Klebsiella

J Thorac Cardiovasc Surg, 1993 Nov, 106(5), 779 - 86
Prolonged survival of orthotopically transplanted heart xenograft in infant baboons; Kawauchi M et al.; Orthotopic concordant xenotransplantation in a juvenile primate model was examined . Eighteen donor rhesus monkeys weighing 2.4 to 3.8 kg (mean 2.9 kg) were matched with juvenile baboons, aged 9 to 19 months (mean 12.7 months) and weighing 3.2 to 4.8 kg (mean 3.9 kg), using ABH blood type and mixed lymphocyte culture . Rhesus monkey hearts were orthotopically transplanted without immunosuppression into six control baboons (group I) . In five baboons (group II), 4 mg/kg per day of antilymphocyte globulin was administered for 3 days before the operation and 5 days after the operation . Splenectomy was also performed, and 18 mg/kg per day of FK 506 was administered orally . Intravenous methotrexate, methylprednisolone, or both were used as rescue therapy . Seven baboons (group III) received the same immunosuppression as those in group II, but an intravenous dose of methotrexate (0.1 to 5 mg) was given twice weekly to suppress the proliferative response as monitored by in vitro immunologic assays . Baboons in group I had a mean survival of 8 days; all died as a result of classic cellular rejection . Baboons in group II had a mean survival of 48.4 days (p < 0.05 versus group I) . Two died during rescue therapy for rejection, and three died of cytomegalovirus infection . Two group II baboons showed mild rejection at autopsy . Baboons in group III had a mean survival of 127 days, and one baboon was still alive after 286 days . Two died of cytomegalovirus infection, one of toxoplasmosis, one of Klebsiella pneumoniae, one of massive micropulmonary embolism, one of renal failure aggravated by ganciclovir . Only two of the baboons that died showed rejection (estimated as mild) at autopsy . The baboon still alive at 286 days had no rejection on myocardial biopsy on the two hundred forty-fourth postoperative day . FK 506 coupled with low-dose maintenance methotrexate and splenectomy has produced prolonged host survival in this xenotransplantation model . Results suggest that concordant xenotransplantation would be a suitable biologic bridge to allotransplantation.

Mol Microbiol, 1993 Nov, 10(3), 615 - 25
Structural variation in the O-specific polysaccharides of Klebsiella pneumoniae serotype O1 and O8 lipopolysaccharide: evidence for clonal diversity in rfb genes; Kelly RF et al.; The O-polysaccharide fraction of the lipopolysaccharide from Klebsiella pneumoniae serotype O8 was found to comprise two galactose-containing homopolymers . Structural analysis, using chemical and high-field nuclear magnetic resonance (NMR) techniques, established that the K . pneumoniae O8 polysaccharides are composed of the linear, disaccharide repeating units {formula: see text} K . pneumoniae O8 mutant RFK-1 was isolated by resistance to phage KO1-2; strain RFK-1 expressed only D-galactan I-OAc . The 1H- and 13C-NMR resonances from this O-polysaccharide indicate that all of the O-acetyl groups within the K . pneumoniae O8 polysaccharide are carried on D-galactan I and O-acetylation occurs only on the beta-D-galactofuranose residues; 60% of the available beta-D-galactofuranose residues are non-acetylated . The O-acetylation of the remaining residues is equally distributed between the O-2 and O-6 positions . The carbohydrate backbone structures in the O8 polysaccharide are identical to D-galactan I and II expressed by K . pneumoniae O1, accounting for the antigenic cross-reaction between strains belonging to serotypes O1 and O8 . However, the O1 polysaccharides are not acetylated and the O-acetyl groups present in the K . pneumoniae serotype O8 polysaccharides provide a structural basis for their recognition as distinct serotypes . The rfb (O-polysaccharide biosynthesis) gene cluster of K . pneumoniae serotype O1 determines the synthesis of D-galactan I . rfbKpO1-specific gene probes were used to examine conservation in the rfb gene clusters of other K . pneumoniae serotypes which produce D-galactan I . Six O1 strains were examined and all showed hybridization with rfbKpO1 probes under conditions of high stringency . Three serotype O2 strains produce D-galactan I and these strains also contained DNA sequences recognized by rfbKpO1 probes under high stringency . The physical maps of these homologous rfb chromosomal regions showed some polymorphism . Surprisingly, the rfbKpO8 region from K . pneumoniae serotype O8 was only recognized by rfbKpO1 probes under low-stringency hybridization conditions, providing evidence for two substantially different clonal groups of rfb genes from K . pneumoniae strains with structurally related O-antigens.

J Biol Chem, 1993 Oct 15, 268(29), 21466 - 9
A bacterial enzyme that catalyzes formation of carbon monoxide; Wray JW et al.; We have isolated and purified an enzyme (E-2) from Klebsiella pneumoniae, which catalyzes the formation of CO from CH3-S-CH2-CH2-CO-C(OH) = CH-O- (III) . This compound is an intermediate in the conversion of 5'-methylthioadenosine to methionine . Concomitant with CO formation, methylthiopropionic acid and formate are produced and O2 is consumed . E-2 also catalyzes the formation of CO, formate, and butyrate from CH3-CH2-CH2-CO-C(OH) = CH-O- (IIIa), the desthio analog of III . Experiments with isotopic IIIa have shown that formate is derived from 1-C, and CO from 2-C . E-2 has a M(r) = 18,500 and requires Mg2+, and no chromophoric cofactor has been detected.

J Biol Chem, 1993 Oct 5, 268(28), 20768 - 71
Two-dimensional, rotational-echo double-resonance NMR of cell culture metabolism; McDowell LM et al.; Two-dimensional, rotational-echo double-resonance 13C NMR, a new solid-state NMR technique, has been used to show that the relative fluxes of the labeled chemical bond of L-{2-13C,15N}serine along four metabolic pathways (direct purine synthesis, direct glycine incorporation into protein, direct non-glycyl incorporation into protein, and nitrogen scrambling with loss of carbon) are 1:2:6:36, respectively, for Klebsiella pneumoniae under conditions of nitrogenase derepression . These determinations were performed on a single sample of lyophilized, double-labeled, intact cells . Analysis of the homogeneity of the distribution of label suggests that the primary role of serine in shortening derepression is in providing specific carbon and nitrogen for RNA synthesis.

Carbohydr Res, 1993 Oct 4, 248, 213 - 23
Klebsiella K43 capsular polysaccharide: primary structure and depolymerisation by a viral-borne endoglycanase; Aereboe M et al.; The capsular polysaccharide of Klebsiella K43 has been studied by glycose analysis, methylation analysis, and NMR spectroscopy, and by bacteriophage depolymerisation of the native polysaccharide . Additional evidence for the structure of the repeating unit came from base-catalysed degradation of the methylated polysaccharide, and from NMR spectroscopic analysis of the lithium-degraded polysaccharide and of the oligosaccharide-alditol derived from the repeating unit oligosaccharide obtained from a bacteriophage degradation . The polysaccharide was shown to have the repeating unit: {formula: see text}

Proc Natl Acad Sci U S A, 1993 Oct 1, 90(19), 8812 - 6
Growth of the cyanobacterium Anabaena on molecular nitrogen: NifJ is required when iron is limited; Bauer CC et al.; The nifJ gene of Klebsiella pneumoniae encodes an oxidoreductase required for the transfer of electrons from pyruvate to flavodoxin, which reduces nitrogenase . The nifJ gene of Anabaena 7120, isolated from a cosmid bank, was found to contain an open reading frame encoding a 1197-aa protein . The deduced amino acid sequence shows 50% identity to the Klebsiella homolog . The nifJ gene in Anabaena 7120 was inactivated by chromosomal interruption . The resulting mutant was unable to grow on medium depleted of both iron and combined nitrogen but grew normally, fixing nitrogen, when iron was present . NifJ transcripts of 2.7 and 4.3 kb are induced by iron depletion irrespective of nitrogen status . One particular stretch of the Anabaena 7120 nifJ gene encodes 12 aa with no complementary matches in the Klebsiella protein . This insert contains five tandem repeats of the heptamer CCCCAGT . These heptamers, as well as heptamers and octamers of other related sequences, have been located in a number of cyanobacterial genomes but are usually not found within the coding region of a gene . The site of the Anabaena 7120 heptamers in the nifJ genes of other filamentous cyanobacteria contains a surprising diversity of repeated sequences, both octamers and heptamers . The corresponding protein inserts range in length from 1 to 21 aa, relative to Klebsiella NifJ.

J Bacteriol, 1993 Oct, 175(19), 6287 - 92
moaR, a gene that encodes a positive regulator of the monoamine regulon in Klebsiella aerogenes; Azakami H et al.; We cloned and sequenced a Klebsiella aerogenes gene (moaR) for activation of arylsulfatase synthesis by tyramine . This gene was cloned by complementation of a K . aerogenes mutant in which tyramine fails to relieve the arylsulfatase repression caused by sulfur compounds . The moaR gene also activated induction of the synthesis of both tyramine oxidase and the 30-kDa protein that is specifically induced by high concentrations of tyramine or catecholamines . The moaR gene on the chromosome of the wild-type strain of K . aerogenes was disrupted by homologous recombination with a plasmid containing the inactivated moaR . The resultant mutant showed the same phenotype as previously isolated atsT mutant strains that are negative for the derepressed synthesis of arylsulfatase . In this mutant strain, tyramine also failed to induce the synthesis of tyramine oxidase or the production of a 30-kDa protein . The moaR gene is capable of encoding a protein of 26,238 Da . The putative MoaR protein has a helix-turn-helix motif in its C terminus . Thus, it seems likely that the MoaR protein regulates the operons by binding to the regulatory region of the monoamine regulon . The MoaR protein is subject to autogenous control, which was shown by use of a moaR'-lacZ transcriptional fusion.

Chest, 1993 Oct, 104(4), 1273 - 4
Thoracoscopic diagnosis of pleurolithiasis after laparoscopic cholecystectomy; Brazinsky SA et al.; We describe a patient with right pleuritic chest pain and an enlarging exudative pleural effusion four months after laparoscopic cholecystectomy . Several radiographic imaging procedures and thoracenteses were nondiagnostic . Thoracoscopy, however, revealed bilious concretions in the parietal pleura . Thoracoscopic drainage, lysis of adhesions, and antibiotic treatment of a Klebsiella pneumoniae pleuritis resulted in relief of symptoms.

J Antimicrob Chemother, 1993 Oct, 32(4), 605 - 9
High-level quinolone resistance amongst clinical isolates of Escherichia coli and Klebsiella pneumoniae from Spain; Alarcon T et al.; A recombinant plasmid containing gyr A encoding wild-type Escherichia coli quinolone susceptible DNA gyrase A subunits has been used as a broad host range gene probe . Strains expressing gyr A-mediated quinolone resistance become susceptible to quinolones upon insertion of the plasmid, whereas the plasmid without gyrA (pLA2917, vector) has no effect . Fifteen highly ciprofloxacin-resistant E . coli and three Klebsiella pneumoniae (MICs 2-64 mg/L) were isolated from clinical specimens in the Hospital de la Princesa, Madrid, Spain . Plasmid pNJR3-2 and pLA2917 were introduced into the clinical isolates by conjugation, and transconjugants selected with tetracycline or kanamycin (for which the plasmids encode resistance) . Ten transconjugants from each mating, the original isolates, the gene probe and vector control were screened for susceptibility to nalidixic acid, ciprofloxacin, ofloxacin, norfloxacin, tetracycline, chloramphenicol, cefoxitin and trimethoprim . Lower MICs of quinolones were seen for the transconjugants of two K . pneumoniae isolates in the presence of the gene probe, suggesting that these isolates harboured mutations in gyr A . Plasmid profiles confirmed the presence of the probe . The susceptibility of the third K . pneumoniae strain and all E . coli isolates were unaffected by insertion of the plasmid, suggesting another mechanism was responsible for quinolone resistance.

J Antimicrob Chemother, 1993 Oct, 32(4), 559 - 70
Extended-spectrum beta-lactamases in Escherichia coli and Klebsiella spp . in European septicaemia isolates; Pornull KJ et al.; Five per cent of Escherichia coli and klebsiella septicaemia isolates from the European Study Group on Antibiotic Resistance (ESGAR) study in 1987 to 1988 showed reduced susceptibility or resistance to cefotaxime, ceftazidime and/oraztreonam . Six of 15 isolates studied were susceptible to cefoxitin and MICs of cefuroxime, cefotaxime, ceftazidime and aztreonam were reduced by clavulanic acid . The isoelectric points of their beta-lactamases were in the range of 5.3-7.6 . DNA hybridization showed that four of these beta-lactamases belonged to the TEM or SHV family . Transfer of cefotaxime resistance by conjugation was seen in two of the strains . Nine strains were resistant to cefoxitin (MIC > 16 mg/L) and MICs of cefuroxime, cefotaxime, ceftazidime and aztreonam were only slightly reduced in the presence of clavulanic acid . All nine strains produced at least one beta-lactamase of chromosomal origin with pI > 8.4, and four of these strains also harboured beta-lactamases with a pI range of 6.6-8.2 . Cefoxitin resistance could be transferred by conjugation in one strain . Thus E . coli and Klebsiella spp . from the ESGAR septicaemia isolates were found to produce extended-spectrum beta-lactamases of both chromosomal and plasmid origin.

Pediatr Infect Dis J, 1993 Oct, 12(10), 840 - 4
Severe Klebsiella infection as a cause of mortality in neonates in Harare, Zimbabwe: evidence from postmortem blood cultures; Nathoo KJ et al.; Postmortem blood cultures were taken from 105 neonates dying at Harare Hospital during a 1-year period . The infants were characterized by prematurity (63% < 37 weeks gestation), low birth weight (60% < 2500 g) and low Apgar score at 1 min (43% < 3) . More than one-half of the infants died within 48 hours of admission . Positive blood cultures within 10 minutes of death occurred in 44% of infants, and Klebsiella sp . were by far the most common isolates . Positive blood cultures were associated with very low birth weight (< 1500 g), and with babies who survived for > 48 hours . Antibodies to human immunodeficiency virus type 1 were found in 40% of the infants, and a high proportion of these had Klebsiella bacteremia . Nearly all the infants had received antibiotic therapy, usually penicillin and gentamicin . Very few babies who had received a cephalosporin had a positive blood culture, and in vitro tests showed that although many organisms were resistant to penicillin and the aminoglycosides, very few showed resistance to the cephalosporins . Our findings suggest that cephalosporins may be useful in treating severe neonatal sepsis, particularly when there is no response to more standard therapy.

Hepatogastroenterology, 1993 Oct, 40(5), 496 - 8
Liver abscess after percutaneous ethanol injection (PEI) therapy for hepatocellular carcinoma . A case report; Okada S et al.; We present a case of liver abscess after percutaneous ethanol injection (PEI) therapy for the treatment of recurrent hepatocellular carcinoma (HCC) . The 56-year-old woman had a past history of cholecystoduodenostomy for cogenital dilatation of the bile duct, and pneumobilia was observed in the intrahepatic bile ducts prior to PEI . The abscess was successfully treated by percutaneous abscess drainage and antibiotic therapy . Klebsiella pneumonia, one of the most common causative organisms of biliary tract infection, was isolated from the abscess . Thus, biliary tract infection related to the previous biliary-enteric anastomosis operation may have been one of the causative factors in the liver abscess in this patient . The rare experience reported here suggests that a careful search for coexistent abscess at the time of PEI is important in HCC patients with biliary-enteric anastomosis, especially in those with pneumobilia.

New Microbiol, 1993 Oct, 16(4), 343 - 50
Plasmid loss from gram-negative bacteria exposed to subinhibitory concentrations of beta-lactam drugs and azithromycin; Saverino D et al.; The stability of F'lac, pW101 and pHSG298 in Escherichia coli K12 exposed to subinhibitory concentrations of beta-lactam antibiotics, amikacin and tetracycline was studied . High molecular weight low copy plasmids (F'lac and pW101) were eliminated from bacteria treated with PBP-3 binding molecules, while a low molecular weight high copy extrachromosomal element (pHSG298) was not . None of the carbapenem antibiotics, mecillinam, amikacin or tetracycline promoted high rate plasmid loss from their hosts . Under the same conditions, plasmid-mediated ampicillin-resistance due to beta-lactamase production was also lost from F'lacTn1-carrying bacteria . In contrast, the high copy R6K plasmid was stably inherited in their hosts with the exception of those organisms treated with cefixime . When the same experiments were performed with a Klebsiella pneumoniae strain induced to form filaments by azithromycin at sub-MICs, F'lacTn1 and pW101 loss was detected, while pHSG298 was stably inherited . These results confirm previous observations that plasmid stability is correlated with cell shape and that recovery is more easily achieved when bacteria undergo an unbalanced division resulting in cell filamentation.

J Wildl Dis, 1993 Oct, 29(4), 620 - 2
Retrospective study of diseases in a captive Lemming Colony; Cutlip RC et al.; Fifty-four ill or nonproductive lemmings (Dicrostonyx spp.) were evaluated for signs, lesions and causes of disease for 5 yr in a domestic colony . Parasitic granulomas caused by Encephalitozoon cuniculi were the most common finding and were seen in 22 lemmings . The disease was characterized by circling and torticollis with granulomas in many tissues, especially the central nervous system . Suppurative otitis occurred in 12 lemmings and was associated with Klebsiella pneumonia infection; circling was the common sign . Hepatic microabscesses were present in seven lemmings but a cause was not identified . Five lemmings had neoplasms and 14 had either suppurative processes, aspermia, or ovarian cysts.

J Clin Microbiol, 1993 Oct, 31(10), 2790 - 3
Study of growth requirements other than cysteine of naturally occurring Escherichia coli and Klebsiella spp . auxotrophic for cysteine; McIver CJ et al.; Cysteine remains the preferred supplement for cultivation of Cys- auxotrophs in vitro . Methionine, which reduced cysteine requirements, and branched-chain amino acids, which decreased cysteine toxicity, were identified as the components of casein hydrolysate responsible for growth enhancement by this additive . Glutathione and DL-homocysteine can be substituted for cysteine . Accumulation of these compounds in patients with renal impairment may favor selection of Cys- strains in vivo.

Eur J Biochem, 1993 Oct 1, 217(1), 395 - 400
Roulette mutagenesis of the FMN-binding site of Klebsiella pneumoniae flavodoxin; Drummond M et al.; A method of randomising specific regions of coding sequences has been devised which utilises the Lac phenotype to identify mutants . Intact genes can be mutagenised, making it unnecessary to reclone the mutations before examining mutant phenotypes . The method has been applied to three residues around the N-terminus of the first alpha helix of the Klebsiella pneumoniae nitrogenase flavodoxin, which are predicted to form part of the phosphate-binding subsite . Surprisingly, most substitutions at Gly12, a highly conserved residue in the chain reversal preceding the alpha helix, appeared to be fairly stable in vivo and were found to retain some function . Substitutions at Lys13, a surface residue which contributes to a patch of positive charge characteristic of the nitrogenase flavodoxins, had no major effect on stability or function . However, most substitutions at Thr14, which is predicted to hydrogen bond to the phosphate of the prosthetic group FMN, were much more destabilising and grossly reduced function . The exceptions were Ala, Cys, Ser and Val, which suggests that the bulk of the residue at this position is critical.

Avian Dis, 1993 Oct-Dec, 37(4), 1136 - 41
Postmortem findings of ostriches submitted to the Oklahoma Animal Disease Diagnostic Laboratory; Terzich M et al.; A review of 121 ostrich necropsies from the files at the Oklahoma Animal Disease Diagnostic Laboratory was conducted . The birds ranged in age from unhatched embryos to 4 years; the majority were less than 3 weeks old . The most common cause of death was ostrich chick fading syndrome (OCFS) . OCFS is characterized by depression, anorexia, and death 3-5 days after onset of clinical signs in ostriches less than 3 weeks old . Escherichia coli and/or Klebsiella pneumoniae were isolated from various organs in these cases, and mortality ranged from 40% to 100% . Other conditions observed were edema in chicks associated with high incubator humidity levels, aspergillosis, leg deformities, and impaction of the proventriculus.

Infect Immun, 1993 Oct, 61(10), 4208 - 16
Genetic exchange of determinants for capsular polysaccharide biosynthesis between Klebsiella pneumoniae strains expressing serotypes K2 and K21a; Ofek I et al.; The production of a capsular polysaccharide (CPS; K antigen) is characteristic of Klebsiella pneumoniae, but CPS structure varies among strains, and many different serotypes are now known . In this study, cps gene clusters encoding the elements of capsular polysaccharide biosynthesis were exchanged by homologous recombination between strains expressing different serotypes . The wild-type K . pneumoniae strains used for genetic exchange were KPA1 (cpsK2), expressing K2 CPS, and KPB1 (cpsK21a), expressing K21a CPS . Plasmid R68.45 was used to mobilize fragments of chromosomal DNA from auxotrophic derivatives of donor strains . Auxotrophic his alleles introduced into recipient strains provided selectable markers to coinherit the adjacent cps gene clusters from donors expressing a heterologous CPS . Each of the capsule-switched recombinants, KPA5 (cpsK21a) and KPB20 (cpsK2), was shown to have a CPS that was immunologically identical to the serotype of the respective donor . The recombinants retained their respective recipient strain background, as evidenced by a genetic marker and demonstration of a distinctive restriction fragment length polymorphism in genomic DNA . KPB1 CPS contained a sequence (mannose-alpha-2-mannose) that binds to a macrophage lectin and may be responsible for their higher susceptibility to macrophage binding and phagocytosis compared with KPA1, whose CPS lacked such sequences . The recombinant strains expressing heterologous cps genes inherited the macrophage-binding phenotype of the donor, thus confirming that relative susceptibility to phagocytosis was determined by the capsule type expressed . KPA1 was highly virulent in a mouse lethality assay, which is a feature typical of K2 strains, whereas KPB1 was not virulent in mice . Recombinant KPA5 retained relatively high virulence in mice, even though it produced the heterologous K21a CPS, which suggests that a virulence factor other than capsule biosynthesis is encoded by the KPA genomic strain background . In contrast, KPB20 gained marginal virulence in the mouse lethality assay through the inheritance and expression of the K2 CPS from the virulent strain . Thus, pathogenesis in K . pneumoniae may be multifactorial . Specific antibody was used to stabilize the CPS on the surface of K . pneumoniae, and the structural organization of the homologous and heterologous capsules was examined by electron microscopy . Recombinant KPB20, expressing heterologous K2 CPS, had a uniform layer of capsule surrounding the organism that was similar to that seen on the surfaces of the parental strains . However, KPA5, expressing the heterologous K21a CPS, was unusual in that the uniform capsular layer was physically separated from the cell wall by approximately 50 nm.(ABSTRACT TRUNCATED AT 400 WORDS)

Biol Chem Hoppe Seyler, 1993 Oct, 374(10), 983 - 92
Molecular basis of the exclusive low-temperature synthesis of an enzyme in E . coli: penicillin acylase; Keilmann C et al.; The enzyme penicillin acylase is synthesized by Escherichia coli only at growth temperatures below 30 degrees C . The biochemical basis of this strict temperature-dependent formation of an enzyme was investigated . When the gene (pac) was under the control of the lacUV5 promoter it showed the same temperature-dependent expression as the chromosomally encoded gene transcribed from its own promoter . This indicates that translation of the pac mRNA rather than transcription of the gene is temperature-dependent . This conclusion could be further confirmed by Northern hybridisation and by analysis of pac-lacZ transcriptional fusions . TnphoA insertion mutagenesis and experiments in which the promoter and 5' sequence encoding the signal peptide of the pac gene was exchanged with those of the cyclodextrin glycosyltransferase gene from Klebsiella oxytoca localised the region of pac mRNA responsible for the temperature-sensitive translation to the 5'-untranslated region and/or the signal peptide . Extension of the 5 nucleotide long spacer separating the Shine-Dalgarno motif from the AUG initiation codon by one or three nucleotides lead to partial or full synthesis of penicillin acylase precursor at 40 degrees C, respectively . The precursor of penicillin acylase formed at 40 degrees C by the mutant variants or when placed under the control of a heterologous upstream region was associated with the membrane but could not be translocated . Taken together these data suggest that transport and translation of the penicillin acylase precursor are coupled and that the short Shine-Dalgarno-AUG distance interferes with a competent interaction between the translation initiation complex and the export system at high temperature . Moreover, evidence was also provided which indicates a direct effect of temperature on the conformation of the precursor and it is proposed that the lack of translation at high temperatures has been selected to prevent the accumulation of transport-incompetent protein locked in the membrane.

