individuals . These studies suggest that a number of different metabolic pathways may be involved in the modification of B27+ AS- lymphocytes by certain enteric antigens. Clin Exp Immunol, 1985 Dec, 62(3), 662 - 71 A reinvestigation of the cross-reactivity between Klebsiella and HLA-B27 in the aetiology of ankylosing spondylitis; Georgopoulos K et al.; The existence of cross-reactivity between Klebsiella antigens and cells from donors who are HLA-B27 positive and exhibit ankylosing spondylitis (AS) has been reinvestigated . Cells and antisera from different laboratories have been tested together using simultaneously microcytoxicity, chromium release and enzyme linked immunosorbent assays (ELISA) . No reproducible interaction has been found . Mitogenic stimulation did not induce cross-reactivity and 'transformation' of B27+AS- cells by Klebsiella culture supernatants failed . Two transformed cell lines from B27+ AS+ donors exhibited specific cross reaction with two anti-Klebsiella antisera but only by chromium release . Immunoprecipitation with these cells and antisera showed the absence of any AS+ -specific antigen . It is concluded that the involvement of Klebsiella in ankylosing spondylitis through simple immunological cross-reactivity or through interaction with HLA-B27 is unlikely. J Bacteriol, 1985 Dec, 164(3), 1386 - 9 Cloning and expression of the Thiobacillus ferrooxidans glutamine synthetase gene in Escherichia coli; Barros ME et al.; The glutamine synthetase (GS) gene glnA of Thiobacillus ferrooxidans was cloned on recombinant plasmid pMEB100 which enabled Escherichia coli glnA deletion mutants to utilize (NH4)2SO4 as the sole source of nitrogen . High levels of GS-specific activity were obtained in the E . coli glnA deletion mutants containing the T . ferrooxidans GS gene . The cloned T . ferrooxidans DNA fragment containing the glnA gene activated histidase activity in an E . coli glnA glnL glnG deletion mutant containing the Klebsiella aerogenes hut operon . Plasmid pMEB100 also enabled the E . coli glnA glnL glnG deletion mutant to utilize arginine or low levels of glutamine as the sole source of nitrogen . There was no detectable DNA homology between the T . ferrooxidans glnA gene and the E . coli glnA gene. Nucleic Acids Res, 1985 Nov 11, 13(21), 7621 - 38 Site-directed mutagenesis of the Klebsiella pneumoniae nifL and nifH promoters and in vivo analysis of promoter activity; Buck M et al.; The role of conserved nucleotides in nitrogen-fixation promoter function has been examined using both oligonucleotide and chemical mutagenesis to introduce base changes in the Klebsiella pneumoniae nifL and nifH promoters . Among ten mutations analysed, including six spontaneous mutations, base changes at -12, -13, -14, and -26, located in previously identified conserved sequences, perturbed the activity of the promoters, demonstrating that these sequences are required for transcription . Not all base changes produced similar strong promoter down phenotypes when the nifL and nifH promoters were compared: activation of the nifH promoter by the nifA gene product was less sensitive to base changes in conserved nucleotides than was activation of the equivalently altered nifL promoter by the nifA or ntrC products . We have found that the nifH promoter can be weakly activated by the ntrC product; this activation shows the same down response to base changes seen with ntrC activation of the nifL promoter . We present evidence that the efficient activation of the nifH promoter by nifA (but not ntrC) can be attributed to specific upstream sequences present in the nifH promoter. Nucleic Acids Res, 1985 Nov 11, 13(21), 7607 - 20 The nucleotide sequence of the nitrogen-regulation gene ntrA of Klebsiella pneumoniae and comparison with conserved features in bacterial RNA polymerase sigma factors; Merrick MJ et al.; The nucleotide sequence of the Klebsiella pneumoniae ntrA gene has been determined . NtrA encodes a 53,926 Dalton acidic polypeptide; a calculated molecular weight which is significantly lower than that determined by SDS polyacrylamide gel analysis . NtrA is followed by another open-reading frame (orf) of at least 75 amino acids . In the spacer region between ntrA and orf there are no apparent transcription termination or promoter sequences and therefore orf may be co-transcribed with ntrA . Previous authors have proposed that NtrA could act as an RNA polymerase sigma factor but the NtrA amino acid sequence does not show a high level of homology to any known sigma factor . However analysis of sequences of five sigma factors from E . coli and B . subtilis has identified two conserved sequences at the C-terminal end of all these polypeptides . These sequences resemble those found in known site-specific DNA-binding domains and may be involved in recognition of conserved -35 and -10 promoter sequences . A similar pair of sequences is present at the C-terminus of NtrA and could play a role in recognition of ntr-activatable promoters. Appl Environ Microbiol, 1985 Nov, 50(5), 1262 - 7 Detoxification of mercury, cadmium, and lead in Klebsiella aerogenes NCTC 418 growing in continuous culture; Aiking H et al.; Klebsiella aerogenes NCTC 418 growing in the presence of cadmium under glucose-, sulfate-, or phosphate-limited conditions in continuous culture exhibited sulfide formation and Pi accumulation as the only demonstrable detoxification mechanisms . In the