Surg Gynecol Obstet, 1993 Sep, 177(3), 279 - 82
Late development of cholangiocarcinoma after the treatment of hepatolithiasis; Chijiiwa K et al.; Although the association of cholangiocarcinoma with intrahepatic calculi (hepatolithiasis) is well recognized, the late development of cholangiocarcinoma after the treatment of hepatolithiasis has not been reported in detail . Of 109 consecutive patients with hepatolithiasis treated during 19 years, eight patients had cholangiocarcinoma, seven of whom had cholangiocarcinoma two to 14 years, with a mean of eight years, after the treatment of hepatolithiasis . Absence of cholangiocarcinoma was confirmed when stones were removed at the time of the initial treatment . The mean age was 56 years, with a female to male ratio of 2:5 . At the time of detecting cholangiocarcinoma, three patients had no gallstones and four had gallstones at the corresponding site to the carcinoma . Cystic dilatation of the intrahepatic bile duct was often observed on the direct cholangiogram . The biles were all infected mainly with Escherichia coli and Klebsiella species . Thus, bile stasis and bacteria infection seems to be the important causative factor, causing cholangiocarcinoma rather than the calculi itself . Because the symptoms only mimic those of cholangitis, the possible presence of cholangiocarcinoma should be considered even after the treatment of hepatolithiasis for early detection and curative resection.

J Infect Dis, 1993 Sep, 168(3), 766 - 9
Salicylate potentiates amikacin therapy in rodent models of Klebsiella pneumoniae infection; Domenico P et al.; In vitro effects of salicylate on Klebsiella pneumoniae include capsule repression and enhanced aminoglycoside activity . The effects of high-dose salicylate on amikacin therapy were assessed in animal models . In a mouse lethality model, amikacin protective doses were reduced up to 12-fold by salicylate doses of 277 mg/kg . Salicylate alone increased the LD50 for K . pneumoniae in mice by nearly 1 log unit . In a lobar pneumonia model, rats received salicylate-amikacin intravenously 3 days after challenge, and bacteria were quantified from lung tissue on day 7 . Rats receiving one-fourth the minimum effective amikacin dose had 1 x 10(6) cfu/g of lung, while no bacteria were detected in those also receiving salicylate (173 mg/kg) . At one-eighth the minimum effective dose of amikacin, salicylate prevented the development of pneumonia . Salicylate potentiation of antimicrobial therapy far surpassed that observed in vitro, yet the potentiation was abolished in neutropenic animals . Thus, salicylate may enhance phagocytosis rather than antibiotic action in therapy.

West Indian Med J, 1993 Sep, 42(3), 115 - 7
Fat embolism syndrome following long bone fractures; Arthurs MH et al.; During the period August, 1979 to December, 1992, 14 patients with the Fat Embolism Syndrome (FES) were admitted to the University Hospital of the West Indies (UHWI) . Two were females and 12 males, their ages ranging from 18 to 78 years, with a median age of 23.5 years . All had lower limb long bone fractures . Clinical features included fever, tachypnoea, confusion and drowsiness . They were all hypoxaemic; 9 required Intensive Care Unit (ICU) admission and, of these, 4 needed ventilatory support . Five patients became comatose, 4 of whom developed decerebrate posturing . There was one death from Klebsiella septicaemia, and 13 patients recovered fully . The FES is a serious life-threatening complication of long bone fractures whether simple or compound, usually occurring within 72 hours of the injury . A high index of suspicion is needed for its prompt detection, and early attempts at maintaining adequate tissue oxygenation must be instituted if serious neurological complications and death are to be avoided.

J Antimicrob Chemother, 1993 Sep, 32(3), 421 - 9
In-vitro evaluation of the four beta-lactamase inhibitors: BRL42715, clavulanic acid, sulbactam, and tazobactam; Muratani T et al.; The in-vitro synergic activities of BRL42715, a new beta-lactamase inhibitor, clavulanic acid, sulbactam, and tazobactam combined with ampicillin, piperacillin, cephalothin, or cefoperazone were tested against various bacteria producing known types of beta-lactamase . BRL42715 showed the best synergistic activity among the inhibitors tested against strains producing penicillinases of type I, II, III, V, and that from Klebsiella pneumoniae, cephalosporinases, and oxyiminocephalosporinases (except that from Klebsiella oxytoca) . Clavulanic acid combined with the beta-lactams tested showed the best synergic activity of the inhibitors against strains producing type IV penicillinase and oxyiminocephalosporinase from K . oxytoca . The 50% inhibitory doses of BRL42715 were superior to those of clavulanic acid against various types of beta-lactamases except for type IV penicillinase and the oxyiminocephalosporinase from K . oxytoca . The inhibitory activity of BRL42715 against cephalosporinases from various bacteria was 10(4) to 10(6)-fold greater than that of clavulanic acid . The synergic effects of BRL42715 and clavulanic acid on the activity of piperacillin were compared against six clinical isolates of bacteria resistant to piperacillin . The synergic activity of BRL42715 was greater than that of clavulanic acid in all six isolates.

Antimicrob Agents Chemother, 1993 Sep, 37(9), 2020 - 3
Novel, plasmid-encoded, TEM-derived extended-spectrum beta-lactamase in Klebsiella pneumoniae conferring higher resistance to aztreonam than to extended-spectrum cephalosporins; Arlet G et al.; A clinical isolate of Klebsiella pneumoniae was more resistant to aztreonam than to cefotaxime and ceftazidime . It produced a clavulanate-susceptible beta-lactamase with an isoelectric point of 6.3 which readily hydrolyzed penicillins, cefotaxime, and ceftazidime, but which hydrolyzed aztreonam poorly . The enzyme was encoded by a gene on a 15-kb plasmid; the gene hybridized with an intragenic DNA probe of blaTEM.

Antimicrob Agents Chemother, 1993 Sep, 37(9), 1989 - 92
Genetically diverse ceftazidime-resistant isolates from a single center: biochemical and genetic characterization of TEM-10 beta-lactamases encoded by different nucleotide sequences; Rasmussen BA et al.; Ceftazidime-resistant isolates of Escherichia coli and Klebsiella pneumoniae produced a plasmid-mediated beta-lactamase with a pI of 5.6 with biochemical characteristics comparable to those of the TEM-10 beta-lactamase . Plasmids from the two strains were nonidentical . Both TEM-10 sequences differed from TEM-1 by substitutions of Ser-162 and Lys-237 . The nucleotide sequences of the two genes were identical except for three silent nucleotide substitutions corresponding to the nucleotide differences in the Tn2 TEM-1 or Tn3 TEM-1 genes . The original TEM-10 plasmid was identical to that found in the E . coli isolate and coded for a gene that corresponded to the TEM-10 beta-lactamase from Tn2.

S Afr Med J, 1993 Sep, 83(9), 643 - 6
Factors associated with airway colonisation and invasion due to Klebsiella spp; Feldman C et al.; The clinical significance of a heavy growth of Klebsiella spp . in sputum was studied in 54 patients . All but 3 patients had significant factors potentially associated with respiratory tract colonisation or invasion . Risk factors identified for colonisation of the airway and for invasive disease were similar . Patients with community-acquired Klebsiella infections were more likely to have underlying chronic respiratory diseases . Prior antibiotic use was a risk factor for nosocomial infections which occurred more commonly with antibiotic-resistant organisms . The most common diagnoses were airway colonisation, acute community-acquired chest infections, and nosocomial chest infections . Primary acute community-acquired pneumonia was uncommon . The sensitivity and specificity of the sputum Gram stain (in the setting of positive sputum cultures) in suggesting the presence of invasive disease due to Klebsiella spp . were 42% and 69% respectively.

Mol Microbiol, 1993 Sep, 9(5), 1107 - 17
The function of the upstream region of the sigma 54-dependent Klebsiella pneumoniae nifL promoter is sensitive to DNA supercoiling; Whitehall S et al.; The positive control protein NTRC activates transcription from the sigma 54-dependent nifL and glnAp2 promoters of Klebsiella pneumoniae by binding to upstream enhancer-like sequences and contacting downstream bound sigma 54-RNA polymerase via looping of the intervening DNA . In contrast to the glnAp2 promoter, the activity of the nifL promoter is very sensitive to changes in DNA supercoiling both in vivo and in vitro . We have shown previously that the downstream elements of the nifL promoter are involved in the supercoiling response . In this study we find that the upstream region of nifL influences the supercoiling response of a hybrid nifL-glnAp2 promoter both in vivo and in vitro, demonstrating that the nifL upstream region also confers supercoiling sensitivity . DNA supercoiling did not appear to influence binding of NTRC to its sites in the nifL upstream region, suggesting that another function of this region, most probably DNA loop formation, is sensitive to changes in DNA topology.

Enzyme Microb Technol, 1993 Sep, 15(9), 756 - 63
Production and characterization of guluronate lyase from Klebsiella pneumoniae for applications in seaweed biotechnology; Ostgaard K et al.; Cultures of Klebsiella pneumoniae fermenting sodium alginate produce an extracellular guluronate-specific alginate lyase . This enzyme production was studied in stirred-tank fermentors . Different alginate substrates gave moderate differences in growth and enzyme yield . Alginates with low guluronic content gave reduced biomass but favored enzyme production . Low molecular weight (down to DPn approximately 270) also favored enzyme production . Excessive depolymerization of substrates occurred during heat sterilization of culture media . The enzyme was characterized by its specificity and sensitivity to pH, salt, and calcium . Improved yields of viable protoplasts were documented for Laminaria digitata (Huds.) Lamour.

Biochim Biophys Acta, 1993 Aug 7, 1164(3), 311 - 8
The in-vivo identification of the MoFe protein (FeMo cofactor) of nitrogenase in Klebsiella pneumoniae and of the Mo-storage protein in Azotobacter vinelandii via the nuclear quadrupole interaction of 99Mo(beta-)99Tc; Mottner P et al.; The expression of the MoFe protein of nitrogenase in Klebsiella pneumoniae was identified in vivo via the nuclear quadrupole interaction (NQI) of 99Mo(beta-)99Tc using perturbed angular correlations of gamma-rays . The NQI parameters were: omega approx . 360 Mrad/s and eta approx . 1 . In addition, the NQI of the 'Mo-storage protein' in Azotobacter vinelandii cells which had been grown in the presence of NH4+ (13 mM), i.e . under conditions of strict repression of nitrogenase synthesis, was determined: omega approx . 190 Mrad/s, eta approx . 0.25 . Under these conditions, the characteristic signal of the MoFe protein (FeMo cofactor) was absent.

Int J Biol Macromol, 1993 Aug, 15(4), 201 - 7
Solution properties of the capsular polysaccharide produced by Klebsiella pneumoniae SK1; Cescutti P et al.; The solution properties of the capsular polysaccharide produced by Klebsiella pneumoniae SK1, SK1-CPS, were investigated by various methods . The SK1-CPS repeating unit is a branched pentasaccharide containing one glucuronic acid as single unit side chain; acetyl groups are present as non-carbohydrate substituents on the uronic acid residue in non-stoichiometric amounts . Chiro-optical, potentiometric, viscometric and rheological measurements have been performed in order to characterize the conformational behaviour of the polymer in water and in aqueous salt solutions . Under the investigated experimental conditions, changes of temperature, ionic strength and pH were shown not to induce any cooperative conformational transition . All the results obtained suggest that the solution conformation of SK1-CPS is a random coil with a certain degree of chain flexibility . The removal of the acetyl substituents apparently does not modify the overall conclusions drawn for the native polymer, except for an incipient tendency to aggregation revealed for high salt conditions.

Proc Natl Acad Sci U S A, 1993 Aug 1, 90(15), 6934 - 8
Properties and purification of an active biotinylated lactose permease from Escherichia coli; Consler TG et al.; A simplified approach for purification of functional lactose permease from Escherichia coli is described that is based on the construction of chimeras between the permease and a 100-amino acid residue polypeptide containing the biotin acceptor domain from the oxaloacetate decarboxylase of Klebsiella pneumoniae {Cronan, J . E., Jr . (1990) J . Biol . Chem . 265, 10327-10333} . Chimeras were constructed with a factor Xa protease site and the biotin acceptor domain in the middle cytoplasmic loop (loop 6) or at the C terminus of the permease . Each construct catalyzes active lactose transport in cells and right-side-out membrane vesicles . Moreover, the constructs are biotinylated in vivo, and in both chimeras, the factor Xa protease site is accessible from the cytoplasmic surface of the membrane . Both biotinylated permeases bind selectively to immobilized monomeric avidin and are eluted with free biotin in a high state of purity, and the loop 6 chimera catalyzes active transport after reconstitution into proteoliposomes . The methodology described should be applicable to other membrane proteins.

Chest, 1993 Aug, 104(2), 627 - 9
Hypersensitivity pneumonitis secondary to Klebsiella oxytoca . A new cause of humidifier lung; Kane GC et al.; A 30-year-old woman developed recurrent episodes of fever, dyspnea, and nonproductive cough after repeated exposure to a home humidifier . The diagnosis of hypersensitivity pneumonitis was confirmed by detection of serum-binding antibodies at significant titer to Klebsiella oxytoca colonizing the humidifier water but not to other potential antigens . This represents a newly recognized cause of hypersensitivity pneumonitis related to exposure to K oxytoca contaminating a commercially available ultrasonic cold air home humidifier . The potential role for these frequently used home humidifier devices in unexplained pulmonary illness is emphasized.

J Bacteriol, 1993 Aug, 175(15), 4907 - 10
The nifY product of Klebsiella pneumoniae is associated with apodinitrogenase and dissociates upon activation with the iron-molybdenum cofactor; Homer MJ et al.; Apodinitrogenase, which lacks the iron-molybdenum cofactor at its active site, is an oligomer that contains an additional protein not found in the active dinitrogenase tetramer . This associated protein in Klebsiella pneumoniae is shown to be the product of the nifY gene . When apodinitrogenase is activated by the addition of the iron-molybdenum cofactor, NifY dissociates from the apodinitrogenase complex . The conditions for this dissociation are described . Finally, there are aspects of the dissociation and insertion process in K . pneumoniae that are different from that in Azotobacter vinelandii.

Mol Microbiol, 1993 Aug, 9(3), 623 - 34
Isolation of a putative fimbrial adhesin from Bordetella pertussis and the identification of its gene; Willems RJ et al.; We report the purification of a minor Bordetella pertussis fimbrial subunit, designated FimD, and the identification of its gene (fimD) . FimD could be purified from the bulk of major fimbrial subunits by exploiting the fact that major subunit-subunit interactions are more stable in the presence of SDS than minor-major subunit interactions . To locate the gene for FimD, internal peptides of FimD were generated, purified and sequenced . Subsequently, an oligonucleotide probe, based on the primary sequence of one peptide, was used to clone fimD . The primary structure of FimD, derived from the DNA sequence of its gene, showed homology with a number of fimbrial adhesins . Most pronounced homology was observed with MrkD, a fimbrial adhesin derived from Klebsiella pneumoniae . These observations suggest that FimD may represent a B . pertussis fimbrial adhesin . With a fimD-specific probe we detected the presence of a fimD homologue in Bordetella parapertussis and Bordetella bronchiseptica but not in Bordetella avium . Cloning and sequencing revealed that the B . parapertussis and B . bronchiseptica fimD product differed from the B . pertussis fimD product in 20 and 1 amino acid residues, respectively . Since B . bronchiseptica is normally not a human pathogen, but causes respiratory disease in a wide range of non-human mammalian species, this may suggest that FimD recognizes a receptor that is well conserved in mammalian species . An in-frame deletion in fimD completely abolished FimD expression and also affected the expression of the major subunits Fim2 and Fim3 suggesting that, in contrast to other adhesins that are minor components of fimbriae, FimD is required for formation of the fimbrial structure.

Antibiot Khimioter, 1993 Aug-Sep, 38(8-9), 44 - 7
{Combined effect of visible light sensitized by chlorine e6 and streptomycin on the obligate pathogenic bacterium Klebsiella rhinoscleromatis}; Fomichev AIu et al.; Chlorine e6-sensitized photodamage of Klebsiella rhinoscleromatis (an obligate pathogen) in the presence of streptomycin was studied . The pathogen is highly resistant to photosensitization characteristic of other gram-negative bacteria . Marked synergism of the bactericidal effect of the chlorine e6-sensitized visible light and streptomycin on K . rhinoscleromatis was observed . It was suggested that the synergistic action was due to the oxidative damage of the cell membranes resulting in higher rates of the streptomycin penetration to the cells and impairment of the intracellular protective mechanisms at the molecular level . The findings are in favour of the further investigation aimed at the development of a comprehensive method of photo- and chemotherapy of scleroma.

Carbohydr Res, 1993 Jul 19, 245(2), 311 - 21
Structural studies of the capsular polysaccharide (S-21) from Klebsiella pneumoniae ATCC 31314; Edebrink P et al.; The structure of the polysaccharide (S-21) elaborated by Klebsiella pneumoniae ATCC 31314 has been investigated . NMR spectroscopy, sugar and methylation analysis, uronic acid degradation, and partial hydrolysis to oligosaccharides were the main methods used . In order to obtain good NMR spectra, the polymer was subjected to non-specific degradation by treatment with fuming hydrochoric acid . It is concluded that S-21 is composed of pentasaccharide repeating units with the following structure . {formula: see text} Approximately 0.7 equivalent of O-acetyl group, distributed over at least three positions, was also present but not located . The carbohydrate backbone in S-21 is identical to that of Klebsiella K30 and K33 capsular polysaccharides.

J Infect Dis, 1993 Jul, 168(1), 164 - 71
Liposomes with prolonged blood circulation and selective localization in Klebsiella pneumoniae-infected lung tissue; Bakker-Woudenberg IA et al.; Studies of two types of small liposomes, differing with respect to their lipid composition in terms of bilayer fluidity, charge, and hydrophilicity of the liposomal surface, were done to evaluate their usefulness for delivery of encapsulated therapeutic agents to sites of infection . The liposomes showed substantial localization in infected lung tissue after intravenous administration . This was demonstrated in a model of unilateral Klebsiella pneumoniae pneumonia in rats, in which the left lung was infected but the right lung of the same animal developed no infection . The degree of localization in the infected left lung was different for the two types of liposomes . For the liposome type with the longer blood residence time, containing a surface coating of polyethylene glycol, localization in the infected left lung was dependent on the liposomal dose, correlated with the intensity of infection, and reached 9% of the injected liposomal dose in severely infected rats.

Curr Genet, 1993 Jul-Aug, 24(1-2), 32 - 7
Expression of the Klebsiella pneumoniae pullulanase-encoding gene in Saccharomyces cerevisiae; Janse BJ et al.; A 3800-base pair (bp) DNA fragment encoding the mature pullulanase from Klebsiella pneumoniae was inserted between two different yeast expression-secretion cassettes and an yeast gene terminator . These cassettes were cloned into an yeast centromeric plasmid YCplacIII and transformed into laboratory strains of Saccharomyces cerevisiae . Transcription initiation signals were derived from the mating pheromone alpha-factor (MF alpha 1p) and alcohol dehydrogenase (ADC1p) gene promoters . Secretion of pullulanase was directed by the leader sequence of the yeast mating pheromone alpha-factor (MF alpha 1s) . Transcription termination was effected by the yeast tryptophan synthase gene terminator (TRP5T) . Southernblot analysis confirmed the presence of pulA in transformed yeasts and Northern-blot analysis revealed the presence of PUL1 mRNA . A pullulan agarose assay indicated the extracellular production of biologically active pullulanase by S . cerevisiae.

Zhonghua Nei Ke Za Zhi, 1993 Jul, 32(7), 467 - 9
{Antibodies to Klebsiella pneumoniae in ankylosing spondylitis}; Yuan GH et al.; This study was performed to evaluate the involvement of Klebsiella pneumoniae (Kp) in ankylosing spondylitis . Serum IgA, IgG and IgM antibodies to Kp were measured with ELISA in 60 patients with ankylosing spondylitis (AS), 28 patients with rheumatoid arthritis (RA) and 45 healthy individuals . A marked elevation of IgA antibody to Kp was detected in the sera of patients with AS, compared to that in patients with RA (P < 0.02) and healthy controls (P < 0.001) . The positive rate of antibodies to Kp was 55% in patients with active AS, significantly higher than that in patients with inactive AS (16.7%, P < 0.01), patients with RA (17.8%, P < 0.05) and healthy controls (4.4%, P < 0.001) . Stool culture for Kp was carried out in 15 of the 60 patients with AS simultaneously, 3 (20%) of them were positive . Our results are in line with the previously published findings suggesting that Kp may play a role in the pathogenesis of AS.

Indian J Pediatr, 1993 Jul-Aug, 60(4), 565 - 72
Clinical profile of klebsiella septicemia in neonates; Gupta P et al.; A detailed clinical study of 51 consecutive cases of neonatal klebsiella septicemia was carried out prospectively over a 20 months period . The incidence was 6.27 per 1000 live births . Majority (85.5%) were either preterms or small for date . Almost fifty percent babies had associated perinatal risk factors . Mean age of onset was 5.7 +/- 2.2 days . General symptoms were the earliest to occur at mean age of 5.7 days followed by respiratory, alimentary, hematological and neurological symptoms at 6.2, 6.3, 6.6 and 7.9 days respectively . About half of the neonates had associated complications; commoner being meningitis (20%), bleeding manifestations and sclerema (17.6% each) and pneumonia (15.7%) . Cefatoxime was found to be the drug of choice (86% sensitivity) . Nine babies (17.6%) died during the study period at a mean age of 9.1 +/- 3.2 days . Mean duration of hospital stay in rest of neonates was 27.9 +/- 12.1 days . Neurological symptoms were commoner in late onset disease . Bleeding manifestations, sclerema and granulocytopenia were seen exclusively in preterms . Presence of respiratory symptoms, bleeding, sclerema, shock and granulocytopenia were identified as poor prognostic factors in neonatal klebsiella septicemia.

Rev Latinoam Microbiol, 1993 Jul-Sep, 35(3), 237 - 43
{Transferrable broad-spectrum beta-lactam resistance in Klebsiella pneumoniae R-3455}; Conde-Bonfil MC et al.; Bacterial resistance to antimicrobial agents is a common problem observed in hospitals . We characterized a clinical isolate of Klebsiella pneumoniae (R-3455) which was resistant to high concentrations of broad spectrum beta-lactams, aminoglycosides, fluoroquinolones, chloramphenicol and tetracycline . Conjugation experiments showed that the multiresistance could be transferred to Escherichia coli J53-2 receptor strain . The transconjugant X-3455 was resistant to all antibacterials assayed in R-3455, except to fluoroquinolones . We found that both strains R-3455 and X-3455 produced a beta-lactamase which was sensitive to clavulanic acid . Southern hybridization and PCR analysis showed the presence of at least, a TEM type beta-lactamase gene in both strains.

Carbohydr Res, 1993 Jun 21, 244(2), 325 - 40
Structural investigation of the capsular polysaccharide produced by a novel Klebsiella serotype (SK1) . Location of O-acetyl substituents using NMR and MS techniques; Cescutti P et al.; The capsular polysaccharide of Klebsiella SK1 was investigated by methylation analysis, Smith degradation, and 1H NMR spectroscopy . The oligosaccharides (P1 and P2) obtained by bacteriophage phi SK1 degradation of the polymer were studied by methylation analysis, and 1D- and 2D-NMR spectroscopy . The resulting data showed that the parent repeating unit is a branched pentasaccharide having a structure identical to the revised structure recently proposed for Klebsiella serotype K8 capsular polysaccharide . {Formula: see text} The 2D-NMR data showed that one third of the glucuronic acid residues in the SK1 polymer are acetylated at O-2, O-3, or O-4 . FABMS studies confirmed the presence of monoacetylated glucuronic acid residues . Thus, the relationship between the Klebsiella K8 and SK1 polymers is akin to that found for Klebsiella polysaccharides K30 and K33, which have been typed as serologically distinct yet their structures differ only in the degree of acetylation.

J Med Microbiol, 1993 Jun, 38(6), 454 - 8
The effects of RU 41.740, a glycoprotein immunomodulating agent derived from Klebsiella pneumoniae, on intra-abdominal abscess formation in mice; Finlay-Jones JJ et al.; The prophylactic and therapeutic efficacies of the immunomodulating agent RU 41.740 (a glycoprotein extract from Klebsiella pneumoniae) were studied in a murine model of intra-abdominal abscess formation with Bacteroides fragilis, Escherichia coli, and bran as an abscess-potentiating agent . Parenteral injection of RU 41.740, either before or after injection of an abscess-inducing mixture (AIM), was associated with significantly diminished incidence and size of abscesses . Abscess incidence and size were significantly decreased by oral administration of RU 41.740 after, but not before, AIM injection . Abscess formation and resolution are the results of complex interactions of host defence mechanisms with bacteria and potentiating agent, and RU 41.740 has been shown previously to activate both macrophage and neutrophil function . These results indicate that activation of non-specific defences may protect against abscess development in chronic sepsis.

Pharmacology, 1993 Jun, 46(6), 341 - 5
The immunomodulating agent RU 41740 complexed with very-low-density lipoproteins enhances human polymorphonuclear neutrophil oxidative metabolism in vitro; Idohou N et al.; Added to human serum in vitro, RU 41740, an immunomodulating agent extracted from Klebsiella pneumoniae, binds selectively to lipoproteins containing apolipoprotein B (low-density lipoproteins and very-low-density lipoproteins, VLDL) and, at higher concentrations, to lipoproteins containing apolipoprotein A (high-density lipoproteins) . The fact that lipoproteins modulate polymorphonuclear neutrophil (PMN) functions led us to suspect that the VLDL-RU 41740 complex might affect PMN functions . In this study, the effect of this complex on PMN superoxide generation was measured in the presence and absence of the classical stimulants formyl-methionyl-leucyl-phenylalanine and phorbol myristate acetate . The VLDL-RU 41740 complex enhanced the stimulating effect of VLDL on quiescent PMN, but not following stimulation with formyl-methionyl-leucyl-phenylalanine . In contrast, it partially counteracted the inhibiting effect exerted by VLDL alone on phorbol myristate acetate stimulation . Such a complex might be formed in vivo during RU 41740 therapy and constitute an important feature in the immunostimulating properties of the drug.

FEMS Immunol Med Microbiol, 1993 Jun, 7(1), 63 - 6
Application of flow cytometry to the study of antiphagocytic properties of Klebsiella pneumoniae capsular polysaccharide; Ruiz C et al.; We used flow cytometry to compare the effects of whole cells and capsular polysaccharides of Klebsiella pneumoniae on the phagocytic ability ot polymorphonuclear leukocytes . Our results showed a light decrease in phagocytic activity in the presence of capsular polysaccharides, but a marked decrease with whole cells . Our findings suggest that the resistance to phagocytosis in these microorganisms is not due exclusively to their capsule, as claimed by other authors.

J Gen Microbiol, 1993 Jun, 139 ( Pt 6), 1307 - 14
Oxygen inhibition of nitrogenase activity in Klebsiella pneumoniae; Kavanagh EP et al.; A purpose-built oxystat has been used to study reversible inhibition of nitrogenase by O2 in the facultative anaerobe Klebsiella pneumoniae . C2H2-reducing activity in samples from either an anaerobic glucose-limited or an O2-limited diazotrophic chemostat culture was completely inhibited by exposure to a dissolved O2 concentration (DOC) of 1.5 microM or above . Subsequently, under anaerobic conditions, C2H2-reducing activity returned in the absence of de novo protein synthesis . The amount of activity returning never reached 100% of the initial anaerobic activity before O2 treatment . The degree of reversibility was inversely proportional to the log of DOC during exposure and was decreased by increasing the time of exposure to O2 (about 60% reversibility occurred after a 20 min exposure to 6 microM-O2) . The failure to obtain complete recovery of activity was apparently not due to inactivation of the very O2-sensitive pyruvate-flavodoxin oxidoreductase (nifJ product) which provides electrons for nitrogenase activity in vivo . Samples from the O2-limited culture behaved similarly to those limited by glucose . Thus, 'training' of the organism to use O2 during growth does not influence the tolerance of nitrogenase to O2 . Since the behaviour towards O2 reported here for K . pneumoniae differs from that known to occur in Azotobacter, the mechanism of protection of nitrogenase from O2 damage may differ in these organisms.

J Vet Med Sci, 1993 Jun, 55(3), 395 - 400
DNA sequence of type 1 fimbrin, Fpul1, gene from a chicken pathogenic Escherichia coli serotype O78; Sekizaki T et al.; The gene encoding type 1 fimbriae of chicken pathogenic Escherichia coli serotype O78 (designated Fpul1) was cloned and the genetic region encoding fimbrial subunit was sequenced . The nucleotide sequence and its deduced amino acid sequence demonstrated that the Fpul1 was a novel variation among E . coli type 1 fimbriae and showed an extensive homology to previously reported Klebsiella pneumoniae type 1 fimbriae . The E . coli K-12 strains carrying the Fpul1 genes did not show the acid-induced autoagglutination, suggesting that the Fpul1 was genetically distinct from the acid-induced autoagglutination.

Protein Sci, 1993 Jun, 2(6), 1042 - 52
Purification and characterization of Klebsiella aerogenes UreE protein: a nickel-binding protein that functions in urease metallocenter assembly; Lee MH et al.; The Klebsiella aerogenes ureE gene product was previously shown to facilitate assembly of the urease metallocenter (Lee, M.H., et al., 1992, J . Bacteriol . 174, 4324-4330) . UreE protein has now been purified and characterized . Although it behaves as a soluble protein, UreE is predicted to possess an amphipathic beta-strand and exhibits unusually tight binding to phenyl-Sepharose resin . Immunogold electron microscopic studies confirm that UreE is a cytoplasmic protein . Each dimeric UreE molecule (M(r) = 35,000) binds 6.05 + 0.25 nickel ions (Kd of 9.6 +/- 1.3 microM) with high specificity according to equilibrium dialysis measurements . The nickel site in UreE was probed by X-ray absorption and variable-temperature magnetic circular dichroism spectroscopies . The data are most consistent with the presence of Ni(II) in pseudo-octahedral geometry with 3-5 histidyl imidazole ligands . The remaining ligands are nitrogen or oxygen donors . UreE apoprotein has been crystallized and analyzed by X-ray diffraction methods . Addition of nickel ion to apoprotein crystals leads to the development of fractures, consistent with a conformational change upon binding nickel ion . We hypothesize that UreE binds intracellular nickel ion and functions as a nickel donor during metallocenter assembly into the urease apoprotein.

Protein Sci, 1993 Jun, 2(6), 1034 - 41
Site-directed mutagenesis of Klebsiella aerogenes urease: identification of histidine residues that appear to function in nickel ligation, substrate binding, and catalysis; Park IS et al.; Comparison of six urease sequences revealed the presence of 10 conserved histidine residues (H96 in the gamma subunit, H39 and H41 in beta, and H134, H136, H219, H246, H312, H320, and H321 in the alpha subunit of the Klebsiella aerogenes enzyme) . Each of these residues in K . aerogenes urease was substituted with alanine by site-directed mutagenesis, and the mutant proteins were purified and characterized in order to identify essential histidine residues and assign their roles . The gamma H96A, beta H39A, beta H41A, alpha H312A, and alpha H321A mutant proteins possess activities and nickel contents similar to wild-type enzyme, suggesting that these residues are not essential for substrate binding, catalysis, or metal binding . In contrast, the alpha H134A, alpha H136A, and alpha H246A proteins exhibit no detectable activity and possess 53%, 6%, and 21% of the nickel content of wild-type enzyme . These results are consistent with alpha H134, alpha H136, and alpha H246 functioning as nickel ligands . The alpha H219A protein is active and has nickel (approximately 1.9% and approximately 80%, respectively, when compared to wild-type protein) but exhibits a very high Km value (1,100 +/- 40 mM compared to 2.3 +/- 0.2 mM for the wild-type enzyme) . These results are compatible with alpha H219 having some role in facilitating substrate binding . Finally, the alpha H320A protein (Km = 8.3 +/- 0.2 mM) only displays approximately 0.003% of the wild-type enzyme activity, despite having a normal nickel content.(ABSTRACT TRUNCATED AT 250 WORDS)

J Hosp Infect, 1993 Jun, 24(2), 123 - 8
Pulsed-field gel electrophoresis for differentiation of hospital isolates of Klebsiella pneumoniae; Poh CL et al.; Restriction enzyme analysis by pulsed-field gel electrophoresis (PFGE) was developed for differentiation of hospital isolates of Klebsiella pneumoniae . Restriction patterns generated by SpeI digestion of genomic DNAs of 36 isolates from patients in two major teaching hospitals established 34 PFGE types . All strains were typable by this technique and the SpeI restriction patterns were reproducible, stable and easy to interpret . As PFGE profiles generated were heterogenous, the incidence of cross-infection appeared to be low in each of the hospitals . The higher discriminatory power of PFGE when compared to conventional restriction endonuclease analysis (REA) suggests that this technique will be very useful for epidemiological investigations of nosocomial K . pneumoniae outbreaks.

Microb Pathog, 1993 Jun, 14(6), 433 - 40
The role of the O-antigen lipopolysaccharide on the colonization in vivo of the germfree chicken gut by Klebsiella pneumoniae; Camprubi S et al.; We isolated lipopolysaccharide and capsular polysaccharide (K antigen)-defective mutants from two Klebsiella pneumoniae parental strains, and compared their ability to colonize in vivo the germfree chicken gut . The high-molecular weight lipopolysaccharide (LPS) (O antigen) was found necessary for the colonization while the capsular polysaccharide (K2 or K29) was not of importance.

Biochem J, 1993 May 15, 292 ( Pt 1), 93 - 8
Klebsiella pneumoniae nitrogenase: pre-steady-state absorbance changes show that redox changes occur in the MoFe protein that depend on substrate and component protein ratio; a role for P-centres in reducing dinitrogen?
Lowe DJ, Fisher K, Thorneley RN.
The pre-steady-state absorbance changes that occur during the first 0.6 s of reaction of the nitrogenase of Klebsiella pneumoniae can be simulated by associating redox changes with the different states of the MoFe protein described by our published kinetic model for nitrogenase {Lowe and Thorneley (1984) Biochem . J . 224, 877-886} . When the substrate is changed, from H+ to C2H2 (acetylene) or N2, or the nitrogenase component protein ratio is altered, these pre-steady-state absorbance changes are affected in a manner that is quantitatively predicted by our model . The results, together with parallel e.p.r . studies, are interpreted as showing that the P-clusters become oxidized when the MoFe protein is in the state where bound N2 is irreversibly committed to being reduced and is protonated to the hydrazido(2-) level.

J Biol Chem, 1993 May 15, 268(14), 10060 - 5
Cation-coupling in chimeric melibiose carriers derived from Escherichia coli and Klebsiella pneumoniae . The amino-terminal portion is crucial for Na+ recognition in melibiose transport; Hama H et al.; The melibiose carrier of Escherichia coli couples sugar transport to H+, Na+, and Li+, while that of Klebsiella pneumoniae utilizes only H+ and Li+ . We made five chimeric carriers derived from the two carriers to identify the region(s) involved in Na+ recognition . The chimeric carriers E2K10, E4K8, E6K6, E8K4, and E10K2 have the amino-terminal 77, 144, 197, 298, and 349 amino acid residues derived from E . coli and the rest derived from K . pneumoniae, respectively . Melibiose accumulation through the chimeric carriers E2K10, E4K8, and E6K6 was strongly stimulated by Na+ and Li+ as is the case with the E . coli carrier . On the other hand, there was very little stimulation with the carriers E8K4 and E10K2 . These results suggest that, 1) the amino-terminal 77 amino acids of the E . coli carrier, which has 5 different and 4 fewer amino acids than the K . pneumoniae carrier, have a crucial role in Na+ recognition in melibiose transport and 2) the carboxyl-terminal half of the carrier also forms a part of the Na+ recognition site which may be distorted in chimeric structures . In contrast with melibiose accumulation, there was very little Na+ stimulation of TMG (methyl-1-thio-beta-D-galactopyranoside) transport and no Na+ stimulation was observed in lactose transport with any of the chimeric carriers, whereas in E . coli Na+ stimulates TMG and lactose transport . These results suggest that there is no universal Na+ recognition site for all the sugar substrates . Instead different parts of the carrier seem to participate in cation recognition for different sugar substrates.

J Clin Microbiol, 1993 May, 31(5), 1379 - 81
Development of an enzyme-linked immunosorbent assay method for typing and quantitation of Klebsiella pneumoniae lipopolysaccharide: application to serotype O1; Alberti S et al.; We describe a method for the typing and quantitation of Klebsiella pneumoniae serotype O1 lipopolysaccharide (LPS) based on inhibition in an enzyme-linked immunosorbent assay of a reaction of known O1 LPS antigen and anti-O1 antibody by unknown LPS extracts . Serotype O1 was found in 32% of the 124 K . pneumoniae clinical isolates tested, showing that this serotype is frequent among the eight O serotypes which have been described previously.

J Bacteriol, 1993 May, 175(10), 2926 - 35
Expression of the nifBfdxNnifOQ region of Azotobacter vinelandii and its role in nitrogenase activity; Rodriguez-Quinones F et al.; The nifBQ transcriptional unit of Azotobacter vinelandii has been previously shown to be required for activity of the three nitrogenase systems, Mo nitrogenase, V nitrogenase, and Fe nitrogenase, present in this organism . We studied regulation of expression and the role of the nifBQ region by means of translational beta-galactosidase fusions to each of the five open reading frames: nifB, orf2 (fdxN), orf3 (nifO), nifQ, and orf5 . Expression of the first three open reading frames was observed under all three diazotrophic conditions; expression of orf5 was never observed . Genes nifB and fdxN were expressed at similar levels . With Mo, expression of nifO and nifQ was approximately 20- and approximately 400-fold lower than that of fdxN, respectively . Without Mo, expression of nifB dropped three- to fourfold and that of nifQ dropped to the detection limit . However, expression of nifO increased threefold . The products of nifB, fdxN, nifO, and nifQ have been visualized in A . vinelandii as beta-galactosidase fusion proteins with the expected molecular masses . The NifB- fusion lacked activity for any of the three nitrogenase systems and showed an iron-molybdenum cofactor-deficient phenotype in the presence of Mo . The FdxN- mutation resulted in reduced nitrogenase activities, especially when V was present . Dinitrogenase activity in extracts was similarly affected, suggesting a role of FdxN in iron-molybdenum cofactor synthesis . The NifO(-)-producing mutation did not affect any of the nitrogenases under standard diazotrophic conditions . The NifQ(-)-producing mutation resulted in an increased (approximately 1,000-fold) Mo requirement for Mo nitrogenase activity, a phenotype already observed with Klebsiella pneumoniae . No effect of the NifQ(-)-producing mutation on V or Fe nitrogenase was found; this is consistent with its very low expression under those conditions . Mutations in orf5 had no effect on nitrogenase activity.

Biochem J, 1993 May 1, 291 ( Pt 3), 709 - 11
Energy transduction by nitrogenase: binding of MgADP to the MoFe protein is dependent on the oxidation state of the iron-sulphur 'P' clusters; Miller RW et al.; Hydrolysis of MgATP to MgADP is essential for nitrogenase action . There is good evidence for binding of both nucleotides to the Fe protein of nitrogenase, but data indicating their binding to the MoFe protein have been controversial {see Miller and Eady (1989) Biochem . J . 263, 725-729} . The binding of MgADP to the MoFe protein of nitrogenase of Klebsiella pneumoniae was investigated by non-equilibrium gel-filtration column methods . No binding of MgADP to the dithionite-reduced protein could be detected . Treatment of the MoFe protein with phenosafranine {midpoint potential (Em) -270 mV} did not affect the activity, and oxidized the 'P' clusters but not the iron-molybdenum cofactor (FeMoco) centres . This oxidized species bound 3.9 mol of MgADP with a binding pattern characteristic of low rates of ligand dissociation . These observations suggest that the variability in published data on nucleotide binding to the MoFe protein is related to poor control of the protein oxidation level . Our data, coupled with the observation that 'P' clusters become oxidized during reduction of N2 {Lowe, Fisher and Thorneley (1993) Biochem . J., in the press}, led us to propose that the ADP binding sites are transiently filled during enzyme turnover by hydrolysis of ATP originally bound to the Fe protein, and that hydrolysis occurs on a bridging site on the MoFe-Fe-protein complex.

Infect Immun, 1993 May, 61(5), 1996 - 2002
Bacteria induce release of platelet-activating factor (PAF) from polymorphonuclear neutrophil granulocytes: possible role for PAF in pathogenesis of experimentally induced bacterial pneumonia; Makristathis A et al.; The role of platelet-activating factor (PAF) as mediator of the endotoxin shock and endotoxin-dependent tissue injury has been examined . The ability of opsonized bacteria to stimulate the release of PAF from human polymorphonuclear neutrophil granulocytes was evaluated by measuring both the activity and the amount of the mediator released in the supernatant of the cell-bacteria reaction in vitro . There was no significant difference between gram-positive and gram-negative bacteria in the ability to release PAF from neutrophils . However, preincubation of the cells with the specific PAF receptor antagonist WEB 2170 decreased release of PAF from the cells . Furthermore, a possible protective effect of the PAF antagonist was examined during experimentally induced pneumonia with Klebsiella pneumoniae in NMRI mice . Oral treatment of mice with WEB 2170, followed by infection with the microorganisms, resulted in a considerable increase in the animals' survival (53 to 73%) compared with the control group (40%); this increase corresponded with a decrease in the CFU per gram of lung tissue . These findings indicate an important role of PAF in the pathogenesis of pneumonia in mice.

Antimicrob Agents Chemother, 1993 May, 37(5), 1061 - 4
Resistance to cefoperazone-sulbactam in Klebsiella pneumoniae: evidence for enhanced resistance resulting from the coexistence of two different resistance mechanisms; Rice LB et al.; We investigated the in vitro activity and the in vivo efficacy of the beta-lactam-beta-lactamase inhibitor combination cefoperazone-sulbactam against an isogenic series of Klebsiella pneumoniae strains . Both cefoperazone and cefoperazone-sulbactam were active in vitro against a susceptible clinical strain, and the combination was highly effective in the treatment of rat intra-abdominal abscesses . Loss of expression of a 39-kDa outer membrane protein resulted in at least a fourfold increase in the MICs of cefoperazone and cefoperazone-sulbactam but did not appreciably affect the in vivo efficacy of either regimen . Introduction of plasmid RP4, which encodes the TEM-2 beta-lactamase, into the susceptible strain resulted in the loss of in vitro activity and in vivo efficacy for cefoperazone . The in vitro activity of cefoperazone-sulbactam against this strain was diminished, but the antibiotic combination remained highly active in vivo . Introduction of RP4 into the strain lacking the 39-kDa outer membrane protein resulted in a fourfold increase in the in vitro MIC of cefoperazone-sulbactam in comparison with the beta-lactamase-producing susceptible strain and resulted in a loss of in vivo efficacy against infections caused by this strain . These results suggest that the combination of different resistance mechanisms, neither of which alone results in substantially diminished cefoperazone-sulbactam efficacy in vivo, can cause in vivo resistance to the beta-lactam-beta-lactamase inhibitor combination in K . pneumoniae.

Mikrobiol Zh, 1993 May-Jun, 55(3), 57 - 63
{The physiological-biochemical properties of Klebsiella pneumoniae strains of different origins}; Semenova EA et al.; While comparatively analyzing the properties of Klebsiella pneumoniae from three studied sources (patients, healthy people, environment), no reliable differences between the comparable strains have been revealed . No differences have been found in the toxin formation, hemagglutinating and antilysozymic activity, adhesivity and invasive properties as well as in the peculiarities of gas exchange, while growing on media with different nitrogen and carbon sources . The studied microorganisms differed only in antibiotic resistance: strains isolated from the environment were sensitive to antibiotics, whereas strains of human origin were resistant to them.

Arch Fr Pediatr, 1993 May, 50(5), 409 - 11
{Severe colectasy in rheumatoid purpura . Probable role of water soluble contrast products}; Berard E et al.; BACKGROUND . About 60-70% of cases of anaphylactoid purpura suffer from abdominal colic, which may be quite severe . Because severe abdominal pain may be difficult to differentiate from a surgical abdomen, repeated examination and radiological studies are often necessary . CASE REPORT . A 4 year 7 month-old girl suffered from abdominal pain, vomiting and the skin rash characteristic of anaphylactoid purpura . Because of worsening of the abdominal manifestations, a Gastrografin enema was given . The result was normal but 48 hours later, the child suffered from acute abdominal colics plus symptoms of shock . X-ray examination revealed a large distension of colon and persistence of the contrast material . Exploratory laparotomy failed to find any perforation or necrosis . The colon was drained and the child was given parenteral nutrition and antibiotics . Blood culture showed Klebsiella pneumoniae . An upper gastrointestinal roentgenogram using meglumine ioxitalamate was performed 19 days later because of recurrence of the abdominal pain . This showed a submucosal hemorrhage in the first small bowel loop . Increased abdominal distension occurred 3 days later and the results of X-rays were similar to those seen the Gastrografin enema . A second enema using meglumine ioxitalamate was performed 3 days later . It showed sigmoid constriction but a second laparotomy failed to confirm this obstruction . The child died a few hours later despite ileostomy and antibiotics . CONCLUSIONS . The severe colectasy seen in this case of anaphylactoid purpura indicates that caution is required in performing enemas in acute digestive complications of this disease . Such X-ray studies should not be repeated and all water-soluble contrast material should be voided as soon as possible.