presence of mercury under similar conditions only HgS formation could be confirmed, by an increased sensitivity to mercury under sulfate-limited conditions, among others . The fact that the cells were most sensitive to cadmium under conditions of phosphate limitation and most sensitive to mercury under conditions of sulfate limitation led to the hypothesis that these inorganic detoxification mechanisms generally depended on a kind of "facilitated precipitation" . The process was coined thus because heavy metals were probably accumulated and precipitated near the cell perimeter due to the relatively high local concentrations of sulfide and phosphate there . Depending on the growth-limiting nutrient, mercury proved to be 25-fold (phosphate limitation), 75-fold (glycerol limitation), or 150-fold (sulfate limitation) more toxic than cadmium to this organism . In the presence of lead, PbS formation was suggested . Since no other detoxification mechanisms were detected, for example, rendering heavy metal ions innocuous as metallo-organic compounds, it was concluded that formation of heavy metal precipitates is crucially important to this organism . In addition, it was observed that several components of a defined mineral medium were able to reduce mercuric ions to elemental mercury . This abiotic mercury volatilization was studied in detail, and its general and environmental implications are discussed. Antimicrob Agents Chemother, 1985 Nov, 28(5), 654 - 9 Relevance of serum protein binding of cefoxitin and cefazolin to their activities against Klebsiella pneumoniae pneumonia in rats; Bakker-Woudenberg IA et al.; An experimental Klebsiella pneumoniae pneumonia in rats was used to study the effect of protein binding of cefoxitin and cefazolin on their therapeutic activity . Both cephalosporins were similar with respect to their antimicrobial activity against the K . pneumoniae in vitro, but they differed in their degree of protein binding, being 34% for cefoxitin and 89 to 93% for cefazolin in uninfected rats and 24 and 71 to 83%, respectively, in infected rats . Various doses of these agents were administered by continuous infusion, which started 5 h after bacterial inoculation and continued for 65 h . Antimicrobial response was evaluated with respect to the numbers of bacteria recovered from lung and blood at the end of treatment . An inhibitory effect of protein binding on the in vivo antimicrobial activity was demonstrated . Cefoxitin was therapeutically effective at a constant plasma level that reached the MIC . To obtain a similar effect with cefazolin the plasma level of that drug had to be increased to a concentration more than three times the MIC. Biochem J, 1985 Nov 1, 231(3), 743 - 53 Electron transfer to nitrogenase in Klebsiella pneumoniae . nifF gene cloned and the gene product, a flavodoxin, purified; Deistung J et al.; The nifF gene of Klebsiella pneumoniae was cloned into a multicopy plasmid in order to construct a strain that synthesizes and retains an elevated concentration of the gene product relative to the wild-type strain . Characterization of the isolated flavodoxin, which serves as an electron donor to nitrogenase, shows unambiguously that it is the product of the nifF gene. Arch Microbiol, 1985 Nov, 143(2), 163 - 8 The functional significance of glucose dehydrogenase in Klebsiella aerogenes; Hommes RW et al.; In order to assess the functional significance of the quinoprotein glucose dehydrogenase recently found to be present in K+ -limited Klebsiella aerogenes, a broad study was made of the influence of specific environmental conditions on the cellular content of this enzyme . Whereas high activities were manifest in cells from glucose containing chemostat cultures that were either potassium- or phosphate-limited, only low activities were apparent in cells from similar cultures that were either glucose-, sulphate- or ammonia-limited . With these latter two cultures, a marked increase in glucose dehydrogenase activity was observed when 2,4-dinitrophenol (1 mM end concentration) was added to the growth medium . These results suggested that the synthesis of glucose dehydrogenase is not regulated by the level of glucose in the growth medium, but possibly by conditions that imposed an energetic stress upon the cells . This conclusion was further supported by a subsequent finding that K+ -limited cells that were growing on glycerol also synthesized substantial amounts of glucose dehydrogenase . The enzyme was found to be membrane associated, and preliminary evidence has been obtained that it is located on the periplasmic side of the cytoplasmic membrane and functionally linked to the respiratory chain . This structural and functional orientation is consistent with glucose dehydrogenase serving as a low impedance energy generating system. South Med J, 1985 Nov, 78(11), 1396 - 8 Progressive destruction of the hip joint due to Klebsiella osteomyelitis; Rawlings CE 3rd et al.; We have described a 52-year-old diabetic man with Klebsiella osteomyelitis of the femoral head and acetabulum . This case illustrates an unusual location of the infection, an absence of a preceding infection with Klebsiella pneumoniae, and a chronic and progressive course of the process over a six-month period. J Bacteriol, 1985 Nov, 164(2), 639 - 45 Structure of two divergent promoters located in front of the gene encoding pullulanase in Klebsiella pneumoniae and