J Bacteriol, 1993 May, 175(9), 2507 - 15
Functional organization of the glnB-glnA cluster of Azospirillum brasilense; de Zamaroczy M et al.; The functional organization of the glnB-A cluster of Azospirillum brasilense, which codes for the PII protein and glutamine synthetase, respectively, was studied with the aid of lacZ fusions, deletion mapping, site-directed mutagenesis, and complementation . It was shown previously by mRNA mapping that the cluster contains two tandemly organized promoters, glnBp1 and glnBp2, of the sigma 70 and sigma 54 types, respectively, upstream of glnB and a third unidentified promoter upstream of glnA . Data obtained with lacZ fusions in the wild-type strain confirmed that cotranscription of glnBA and transcription of glnA alone were oppositely regulated by the cell N status . Quantification of promoter activities showed a high level of transcription from glnBp1p2 and a low level from glnAp under conditions of nitrogen limitation . The opposite situation prevails under conditions of nitrogen excess . As a consequence, PII polypeptide synthesis is increased under conditions of nitrogen fixation, which strongly suggests that PII plays an important role under these conditions . Null mutant strains of glnB, ntrB-ntrC, nifA, and point mutant strains in glnA were analyzed . NtrB and NtrC are not involved in the regulation of glnBA expression, in contrast to PII and glutamine synthetase . Glutamine synthetase probably acts by modulating the intracellular N status, and PII acts by modifying the properties of an unidentified regulator which might be a functional homolog of NtrC . In addition, a Nif- null mutant strain of glnB was characterized further . A Nif+ phenotype was restored to the strain by nifA from Klebsiella pneumoniae but not by nifA from A . brasilense . This mutant strain is not impaired in NifA synthesis, which is relatively independent of the growth conditions in A . brasilense . It is therefore most likely that PII is required for NifA activation under conditions of nitrogen fixation . Deletion mapping and site-directed mutagenesis showed glnAp was located within a 45-bp DNA fragment upstream of the mRNA start site, dissimiar to previously described consensus sites for sigma factors.

Res Microbiol, 1993 May, 144(4), 259 - 69
Construction and first characterization of two reciprocal hybrids between LamB from Escherichia coli K12 and Klebsiella pneumoniae; Werts C et al.; The LamB proteins from Klebsiella pneumoniae and Escherichia coli K12 were previously shown to be highly homologous . The most conserved parts correspond to the N-proximal third and to the transmembranous portions of the molecule, while the variability occurred essentially within regions exposed to the cell surface or to the periplasm . Since the two proteins displayed identical in vitro trimer stability and in vivo pore properties, we tested whether the N-terminal parts of the two proteins could be exchanged and still allow the formation of stable and functional maltoporins . For that purpose, we expressed the LamB protein from K . pneumoniae in E . coli K12, and constructed two reciprocal hybrids between LamB from E . coli K12 and LamB from K . pneumoniae . The first hybrid (LamBE.c.-K.p.) is composed of residues 1 to 183 from LamBE.c . followed by residues 184 to 404 from LamBK.p . The second one comprises residues 1 to 183 from LamBK.p., followed by residues 184 to 421 from LamBE.c . (LamBK.p.-E.c.) . Both hybrid proteins were correctly incorporated in the outer membrane of E . coli K12 . Like the two parental LamB proteins, the two hybrids could be purified by affinity chromatography on a starch-sepharose column . The LamBE.c.-K.p . hybrid formed highly stable trimers, but was strongly impaired in its in vivo maltose transport function (15% of the wild-type level) . The trimers formed by LamBK.p.-E.c . hybrid were less stable, but could be detected on the surface of intact cells by four anti-LamBE.c . monoclonal antibodies . This hybrid was also affected in its in vivo maltose transport function (30% of the wild-type level) . As expected from the location of the residues critical for phage adsorption, both proteins had lost the phage receptor activity of the E . coli K12 LamB protein . We also examined whether LamBE.c . could form heterotrimers with LamBK.p., LamBK.p.-E.c., and LamBE.c.-K.p . In no case were heterotrimers detected, indicating that both terminal parts of the LamB protein are involved in homotrimer formation . All these data suggest that trimer formation and activity involve rare variable residues in the conserved regions and/or variable regions.

New Microbiol, 1993 Apr, 16(2), 165 - 70
Peptidoglycan synthesis and its fine chemical composition in dividing and not dividing Klebsiella pneumoniae cocci; Canepari P et al.; Peptidoglycan synthesis and its fine chemical composition were studied in dividing and in non-dividing Klebsiella pneumoniae cocci and compared with rods . The beta-lactam mecillinam, a specific inhibitor of lateral wall elongation which causes rod-to-sphere transition in rods, showed 50% inhibition of the peptidoglycan in normal rods of the parent Mir A12 only if added at an early stage of the cell cycle and no effect if added later or during septation . In the rods of the mutant Mir M7, mecillinam was shown to inhibit 50% of peptidoglycan synthesis until rods become cocci, and thereafter to be absolutely devoid of effects . On the contrary, piperacillin, a specific inhibitor of septum formation, was active on all strains regardless of their cell shape, only if added at 20 and removed at 40 min of the cell cycle . As regards the analysis of peptidoglycan fine chemical composition, bacteria dividing as cocci showed alterations in the muropeptide composition consisting in a 50-fold increase in the tetramer family . This alteration was not seen in the cocci that did not divide as such . These results confirm our previous claim that septum formation and lateral wall elongation are mutually exclusive in normal rods and that septum formation requires the synthesis of a peptidoglycan of different chemical composition.

New Microbiol, 1993 Apr, 16(2), 135 - 40
The reshaping process of Klebsiella pneumoniae cells after removal of mecillinam, an antibiotic that causes transition from rod to coccal shape; Satta G et al.; The process of bacterial morphogenesis that leads to rod shape formation was studied in synchronous cells during the reshaping process after removal of mecillinam, a beta-lactam antibiotic which, by specifically inhibiting lateral wall formation of rods, cause rod-to-sphere transition in Gram-negative rods . The addition of mecillinam for 50 min of the cell cycle made the cells to skip a division, while the addition of the antibiotic for 30 min (or less), allowed the cells to divide regularly . In order to study the interplay between lateral wall elongation and septum formation in reacquisition of rod shape, we evaluated the effect of re-adding mecillinam or adding piperacillin, a specific inhibitor of septum formation, at various stages of the reshaping process . It was found that mecillinam was active only when added within the first 30 min of the reshaping process, while piperacillin was active only after 30 min when the cells were close to starting to divide again . These findings provide further support for our previous proposal that, in bacterial rods, elongation and septation are two alternating and competing events of the cell cycle, and are linked to each other in such a way as to force bacterial rods to grow to a given length.

Mol Microbiol, 1993 Apr, 8(1), 187 - 98
Identification of a nitrogen-regulated promoter controlling expression of Klebsiella pneumoniae urease genes; Collins CM et al.; Synthesis of urease by Klebsiella species is known to be induced when the nitrogen source of the growth medium is limiting, suggesting that urease gene expression is controlled by the nitrogen regulatory (ntr) system . This study showed that K . pneumoniae with mutations in either ntrA or ntrC, two integral components of the ntr system, were phenotypically urease-negative . These mutants could be complemented back to a urease positive phenotype with recombinant plasmids encoding the corresponding ntr gene . A series of ure-lacZYA transcriptional fusions, in conjunction with primer extension analysis, identified a DNA region that encoded a nitrogen-regulated promoter . This promoter region controlled transcription of ureD, the first gene in the Klebsiella pneumoniae urease gene cluster, and ureA, a gene that resides immediately downstream of ureD . A high level of transcription from the ureD promoter required NAC, a recently characterized member of the nitrogen regulatory cascade . NAC is a Lys R-like transcriptional regulator that can act at sigma 70 promoters; expression from nac itself is dependent upon NTRA . Therefore, expression of K . pneumoniae urease was dependent upon the nitrogen regulatory cascade, and transcription of at least two urease genes was from a promoter that was positively regulated by NAC.

Mol Gen Genet, 1993 Apr, 238(3), 369 - 82
Nucleotide sequence and genetic analysis of the Rhodobacter capsulatus ORF6-nifUI SVW gene region: possible role of NifW in homocitrate processing; Masepohl B et al.; DNA sequence analysis of a 3494-bp HindIII-BclI fragment of the Rhodobacter capsulatus nif region A revealed genes that are homologous to ORF6, nifU, nifS, nifV and nifW from Azotobacter vinelandii and Klebsiella pneumoniae . R . capsulatus nifU, which is present in two copies, encodes a novel type of NifU protein . The deduced amino acid sequences of NifUI and NifUII share homology only with the C-terminal domain of NifU from A . vinelandii and K . pneumoniae . In contrast to nifA and nifB, which are almost perfectly duplicated, the predicted amino acid sequences of the two NifU proteins showed only 39% sequence identity . Expression of the ORF6-nifUISVW operon, which is preceded by a putative sigma 54-dependent promoter, required the function of NifA and the nif-specific rpoN gene product encoded by nifR4 . Analysis of defined insertion and deletion mutants demonstrated that only nifS was absolutely essential for nitrogen fixation in R . capsulatus . Strains carrying mutations in nifV were capable of very slow diazotrophic growth, whereas ORF6, nifUI and nifW mutants as well as a nifUI/nifUII double mutant exhibited a Nif+ phenotype . Interestingly, R . capsulatus nifV mutants were able to reduce acetylene not only to ethylene but also to ethane under conditions preventing the expression of the alternative nitrogenase system . Homocitrate added to the growth medium repressed ethane formation and cured the NifV phenotype in R . capsulatus . Higher concentrations of homocitrate were necessary to complement the NifV phenotype of a polar nifV mutant (NifV-NifW-), indicating a possible role of NifW either in homocitrate transport or in the incorporation of this compound into the iron-molybdenum cofactor of nitrogenase.

J Dairy Sci, 1993 Apr, 76(4), 978 - 82
Growth responses of coliform bacteria to recombinant bovine cytokines; Hogan JS et al.; Growth responses of 10 coliform isolates to recombinant bovine cytokines were measured in vitro . Six Escherichia coli and four Klebsiella pneumoniae isolates obtained from bovine IMI were tested for growth responses to recombinant bovine interleukin-1 beta, interleukin-2, and interferon-gamma . Cytokines were tested at 10(4), 10(3), 10(2), and 10 U/ml of media . Media used were a synthetic tissue culture medium, a chemically defined synthetic bacterial growth medium, and UHT sterilized milk . Bacterial counts in the synthetic tissue culture medium and UHT milk increased slightly as concentration of interferon-gamma in the media increased . Recombinant bovine interferon-gamma increased bacterial populations during the log growth phase but did not affect the number of bacteria in stationary growth phase . Bacterial growth responses were not related to either interleukin-2 or interleukin-1 beta concentrations in any of the three media . Bacterial growth responses to cytokines were not related to differences in either serum susceptibility, growth of isolates in dry cow secretion, duration of IMI from which isolates were obtained, or bacterial species.

Eur J Biochem, 1993 Apr 1, 213(1), 445 - 53
Structural determination of the capsular polysaccharide produced by Klebsiella pneumoniae serotype K40 . NMR studies of the oligosaccharide obtained upon depolymerisation of the polysaccharide with a bacteriophage-associated endoglycanase; Cescutti P et al.; The Klebsiella pneumoniae K40 capsular polysaccharide has been isolated and investigated by use of methylation analysis, specific degradations and NMR spectroscopy . The polysaccharide was depolymerised by a bacteriophage-associated endogalactosidase, and the resulting oligosaccharide was characterised by one-dimensional and two-dimensional NMR spectroscopy and direct chemical ionisation MS . The repeating unit of the K40 capsular polysaccharide was shown to be a linear hexasaccharide with the composition-->3)- alpha-L-Rhap-(1-->2)-alpha-L-Rhap-(1-->4)-alpha-D-GlcpA++ +-(1-->2-)- alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->3)-alpha-D-Galp-(1--> (Rha, rhamnose).

J Bacteriol, 1993 Apr, 175(7), 2162 - 7
Characterization of an Escherichia coli aromatic hydroxylase with a broad substrate range; Prieto MA et al.; The hpaB gene encoding an aromatic hydroxylase of Escherichia coli ATCC 11105, a penicillin G acylase-producing strain, has been cloned and expressed in E . coli K-12 . This gene was located near the pacA gene coding for penicillin G acylase . The hydroxylase has a molecular mass of 59,000 Da, uses NADH as a cosubstrate, and was tentatively classified as a 4-hydroxyphenylacetic acid hydroxylase, albeit it exhibited a rather broad substrate specificity acting on different monohydric and dihydric phenols . E . coli W, C, and B as well as Klebsiella pneumoniae M5a1 and Kluyvera citrophila ATCC 21285 (a penicillin G acylase-producing strain) but not E . coli K-12 contained sequences homologous to hpaB . Our results support the hypothesis that hpaB is a component of the 4-hydroxyphenylacetic acid degradative pathway of E . coli W.

J Bacteriol, 1993 Apr, 175(7), 2116 - 24
The product of the Klebsiella aerogenes nac (nitrogen assimilation control) gene is sufficient for activation of the hut operons and repression of the gdh operon; Schwacha A et al.; In Klebsiella aerogenes, the formation of a large number of enzymes responds to the quality and quantity of the nitrogen source provided in the growth medium, and this regulation requires the action of the nitrogen regulatory (NTR) system in every case known . Nitrogen regulation of several operons requires not only the NTR system, but also NAC, the product of the nac gene, raising the question of whether the role of NAC is to activate operons directly or by modifying the specificity of the NTR system . We isolated an insertion of the transposon Tn5tac1 which puts nac gene expression under the control of the IPTG-inducible tac promoter rather than the nitrogen-responsive nac promoter . When IPTG was present, cells carrying the tac-nac fusion activated NAC-dependent operons and repressed NAC-repressible operons independent of the nitrogen supply and even in the absence of an active NTR system . Thus, NAC is sufficient to regulate operons like hut (encoding histidase) and gdh (encoding glutamate dehydrogenase), confirming the model that the NTR system activates nac expression and NAC activates hut and represses gdh . Activation of urease formation occurred at a lower level of NAC than that required for glutamate dehydrogenase repression, and activation of histidase formation required still more NAC.

J Bacteriol, 1993 Apr, 175(7), 2107 - 15
The nac (nitrogen assimilation control) gene from Klebsiella aerogenes; Schwacha A et al.; The Klebsiella aerogenes nac gene, whose product is necessary for nitrogen regulation of a number of operons, was identified and its DNA sequence determined . The nac sequence predicted a protein a 305 amino acids with a strong similarity to members of the LysR family of regulatory proteins, especially OxyR from Escherichia coli . Analysis of proteins expressed in minicells showed that nac is a single-gene operon whose product has an apparent molecular weight of about 32 kDa as measured in sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Immediately downstream from nac is a two-gene operon, the first gene of which encodes another member of the LysR family . Upstream from nac is a tRNAAsn gene transcribed divergently from nac . About 60 bp upstream from the nac open reading frame lies a sequence nearly identical to the consensus for sigma 54-dependent promoters, with the conserved GG and GC nucleotides at -26 and -14 relative to the start of transcription . About 130 bp farther upstream (at -153 relative to the start of transcription) is a sequence nearly identical to the transcriptional activator NTRC-responsive enhancer consensus . Another weaker NTRC-binding site is located adjacent to this site (at -133 relative to the start of transcription) . Thus, we propose that nac is transcribed by RNA polymerase carrying sigma 54 in response to the nitrogen regulatory (NTR) system . A transposon located between the promoter and the nac ORF prevented NTR-mediated expression of nac, supporting this identification of the promoter sequence . The insertion of over 5 kb of transposon DNA between the enhancer and its target promoter had only a weak effect on enhancer-mediated regulation, suggesting that enhancers may be able to act at a considerable distance on the bacterial chromosome.

Biochem J, 1993 Apr 1, 291 ( Pt 1), 309 - 14
Klebsiella pneumoniae nitrogenase MoFe protein: chymotryptic proteolysis affects function by limited cleavage of the beta-chain and provides high-specific-activity MoFe protein; Fisher K et al.; Proteinase treatment with chymotrypsin has been used to probe the structure of native Klebsiella pneumoniae nitrogenase MoFe protein (Kp1) . Reaction with chymotrypsin did not bleach Kp1, suggesting that it did not destroy the metal centres, and the Mo and Fe contents of Kp1 were unchanged . High ratios of chymotrypsin to Kp1 (1:1 by mass) cleaved the beta-chain of Kp1 to give 44 and 14 kDa polypeptides, which N-terminal amino acid sequence analysis showed to be derived from cleavage at residue beta-Phe124 . A mutant MoFe protein, Kp1Met-124, in which beta-Phe124 is replaced by methionine, was not cleaved by chymotrypsin . Under non-denaturing conditions, the 'nicked' beta-chain of the wild-type protein remained associated with the alpha-chain . The alpha-chain was not cleaved by the proteinase treatment . Fission of the wild-type beta-chain was accompanied by loss of enzyme activity, loss of intensity of the g = 3.7 e.p.r . signal derived from dithionite-reduced FeMoco and by changes in the visible spectrum . The e.p.r . spectra of potassium ferricyanide-oxidized native and digested Kp1 show differences in the signals between g = 1.6 and 2.0 . After prolonged treatment, the final specific activity of Kp1 was about 25 +/- 5% of the initial activity . This corresponded to 25 +/- 5% of the beta-chain which was resistant to proteolytic action . Brief treatment of Kp1 with a lower concentration of chymotrypsin (chymotrypsin/Kp1 ratio = 1:10 by mass, for 10 min) preferentially cleaved high-molecular-mass polypeptides that routinely contaminate preparations of Kp1 prepared by standard procedures . Treatment with chymotrypsin followed by gel filtration to remove the proteinase and cleaved protein fragments can therefore be used to increase significantly the specific activity of Kp1 preparations and remove contaminating activities, such as the ATPase activity of myokinase.

Pathol Biol (Paris), 1993 Apr, 41(4), 343 - 8
{Determination of the isoelectric point of beta-lactamases isolated from 67 Klebsiella oxytoca strains and phenotype behaviour against eight beta-lactam antibiotics}; Chardon H et al.; Klebsiella oxytoca is naturally resistant to aminopenicillins and carboxypenicillins by production of a chromosomal beta-lactamase but susceptible to third generation cephalosporins . The third generation cephalosporins activity may be reduced by: overproduction of the chromosomally encoded beta-lactamase or an extended-spectrum beta-lactamase . These activity modification are rarely found in the hospital of Aix-en-Provence (France) . The activity modification rate of K . oxytoca resistant by one of these mechanisms between 1986 and 1991 are 3.6% for chromosomally encoded beta-lactamase overproduction and 0.7% for acquisition of an extended-spectrum beta-lactamase . We have determined the isoelectric point (pI) by isoelectrofocusing of the beta-lactamases isolated from 67 K . oxytoca and the activity of 8 beta-lactams has been studied by disk-diffusion . 51 wild strains, 14 overproducing strains (including 2 in vitro mutants) and 2 strains with extended-spectrum beta-lactamase were studied . For every wild strain, we observed only one band except for two strains with two bands (pl 5.4 + 6.3 and pl 5.6 + 7.7) . The isoelectric points for the other strains are comprised between pl 5.25 and pl 8.2: 22 pl 7.7; 13 pl 5.25; 4 pl 5.6; 4 pl 6.3; 2 pl 6.6; 2 pl 8.1 and 2 pl 8.2 . In the strains with chromosomally encoded beta-lactamase overproduction we observed several bands in each extract and only the major band was considered . The isoelectric point of in vitro mutant strains with beta-lactamase overproduction was the same that the wild strains . We observed 3 pl: 5.25 - 1 pl: 5.6 - 3 pl 6.3 and 5 pl 7.7.(ABSTRACT TRUNCATED AT 250 WORDS)

Proc Natl Acad Sci U S A, 1993 Mar 15, 90(6), 2266 - 70
Activity of purified NIFA, a transcriptional activator of nitrogen fixation genes; Lee HS et al.; The NIFA protein activates transcription of nitrogen fixation (nif) operons by the sigma 54-holoenzyme form of RNA polymerase . We purified active NIFA from Klebsiella pneumoniae in the form of a maltose-binding protein (MBP)-NIFA fusion; proteolytic release of MBP yielded inactive and insoluble NIFA . MBP-NIFA activated transcription from the nifHDK promoter in a purified transcription system . Like the related transcriptional activator NTRC, MBP-NIFA catalyzed the ATP-dependent isomerization of closed complexes between sigma 54-holoenzyme and a promoter to open complexes . MBP-NIFA had a broader nucleotide specificity than NTRC, being able to utilize pyrimidine in addition to purine nucleoside triphosphates . Both MBP-NIFA and a purified C-terminal fragment of NIFA bound to the upstream activation sequence for the nifHDK promoter, as assessed by DNAse I footprinting . When assays were performed at 37 degrees C instead of the usual 30 degrees C, transcriptional activation, open complex formation, and DNA binding by MBP-NIFA were all abolished, consistent with the known heat lability of NIFA . However, the purified C-terminal fragment of NIFA still bound the upstream activation sequence at 37 degrees C, indicating that the function of the helix-turn-helix DNA-binding motif is not inherently heat-labile.

Vopr Med Khim, 1993 Mar-Apr, 39(2), 58 - 62
{Induction of Ca2+-dependent chemiluminescent response of peritoneal macrophages with bacterial lipopolysaccharides}; Uvarov VD et al.; A lipopolysaccharide preparation obtained from Klebsiella pneumonia was shown to affect primarily the peritoneal macrophages after intraperitoneal administration . Transition of the macrophages to a new metabolic state in response to opsonized zymosan was responsible for a distinct increase in the rate of luminol-dependent chemoluminescence . At the same time, the macrophages produced chemiluminescence was only slightly increased in the presence of Ca2+ ionophore A23187 . These dissimilar alterations observed suggest that the primary reaction of macrophages in response to the lipopolysaccharide occurred with the increase in content of intracellular free calcium . Administration of finoptin (calcium influx inhibitor) into suspension of macrophages caused a decrease in the rate of chemiluminescence response.

Mol Gen Genet, 1993 Mar, 237(3), 400 - 6
The Azotobacter vinelandii nifL-like gene: nucleotide sequence analysis and regulation of expression; Raina R et al.; The nucleotide sequence of the Azotobacter vinelandii nifL-like gene (Av-nifL) was determined . The 1.9 kb sequence shows an open reading frame (ORF) of 1577 bp which encodes a polypeptide of 519 amino acids, with a calculated molecular weight of 57,793 . Av-nifL has about 50% homology with the Klebsiella pneumoniae nifL gene (Kp-nifL) at the nucleotide level and a little more than 52% homology at the amino acid level . The N-terminal regions show more homology than the C-terminal regions . As is the case in K . pneumoniae, Av-nifL is located just upstream of the A . vinelandii nifA gene (Av-nifA) and both genes constitute an operon . The expression of Av-nifL, however, seems to be independent of NtrA and NtrC . Furthermore, Av-nifL expression is not autogenously regulated by NifA, unlike the case in K . pneumoniae . The expression of an Av-nifL::lacZ fusion in A . vinelandii is inhibited by novobiocin and coumermycin A, which are inhibitors of DNA gyrase.

Mol Microbiol, 1993 Mar, 7(6), 1007 - 21
The Klebsiella pneumoniae nifJ promoter: analysis of promoter elements regulating activation by the NifA promoter; Charlton W et al.; The nifJ and nifH promoters of Klebsiella pneumoniae are divergently transcribed sigma 54-dependent promoters that are positively activated by the NifA protein . NifA binds to upstream activator sequences (UASs), usually located 60-200 bp upstream of the start of transcription . Bound NifA is presented to the RNA polymerase-sigma 54 complex (E sigma 54) via DNA loop formation, mediated by the binding of integration host factor protein (IHF) between E sigma 54 and NifA . The nifJ promoter sequence contains three potential NifA binding sites (UAS1, 2 and 3) and two potential RNA polymerase-sigma 54-binding sites (downstream promoter elements, DPEs 1 and 2) . DPE2 is located 420 bp into the coding region and DPE1 overlaps UAS1 by 5 bp . Mutational and footprinting analyses have shown efficient activation of the nifJ promoter requires that NifA is bound at UAS 2 and 3 . Transcription is initiated at DPE1 . Only a weak interaction of NifA with the UAS overlapping DPE1 was detected . Footprints demonstrated that E sigma 54 forms a closed complex at DPE1 but not DPE2 and that bound E sigma 54 closely approaches the -15 region of DPE1 . Stimulation of nifJ promoter activity by IHF was not as great as that observed for other nif promoters . In the absence of IHF nifH promoter sequences stimulated activation of the nifJ promoter . This appeared to require NifA bound at the nifH UAS . Thus, one additional role of IHF may be to partition NifA between the two promoters by constraining the topology of the DNA.

Can J Microbiol, 1993 Mar, 39(3), 351 - 4
Accumulation of factor F395 in nifNE mutants of Klebsiella pneumoniae; Downs DM et al.; The nifNE gene products of Klebsiella pneumoniae are required for the in vivo and in vitro synthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase . Derepression of nifNE mutants for nitrogenase resulted in the accumulation of a small molecule, factor F395 . Factor F395 is protein associated in vivo . We report here initial spectral characterization of this factor.

Clin Infect Dis, 1993 Mar, 16(3), 441 - 2
Infection due to Klebsiella rhinoscleromatis in two patients infected with human immunodeficiency virus; Paul C et al.; Two cases of rhinoscleroma in patients infected with the human immunodeficiency virus (HIV) who had stayed in an area of endemic Klebsiella rhinoscleromatis are reported . One of the patients presented with oropharyngeal lesions, an unusual clinical picture . Both patients suffered from a major cellular immune deficiency . The importance of Klebsiella rhinoscleromatis infection in AIDS-related oropharyngeal pathology and the possible treatment of such infection in HIV-positive patients are not yet clearly established.

Infect Immun, 1993 Mar, 61(3), 926 - 32
The interleukin-1 receptor antagonist can either reduce or enhance the lethality of Klebsiella pneumoniae sepsis in newborn rats; Mancilla J et al.; Klebsiella pneumoniae, a worldwide cause of nosocomial infections, is one of the most common causes of death in newborns in nurseries . In this study, we investigated the role of interleukin-1 (IL-1) in an experimental animal model of neonatal sepsis, using a natural antagonist of IL-1 receptors, the IL-1 receptor antagonist (IL-1Ra), to block IL-1's effects in neonatal Klebsiella sepsis in the absence of antibiotic treatment . Newborn Wistar-Kyoto rats were injected intraperitoneally with a single dose (10 mg/kg) of either IL-1Ra (n = 43) or human serum albumin as a control (n = 40) . At the same time, a 50% lethal dose of K . pneumoniae was injected subcutaneously . No antibiotics were given at any time . After 10 days, survival was 60% for the albumin group and 80% for the IL-1Ra group (P < 0.01) . IL-1Ra treatment also afforded protection when the dose of bacteria was increased sixfold (P < 0.01) . There were two episodes of leukopenia in the control group, which were suppressed by IL-1Ra (P < 0.01 and P < 0.001) . IL-1 and IL-6 levels were lower in the IL-1Ra-treated group (P < 0.05 and P < 0.001, respectively) . No differences between the two groups were observed in the number of bacteria in cultures of the blood, lungs, liver, or spleen . When IL-1Ra (10 mg/kg) was given both at time zero and 24 h after bacterial challenge, lethality was significantly increased (P < 0.01) . Single doses of IL-1Ra of from 20 to 40 mg/kg progressively increased lethality compared with controls (P < 0.01) in both Wistar-Kyoto and Sprague-Dawley strain rats . In the same model, low doses of IL-1 itself (0.4 ng per rat), given 24 h prior to bacterial challenge, afforded protection (P < 0.001) . These studies suggest that, in the absence of antibiotics, partial blockade of IL-1 receptors improves survival, whereas a longer or greater blockade increases lethality in newborn rats infected with K . pneumoniae.

Infect Immun, 1993 Mar, 61(3), 852 - 60
C1q binding and activation of the complement classical pathway by Klebsiella pneumoniae outer membrane proteins; Alberti S et al.; The mechanisms of killing of Klebsiella pneumoniae serum-sensitive strains in nonimmune serum by the complement classical pathway have been studied . The bacterial cell surface components that bind C1q more efficiently were identified as two major outer membrane proteins, presumably the porins of this bacterial species . These two outer membrane proteins were isolated from a representative serum-sensitive strain . We have demonstrated that in their purified form, they bind C1q and activate the classical pathway in an antibody-independent manner, with the subsequent consumption of C4 and reduction of the serum total hemolytic activity . Activation of the classical pathway has been observed in human nonimmune serum and agammaglobulinemic serum (both depleted in factor D) . Binding of C1q to other components of the bacterial outer membrane, in particular the rough lipopolysaccharide, could not be demonstrated . Activation of the classical pathway by this lipopolysaccharide was also much less efficient than activation by the two outer membrane proteins . The antibody-independent binding of C1q to serum-sensitive strains was independent of the presence of capsular polysaccharide, while strains possessing lipopolysaccharide O antigen bind less C1q and are resistant to complement-mediated killing.

Kidney Int, 1993 Mar, 43(3), 592 - 600
Binding of bacterial adhesins to rat glomerular mesangium in vivo; Miettinen A et al.; Two well characterized bacterial adhesins, the O75X fimbriae of Escherichia coli and the type-3 fimbriae of Klebsiellae, with in vitro affinities to type IV and V collagens, respectively, were used to test whether bacterial components with affinity for glomerular matrix could bind to glomeruli in vivo . The purified fimbrial proteins were injected into rats, and kidney samples were studied by immunofluorescence at two hours to nine months postinjection . The O75X, but not the type-3 fimbriae, formed mesangial deposits that persisted for months . Preincubation of the O75X fimbriae with type IV collagen significantly reduced the glomerular binding . The fimbrial deposits were extracellular, as anti-O75X IgG injected into rats bound to glomeruli . Proteinuria or histological damage could not be detected even after passive or active immunizations of the rats . The results demonstrate that bacterial adhesins may bind in vivo to and persist in glomeruli by their specific affinities . The results also indicate that additional factors provided by the bacteria or the host are needed for glomerular damage to take place.

Biochemistry, 1993 Feb 23, 32(7), 1734 - 9
On the mechanism of sodium ion translocation by oxaloacetate decarboxylase of Klebsiella pneumoniae; Dimroth P et al.; Proteoliposomes reconstituted with purified oxaloacetate decarboxylase of Klebsiella pneumoniae catalyzed the uptake of Na+ ions upon oxaloacetate decarboxylation . The degree of coupling between the chemical and the vectorial reaction is dependent on the reconstitution conditions, and with the best preparations approaches a stoichiometry of two Na+ ions per decarboxylation of one oxaloacetate . This coupling ratio is observed only in the absence of a delta mu Na+, immediately after oxaloacetate addition . The ratio gradually declines during development of the electrochemical Na+ ion gradient and becomes zero in the steady state . The Na+ pump, however, continued to decarboxylate oxaloacetate and to catalyze Na+ influx at the apparent stoichiometry of two Na+ ions per decarboxylation event . During the steady state, this influx must be compensated by Na+ efflux of the same size . The efflux is catalyzed by the Na+ pump upon oxaloacetate decarboxylation, because in the absence of the substrate the efflux rate dropped to less than 10% . Proteoliposomes loaded with Na2SO4 catalyzed a bicarbonate-dependent uptake of 22Na+ that was completely abolished after incubation with avidin . These results suggest coupling of Na+ translocation to the carboxylation/decarboxylation of the biotin prosthetic group without the requirement for the oxaloacetate/pyruvate interconversion . The oxaloacetate-dependent transport of Na+ into proteoliposomes was inhibited by the additional presence of the beta + gamma subunits of oxaloacetate decarboxylase . A model of Na+ translocation by oxaloacetate decarboxylase based on these experimental results is proposed.

Thromb Haemost, 1993 Feb 1, 69(2), 98 - 102
Reduction of mortality with antithrombin III in septicemic rats: a study of Klebsiella pneumoniae induced sepsis; Dickneite G et al.; Experimental gram-negative sepsis was induced in the rat by Klebsiella pneumoniae . Although bacteria are susceptible to the treatment with the antibiotic Tobramycin, DIC could not be prevented . DIC was manifested by a leuko- and thrombocytopenia, decreases in fibrinogen and AT III and an increase of the aPTT . In this model the therapeutic treatment with human AT III was evaluated . To determine the optimal concentration of AT III a prestudy in a LPS induced DIC in the rat was performed . It was shown that a bolus i.v . injection of 500 U/kg improved survival and DIC, and was thus chosen for the Klebsiella sepsis model . The infectious load was adjusted to yield a mortality rate of 90-100% in the untreated Klebsiella group and a reduction to about 40-50% of the mortality rate by Tobramycin . It was found that AT III reduced mortality in the Klebsiella induced sepsis not only when given prophylactically but was effective even when administrated in a late stage of the DIC, i.e . 3 or 5 h post infection.

J Clin Microbiol, 1993 Feb, 31(2), 179 - 84
Molecular epidemiology of plasmid spread among extended broad-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates in a pediatric hospital; Bingen EH et al.; Over a 12-month period, 43 children in eight different wards of our hospital (Hopital Robert Debre) were infected or colonized with Klebsiella pneumoniae strains producing extended broad-spectrum beta-lactamases . The epidemiology of the outbreak was studied by a molecular approach including the determination of the beta-lactamase physicochemical parameters and plasmid profiles, as well as analysis of the restriction fragment length polymorphisms of the rDNA regions (ribotyping) . The last approach produced 12 and 5 different patterns with EcoRI and HindIII, respectively, thus identifying 15 different ribotypes among the 43 clinical K . pneumoniae strains . However, 60% of the strains in six wards belonged to only two ribotypes, whereas nine ribotypes were observed only once . Twelve isolates from different wards that were representative of the eight most common ribotypes showed four different beta-lactamase isoelectric focusing patterns and seven different plasmid profiles by direct analysis or after EcoRI digestion . Thus, at least two genetically unrelated strains in the same ward were found to have the same plasmid content . Our results show the complexity of the outbreak, which was associated with patient-to-patient cross-contamination with several epidemic strains with different plasmid contents, interspersed sporadic cases with nonepidemic strains, and the possible spread of a plasmid . The combination of plasmid profile analysis and ribotyping therefore seems to be powerful at deciphering the details of such outbreaks.

J Protein Chem, 1993 Feb, 12(1), 51 - 6
Diethylpyrocarbonate reactivity of Klebsiella aerogenes urease: effect of pH and active site ligands on the rate of inactivation; Park IS et al.; Reaction of Klebsiella aerogenes urease with diethylpyrocarbonate (DEP) led to a pseudo-first-order loss of enzyme activity by a reaction that exhibited saturation kinetics . The rate of urease inactivation by DEP decreased in the presence of active site ligands (urea, phosphate, and boric acid), consistent with the essential reactive residue being located proximal to the catalytic center . The pH dependence for the rate of inactivation indicated that the reactive residue possessed a pKa of 6.5, identical to that of a group that must be deprotonated for catalysis . Full activity was restored when the inactivated enzyme was treated with hydroxylamine, compatible with histidinyl or tyrosinyl reactivity . Spectrophotometric studies were consistent with DEP derivatization of 12 mol of histidine/mol of native enzyme . In the presence of active site ligands, however, approximately 4 mol of histidine/mol of protein were protected from reaction . Each protein molecule is known to possess two catalytic units; hence, we propose that urease possesses at least one essential histidine per catalytic unit.

Arch Biochem Biophys, 1993 Feb 1, 300(2), 694 - 8
Evidence that the enzyme catalyzing the conversion of guanosine diphosphate D-mannose to a 4-keto sugar nucleotide intermediate requires nicotinamide adenine dinucleotide phosphate; Yamamoto K et al.; The first enzyme in the formation of GDP-L-fucose from GDP-D-mannose, which forms a GDP-4-keto sugar intermediate, was purified to homogeneity from cell extracts of Klebsiella pneumoniae . During purification, the enzyme was found to be highly activated by NADP . It was proven that the pyridine nucleotide coenzyme of the enzyme was NADP, not NAD, which differs from previously accepted information . NAD had no effect on enzyme activity . The product of the enzyme reaction with NADP as coenzyme was separated from other nucleotides by high-performance liquid chromatography, and using ion spray liquid chromatography/mass spectrometry the mass was determined for the first time, as 587, which is same as the calculated mass of GDP-4-keto-6-deoxy-D-mannose.

Bull World Health Organ, 1993, 71(2), 183 - 8
Antibiotic therapy for bacterial meningitis in children in developing countries; Kumar P et al.; We carried out a study to investigate the effectiveness of chloramphenicol alone as a treatment for bacterial meningitis . A total of 70 consecutive children aged > 3 months with bacterial meningitis, who had been admitted to the paediatric hospital of the All India Institute of Medical Sciences, were randomized to receive chloramphenicol alone or chloramphenicol + penicillin . The two groups were matched with each other . Treatment failure occurred with three (9%) patients in the chloramphenicol-alone group and with four (12.1%) patients in the combination therapy group (P > 0.05) . The mean duration of intravenous therapy, the number of intravenous cannulae used per patient, and the incidence of thrombophlebitis were significantly higher for the group that received the combination therapy . Also, the cost of using chloramphenicol + penicillin was four times higher than that of chloramphenicol alone . Hence, chloramphenicol alone was as effective as chloramphenicol + penicillin and much cheaper and more convenient to usePIP: A prospective, randomized controlled trial was performed from January 1988 to August 1989 involving 66 (33 in each group) children over 3 months of age diagnosed with bacterial meningitis in the pediatric hospital of the All India Institute of Medical Sciences . Children were administered chloramphenicol alone intravenously at a dose of 100 mg.kg-1.day-1 in 4 divided doses . Those who received chloramphenicol + penicillin were given 100 mg.kg-1.day-1 of chloramphenicol and 300,000-400,000 IU.kg-1.day-1 of crystalline penicillin in 6-hourly doses intravenously . Chloramphenicol was given orally after 3-5-days' treatment, if there was an improvement in sensorium and no vomiting . The antibiotics were prescribed for 10-14 days . The cell count ranged from 525 to 16,000 per mcl, while the protein level varied from 53 to 1000 mg.dl . The CSF glucose level as a proportion of the blood glucose ranged from 0 to 69% . There were 3 deaths (4.5%): all in the chloramphenicol + penicillin group . 1 death occurred within 4 hours of admission from Waterhouse-Friderichsen syndrome; in 1 fatal case, the causative agent was a Klebsiella pneumoniae strain resistant to both chloramphenicol and penicillin . Treatment failure (deaths + change of treatment) was recorded for 3 patients (9%) in the chloramphenicol-alone group and for 4 patients (12%) in the chloramphenicol + penicillin group (P 0.05) . Intravenous therapy was continued for 4.27 + or - 1.01 days in the chloramphenicol-alone group, while it was required for 10.3 + 1.99 days in the chloramphenicol + penicillin group )P 0.01) . Significant thrombophlebitis occurred in 17 patients (58.6%) in the combination group but only in 1 patient (3.3%) in the chloramphenicol-alone group (P 0.001) . Drug fever occurred in 3 patients in the combination group and in 1 patient in the chloramphenicol-alone group . After a week of therapy, none of the patients had a total leukocyte count 4000 .

Pediatr Med Chir, 1993 Jan-Feb, 15(1), 53 - 5
{Cholestasis associated with sepsis in newborns}; De Ritis L et al.; Cholestatic jaundice is a well-known complication of gram-negative bacterial infections in the neonates . Newborn and premature infants are particularly vulnerable to cholestasis because of immaturities in bile forming mechanisms . The Authors describe two premature infants who developed cholestasis in the course of a proved bacterial infection by Klebsiella pneumoniae . The main clinical aspects of this association are illustrated and the recent concepts of pathogenetic mechanism of intrahepatic cholestasis is discussed.

Arch Microbiol, 1993, 159(3), 276 - 81
Characterisation of mutations in the Klebsiella pneumoniae nitrogen fixation regulatory gene nifL which impair oxygen regulation; Sidoti C et al.; The nifL gene product of Klebsiella pneumoniae inhibits the activity of the positive activator protein NifA in response to increased levels either of fixed nitrogen or of oxygen in the medium . In order to demonstrate that the responses to these two effectors are discrete we have subjected nifL to hydroxylamine mutagenesis and isolated nifL mutants that are impaired in their ability to respond to oxygen but not to fixed nitrogen . Two such mutations were sequenced and shown to be single base pair changes located in different parts of nifL . The amino acid sequence of NifL shows limited homology to the histidine protein kinases which comprise the sensing component of bacterial two-component regulatory systems . In the light of the location of one of the oxygen-insensitive mutations (Leu294Phe) we have reassessed this homology and we suggest that the Gln273-Leu317 region of NifL may facilitate interactions between NifL and NifA.

Int Surg, 1993 Jan-Mar, 78(1), 40 - 5
Diagnostic and therapeutic strategies of pyogenic liver abscess; Shimada H et al.; The infectious routes and etiologies of 26 cases with pyogenic liver abscess were portal spread in one, hematogenous in three, biliary in 12, transarterial embolization (TAE) in three, posthepatectomy in one and cryptogenic in five cases . Portal and hematogenous cases tend to show solitary and cystic pattern on echogram, and the majority of the bacteria detected was Klebsiella . While most biliary cases show multiple and cystic with tumor pattern on echogram, and an unhomogeneous low density in CT feature, anaerobic bacteria and candida were isolated only from the biliary or TAE cases . Most cases could be cured completely by the various kinds of abscess drainage, but two TAE cases with PTAD (percutaneous transhepatic abscess drainage) and two biliary cases with PTBD (PT-biliary drainage) and PTAD died due to a delay in establishing a diagnosis and to the severity of the condition . An early diagnosis followed by PTAD or PTBD were thought to be of prime importance.

J Biochem (Tokyo), 1993 Jan, 113(1), 93 - 6
Interaction between pullulanase from Klebsiella pneumoniae and cyclodextrins; Iwamoto H et al.; The interaction between pullulanase from Klebsiella pneumoniae and alpha-, beta-, and gamma-cyclodextrins and 6-O-alpha-glucosyl-alpha-cyclodextrin and 6-O-alpha-glucosyl-beta-cyclodextrin was examined by means of inhibition studies of the enzyme activity, UV difference spectroscopy, and flow calorimetry . All the above cyclodextrins were found to be competitive inhibitors, but beta-cyclodextrin and 6-O-alpha-glucosyl-beta-cyclodextrin showed strong inhibition, the inhibitor constants being two orders of magnitude less than those of alpha- and gamma-cyclodextrins . The difference spectra of beta-cyclodextrin were slightly but significantly different from those of the other cyclodextrins, showing blue shift of a few nanometers . Moreover, only beta-cyclodextrin has a positive entropy change upon binding with the enzyme; all the other cyclodextrins have negative values . These results show that the binding mode of beta-cyclodextrin is subtly different from those of alpha- and gamma-cyclodextrins.

Kansenshogaku Zasshi, 1993 Jan, 67(1), 76 - 80
{A case of liver abscess associated with endophthalmitis caused by Klebsiella pneumoniae}; Hidaka T et al.; A 54-year-old female was admitted to our hospital because of a spiking fever, right hypochondriac pain, right orbital pain and visual disturbance . Before admission she was treated with systemic antibiotics infusion for a diagnosis of liver abscess at the other hospital and the liver abscess almost diminished for a while . With the diagnosis of liver abscess and endophthalmitis, liver drainage and evisceration were carried out . The culture of pus from the eye and liver yielded K . pneumoniae . After liver drainage, evisceration, and direct injection of antibiotics into the eye, inflammatory findings tended to improve . Seven cases of metastatic K . pneumoniae endophthalmitis have been reported so far in Japan . The cases had liver abscess as the primary disease and 3 cases had bilateral endophthalmitis . Five cases with liver abscess survived except one who died of sepsis, but unfortunately, all cases became blind in the affected eyes . The prognosis of bacterial endophthalmitis, especially associated with K . pneumoniae liver abscess, is poor and as the outcome could appear to depend on time when treatment is started, a more aggressive diagnostic approach is required . Moreover systemic antibiotic infusion alone is inadequate for treatment of liver abscess and endophthalmitis, and liver drainage, evisceration and intravitreal injection of antibiotics must be given in early stage.

Infection, 1993 Jan-Feb, 21(1), 18 - 22
Spread of Klebsiella pneumoniae producing SHV-5 beta-lactamase among hospitalized patients; Bauernfeind A et al.; The first outbreak of infections caused by an SHV-5 producing strain of Klebsiella pneumoniae is reported . Within a period of 1 year and 9 months, multiresistant K . pneumoniae strains caused severe infections, mostly of the lower respiratory tract, in 22 patients . The strains were resistant to penicillins, third-generation cephalosporins, aztreonam, chloramphenicol, tetracycline and co-trimoxazole . The resistance determinants were transferable to Escherichia coli . All isolates produced a beta-lactamase with a pI of 8.2 . Ceftazidime was hydrolyzed at this band . These characteristics, together with the resistance phenotype, are identical to those of a reference strain producing the beta-lactamase SHV-5 . The K . pneumoniae strains of all patients were identical in their capsular serotype (K1), plasmid pattern and plasmid fingerprint after digestion with Dra I restriction endonuclease . We conclude that this outbreak was caused by the spread of one clone of K . pneumoniae producing SHV-5 beta-lactamase among patients of different wards . Our results indicate a real risk for failure of therapy by third-generation cephalosporins in intensive care patients due to SHV-5 producing pathogens.

J Antimicrob Chemother, 1993 Jan, 31(1), 29 - 35
A survey of beta-lactamases from 618 isolates of Klebsiella spp; Reig R et al.; beta-Lactamases from 618 isolates of Klebsiella spp . (487 Klebsiella pneumoniae and 131 Klebsiella oxytoca) were identified by analytical isoelectric focusing . The strains were consecutive isolates from 618 different patients in five hospital centres between 1980 and 1984 . Most of the chromosomal beta-lactamases from the K . pneumoniae isolates had a pI of 7.1, and most of the K . oxytoca isolates had chromosomal beta-lactamases of pIs 7.8, 5.3 and 5.5 . Plasmid-mediated beta-lactamases were produced by 91.8% of K . pneumoniae isolates and 9.9% of K . oxytoca isolates . The beta-lactamase types identified most frequently were SHV-1 in the former species, and TEM-1 in the latter species . Of the K . pneumoniae isolates, 101 produced both of these plasmid-mediated beta-lactamases, while one strain produced an extended-spectrum SHV-like beta-lactamase . Of the 131 K . oxytoca isolates, 20 were hyper-producers of chromosomal beta-lactamase . The susceptibility of this genus to beta-lactam antibiotics is controlled almost exclusively by beta-lactamases, the production of which may differ from one strain to another; thus the empirical therapeutic use of beta-lactam antibiotics is not recommended for this genus.