positively regulated by the malT product; Chapon C et al.; Pullulanase is an extracellular starch-debranching enzyme produced by Klebsiella pneumoniae . When its structural gene, pulA, is introduced into Escherichia coli, it is controlled by malT, the positive regulator gene of the maltose regulon . Characterization of the region 5' to pulA and of the beginning of the gene described herein demonstrate that (i) pullulanase is probably a lipoprotein; (ii) an additional malT-controlled promoter (the malX promoter) lies adjacent to the pulA promoter and is oriented in the opposite direction; (iii) in common with the three previously described malT-controlled promoters, the pulA and malX promoters have a conserved hexanucleotide (consensus sequence, 5'-GGATGGA) 35 base pairs upstream from the transcription initiation site; and (iv) upstream from this conserved hexanucleotide the pulA and malX promoters differ from the other mal promoters in that they lack any detectable binding site for the cyclic AMP-binding protein. J Bacteriol, 1985 Nov, 164(2), 633 - 8 Characterization and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae; Michaelis S et al.; Some strains of Klebsiella pneumonia secrete pullulanase, a debranching enzyme which produces linear molecules (maltodextrins, amylose) from amylopectin and glycogen . pulA, the structural gene for pullulanase, was introduced into Escherichia coli, either on a multiple-copy-number plasmid or as a single copy in the chromosome . When in E . coli, pulA was controlled by malT, the positive regulatory gene of the maltose regulon . Indeed, pulA expression was undetectable in a malT-negative mutant and constitutive in a malTc strain . Furthermore, the plasmid carrying pulA titrated the MalT protein . When produced in E . coli, pullulanase was not localized in the same way as in K . pneumoniae . In the latter case it was first exported to the outer membrane, with which it remained loosely associated, and was then released into the growth medium . In E . coli the enzyme was distributed both in the inner and the outer membranes and was never released into the growth medium. Immunology, 1985 Nov, 56(3), 571 - 3 Adjuvant actions of linear mannan-possessing lipopolysaccharide (LPS) in LPS-resistant C3H/HeJ mice; Ohta M et al.; Immunopotentiation has been demonstrated when Klebsiella O3 lipopolysaccharide (KO3 LPS), which possesses a linear mannan as the O-specific side chain, was injected subcutaneously into endotoxin resistant C3H/HeJ mice together with soluble protein antigens . The LPS exhibited significantly positive adjuvant effects on antibody responses in vivo after secondary antigen challenge and on delayed-type hypersensitivity (DTH) reactions against protein antigens . However, KO3 LPS was not a polyclonal B cell activator (PBA) in C3H/HeJ mice nor mitogenic in cultures of spleen cells of C3H/HeJ . Thus, the activity of the LPS in C3H/HeJ mice is confined to the potentiation of T-dependent immune responses . The contribution of the mannan O side chain to the adjuvant action of KO3 LPS was suggested. Cell Immunol, 1985 Oct 15, 95(2), 330 - 9 Depression of afferent arc of the in vivo cytotoxic T-cell immunity by bacterial lipopolysaccharides; Mizoguchi K et al.; The afferent arc of the in vivo cytotoxic T-cell immunity assessed by second set rejection of ascitic allogeneic tumors was shown to be depressed by bacterial lipopolysaccharide (LPS) that was administered simultaneously with or 1 day before injection of allogeneic spleen cells as stimulators . Two different LPSs from Escherichia coli O55 and Klebsiella O3 displayed similar activities whereas dextran sulfate, concanavalin A, or poly A:U was not effective . Stimulator activities of allogeneic cells was not directly modified by LPS . Any definite suppressor activity on afferent or efferent arc of the T-cell response was not demonstrable in mice receiving LPS and allogeneic cells . Further, the LPS effect for immune depression was not diminished by whole body X-ray irradiation to the recipient at 300 R, which ablated the B-cell reactivity to LPS for polyclonal activation, or by treatment of the recipient with carrageenan, a known toxic agent to macrophages . It was suggested from these results that LPS suppresses the cytotoxic T-cell immunity by modulating responder T cells to be temporarily refractory to the allogeneic stimulus rather than by activating suppressor cells such as radiation-sensitive lymphocytes and carrageenan-sensitive macrophages. Carbohydr Res, 1985 Oct 1, 142(1), 85 - 92 Structural investigation of the capsular polysaccharide of Klebsiella serotype K 49; Joseleau JP; The structure of the capsular polysaccharide from Klebsiella type K 49 was investigated by 1H- and 13C-n.m.r . spectroscopy of the original, carboxyl-reduced, and Smith-degraded polysaccharides . Methylation of the original K 49 and derivatives showed that the polysaccharide consists of a tetrasaccharide repeating-unit having D-galacturonic as a single lateral substituent . All of the sugars have the alpha-D-configuration . This conclusion is in agreement with measurements of spin-lattice relaxation-times for the anomeric proton . O-Acetyl groups are located on galacturonic acid, but do not occupy a unique position . (Formula: see text). J Hyg (Lond), 1985 Oct, 95(2), 265 - 76 Biochemical and serological investigations on clinical isolates of klebsiella; Simoons-Smit AM et al.; A series of 925 clinical isolates of klebsiella was examined by serological and biochemical typing . To perform serological typing (capsular swelling) 77 capsular antisera were prepared, tested against the type strains and grouped in 13 pools . With this serotyping method 80% of the cultures were typable and 63 distinct types could be recognized . All strains were typable biochemically by means of the numerical coding system of the API-20E system supplemented by digits derived from 15 additional conventional biochemical tests . With the API-20E system 24 different biotypes could be distinguished whereas the combination of API-20E and the 15 additional tests produced 93 biotypes . Maximum discrimination of strains was achieved by the combination of serological and biochemical typing (256 bioserotypes) . The reproducibility, typability and discriminating power of the biotyping system was not inferior to serotyping . For epidemiological purposes biotyping can replace serotyping of Klebsiella species, especially in laboratories less well equipped. Infect Immun, 1985 Oct, 50(1), 225 - 30 Purification and vaccine potential of Klebsiella capsular polysaccharides; Cryz SJ Jr et al.; Capsular polysaccharide (CPS) from 18 Klebsiella strains of different capsular types was isolated and characterized . Purified CPSs were composed primarily of carbohydrate with trace quantities of protein, nucleic acids, and lipopolysaccharide . All CPSs were of a high molecular weight, possessing a Kd of 0.01 to 0.11 as determined by gel filtration over Sepharose CL-4B . Low levels of lipopolysaccharide present in all preparations were responsible for the highly pyrogenic nature of one-half of the CPS preparations . Treatment of capsular material with dilute NaOH in 95% ethanol markedly reduced the pyrogenicity of all preparations and had a negligible effect on their molecular weight . The immunogenicity of the various native CPSs for mice varied considerably from serotype to serotype, but all evoked an anticapsular immunoglobulin G response . Five of 18 NaOH-treated polysaccharides were significantly (P less than 0.05) less immunogenic than their native counterparts . Human immunoglobulin G prepared from volunteers immunized with either native or NaOH-treated KP1-0 capsular polysaccharide was equally effective at preventing experimental fatal Klebsiella pneumoniae burn wound sepsis in mice. Appl Environ Microbiol, 1985 Oct, 50(4), 924 - 9 Butanediol production from cellulose and hemicellulose by Klebsiella pneumoniae grown in sequential coculture with Trichoderma harzianum; Yu EK et al.; The bioconversion of cellulose and hemicellulose substrates to 2,3-butanediol by a sequential coculture approach was investigated with the cellulolytic fungus Trichoderma harzianum E58 and the fermentative bacterium Klebsiella pneumoniae . Vogel medium optimal for the production of the cellulolytic and xylanolytic enzymes of the fungus was found to be inhibitory to butanediol fermentation . This inhibition appeared to be due to a synergistic effect of various ingredients, particularly the salts, present in the fungal medium . The removal or replacement of such ingredients from Vogel medium led to the relief of fermentation inhibition, but the treatments also resulted in a significant decrease in fungal enzyme production . Resting cells of K . pneumoniae could be used for butanediol production in the fungal medium, indicating that the inhibitory effect on solvent production under such conditions was due to the indirect result of growth inhibition of the bacterial cells . The resting-cell approach could be combined with a fed-batch system for the direct conversion of 8 to 10% (wt/vol) of Solka-Floc or aspenwood xylan to butanediol at over 30% of the theoretical conversion efficiencies. Biochem J, 1985 Oct 1, 231(1), 25 - 30 Ring cleavage and degradative pathway of cyanuric acid in bacteria; Cook AM et al.; The degradative pathway of cyanuric acid {1,3,5-triazine-2,4,6(1H,3H,5H)-trione} was examined in Pseudomonas sp . strain D . The bacterium grew with cyanuric acid, biuret, urea or NH4+ as sole source of nitrogen, and each substrate was entirely metabolized concomitantly with growth . Enzymes from strain D were separated by chromatography on DEAE-cellulose and three reactions were examined . Cyanuric acid (1 mol) was converted stoichiometrically into 1.0 mol of CO2 and 1.1 mol of biuret, which was conclusively identified . Biuret (1 mol) was converted stoichiometrically into 1.1 mol of NH4+, about 1 mol of CO2 and 1.0 mol of urea, which was conclusively identified . Urea (1 mol) was converted into 1.9 mol of NH4+ and 1.0 mol of CO2 . The reactions proceeded under aerobic or anoxic conditions and were presumed to be hydrolytic . Data indicate that the same pathway occurred in another pseudomonad and a strain of Klebsiella pneumoniae. Biochem Pharmacol, 1985 Oct 1, 34(19), 3433 - 8 Diclofenac sodium, a negative chemokinetic factor for neutrophil locomotion; Perianin A et al.; Diclofenac sodium, a non steroidal anti-inflammatory agent, was studied for its influence on the locomotion of human polymorphonuclear neutrophils (PMN), in an attempt to define the mechanism governing the drug's anti-inflammatory properties . PMN locomotion was measured by the agarose technique under two conditions of stimulation of cell migration: in the presence of a gradient of stimuli (chemotaxis) and in the presence