Microbios, 1993, 73(294), 59 - 60
Types of bacteria found in the canals of Amsterdam; Brook JZ et al.; The bacteriological quality of the canal system in Amsterdam was assessed by determination of the quantity and quality of the water . Eight samples of water from different canals were studied . No Gram-positive bacteria were found . The number of Gram-negative bacteria varied from 720-1,820 (average 1,250) organisms per ml of water . The predominant bacteria were: Pseudomonas sp., Klebsiella pneumoniae and Escherichia coli . These data illustrate the high number of potential pathogens in the water canal system in Amsterdam.

EMBO J, 1993 Jan, 12(1), 271 - 8
Stable periplasmic secretion intermediate in the general secretory pathway of Escherichia coli; Poquet I et al.; The secretion of the Klebsiella oxytoca cell surface lipoprotein pullulanase involves translocation across the cytoplasmic and outer membranes of the Gram-negative bacterial cell envelope . A variant of pullulanase was created by fusing the signal peptide-encoding 5' region of the Escherichia coli gene for periplasmic MalE protein to the 3' end of the pulA gene encoding almost the entire mature part of pullulanase . When produced in E . coli carrying the malE-pulA gene fusion on a high copy number plasmid and the complete set of genes specifically required for pullulanase secretion on a second plasmid, the hybrid protein differed from wild-type pullulanase as follows: (i) it was not fatty-acylated; (ii) it was apparently processed by LepB signal peptidase rather than by LspA lipoprotein signal peptidase; (iii) it was released into the periplasm and was only slowly transported across the outer membrane, and (iv) it was released directly into the medium rather than via the usual surface-anchored intermediate . The hybrid protein was secreted more rapidly when malE-pulA was expressed from a low copy number plasmid . The two steps in the secretion pathway could be totally uncoupled by expressing first the malE-pulA gene fusion and then the cognate secretion genes . These results show that fatty-acylation of wild-type PulA is not essential for secretion but may improve its efficiency when large amounts of the protein are produced, that the two steps in secretion can occur quite independently and that the periplasmic intermediate can persist for long periods under certain circumstances.

Diagn Microbiol Infect Dis, 1993 Jan, 16(1), 35 - 41
Plasmid DNA analysis, biotyping, and antimicrobic susceptibility as subtyping tests for Klebsiella pneumoniae and Klebsiella oxytoca; Hartstein AI et al.; We compared plasmid DNA analysis, biotyping by Vitek, and disk diffusion antimicrobic susceptibility as subtyping tests of Klebsiella pneumoniae and Klebsiella oxytoca . The 92 tested isolates were from alternate, culture-positive patients over 6 months . No outbreak or cluster of infections was recognized during this interval . Plasmid DNA was detected in 85% of the isolates . Each isolate except one had a reproducible absence of plasmid DNA or a reproducible plasmid DNA profile on repetitive testing . Restriction endonuclease enzyme analysis of plasmid DNA was necessary to distinguish differences among some isolates that had only large plasmids . Isolates with only large plasmids represented 18% of the collection . Of the 78 isolates with plasmid DNA, all but two were considered different from one another by plasmid DNA analysis . Biotyping and antimicrobic susceptibility testing were not highly reproducible . In addition, biotyping did not demonstrate a sufficient variety of patterns among the isolates for subtyping purposes . We conclude that plasmid DNA analysis is very useful as a subtyping test for isolates of K . pneumoniae and K . oxytoca . Neither biotyping nor antimicrobial susceptibility as performed in our laboratory had sufficient discriminatory power and reproducibility for subtyping these organisms.

Arch Biochem Biophys, 1993 Jan, 300(1), 142 - 7
4-Hydroxybenzoate uptake in Klebsiella pneumoniae is driven by electrical potential; Allende JL et al.; The uptake of 4-hydroxybenzoic acid (4-HBA) in intact cells of a mutant of Klebsiella pneumoniae was investigated . Uptake of 4-HBA was shown to be an inducible system . This uptake system, at pH 7.0, has a high affinity for its substrate (apparent Kt = 13 microM) and a maximal velocity of 27.6 nmol min-1 mg protein-1 . Competition studies with various structural analogs indicated a very narrow specificity of the 4-HBA uptake system . The transport system has been inhibited by inhibitors of energy metabolism and its activity has not been detected in the crude shock extracts . The effect of two ionophores, nigericin and valinomycin, on 4-HBA uptake with respect to the external pH has been studied . All observations indicate that 4-HBA uptake is active and energized by the membrane potential.

Arch Microbiol, 1993, 159(4), 386 - 91
Gluconate metabolism of Klebsiella pneumoniae NCTC 418 grown in chemostat culture; Simons JA et al.; The metabolism of gluconate by Klebsiella pneumoniae NCTC 418 was studied in continuous culture . Under all gluconate-excess conditions at low culture pH values (pH 4.5-5.5) the majority (70-90%) of the gluconate metabolized was converted to 2-oxogluconate via gluconate dehydrogenase (GADH), although specific 2-oxogluconate production rates under potassium-limited conditions were significantly lower than under other gluconate-excess conditions . At high culture pH values, metabolism shifted towards production of acetate . Levels of GADH were highest at low culture pH values and synthesis was stimulated by the presence of (high concentrations of) gluconate . An increase in activity of the tricarboxylic acid cycle was accompanied by a decrease in GADH activity in vivo and in vitro, suggesting that the GADH serves a role as an alternative energy-generating system . Anaerobic 2-oxogluconate production was found to be possible in the presence of nitrate as electron acceptor . Levels of gluconate kinase were highest when K . pneumoniae was grown under gluconate-limited conditions . Under carbon-excess conditions, levels of this enzyme correlated with the intracellular catabolic flux.

Diagn Microbiol Infect Dis, 1993 Jan, 16(1), 25 - 9
Increased frequency of large R-plasmids in Klebsiella pneumoniae colonizing patients with spinal cord injury; Montgomerie JZ et al.; From 1978 to 1988 strains of gentamicin-susceptible (Gms) and gentamicin-resistant (Gmr) Klebsiella pneumoniae were saved from annual surveillance cultures of the perineal region of patients with spinal cord injury (SCI) . Of 38 strains selected for further study (24 Gms and 14 Gmr), there were 23 different serotypes (two nontypable) . Fourteen Gms as well as 14 Gmr strains displayed no common plasmid patterns, but all contained a large plasmid of 168-208 kb . Among the 14 Gmr strains, nine had large conjugative plasmids of approximately the same size (166-193 kb), which conferred to a susceptible Escherichia coli host an identical resistance pattern: ampicillin, chloramphenicol, gentamicin, piperacillin, trimethoprim-sulfamethoxazole, tetracycline, and tobramycin . Of the nine transconjugants, eight contained a single plasmid . One transconjugant contained a 168- and 80-kb plasmid . Restriction endonuclease digestion patterns of the R-plasmids revealed minimal similarity . We conclude that, during a 10-year period, different large R-plasmids have spread among multiple serotypes of K . pneumoniae in spinal cord injury (SCI) patients in one rehabilitation hospital . We hypothesize that other genes located on large, R-, and non-R-plasmids may confer an additional advantage for colonization by K . pneumoniae in SCI patients.

Autoimmunity, 1993, 14(3), 231 - 6
Experimental murine model for autoimmune myocarditis using Klebsiella pneumoniae O3 lipopolysaccharide as a potent immunological adjuvant; Kato N et al.; Experimental autoimmune myocarditis could be produced in mice by repeated injection of syngeneic heart extract together with Klebsiella pneumoniae O3 lipopolysaccharide (KO3 LPS) as a powerful adjuvant . Histological changes in the cardiac lesions were characterized by infiltration with mononuclear cells in the myocardium, degeneration and loss of myocardial fibers, and replacement of granulation tissues . No such cardiac lesions were produced in mice receiving injections of heart extract alone or KO3 LPS alone . Development of the autoantibody and the delayed type-hypersensitivity (DTH) against syngeneic heart extract was found in mice immunized repeatedly with the mixture of heart extract and KO3 LPS . Moreover, definite cardiac lesions were produced in normal recipient mice by transfer of sensitized spleen cells from hyperimmunized mice . Therefore, it was suggested that those cardiac lesions were caused by the autoimmune mechanism . Our methodology provided a new experimental murine model for autoimmune myocarditis.

Med Dosw Mikrobiol, 1993, 45(2), 167 - 71
{Usefulness of plasmids as epidemiologic markers for typing of Klebsiella}; Kurlenda J et al.; The study was aimed at evaluation of utility of plasmids as epidemiological markers for interspecific differentiation of 200 strains of Klebsiella isolated from patients with generalized infections and hospitalized at intensive care ward of the Gdansk Medical Academy . Strains isolated from the ward environment were also investigated . In the first phase of this study classical methods of typing were used, such as determination of the biochemical, bacteriocin (susceptibility to 8 bacteriocins) and phage types, and MIC for 9 antibiotics . Basing on these methods it was found that 41 types of Klebsiella pneumoniae, 13 types of Klebsiella oxytoca and 1 type of Klebsiella ozenae occur . Six groups of bacteria were differentiated in the group which had the same type . These strains were investigated for plasmid and restrictive patterns . Identity of types occurring in individual groups was detected . For one of them, a restrictive pattern was determined by cleavage of plasmid DNA by a restrictive Eco RV enzyme . In remaining groups such pattern was not found, both after treatment with Eco RV and Hha I enzyme and this may be associated with modification of the plasmid DNA . Strains belonging to one of these groups did not contain plasmids in their cells.

Antonie Van Leeuwenhoek, 1993, 63(3-4), 315 - 21
Control and regulation of metabolic fluxes in microbes by substrates and enzymes; van Dam K et al.; The control of enzymes and substrates on the flux through microbial metabolic pathways can be quantified in terms of flux control coefficients . In pathways involving group transfer, the summation theorem for flux control by the enzymes has to be modified: the sum of control by all enzymes is between 1 and 2 . The phosphoenolpyruvate:glucose phosphotransferase system is such a pathway . Experimental determination of the control by the enzymes in this pathway is under way . The control of the enzymes on the glycolytic flux in yeast is low, with the possible exception of the uptake step . In Klebsiella pneumoniae potassium and ammonium ions can simultaneously be 'limiting', (i.e . have significant control on growth) at pH6, but not at pH8 . This may be due to the fact that at pH8 the high-affinity potassium uptake system is absent.

Am J Nephrol, 1993, 13(4), 278 - 80
Bacteremia complicating peritonitis in peritoneal dialysis patients; Morduchowicz G et al.; Bacteremia is a rare complication of peritonitis in end-stage renal failure (ESRF) patients treated by peritoneal dialysis . Three of our ESRF patients on peritoneal dialysis developed bacteremia during a peritonitis episode (1/19 peritonitis episodes) . In 2 cases, the responsible organism was Escherichia coli and peritonitis was most likely associated with infection of the biliary tract . The 3rd patient had a perforation of the colon and Klebsiella spp . was the infective organism . Only the last patient survived but had to be transferred to hemodialysis . Bacteremia during peritonitis is infrequent in peritoneal dialysis patients and it appears to be related to other intra-abdominal events.

Nucleic Acids Symp Ser, 1993, (29), 131 - 2
Enzymology and gene technology of Streptomyces nucleotide 3'-pyrophosphokinase-2',3'-cyclic monophosphokinase (PPKase); Ezaki S et al.; Streptomyces extracellular nucleotide 3'-pyrophosphokinase-2',3'-cyclic monophosphokinase transfers 5'-beta,gamma-pyrophosphate group from ATP, some ATP derivatives and dATP to a variety of nucleot(s)ides at the 3'-OH site and synthesizes the respective 3'-pyrophosphoryl nucleotides . The enzyme also utilizes A-5'-pn (n = 3-5)-5'-N, transferring their adenosine 5'-pyrophosphoryl group and concomitantly eliminating the AMP moiety therefrom, leading to the synthesis of 2',3'-cyclic monophosphoryl nucleotides . The enzyme gene and its neighbouring up- and downstream sequences were analyzed . Streptomyces and enteric bacteria E . coli and Klebsiella pneumoniae transformants were constructed by incorporation of the pyrophosphokinase gene-recombined expression plasmids . Various effects of the treatments--intracellular synthesis of 3'-pyrophosphoryl nucleotides, retarded cellular growth, stimulated N2 fixation by K . pneumoniae and so on--were seen.

Indian J Pediatr, 1993 Jan-Feb, 60(1), 25 - 7
Outbreak of neonatal septicemia with multidrug resistant Klebsiella pneumoniae; Banerjee M et al.; A nosocomial outbreak of neonatal septicemia due to K . pneumoniae occurred in nursery during June-July, 1991 . Klebsiella pneumoniae (Klebocin type 314) was recovered from blood of 33 (70.2%) of 47 neonates with septicemia . Multiple drug resistance was observed in all the cases . The same strain of K . pneumoniae was recovered from the neonates and environment of nursery and labour room as well . The outbreak was attributable to environmental dissemination.

Vet Hum Toxicol, 1993, 35 Suppl 1, 31 - 6
Human studies to measure the effect of antibiotic residues; Elder HA et al.; This epidemiological study compares the frequency of resistant bacteria in stool microflora among vegetarians and nonvegetarians over a 12 month period . Two well characterized vegetarian populations (one in Boston, MA and the other in Loma Linda, CA) as well as appropriate controls were studied . No apparent differences in the prevalence of antibiotic resistance in the microflora were noted; however, vegetarians had a significantly greater incidence of multi-antibiotic resistance . E . coli of the same API biotype had the same frequency of antibiotic resistance in both vegetarians and nonvegetarians . Quantitative studies showed similar percents of tetracycline resistant facultative isolates and of "bacteroides." Klebsiella were more common in the stool of the nonvegetarians . As shown in previous studies, exposure to animal products either as meat eaters or production workers in a poultry abattoir was not associated with an increased incidence of resistant bacterial flora or infections caused by resistant strains.

Med Dosw Mikrobiol, 1993, 45(3), 295 - 9
{Epidemiologic investigation of infection with Klebsiella using phage typing as a method}; Przondo-Mordarska A et al.; In epidemiological investigations of infections with Klebsiella carried on by us for many years, a lysotyping method was used with a success . The results which are now being presented deal with 638 strains of Klebsiella pneumoniae and Klebsiella oxytoca originating from patients treated in hospitals in Wroclaw, Krakow, Gdansk and Sosnowiec during the years 1986-1992 . Susceptibility of strains to the standard set of 14 phages used since ten years decreased significantly, when compared with previously typed strains of Klebsiella . Out of 312 susceptible strains of Klebsiella, majority (302) represented 47 phage types described previously . Sporadic appearance of new phage types was observed . Some phagotypes (37, 47, 112, 117, 119, 142, 153 and 157) were represented most frequently and concerned 55% of susceptible strains . High repeatability of phage types in consecutive groups of typed strains suggests a need for continuation of this method for typing Klebsiella . Decreased susceptibility of strains to standard set of phages requires introduction of new typing bacteriophages.

Rev Latinoam Microbiol, 1993 Jan-Mar, 35(1), 109 - 15
{Capsular polysaccharide of Klebsiella pneumoniae . II . Immunogenic properties}; Alcantar-Curiel MD et al.; It has been considered that the polysaccharide capsular material from microorganisms such as Klebsiella pneumoniae induces mainly thymus-independent humoral immunity, nevertheless studies done with Bacteroides fragilis have shown the participation of cellular immune effector mechanisms . Works done to define the immunogenic role of the capsular polysaccharide of Klebsiella pneumoniae indicate that both active and passive immunization with these antigens confer protection against infections with this bacteria . Several studies propose the use of vaccines prepared with one or various capsular serotypes of Klebsiella pneumoniae in immunoprophylaxis or immunotherapy.

Fiziol Zh, 1993 Jan-Feb, 39(1), 73 - 8
{Effect of Klebsiella Pneumoniae bacterial colonization of the gastrointestinal tract on the immune processes in mice after weaning}; Chernyshova LI et al.; The influence of intestinal colonization by the Klebsiella pneumonia strain 50/59 on the characteristics of local and humoral immunity has been studied on CBA mice after weaning . Bacteria have been introduced intragastrally in doses of 1 x 10(6) bacteria per mice . Observations have been performed in dynamics during 11 days . They have shown that intestinal colonization induces significant elevation of the concentration of IgA in serum, intestinal content and percentage of splenocytes, expressing receptors of IgA, IgM, CD4 on the 7th day . On the 11th day a significant decrease in these parameters was observed . A conclusion is made that intestinal colonization is not indifferent for the whole organism and induces complex influences on its immune system.

J Hosp Infect, 1993 Jan, 23(1), 27 - 34
Cross infection in an intensive care unit by Klebsiella pneumoniae from ventilator condensate; Gorman LJ et al.; Klebsiella pneumoniae serotype K28 was cultured from six patients over 5 weeks in a general Intensive Care Unit . Colonized condensate in the ventilator expiratory water traps was the probable source of the organism, and hand carriage the vehicle of transmission . Although the cross-infection hazard of ventilator tubing condensate is recognized, there is no report in the literature of an outbreak caused by such fluid . Ventilator tubing condensate should be viewed as contaminated clinical waste and dealt with accordingly.

Biotherapy, 1993, 7(1), 71 - 8
Tissue distribution and cellular distribution of liposomes encapsulating muramyltripeptide phosphatidyl ethanolamide . Tissue and cellular distribution of LE-MTPPE; Melissen PM et al.; In previous studies it was shown that administration of liposome-encapsulated MTPPE (LE-MTPPE) led to resistance against Klebsiella pneumoniae infection . To get more insight in the cell types that are involved in this by LE-MTPPE induced antibacterial resistance, the tissue distribution of liposomes encapsulating MTPPE and the distribution over the cells in the main target organs were investigated . After intravenous injection of the liposomes in mice a substantial amount was recovered from liver and spleen and a smaller amount from the lung . In the liver 83% of the liposomes was taken up by the macrophages . In the spleen also most liposomes were taken up by macrophages of the red and white pulp as well as by dendrocytes . The liver and spleen were also the organs in which, after intravenous inoculation, K . pneumoniae was trapped . It was observed that cells containing LE-MTPPE often had not taken up bacteria . Most bacteria, about 73%, were found in cells not containing liposomes . The capacity of the liposome-containing cells to take up bacteria did not change with time . This suggests that the by LE-MTPPE immunostimulating effect is due to the production of cytokines by the cells that take up LE-MTPPE . These cytokines might stimulate other cells to the killing of bacteria.

Ann Trop Paediatr, 1993, 13(3), 285 - 9
Klebsiella septicaemia, osteomyelitis and septic arthritis in neonates in Ibadan, Nigeria; Adeyemo AA et al.; An outbreak of skeletal infections associated with neonatal Klebsiella septicaemia seen over a 6-month period at the Special Care Baby Unit, University College Hospital, Ibadan is reported . It involved 12 neonates, and the significant antecedent events included perinatal asphyxia, fetal distress and prolonged rupture of membranes . All the babies had septic arthritis and ten cases had osteomyelitis in addition: multiple joint involvement occurred in 50% of cases . All the babies exhibited severe systemic disturbance and the Klebsiella isolated demonstrated multiple antibiotic resistance . The epidemic coincided with a period of severe water shortage which affected the hospital . The probable nosocomial acquisition of the infection is highlighted.

Microbiol Immunol, 1993, 37(8), 601 - 6
Isolation of rfb gene clusters directing the synthesis of O polysaccharides consisting of mannose homopolymers and serological analysis of lipopolysaccharides; Saeki A et al.; Four serotypes of two genera, Escherichia coli O8 and O9 and Klebsiella O3 and O5, produce the O polysaccharides consisting of mannose homopolymers . Previously, we reported the isolation and expression of E . coli O9 rfb in E . coli K-12 strains (Kido et al, J . Bacteriol., 171: 3629-3633, 1989) . In this study, R' plasmids carrying his-rfb region of the other three strains were isolated and expressed in E . coli K-12 strain . Serological study of lipopolysaccharides (LPS) synthesized in E . coli K-12 strain was carried out . His-linked rfb genes from E . coli O9 and Klebsiella O3 directed the synthesis of O polysaccharides with the same antigenicity as those of the parental strains in E . coli K-12 strain . On the other hand, rfb genes from E . coli O8 and Klebsiella O5 directed the synthesis of O polysaccharides which were antigenically not identical but partially common to those of the parental strains . A rough strain derived from E . coli O8 synthesized LPS which showed the identical antigenicity as the wild strain when the his-rfb region of E . coli O8 was introduced . The results suggest that some genes located distantly from his are additionally required to complete the synthesis of O polysaccharides of E . coli O8 and Klebsiella O5.

Biochim Biophys Acta, 1992 Dec 28, 1160(3), 251 - 61
Specific binding sites on human phagocytic blood cells for Gly-Leu-Phe and Val-Glu-Pro-Ile-Pro-Tyr, immunostimulating peptides from human milk proteins; Jaziri M et al.; Two immunostimulating peptides were isolated from human milk proteins by enzymatic digestion, the tripeptide GLF and the hexapeptide VEPIPY . These peptides increased the phagocytosis of human and murine macrophages and protected mice against Klebsiella pneumoniae infection . The present study showed that this activity may be correlated to the presence of specific binding sites on human blood phagocytic cells . The receptor molecules implicated were different for the two peptides . {3H}GLF specifically bound to PMNL and monocytes, whereas {3H}VEPIPY only bound to monocytes . The leukemic promyelocytic cell line HL-60 differentiated into granulocytes or into macrophages (depending on inducer used) coroborated these results . Specific binding of {3H}GLF on plasma membrane preparations of human PMNL (20 degrees C) was saturable and Scatchard analysis indicated two classes of binding sites: high-affinity sites of Kd 2.3 +/- 1.0 nM and Bm 60 +/- 9 fmol/mg protein and low-affinity sites of Kd 26.0 +/- 3.5 nM and Bm 208 +/- 45 fmol/mg protein . {3H}GLF binding was inhibited in a concentration-dependent manner by various analogous peptides, such as LLF, GLY, LLY and RGDGLF, but not by RGD, RGDS, VEPIPY and the chemotactic peptide f-Met-Leu-Phe (f-MLF) . Only at high concentrations the direct analog MLF competed with labeled GLF . An important inhibitory effect was also observed with C1q component of the complement whereas C3 and BSA were uneffective . Specific binding of {3H}VEPIPY on monocyte membranes (20 degrees C) was saturable and Scatchard analysis was consistent with one class of binding sites of Kd 3.7 +/- 0.3 nM and Bm 150 +/- 6 fmol/mg protein.