of various amounts of stimuli incorporated in the gel (chemokinesis) . At concentrations below 10 micrograms/ml, diclofenac in the gel reduced, in a dose-dependent manner, the directed locomotion of PMN induced by a gradient of C5a-activated serum, peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) or Klebsiella pneumoniae culture supernatant (KPCS) . Diclofenac also inhibited the random locomotion of unstimulated PMN, as well as the PMN chemokinetic activity induced by various amounts of FMLP or activated serum . Inhibition of PMN locomotion by diclofenac decreased when the concentration of the stimulant was raised; this inhibition was inversely related to the concentration of heat-inactivated fetal calf serum in the medium . The directed locomotion and chemokinesis of PMN, induced by FMLP were also reduced in PMN preincubated with diclofenac before migration, suggesting a direct cellular effect of diclofenac . On the other hand, diclofenac did not affect the changes in shape induced in floating PMN by FMLP or activated serum . The observation that diclofenac did not alter the ingestion rate of bacteria by PMN indicates that this drug is not cytotoxic for PMN . Consequently, diclofenac reduces PMN locomotion by interfering with the PMN chemokinetic activity . Diclofenac is an anti-inflammatory drug possessing the original property of acting as a negative chemokinetic agent, for migration of both stimulated and unstimulated PMN . It should therefore be a useful tool for analyzing the elements controlling PMN locomotion speed. J Bacteriol, 1985 Oct, 164(1), 479 - 83 Inactivation of glycerol dehydrogenase of Klebsiella pneumoniae and the role of divalent cations; Johnson EA et al.; Anaerobically induced NAD-linked glycerol dehydrogenase of Klebsiella pneumoniae for fermentative glycerol utilization was reported previously to be inactivated in the cell during oxidative metabolism . In vitro inactivation was observed in this study by incubating the purified enzyme in the presence of O2, Fe2+, and ascorbate or dihydroxyfumarate . It appears that O2 and the reducing agent formed H2O2 and that H2O2 reacted with Fe2+ to generate an activated species of oxygen which attacked the enzyme . The in vitro-oxidized enzyme, like the in vivo-inactivated enzyme, showed an increased Km for NAD (but not glycerol) and could no longer be activated by Mn2+ which increased the Vmax of the native enzyme but decreased its apparent affinity for NAD . Ethanol dehydrogenase and 1,3-propanediol oxidoreductase, two enzymes with anaerobic function, also lost activity when the cells were incubated aerobically with glucose . However, glucose 6-phosphate dehydrogenase (NADP-linked), isocitrate dehydrogenase, and malate dehydrogenase, expected to function both aerobically and anaerobically, were not inactivated . Thus, oxidative modification of proteins in vivo might provide a mechanism for regulating the activities of some anaerobic enzymes. FEBS Lett, 1985 Sep 23, 189(2), 250 - 4 A novel S = 3/2 EPR signal associated with native Fe-proteins of nitrogenase; Hagen WR et al.; In addition to their g = 1.94 EPR signal, nitrogenase Fe-proteins from Azotobacter vinelandii, Azotobacter chroococcum and Klebsiella pneumoniae exhibit a weak EPR signal with g approximately equal to 5 . Temperature dependence of the signal was consistent with an S = 3/2 system with negative zero-field splitting, D = -5 +/- 0.7 cm-1 . The ms = +/- 3/2 ground state doublet gives rise to a transition with geff = 5.90 and the transition within the excited ms = +/- 1/2 doublet has a split geff = 4.8, 3.4 . Quantitation gave 0.6 to 0.8 spin . mol-1 which summed with the spin intensity of the S = 1/2 g = 1.94 line to roughly 1 spin/mol . MgATP and MgADP decreased the intensity of the S = 3/2 signal with no concomitant changes in intensity of the S = 1/2 signal. Biochem J, 1985 Sep 15, 230(3), 569 - 78 Ribitol dehydrogenase of Klebsiella aerogenes . Sequence and properties of wild-type and mutant strains; Dothie JM et al.; Evidence is presented for the sequence of 249 amino acids in ribitol dehydrogenase-A from Klebsiella aerogenes . Continuous culture on xylitol yields strains that superproduce 'wild-type' enzyme but mutations appear to have arisen in this process . Other strains selected by such continuous culture produce enzymes with increased specific activity for xylitol but without loss of ribitol activity . One such enzyme, ribitol dehydrogenase-D, has Pro-196 for Gly-196 . Another, ribitol dehydrogenase-B, has a different mutation. Biochem J, 1985 Sep 15, 230(3), 579 - 85 Ribitol dehydrogenase of Klebsiella aerogenes . Sequence of the structural gene; Loviny T et al.; The ribitol dehydrogenase gene was cloned from wild-type Klebsiella aerogenes and also from a transducing phage lambda prbt which expresses the rbt operon constitutively . The coding sequence for 249 amino acids is separated from the following D-ribulokinase gene by 31 base pairs containing three stop codons, one of which overlaps the ribosome binding site for D-ribulokinase . Three residues in the amino acid sequence differ from that predicted from the DNA sequence: Asp-212 for Asn-212 is probably a protein sequencing error, but -Ala-Val- for -Ser-Ser- at 146-147 appears to be a 'neutral mutation' that may have arisen during prolonged chemostat selection of a strain that superproduces the enzyme from which the protein sequence was determined. J Gen Microbiol, 1985 Sep, 131 ( Pt 9), 2313 - 8 Three immunity types of klebicins which use the cloacin DF13 receptor of Klebsiella pneumoniae; Cooper PC et al.; We have investigated a group of bacteriocinogenic strains used in the epidemiological investigation of Klebsiella infections . Transfer of plasmids from these strains to laboratory strains allowed the identification of three klebicins which use the cloacin DF13 receptor in Klebsiella, but are of three distinct immunity types . These klebicins use the ferric-aerobactin receptor determined by ColV-K30 in Escherichia coli, which is also used by cloacin DF13 . We propose to call them group A klebicins, of immunity types A1, A2 and A3 . On the basis of immunity, cloacin DF13 also belongs to the klebicin A1 group. J Bacteriol, 1985 Sep, 163(3), 858 - 62 Regulatory region of the divergent Klebsiella pneumoniae lac operon; Buvinger WE et al.; The chromosomal DNA that lies between the lacI and lacZ genes of Klebsiella pneumoniae constitutes a 196-base pair intercistronic region that contains regulatory sequences for both genes . The probable locations of specific regulatory elements for both lacI and lacZ genes were determined by analogy with the corresponding Escherichia coli sequences . A recombinational event in ancestral DNA evidently has inverted the transcriptional direction of lacI in K . pneumoniae relative to the transcriptional direction of lacI in E . coli . One end of the inversion was located within a 19-base pair sequence in the K . pneumoniae regulatory region . Sequences partially homologous to these 19 base pairs were found in two locations on either side of the E . coli lacI gene . The nucleotide sequence of the lac regulatory region in K . pneumoniae exhibits more than one possibility for folded tertiary structures . The spatial relationships of transcriptional binding sites differ in two possible structures . Associations of regulatory and transcriptional proteins with the DNA might affect conformation of the regulatory sequences and, as a consequence, transcription of the lac genes. J Bacteriol, 1985 Sep, 163(3), 850 - 7 Nucleotide sequence of Klebsiella pneumoniae lac genes; Buvinger WE et al.; The nucleotide sequences of the Klebsiella pneumoniae lacI and lacZ genes and part of the lacY gene were determined, and these genes were located and oriented relative to one another . The K . pneumoniae lac operon is divergent in that the lacI and lacZ genes are oriented head to head, and complementary strands are transcribed . Besides base substitutions, the lacZ genes of K . pneumoniae and Escherichia coli have suffered short distance shifts of reading frame caused by additions or deletions or both during evolutionary divergence from a common ancestral gene . Relative to corresponding E . coli sequences, the nucleotide sequences of the lacZ and lacY genes are 61 and 67% conserved, and the lacI genes are 49% conserved . A comparison of both nucleotide and amino acid sequences revealed that the K . pneumoniae and E . coli lacI genes and lac repressor proteins each are related to the galR gene and gal repressor of E . coli to about the same extent . In terms of evolutionary relationships, the divergence of the forerunner of the galR gene from an ancestral lac repressor gene preceded separation and differentiation of the K . pneumoniae and E . coli lac repressor genes. J Bacteriol, 1985 Sep, 163(3), 1285 - 7 Mol- mutants of Klebsiella pneumoniae requiring high levels of molybdate for nitrogenase activity; Imperial J et al.; Mol- mutants of Klebsiella pneumoniae requiring high levels of molybdate for nitrogenase and nitrate reductase activity were characterized . The effects of mol mutations on nitrogenase activity were very similar to those caused by nifQ mutations . Mol- mutants of K . pneumoniae appear to be equivalent to ChlD- mutants of Escherichia coli. J Bacteriol, 1985 Sep, 163(3), 1296 - 9 Regions on plasmid pCU1 required for the killing of Klebsiella pneumoniae; Thatte V et al.; Plasmid pCU1 was Kik+ (promotes killing of Klebsiella pneumoniae) . All Tn5 insertions within the tra region of pCU1 were Kik- . Two other regions, kikA and kikB, were needed . They may be separated on different plasmids, but both must be mobilized into Klebsiella pneumoniae . Establishment of one kik region in K . pneumoniae followed by receipt of the second did not lead to killing . Kik was therefore intracellular and required concerted and transient action of both regions. FEBS Lett, 1985 Aug 5, 187(2), 237 - 9 Energy expenditure for cyclic retention of NH3/NH4+ during N2 fixation by Klebsiella pneumoniae; Kleiner D; The permeability coefficient for NH3 through bacterial membranes was determined to be around 2 X 10(-3) cm X s-1 . This value was used to calculate the outward diffusion of NH3 from intracellular pools and the energy costs for reabsorption as NH4+ . For an N2-fixing continuous culture of Klebsiella pneumoniae an energy expenditure of around 4 ATP per NH3 produced was calculated, thus increasing significantly the energy requirement for N2 fixation in vivo. Jpn J Antibiot, 1985 Aug, 38(8), 2313 - 37 {Fundamental and clinical study on cefpiramide in obstetrics and gynecology . Obstetrics and Gynecology Study Group for Cefpiramide}; Takase Z et al.; Fundamental and clinical studies on a new cephalosporin antibiotic, cefpiramide (CPM), was carried out under a joint study program, in