Ann Rheum Dis, 1992 Dec, 51(12), 1296 - 300
Systemic and mucosal antibodies to Klebsiella in patients with ankylosing spondylitis and Crohn's disease; O'Mahony S et al.; Whole gut lavage fluid is a useful source of material for the study of intestinal immunity and inflammation in humans . Systemic and mucosal antibodies to Klebsiella pneumoniae were measured by enzyme linked immunosorbent assay (ELISA) in serum samples and whole gut lavage fluid from 14 patients with ankylosing spondylitis, 14 with Crohn's disease, and 16 immunologically normal controls . As the concentration of IgG in whole gut lavage fluid reflects disease activity in Crohn's disease, this approach was used to detect intestinal inflammation in patients with ankylosing spondylitis who also had disease activity and use of non-steroidal anti-inflammatory drugs (NSAIDs) recorded . Small intestinal permeability to cellobiose and mannitol was also studied . In serum samples, levels of IgA antibody to klebsiella were high in patients with Crohn's disease and in patients with active ankylosing spondylitis, and were significantly correlated with the erythrocyte sedimentation rate in patients with ankylosing spondylitis . Levels of IgG antibody to klebsiella were also high in patients with Crohn's disease . Studies of whole gut lavage fluid showed similar levels of IgA antibody to klebsiella in the three study groups, but levels of whole gut lavage fluid IgM and IgG antibodies to klebsiella were high in patients with Crohn's disease . Levels of IgG in whole gut lavage fluid were high in patients with Crohn's disease but in only one patient with ankylosing spondylitis, though the cellobiose/mannitol permeability ratio was abnormal in eight of 13 patients with ankylosing spondylitis . It is concluded that high levels of serum IgA antibody to klebsiella are not specific to ankylosing spondylitis, and that there is no evidence of an abnormal intestinal IgA antibody response to klebsiella in patients with ankylosing spondylitis.

J Bacteriol, 1992 Dec, 174(23), 7784 - 90
Cloning of the Escherichia coli sor genes for L-sorbose transport and metabolism and physical mapping of the genes near metH and iclR; Wehmeier UF et al.; The sor genes for L-sorbose (Sor) degradation of Escherichia coli EC3132, a wild-type strain, have been cloned on a 10.8-kbp fragment together with parts of the metH gene . The genes were mapped by restriction analysis, by deletion mapping, and by insertion mutagenesis with Tn1725 . Seven sor genes with their corresponding gene products have been identified . They form an operon (gene order sorCpCDFBAME) inducible by L-sorbose, and their products have the following functions: SorC (36 kDa), regulatory protein with repressor-activator functions; SorD (29 kDa), D-glucitol-6-phosphate dehydrogenase; SorF and SorB (14 and 19 kDa, respectively), and SorA and SorM (27 and 29 kDa, respectively), two soluble and two membrane-bound proteins, respectively, of an L-sorbose phosphotransferase transport system; SorE (45 kDa), sorbose-1-phosphate reductase . The sor operon from E . coli EC3132 thus is identical to the operon from Klebsiella pneumoniae KAY2026 . On the basis of restriction mapping followed by Southern hybridization experiments, the sor genes were mapped at 91.2 min on the chromosome, 3.3 kbp downstream of the metH-iclR gene cluster, and shown to be transcribed in a counterclockwise direction . The chromosomal map of the Sor+ strain EC3132 differs from that of the Sor- strain K-12 in approximately 8.6 kbp.

J Vet Med Sci, 1992 Dec, 54(6), 1145 - 9
Isolation and characterization of type 1 fimbriae from a chicken pathogenic Escherichia coli serotype O78; Sekizaki T et al.; Type 1 fimbriae from chicken pathogenic Escherichia coli strain PDI-386 (serotype O78) was purified and characterized . Because of the acid-induced autoagglutination (T . Sekizaki, Y . Nakasato, and I . Nonomura, J . Vet . Med . Sci . 54, 493-499, 1992), the fimbriae could be easily purified by repeating acid sedimentation, washing, and dissolving in buffer (pH 8.0) . In electron microscopy, the purified fimbriae showed a filament of 8 nm in diameter and 10 microns in average length . The molecular mass of the protein subunit of the purified fimbriae estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 19,000 daltons . The amino acid composition and its NH2-terminal sequence were similar to the previously described one of the Klebsiella pneumoniae type 1 fimbriae . Moreover, there was an immunological relatedness between them . These results indicated that a molecular diversity found between the fimbriae of E . coli and that of K . pneumoniae has already been existed among chicken pathogenic E . coli strains.

J Clin Gastroenterol, 1992 Dec, 15(4), 329 - 31
Injection sclerotherapy as a cause of Klebsiella pneumoniae peritonitis; Trotter JF et al.; We report a case of Klebsiella pneumoniae peritonitis after endoscopic sclerotherapy and discuss its pathogenesis and risk factors . We also review previous cases in the literature and make recommendations for prophylactic therapy . Endoscopists should be aware of peritonitis as a possible complication of endoscopic sclerotherapy.

J Biol Chem, 1992 Nov 25, 267(33), 24007 - 16
Electrophoretic studies on the assembly of the nitrogenase molybdenum-iron protein from the Klebsiella pneumoniae nifD and nifK gene products; White TC et al.; The electrophoretic properties of the molybdenum-iron (MoFe) protein component of nitrogenase and an iron-molybdenum cofactor (FeMoco)-reactivatable apoMoFe protein from Klebsiella pneumoniae were examined under anaerobic ({O2} < 5 ppm), nondenaturing conditions . In wild type K . pneumoniae extracts, two immunoreactive species migrating more slowly than purified MoFe protein were detected using anti-MoFe protein antibodies . The uppermost species comigrates with the apoMoFe protein produced by a K . pneumoniae mutant unable to synthesize FeMoco (UN106) and by Escherichia coli harboring the plasmids pVL222+pVL15 (nifHDKTYUSWZM+A) . In vitro FeMoco titration of the UN106 and pVL222+pVL15 extracts increases the electrophoretic mobility of the apoMoFe protein to that of purified MoFe protein in a two-step process giving rise to a species of intermediate mobility between the apo- and holoMoFe proteins . Two-dimensional gel electrophoresis showed that a 20-kDa peptide is associated with the apoMoFe protein and with the intermediate species, but not with the holoMoFe protein . N-terminal sequencing identified this associated peptide as the nifY gene product, which we propose is acting as a temporary enforcer of the apoMoFe protein structure required for cofactor binding that is released upon FeMoco activation . This FeMoco-induced mobility shift was used to characterize the mutant apoMoFe proteins produced in E . coli as a result of deleting the various nitrogen fixation (nif) genes from the plasmid pVL222 . E . coli extracts bearing plasmids deleted in nifH, nifS, nifTYUM, or nifWZM exhibit less than 10% of the apoMoFe protein activity of derepressed UN106 and contain an immunoreactive species whose electrophoretic mobility is increased upon addition of FeMoco from that of apoMoFe protein to that of holoMoFe protein in a single step . Anaerobic nondenaturing gel electrophoresis of 55Fe-labeled E . coli extracts followed by autoradiography showed that these inactive apoMoFe species do not contain iron, indicating that the P-clusters are absent . We therefore propose that NifH, S, U, W, Z, and M are all involved, to varying degrees, in P-cluster assembly . In addition, the presence of the P-clusters does appear to be necessary for the two-step FeMoco activation of the apoMoFe protein to occur.

Immunobiology, 1992 Nov, 186(3-4), 183 - 98
Binding of a membrane proteoglycan from Klebsiella pneumoniae and its derivatives to human leukocytes; Hmama Z et al.; The binding of a membrane proteoglycan from a non-encapsulated strain of Klebsiella pneumoniae (Kp-MPG) and four derivatives thereof, to human leukocytes, was investigated by indirect immunofluorescence using biotinylated F(ab')2 fragments of anti-Kp-MPG antibodies and the streptavidin-phycoerythrin amplification system in flow cytometry . Four Kp-MPG derivatives were studied: 1/ an acylpoly(1,3)galactoside (APG), 2/ an APG preparation submitted to acid hydrolysis which removed all fatty acids, but left intact the galactose chain of APG (GC-APG), 3/ a preparation obtained by mild alkaline hydrolysis, containing additional ester-linked C14 and C16 fatty acids bound to the APG molecule (EFA-APG) and 4/ a polymer of the latter compound (APG pol) . Kp-MPG, APG and EFA-APG were shown to bind exclusively to monocytes at the lowest concentrations (from 0.15 to 3 microM APG) . At higher concentrations, these compounds interacted with polymorphonuclear leukocytes, and with lymphocyte subsets in the following decreasing order: B cells, NK cells, CD8+ and CD4+ lymphocytes . Neither APG pol or GC-APG nor K . pneumoniae smooth LPS showed significant binding to leukocytes . However Kp-LPS treated by drastic alkaline hydrolysis displayed binding properties similar to those of APG . Removal of the ester-linked C14 and C16 fatty acids from EFA-APG did not affect the binding of the molecule . The capacity of cells from the myelomonocytic lineage to bind Kp-MPG and APG was very low in phenotypically immature cell lines (HL60 and U937) as compared with monocytes or polymorphonuclear cells . Treatment of U937 cells with interferon-gamma up-regulated their APG binding capacity along with the expression of the integrin CD 11 b and the CD 14 molecule, whereas monocytes exposed to interferon-gamma showed an increased binding of APG associated with an elevated expression of the galactose specific lectin Mac-2 . The data demonstrate a preferential binding of Kp-MPG and APG to cells of the monocyte/macrophage lineage . APG binding does not involve the poly (1,3) galactose chain and the ester-linked C14 and C16 fatty acids but requires the presence of the hydrophobic part of the molecule.

APMIS, 1992 Nov, 100(11), 1008 - 14
Nationwide spread of Klebsiella oxytoca K55 in Swedish neonatal special care wards; Tullus K et al.; In a nationwide survey of invasive bacterial infections in Swedish neonates, 36% of Klebsiella spp . were Klebsiella oxytoca serotype K55 . This unexpectedly high proportion of K55 infections was due to clusters of infection in neonatal special care wards, and at first seemed attributable to nosocomial spread of a K . oxytoca strain of high virulence . Factors predisposing infants to infection were, however, found irrespective of whether the infecting strain was of serotype K55 or not . Additionally, the prevalence rates of a potential virulence factor, siderophore production, were similar among the two groups of strains . During the same period of time a K . oxytoca K55 with similar biochemical phenotype and drug resistance pattern was found to be spread among the neonates in 12 of 22 neonatal wards in Sweden . The increased proportion of invasive neonatal K . oxytoca K55 infections thus seemed to reflect a high rate of colonization rather than an increased virulence of the K55 strain.

J Bacteriol, 1992 Nov, 174(22), 7185 - 93
A locus that contributes to colonization of the intestinal tract by Bacteroides thetaiotaomicron contains a single regulatory gene (chuR) that links two polysaccharide utilization pathways; Cheng Q et al.; Previously, we isolated two Tn4351-generated mutants of Bacteroides thetaiotaomicron (46-1 and CS3) that were unable to grow either on heparin or on chondroitin sulfate . This phenotype was unexpected, since the heparin and chondroitin sulfate utilization pathways had appeared from earlier studies to be independent of each other . Mutants 46-1 and CS3 were also of interest because both were unable to compete successfully with wild-type B . thetaiotaomicron in the intestinal tracts of germfree mice . Thus, both appeared to have a colonization defect . We have now cloned the chromosomal locus in which the transposon insertions in 46-1 and CS3 occurred . Southern blot analysis showed that the Tn4351 insertions in 46-1 and CS3 were about 100 bp apart . Using complementation and insertional mutagenesis, we localized the region affected by the 46-1 and CS3 insertions to within 2.5 kbp . This DNA segment was sequenced and found to contain a 401-codon open reading frame (ORF1) and the N-terminal segment of a second open reading frame (ORF2), which was downstream of ORF1 and transcribed in the same direction . The deduced amino acid sequence of ORF1 showed significant homology to that of a putative positive regulator of an arylsulfatase gene in Klebsiella aerogenes . ORF2 was at least 381 amino acids long and did not exhibit homology to any proteins in the data bases searched . Transposon insertions in both mutants 46-1 and CS3 disrupted ORF1 . The results of insertional mutagenesis and complementation experiments indicated that ORF2 was not essential for growth on chondroitin sulfate or heparin . Thus, the chondroitin sulfate-negative and heparin-negative phenotypes of 46-1 and CS3 appear to be due to the interruption of a regulatory gene encoded by ORF1 and not to a polar effect of the insertions on a downstream gene(s) . The gene encoding ORF1 has been designated chuR, for regulation of chondroitin sulfate and heparin utilization . Transcriptional fusion studies showed that the expression of chuR occurred at the same level under inducing and noninducing conditions, in contrast to the regulated expression of structural genes of the chondroitin sulfate utilization system . chuR was not autoregulated, nor was its expression affected by a mutation (46-4) that eliminated the expression of all chondroitin sulfate utilization genes but did not affect the utilization of heparin.

J Am Vet Med Assoc, 1992 Nov 1, 201(9), 1419 - 24
Gram-negative bacterial infection in neonatal New World camelids: six cases (1985-1991); Adams R et al.; Gram-negative bacterial infections were documented in 6 neonatal New World camelids (5 Ilamas and 1 alpaca) . The organisms isolated from blood before death or from multiple organs after death were Escherichia coli (n = 3), Actinobacillus sp (n = 1), and Klebsiella pneumoniae (n = 1) . Only 2 crias survived, and 1 became blind secondary to retinal detachment and ocular inflammation, which developed after treatment for bacterial infection . Abnormal events during the perinatal period (prematurity, dystocia, cesarean section, weak at birth) were reported in all 6 crias . Signs of depression, convulsions, and/or coma were observed in all animals . Diarrhea and respiratory distress were also noticed in the 3 crias that died shortly after admission . Serum immunoglobulins were assessed, but without the benefit of a stall-side test specific for Ilama immunoglobulins . All crias were suspected to have poor transfer of maternal immunoglobulins . Hemograms and serum biochemical values prior to the initiation of treatment were obtained on 5 of the 6 crias . Total nucleated cells ranged from 1,400 to 23,100 cells/microliter . Four of the 5 crias has a left shift, and 2 crias had toxic neutrophils . Serum glucose concentrations, measured in 5 of 6 crias, ranged from 83 to 293 mg/dl . Serum creatinine values were high in 2 of 5 crias, 1 of which had acute tubular necrosis . Three crias with high serum electrolyte (sodium, chloride, or potassium) values subsequently died . Arterial blood gas values were assessed in 3 crias, 1 of which had respiratory alkalosis and mild hypoxemia.

Biotechniques, 1992 Nov, 13(5), 750 - 4
A visual method for rapid screening of xylose-fermenting ethanolic strains of Klebsiella pneumoniae; Banerjee M; A simple and rapid screening method for selecting hyper-ethanolic strains of Klebsiella pneumoniae is described . The method involves a novel biological screening marker, namely, the yeast Candida ethanothermophilum . The screening marker was seeded on an agar plate to the surface of which agar blocks, each containing a colony of K . pneumoniae, were subsequently fixed . This seeded plate lacked sources of carbon and energy . Ethanol formed in the agar blocks by the K . pneumoniae colonies diffused into the seeded medium and served as a carbon and energy source for the ethanotrophic yeasts . Colonies of yeasts appeared around the agar blocks in regions of ethanolic diffusion . Hyper-ethanolic strains of K . pneumoniae were thus selected on the basis of the number of colonies of the screening marker that appeared around the blocks containing the ethanolic colonies of K . pneumoniae.

Infect Immun, 1992 Nov, 60(11), 4891 - 7
Roles of peripheral leukocytes and tissue macrophages in antibacterial resistance induced by free or liposome-encapsulated muramyl tripeptide phosphatidylethanolamide; Melissen PM et al.; Administration of free muramyl tripeptide phosphatidylethanolamide (MTPPE) or liposome-encapsulated MTPPE (LE-MTPPE) in a twofold-lower dose at 24 h before bacterial inoculation resulted in clearance of intravenously inoculated Klebsiella pneumoniae by tissue macrophages, whereas in control mice, bacteria were not effectively cleared from the blood . In addition, MTPPE and LE-MTPPE led to increased numbers of leukocytes in the blood, which could compensate for the leukopenia in mice resulting from infection with K . pneumoniae . In an attempt to elucidate the relative contributions of the activation of tissue macrophages and the recruitment of leukocytes to the antibacterial resistance induced by MTPPE and LE-MTPPE, mice were infected intraperitoneally with K . pneumoniae . In these MTPPE- and LE-MTPPE treated mice, intraperitoneal influx of leukocytes and the phagocytic capacity of leukocytes were not higher than in untreated control mice . However, MTPPE- and LE-MTPPE-treated mice survived much longer; eventually 33% of the LE-MTPPE-treated mice survived, whereas all untreated control mice died as a result of bacterial septicemia . This prevention of early death appeared to be the result of an increased clearance of bacteria from the blood by activated tissue macrophages . It was observed that depletion of these tissue macrophages in liver and spleen abrogates the effect of LE-MTPPE treatment, indicating that tissue macrophages are of major importance in the LE-MTPPE-induced resistance against K . pneumoniae infection.

Infect Immun, 1992 Nov, 60(11), 4953 - 6
Novel adjuvant action of lipopolysaccharides that possess mannose homopolysaccharides as O-specific polysaccharides on immune responses to nonimmunogenic autoantigens in mice; Yokochi T et al.; The adjuvant action of various lipopolysaccharides on immune responses to syngeneic tissue extract in mice was examined . Only lipopolysaccharides possessing the linear mannose homopolysaccharides as O-specific polysaccharides exhibited definite adjuvant action on immune responses to the autoantigens . The intensity of this adjuvant activity of lipopolysaccharide from Klebsiella O3 seemed to be the strongest.

J Hosp Infect, 1992 Nov, 22(3), 251 - 5
Pseudobacteraemia with multiply-resistant Klebsiella pneumoniae resulting from contamination from the blood gas machine on a neonatal unit; Jumaa P et al.; Klebsiella pneumoniae with an unusual antibiotic susceptibility pattern was isolated from blood cultures of seven unwell premature babies on the Special Care Baby Unit . Although the organism was sensitive to cefuroxime it was resistant to ceftazidime . It was also resistant to gentamicin, tobramycin, netilmicin, piperacillin and aztreonam but sensitive to ciprofloxacin, imipenem and amikacin . On extensive investigation to trace the source, a K . pneumoniae with the same susceptibility pattern as that obtained from blood cultures was isolated from the probe and probe cover on the blood gas machine but not from any other environmental samples or clinical specimens . Where clinically indicated, antibiotics were used to treat these babies, with success . The difficulty, however, of differentiating between true septicaemia and pseudobacteraemia could be enormous as withholding treatment will have disastrous consequences in genuine cases of bacteraemia.

Microb Pathog, 1992 Nov, 13(5), 371 - 9
Interaction of Klebsiella capsule type 7 with human polymorphonuclear leucocytes; Podschun R et al.; Klebsiella serotype K7 is found among the capsule types that are most prevalent in respiratory tract isolates . To evaluate the significance of the K7 antigen in bacteria-leucocyte interactions, K7-encapsulated Klebsiella pneumoniae strains and their non-capsulate mutants were investigated . The K7 isolates were compared to K2-capsulate strains and their respective K- derivatives . K7-capsulate bacteria were less hydrophilic, and more readily phagocytosed and killed by human polymorphonuclear leucocytes (PMNL) than K2 strains . Loss of the K7 antigen resulted in increased surface hydrophobicity but did not affect phagocytosis and killing, whereas loss of the K2 capsule caused greater susceptibility to the phagocytic and killing action of PMNL . Both the K7 and K2 antigen stimulated the extracellular release of lysozyme from neutrophils but not of myeloperoxidase, indicating degranulation of only secondary granules . All K- mutants induced the release of both lysozyme and myeloperoxidase . Our results suggest that, in contrast to the K2 antigen, the K7 capsular polysaccharide does not confer antiphagocytic properties on bacteria . However, the K7 antigen is able to impede the extracellular release of primary granule enzymes.

J Assoc Physicians India, 1992 Nov, 40(11), 767 - 8
Klebsiella pneumonia with lung abscess in a male child; Majumdar AK; An unusual presentation of Klebsiella pneumonia with lung abscess in a male child with no apparent predisposing factor is described.

Clin Exp Rheumatol, 1992 Nov-Dec, 10(6), 583 - 7
Effect of the substitution of critical residues on the allorecognition of HLA-B27; Ozen S et al.; The three-dimensional structure of the HLA class I molecules has highlighted the importance of the "groove" formed by the helices . We used site-directed mutagenesis to construct a series of HLA-B27 mutants with different substitutions at the sites of the conserved amino acid residues of HLA-B27 subtypes, specifically residue 77 which is thought to be critical to the binding site of the molecule, and a residue at the CD8 binding site . We formed an anti-B27 CTL line and derived six anti-B27 clones . Each of the six clones showed a different pattern of reaction, reflecting the diversity of the epitopes recognized . All nine mutants were effective in altering allorecognition by HLA-B27 specific CTL, although positions 45 and 77 caused the most drastic effect . The residue in position 77 is also the last amino acid of the peptide sequence shared with Klebsiella . Our results highlight the importance of certain epitopes in allorecognition that may have important implications for the immunotherapy of autoimmune diseases.

J Mol Biol, 1992 Oct 5, 227(3), 934 - 7
Preliminary crystallographic studies of urease from jack bean and from Klebsiella aerogenes; Jabri E et al.; Ureases from both jack bean (Canavalia ensiformis) seeds and Klebsiella aerogenes have been crystallized by the hanging drop method . The plant-derived urease crystals are regular octahedra analogous to those obtained by Sumner . Preliminary X-ray diffraction studies show that the crystals belong to the cubic space group F4(1)32, with a = 364 A, and appear to contain one or two subunits in the asymmetric unit . Using a synchrotron source, the crystals diffract to near 3.5 A resolution . Crystals of urease from K . aerogenes belong to the cubic space group I23 or I2(1)3, with a = 170.8 A and appear to contain a single catalytic unit per asymmetric unit . The crystals diffract to better than 2.0 A resolution and are well suited for structural analysis.

J Biol Chem, 1992 Oct 5, 267(28), 20024 - 7
Site-directed mutagenesis of the active site cysteine in Klebsiella aerogenes urease; Martin PR et al.; Cysteine 319 in the large subunit of Klebsiella aerogenes urease was identified as an essential catalytic residue based on chemical modification studies (Todd, M.J., and Hausinger, R.P . (1991) J . Biol . Chem . 266, 24327-24331) . Through site-directed mutagenesis, this cysteine has been changed independently to alanine, serine, aspartate, and tyrosine . None of these mutations (C319A, C319S, C319D, and C319Y, respectively) affected the size or level of synthesis of the urease subunits as monitored by polyacrylamide gel electrophoresis . The wild type enzyme and each of the mutant proteins was purified and their properties were compared . The C319Y protein possessed no detectable activity, while activity was reduced in C319A, C319S, and C319D to 48, 4.5, and 0.03% of wild type levels under normal assay conditions . All of the active mutants had a small increase in Km when compared to the wild type value . The active mutants displayed a greatly reduced sensitivity to inactivation by iodoacetamide in comparison to the wild type enzyme, confirming our previous assignment of the essential cysteine to this residue based on active site peptide mapping . In contrast to the wild type enzyme, inactivation of the mutant proteins was not affected by the presence of the competitive inhibitor phosphate, suggesting that the remaining slow rate of iodoacetamide inactivation is due to modification away from the active site . The pH dependence of urease activity was substantially altered in the active mutants with C319S and C319D showing a pH optimum near 5.2, and C319A near 6.7, compared to the pH 7.75 optimum of wild type urease . These data are consistent with Cys-319 facilitating catalysis at neutral and basic pH values by participating as a general acid.