order to evaluate the usefulness of the drug in treating infection of the female genital organs . The results obtained were as follows: CPM was readily transported to female genital organ tissues, and the concentrations of the drug exceeded 35 micrograms/g in various organ tissues in about 1 hour, following intravenous injection of 1 g . A level of more than 2 micrograms/g was maintained even 14 hours after the injection . The transport of CPM to various tissues was also studied following intravenous drip infusion of 1 g for 1 hour . The concentrations in tissues were slightly low but similar to those following intravenous injection . The peak concentration of the drug in the dead space exudate was 3.1-20.4 micrograms/ml, following intravenous injection and intravenous drip infusion of 1 g . The MIC80 of CPM were 3.13-12.5 micrograms/ml against S . aureus, Klebsiella sp., P . mirabilis and P . aeruginosa . Clinical effects of CPM were analyzed in 158 patients, including 56 cases with intrauterine infection, 37 cases with intrapelvic infection, 22 cases with external genital infection, 31 cases with adnexitis, 6 cases with postoperative wound infection and 6 cases with other infections . Excellent response was seen in 28 cases (17.7%), good response in 120 (75.9%) poor response in 10 (6.3%) . The rate of response was calculated as 93.7% . Safety of the drug was analyzed in 258 patients, and side effects occurred in 4 (1.6%) . Of these 4 patients, rash was in 1 patient, heat sensation in 1 patient, nausea in 1 patient and rash accompanying edema in 1 patient . Abnormal values in clinical laboratory findings were seen in 7 patients . Elevations of transaminase were seen in 5 patients and decrease of platelet was seen in a patient, and then elevations of transaminase with decrease of platelet was seen in a patient, and no other changes of particular note appeared. Jpn J Pharmacol, 1985 Aug, 38(4), 355 - 60 Comparative studies on antitumor activity of Klebsiella O3 lipopolysaccharide and its polysaccharide fraction in mice; Hasegawa T et al.; The antitumor activity of the polysaccharide fraction (OPS) obtained by the acid hydrolysis of Klebsiella O3 lipopolysaccharide (KO3 LPS) isolated from the culture supernatant of the decapsulated mutant strain LEN-1 (03: K1-) against both allogeneic tumor and syngeneic tumor systems in mice was compared with that of KO3 LPS . OPS prolonged the life span of MM2-bearing C3H/He mice by intraperitoneal (i.p.) pre- and post-treatment at the doses of 100 and 1000 mg/kg . However, large amounts of OPS were needed to show the antitumor activity as compared with KO3 LPS . OPS showed no growth inhibitory activity against Meth-A sarcoma in BALB/c mice by i.p., intravenous (i.v.) or intratumoral (i.t.) administration . When 1000 mg/kg of OPS was i.p . administered once a day for 10 days, OPS significantly inhibited the tumor growth of Sarcoma-180 solid type tumor . On the other hand, KO3 LPS significantly suppressed the growth of Meth-A tumor by i.t . administration at the doses of 0.3 and 1.0 mg/kg and showed complete regression in 8 and 9 out of 10 mice, respectively . In MM2 tumor, KO3 LPS also showed complete regression in all mice post-treated by i.p . administration at the dose of 1.0 mg/kg . These results suggest that OPS has antitumor activity on the tumors used in this study, but the activity was less than that of KO3 LPS. Antimicrob Agents Chemother, 1985 Aug, 28(2), 195 - 9 Protein antigens of encapsulated Klebsiella pneumoniae surface exposed after growth in the presence of subinhibitory concentrations of cephalosporins; Kadurugamuwa JL et al.; It recently has been reported by us that cephalosporins, at a concentration below that influencing growth rate, reduced the production of enterochelin and capsule formation of iron-depleted Klebsiella pneumoniae . We now report on the antigenicity of the outer membrane components and surface-exposed protein antigens of iron-depleted cells grown in the presence or absence of cephalosporins . All major outer membrane proteins, including iron-regulated membrane proteins, were immunogenic . Encapsulated K . pneumoniae grown in antibiotic-free media had three protein antigens (60, 35.5, and 32.5 kilodaltons) exposed on the surface that were accessible to antibodies . Growth of the same cultures in the presence of subinhibitory concentrations of cephalosporins resulted in the exposure of a greater number of protein antigenic determinants, including iron-regulated membrane proteins, which become readily accessible to antibodies . It was also found that immunoblotting was generally more sensitive than conventional staining of the acrylamide gel with Coomassie blue in the detection of proteins. Chemioterapia, 1985 Aug, 4(4), 324 - 8 Immunopotentiation in infectious disease, II . Effect of bestatin on experimental infection; Ormrod DJ et al.; A two-stage protocol designed to evaluate putative immunomodulators for use in infectious disease has been proposed . In this report the effect of bestatin on a series of clinically relevant, and sub-lethal infections is described . Pyelonephritis, peritonitis and bacteremia were induced with Escherichia coli, while Klebsiella pneumoniae was used to produce a lung infection . Bestatin had no effect on the course of these infections . In a further experiment we assessed the effect of combined bestatin/antibiotic therapy on the course of renal infection . Again no effect was observed . These findings are consistent with the known immunomodulatory properties of bestatin . This methodology will be used to evaluate selected agents for their potential in infectious disease and should lead to useful clinical protocols. J Infect Dis, 1985 Aug, 152(2), 373 - 8 Therapeutic efficacy of continuous versus intermittent administration of ceftazidime in an experimental Klebsiella pneumoniae pneumonia in rats; Roosendaal R et al.; An experimental Klebsiella pneumoniae pneumonia in rats was used to study the influence of continuous or of intermittent (8-hr intervals) administration of ceftazidime on therapeutic efficacy . Antimicrobial response was evaluated with respect to the calculated total daily dose that protected 50% of the animals from death (PD50) until 16 days after termination of a four-day treatment . When antibiotic treatment was started 5 hr after bacterial inoculation, the PD50 values after continuous and after intermittent administration of ceftazidime were 0.36 and 1.42 mg/kg per day, respectively (P less than .001) . With a delay in the administration of the antibiotic to 34 hr after inoculation, the respective PD50 values were 1.08 and 13.06 mg of ceftazidime/kg per day (P less than .001) . These studies show an improved therapeutic efficacy that increased with a delay in treatment when ceftazidime was administered by continuous infusion as compared with administration at 8-hr intervals . Continuous administration of PD50 doses of ceftazidime resulted in serum levels that were constantly below the MIC of the infecting Klebsiella strain. Antimicrob Agents Chemother, 1985 Aug, 28(2), 302 - 7 Evolution of plasmid-coded resistance to broad-spectrum cephalosporins; Kliebe C et al.; A clinical isolate of Klebsiella ozaenae with transferable resistance to broad-spectrum cephalosporins produces a beta-lactamase determined by plasmid pBP60 . The beta-lactamase had the same isoelectric point as SHV-1 (7.6) . From heteroduplex analysis, an extensive homology between the two bla genes could be deduced; therefore, the new beta-lactamase was designated SHV-2 . Enzymatic studies revealed that SHV-2 was able to hydrolyze broad-spectrum cephalosporins due to an increased affinity of these compounds for the enzyme . The assumption that SHV-2 is a natural mutant of SHV-1 was strongly supported by the isolation of a laboratory mutant of SHV-1 that showed activities similar to those of SHV-2. Infect Control, 1985 Aug, 6(8), 310 - 3 Klebsiella oxytoca isolates in a general hospital; Alvarez S et al.; Primary infections caused by Klebsiella species are uncommon, but the organism is an important nosocomial pathogen . We report the predisposing factors, clinical features and outcome of 44 hospitalized patients in whom Klebsiella oxytoca was isolated . Twenty-one (48%) isolates were community-acquired and 23 (52%) were considered nosocomial in origin . Most of the patients were elderly males with serious underlying diseases . There were significant differences between those patients who acquired Klebsiella oxytoca in the hospital and in the community . Nosocomially acquired organisms were associated with a higher mortality (52% vs . 24%) (p less than 0.05), a higher incidence of infection vs . colonization (83% vs . 57%) (p less than 0.05), and a higher percentage of cases of pneumonia (43% vs . 19%) (p less than 0.05) . The clinical features, the hospital service and the patients' underlying diseases were similar when patients who died and those who survived were compared . Patients who died were exposed to antibiotics more often prior to the positive culture with K . oxytoca (p less than 0.05) . K . oxytoca is a significant pathogen in hospitalized elderly patients . It is likely to cause infections, especially pneumonia, and carry a high mortality . The organism can become endemic within the hospital setting with continued carriage and nosocomial spread. Carbohydr Res, 1985 Jul 15, 140(2), 263 - 75 A structural investigation of the capsular polysaccharide of Klebsiella K14; Dutton GG et al.; The structure of the capsular polysaccharide isolated from Klebsiella serotype K14 has been investigated employing a combination of chemical and spectroscopic methods . The repeating structure is shown to be of the "4 + 1 + 1" type, and it carries a 1-carboxyethylidene acetal substituent at positions 4 and 6 of a terminal glucose residue . The polysaccharide is one of a group of only three Klebsiella polysaccharides that have been found to contain a galactofuranose residue in the repeating unit . The repeating unit has the following structure: (Formula: see text). J Antibiot (Tokyo), 1985 Jul, 38(7), 941 - 7 Therapeutic efficacy of beta-lactam and aminoglycoside antibiotics on experimental pneumonia caused by Klebsiella pneumoniae B-54 in diabetic mice; Obana Y et al.; The therapeutic efficacy of six beta-lactam and aminoglycoside antibiotics were compared in diabetic mice with experimentally induced Klebsiella pneumoniae pneumonia . beta-Lactams caused a reduction in the numbers of bacteria, with clearance of bacteria from the lungs of diabetic and normal mice . The effect in diabetic mice, however, was very poor . In contrast thereto, no remarkable difference was seen between diabetic and normal mice when treated with aminoglycosides . The concentration of the test antibiotics to the lungs in diabetic mice was lower than in