AJR Am J Roentgenol, 1992 Oct, 159(4), 811 - 5
Isolation of the fourth ventricle causing transtentorial herniation: neurosonographic findings in premature infants; Hall TR et al.; OBJECTIVE . Significant posthemorrhagic enlargement of the fourth ventricle occurs only in a small minority of patients . Although entrapment or isolation of any ventricle can occur, the fourth ventricle is the most common site . This study was undertaken to better understand enlargement of the fourth ventricle after intraventricular hemorrhage and the neurosonographic features of isolation and transtentorial herniation . MATERIALS AND METHODS . The sonographic records of 1535 premature neonates were reviewed for evidence of intracranial hemorrhage and ventriculomegaly . Cranial sonography was performed within the first 3 days of life in all neonates born in our hospital and within the first week of admission for transferred neonates . Follow-up sonograms were obtained at weekly intervals until discharge from the nursery . Intracranial hemorrhages were classified as severe when the hemorrhage was associated with hydrocephalus, intraparenchymal hemorrhage, or both . RESULTS . Intracranial hemorrhage occurred in 761 (50%) of 1535 patients, with severe hemorrhages in 229 . Ten had massive enlargement of the fourth ventricle with isolation in six of the 10 after shunt placement . Isolation of the fourth ventricle causing transtentorial herniation occurred in three . Of the patients with isolation of the fourth ventricle, two had associated infection and ventriculitis caused by Klebsiella oxytoca and one had Candida ventriculoencephalitis . CONCLUSION . The early recognition of the neurosonographic features of an enlarged fourth ventricle causing transtentorial herniation has important therapeutic implications . Although isolation can occur after intraventricular hemorrhage alone, infection and ventriculitis are significant factors in the development of an isolated fourth ventricle.

J Biochem (Tokyo), 1992 Oct, 112(4), 456 - 60
Purification and characterization of novel sulfotransferase obtained from Klebsiella K-36, an intestinal bacterium of rat; Kim DH et al.; A novel type of sulfotransferase was purified from Klebsiella K-36, an intestinal bacterium of rat . The enzyme (M(r) 160,000) is composed of two subunits (M(r) 73,000) with pI and optimal pH values of 5.3 and 10-10.5, respectively . The apparent Km for PNS (p-nitrophenyl sulfate) using phenol as an acceptor and that for phenol using PNS as a donor substrate were determined to be 0.11 and 0.66 mM, respectively . The enzyme is activated by magnesium ion and inhibited by EDTA.

Med Klin (Munich), 1992 Oct, 87 Suppl 1, 53 - 5
{Pentamidine inhalation in prevention of pneumocystis carinii pneumonia in treatment of rejection with monoclonal antibody Orthoclone (OKT-3)}; Barenbrock M et al.; The use of the monoclonal antibody OKT-3 (Orthoclone) is associated with an increased risk of pneumocystis carinii pneumonia . In a retrospective study, the efficiency of a prophylactic inhalation of pentamidine during acute renal allograft rejection therapy with OKT-3 was investigated . From July 1988 until October 1989 32 renal transplanted patients with acute rejection refractory to steroids had been treated with OKT-3 . Twelve of the patients developed a pneumonia (four pneumococcus, one klebsiella, one cytomegalovirus), in six cases, a pneumocystis carinii infection was diagnosed in the bronchial lavage . Four of these patients with pneumocystis carinii pneumonia died despite high dose treatment with cotrimoxazole . From November 1989, a prophylactic inhalation of pentamidine was performed during acute renal allograft rejection therapy with OKT-3 . From 33 patients, in eleven cases, a pneumonia was diagnosed (three pneumococcus, one klebsiella, two legionella, three cytomegalovirus, one candida), one patient developed a pneumocystis carinii pneumonia, which was successfully treated with cotrimoxazole . No patient in this group died because of pulmonary infection . The results suggest, that a prophylactic inhalation of pentamidine in severely immunosuppressed solid organ transplant recipients can prevent pneumocystis carinii pneumonia.

Ethiop Med J, 1992 Oct, 30(4), 197 - 205
An outbreak of gentamicin-resistant Klebsiella bacteraemia at a children's hospital; Moss W; An outbreak of Klebsiella bacteraemia at the Ethio-Swedish Childrens' Hospital, Addis Abeba, Ethiopia is described . The epidemic occurred between February 1988 and February 1990 and appeared to originate in the neonatal nursery and subsequently spread to involve other wards in the hospital . Resistance to several antibiotics, including gentamicin, was common . The importance of effective surveillance and the early institution of control measures for the prevention of nosocomial infection are discussed.

Ann Otol Rhinol Laryngol, 1992 Oct, 101(10), 879 - 82
Rhinoscleroma and rhinosporidiosis; Batsakis JG et al.; For North American otolaryngologists, rhinoscleroma and rhinosporidiosis are moving more into the differential diagnosis of chronic inflammatory diseases of the upper airway, particularly the nasal cavity . This change is occasioned by the influx of immigrants from parts of the world to which the diseases are endemic . Diagnosis is established by identification of the causal organisms, the readily seen Rhinosporidium seeberi for rhinosporidiosis and the more elusive Klebsiella rhinoscleromatis for rhinoscleroma.

J Am Geriatr Soc, 1992 Oct, 40(10), 996 - 1000
Community-acquired bacteremia in the elderly: a prospective study of 121 cases; Whitelaw DA et al.; OBJECTIVE: To describe community-acquired bacteremia in the elderly and correlate clinical and laboratory findings with outcome . DESIGN: Prospective study of consecutive cases . SETTING: Large community-based teaching hospital . PATIENTS: One hundred and twenty-one elderly patients aged 65 to 89 years, seen between February 1, 1986 and January 31, 1988 . MAIN OUTCOME MEASURES: Bacteriological cultures, symptoms and signs, laboratory findings, and mortality . RESULTS: Gram-negative organisms accounted for 65 (54%) cases and Gram-positive organisms for 47 (39%) cases, while nine (7%) cases were polymicrobial . E . coli (39%), Klebsiella sp . (8%), S . pneumoniae (14%), and S . aureus (12%) were the most commonly isolated organisms . The overall mortality was 38% . A poor prognosis was associated with confusion as a presenting symptom (P < 0.0003), hypotension (P < 0.0003), and inappropriate or delayed treatment (P < 0.02) . A good prognosis was associated with E . coli as the pathogen (P < 0.0003) and prompt, appropriate antibiotic therapy . CONCLUSION: Community-acquired bacteremia in the elderly has a high mortality rate . Early recognition and prompt, appropriate treatment are critical in reducing the mortality.

J Bacteriol, 1992 Oct, 174(19), 6298 - 302
Molybdenum cofactor (chlorate-resistant) mutants of Klebsiella pneumoniae M5al can use hypoxanthine as the sole nitrogen source; Garzon A et al.; Selection for chlorate resistance yields mol (formerly chl) mutants with defects in molybdenum cofactor synthesis . Complementation and genetic mapping analyses indicated that the Klebsiella pneumoniae mol genes are functionally homologous to those of Escherichia coli and occupy analogous genetic map positions . Hypoxanthine utilization in other organisms requires molybdenum cofactor as a component of xanthine dehydrogenase, and thus most chlorate-resistant mutants cannot use hypoxanthine as a sole source of nitrogen . Surprisingly, the K . pneumoniae mol mutants and the mol+ parent grew equally well with hypoxanthine as the sole nitrogen source, suggesting that K . pneumoniae has a molybdenum cofactor-independent pathway for hypoxanthine utilization.

Indian J Biochem Biophys, 1992 Oct, 29(5), 411 - 4
Studies on induction of metachromasy in cationic dye pinacyanol chloride by Klebsiella K7 capsular polysaccharide; Mitra A et al.; The acidic capsular polysaccharide isolated from Klebsiella K7 induced metachromasy in the cationic dye pinacyanol chloride indicating its chromotropic character . Interaction of the biopolymer with the cationic dye was studied by visible absorption spectrophotometry, and thermodynamic parameters of the interaction evaluated . The polymer induced a metachromatic blue shift in the spectrum from 600 nm to 495 nm . The spectral changes were studied during interaction of the dye with the polymer at different polymer/dye molar ratios (P/D = 0 approximately 50) . Effects of co-solvents on the stability of the dye-polymer compound were studied . A complete reversal of metachromasy was observed upon addition of different alcohols and urea solution . Thermodynamic parameters obtained from the spectral data indicated chromotropic character of the polymer in interacting with the cationic dye molecules in solution.

Afr J Med Med Sci, 1992 Oct, 21(1), 73 - 8
Cross reactivity between Klebsiella pneumoniae and ocular tissue; Ashaye AO et al.; Klebsiella pneumoniae has been implicated as a possible aetiological agent in ankylosing spondylitis and acute anterior uveitis . Cross-reactivity between antigens of klebsiella and bovine vitreous has been reported . In the present study sera from rabbits immunised with Klebsiella pneumoniae was tested against fresh guinea pig and human ocular tissues using immunodiffusion and immunofluorescent methods . No cross-reactivity between klebsiella and the ocular tissues used could be demonstrated by these techniques.

Schweiz Med Wochenschr, 1992 Sep 19, 122(38), 1414 - 23
{Decrease in general health status, febrile state and sinusitis in renal insufficiency}; de Torrente A et al.; A 59-year old man, dialysed for 9 years because of chronic renal failure of unknown etiology, was also suffering from a hypertensive cardiopathy and anemia . He has received more than 70 units of blood over the past few years . An elevated blood aluminium level prompted treatment with desferrioxamine for 3 years . Following episodes of melena, an intestinal-wall lesion, located 8 cm above the ileo-caecal valve was discovered . Blood serotonin levels were elevated; biopsy of the liver revealed tumor cells with round-shaped nuclei which stained positive for synaptophysin, findings compatible with a carcinoid tumor or a pancreatic islet-cells tumor . Considering the whole clinical situation, aggressive diagnostic of therapeutic measures were not pursued any further . The patient lost 8 kg over the next few months and was finally hospitalised with fever . Physical examination revealed ascites, edema and a gingival abscess which was drained . Blood cultures grew Klebsiella pneumoniae . Antibiotics were prescribed . A month later the patient complained again of fever accompanied by myalgias, edema and pain of the eyelids, mainly on the right side, pain in the maxillary area and on right eye motion . ENT examination revealed a necrotic lesion of the right middle turbinate which on histology was diagnosed as acute purulent rhinitis without granuloma or vasculitis . A CT-scan demonstrated mucosal thickening of the left sphenoidal, ethmoidal, frontal and maxillary sinuses without any osteolytic lesions . The patient suffered a sudden right eye blindness with signs of retinal ischemia accompanied by an elevated sedimentation rate of 130 mm . Steroid treatment for suspicion of a temporal arteritis was introduced . 2 days later the patient was rehospitalized in a semi-comatose condition . No new information was gained from the usual laboratory investigations and the physical examination was comparable to the previous hospital stay . Blood cultures remained sterile . He died a few hours later . Discussion: The discussant, A . de Torrente MD, accepts the diagnosis of a hypertensive cardiopathy, renal failure of unknown origin and a carcinoid of the terminal ileum with hepatic metastases . The role of iron overload as a contributory factor for the cardiac disease is discussed . More problematic are the manifestations of the terminal disease with involvement of the sinuses, the eyes and the mouth . The "lethal midline granuloma" (a syndrome and not a disease entity) covers many different diseases which can be considered in this case: Wegener's granuloma, various lymphomas, parasitic, bacteria- and fungus-induced diseases.(ABSTRACT TRUNCATED AT 400 WORDS)

Biochem Biophys Res Commun, 1992 Sep 16, 187(2), 1048 - 54
Cloning and properties of a cyanide hydratase gene from the phytopathogenic fungus Gloeocercospora sorghi; Wang P et al.; The Cht gene encoding cyanide hydratase (CHT, EC 4.2.1.66), which detoxifies HCN and is thought to be important in fungal infection of cyanogenic plants, has been cloned from the phytopathogenic fungus Gloeocercospora sorghi . The gene was isolated by screening an expression library of G . sorghi using a CHT-specific antibody and using one of the positive cDNA clones as a probe in Southern hybridization to identify a 3.1 kb PstI genomic fragment . This PstI fragment expressed CHT activity when transformed into Aspergillus nidulans, a fungus that normally lacks CHT activity . Sequence analysis identified a single open reading frame of 1,107 base pairs which encodes a polypeptide of 40,904 daltons . The deduced amino acid sequence of CHT shares 36.5% identity to a nitrilase from the bacterium Klebsiella pneumoniae subsp . ozaenae.

J Biol Chem, 1992 Sep 15, 267(26), 18371 - 6
Primary structure and characteristics of the melibiose carrier of Klebsiella pneumoniae; Hama H et al.; The melB gene coding for the melibiose carrier of Klebsiella pneumoniae was cloned and sequenced . There were two potential translation initiation sites . It was predicted that the melibiose carrier consists of 471 (or 467) amino acid residues . Seventy-eight percent of the 471 amino acids were identical to the Escherichia coli melibiose carrier . Sugar transport characteristics were studied using an E . coli mel- mutant expressing cloned K . pneumoniae melB gene . Accumulation of melibiose via the K . pneumoniae melibiose carrier was not stimulated by adding NaCl or LiCl which stimulates melibiose accumulation via the E . coli melibiose carrier . Lactose was accumulated only in the presence of LiCl . TMG (methyl-1-thio-beta-D-galactopyranoside) was accumulated in the absence of added NaCl or LiCl . The accumulation was stimulated by LiCl but not by NaCl . Rapid H+ uptake was observed when melibiose or TMG was added to cell suspensions . These results suggest that the preferred cation couplings via K . pneumoniae melibiose carrier are H(+)-melibiose, Li(+)-lactose, and H+/Li(+)-TMG . This coupling spectrum is quite different from that of the E . coli melibiose carrier . It is of special interest that the K . pneumoniae melibiose carrier seems to be lacking the ability to recognize Na+ which is a preferred coupling cation of the E . coli melibiose carrier for all known sugar substrates . Further investigation of these two carriers may give us insight into the Na+ recognition site.

Appl Environ Microbiol, 1992 Sep, 58(9), 2918 - 27
Physiological studies of chloramine resistance developed by Klebsiella pneumoniae under low-nutrient growth conditions; Stewart MH et al.; This study investigated the physiological mechanisms of resistance to chloramines developed by Klebsiella pneumoniae grown in a nutrient-limited environment . Growth under these conditions resulted in cells that were smaller than cells grown under high-nutrient conditions and extensively aggregated . Cellular aggregates ranged from 10 to more than 10,000 cells per aggregate, with a mean population aggregate size of 90 cells . This aggregation may have been facilitated by the presence of extracellular polymer material . By using glucose as a reference of capsule content, it was determined that growth under low-nutrient conditions produced cells with 8 x 10(-14) to 41 x 10(-14) g of carbohydrate per cell, with a mean +/- standard deviation of 27 x 10(-14) +/- 16 x 10(-14) g of carbohydrate per cell . In comparison, growth under high-nutrient conditions resulted in 2.7 x 10(-14) to 5.9 x 10(-14) g of carbohydrate per cell, with a mean and standard deviation of 4.3 x 10(-14) +/- 1.2 x 10(-14) g of carbohydrate per cell . Cell wall and cell membrane lipids also varied with growth conditions . The ratio of saturated to unsaturated fatty acids in cells grown under low-nutrient conditions was approximately five times greater than that in cells grown under high-nutrient conditions, suggesting possible differences in membrane permeability . An analysis of sulfhydryl (-SH) groups revealed no quantitative difference with respect to growth conditions . However, upon exposure to chloramines, only 33% of the -SH groups of cells grown under low-nutrient conditions were oxidized, compared with 80% oxidization of -SH groups in cells grown under high-nutrient conditions . The reduced effectiveness of chloramine oxidization of -SH groups in cells grown under low-nutrient conditions may be due to restricted penetration of chloramines into the cells, conformational changes of enzymes, or a combination of both factors . The results of this study suggest that chloramine resistance developed under low-nutrient growth conditions may be a function of multiple physiological factors, including cellular aggregation and protection of sulfhydryl groups within the cell.

J Gen Microbiol, 1992 Sep, 138 ( Pt 9), 1921 - 7
Biological activities and chemical composition of a cytotoxin of Klebsiella oxytoca; Minami J et al.; A low-molecular-mass cytotoxin produced by Klebsiella oxytoca isolated previously from patients with antibiotic-associated haemorrhagic enterocolitis was purified, and its biological and chemical properties were elucidated . The toxin inhibited the syntheses of DNA and RNA by HEp-2 cells dose-dependently, whereas protein synthesis was only slightly inhibited, as measured by the incorporation of radioactive precursors . When synchronously cultured HEp-2 cells were examined in the presence of cytotoxin, inhibition of DNA synthesis occurred promptly within 5 h, but cell-rounding, the earliest visible morphological change, was not observed until 6 h after exposure . The intracellular levels of ATP decreased with an approximately similar time course . These results suggest that cytotoxicity toward HEp-2 cells is primarily due to the inhibitory effect of the cytotoxin on nucleic acid synthesis, possibly on DNA synthesis . Cell rounding and cell death were induced even in the absence of the cytotoxin after incubation with the cytotoxin for 6 h . The cytotoxin was heat-labile, cytotoxic activity decreasing to 50% of the initial level on heating at 70 degrees C for 20 min . Plasmids were extracted from three strains of K . oxytoca producing the cytotoxin and analysed by agarose gel electrophoresis . Two strains possessed plasmids of different sizes, but one strain possessed no plasmid, indicating that the cytotoxin is probably chromosomally encoded . Analysis by NMR and FAB-mass-spectrometry revealed that the molecular mass of the cytotoxin should be 217.1062 Da (exact mass), its molecular formula being C8H15O4N3.

FEMS Microbiol Lett, 1992 Sep 1, 75(1), 1 - 5
Incidence of two virulence factors (aerobactin and mucoid phenotype) among 190 clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamase; Vernet V et al.; Because outbreaks of multiple-resistant Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases were recently observed in French hospitals, the presence of virulence factors was examined for (i) phenotype by bioassay for aerobactin production and by culture for the mucoid phenotype, and (ii) genotype using intragenic probes of respectively 2-kb BglII and 235-bp BamHI-BglII fragments and dot-blotting among 190 unreplicated K . pneumoniae clinical isolates issued from 25 French hospitals and producing different types of extended-spectrum beta-lactamases (TEM-related enzymes: TEM-3, TEM-4, CAZ-1, CAZ-2, TEM-8, or SHV-related enzymes: SHV-2, SHV-3, SHV-4) . Only 3.7% and 7% of K . pneumoniae isolates produced aerobactin and mucoid phenotypes respectively, unrelated to type of beta-lactamase . Only 2% had both factors . No discordance was reported according to the detection method tested . The low prevalence of such virulence factors seems to indicate they were not involved in dissemination of nosocomial K . pneumoniae isolates producing an extended-spectrum beta-lactamase.

Antimicrob Agents Chemother, 1992 Sep, 36(9), 1981 - 6
Transposition of the gene encoding a TEM-12 extended-spectrum beta-lactamase; Heritage J et al.; An isolate of Klebsiella oxytoca from the blood culture of a child with leukemia was found to produce two beta-lactamases, at least one of which conferred resistance to ceftazidime . Genes encoding both enzymes were located on a single self-transmissible 100-kb plasmid, pOZ201 . This plasmid was introduced into Escherichia coli UB5201 (pACYC184), and the gene encoding one beta-lactamase was transposed onto plasmid pACYC184 by exploiting a gene dosage effect . The transposable gene was found to encode a TEM-12 enzyme as determined by nucleotide sequencing . This gene was subsequently transposed onto plasmid pUB307 . The transposable element encoding the TEM-12 enzyme has been designated Tn841 . Both plasmids pACYC184::Tn841 and pUB307::Tn841 were shown to encode a beta-lactamase with the same isoelectric point and substrate profile as the TEM-12 beta-lactamase . Transposon Tn841, at approximately 7 kb, is larger than TnA (4.8 kb) and transposes at a lower frequency . Although it produced a resolvase which can complement the resolvase of Tn3, its transposase function was not able to complement the transposition of a TnA element which lacked transposase . The occurrence of a gene encoding an extended-spectrum beta-lactamase on a transposable element in a clinically significant bacterium is potentially a cause for concern for the spread of resistance to the extended-spectrum cephalosporins.

Antimicrob Agents Chemother, 1992 Sep, 36(9), 1883 - 9
Piperacillin, tazobactam, and gentamicin alone or combined in an endocarditis model of infection by a TEM-3-producing strain of Klebsiella pneumoniae or its susceptible variant; Mentec H et al.; The efficacy of tazobactam, a beta-lactamase inhibitor, in combination with piperacillin, was studied in vitro and in rabbit experimental endocarditis due to a Klebsiella pneumoniae strain (KpR) producing an extended-spectrum beta-lactamase, TEM-3, or its nonproducing variant (KpS) . In vitro, piperacillin was active against KpS (MIC = 4 micrograms/ml, MBC = 8 micrograms/ml with 10(7)-CFU/ml inoculum) but not against KpR (MIC = MBC = 256 micrograms/ml) . Tazobactam (1 microgram/ml) restored the activity of piperacillin against KpR (MIC = 2 micrograms/ml, MBC = 4 micrograms/ml) . Gentamicin was active against both strains (MIC = 0.25 and 0.5 micrograms/ml for KpS and KpR, respectively) . The piperacillin-tazobactam-gentamicin combination was synergistic in vitro . The piperacillin/tazobactam ratio in plasma and in vegetations was always lower than the 4/1 injected dose ratio . In vivo, piperacillin (300 mg/kg of body weight four times a day {QID}) was active against KpS but not against KpR . Tazobactam (75 mg/kg QID) was able to restore the in vivo effect of piperacillin (300 mg/kg QID) against KpR (-3.0 log10 CFU/g of vegetation versus that of controls) . Gentamicin (4 mg/kg twice a day {BID}) was active against both strains . Compared with controls, the combination of gentamicin plus piperacillin against KpS (-5.6 log10 CFU/g of vegetation), and the gentamicin-piperacillin-tazobactam combination against KpR (-4.4 log10 CFU/g of vegetation) achieved the greatest decrease in bacterial counts in vegetations and were the only regimens that significantly increased the proportion of sterile vegetations . It is concluded that (i) tazobactam was able to restore the effect of piperacillin against a TEM-3 extended-spectrum Beta-lactamase-producing strain of K . pneumoniae, both in vitro and in a severe experimental infection with high inoculum, when used in a 4/1 piperacillin/tazobactam dose ratio; (ii) gentamicin alone was effective because of the high peak/MBC ratio in plasma; (iii) piperacillin-tazobactam-gentamicin, probably because of the effect of gentamicin in reducing bacterial inoculum in vivo, as stressed by the results obtained by piperacillin-gentamicin against KpS, may be the most effective regimen against KpR.

Chest, 1992 Sep, 102(3), 670 - 6
Scintigraphy with J001 macrophage targeting glycolipopeptide . A new approach for sarcoidosis imaging; Diot P et al.; Scintigraphy with radiolabeled J001 as a ligand for macrophage targeting is a new approach for sarcoidosis imaging . J001 is a fully characterized acylated peptido-poly (1,3) galactoside isolated from Klebsiella membrane proteoglycans and able to bind electively recruited macrophages . Its physiochemical properties allow rapid absorption by the respiratory tract when this agent, labeled by 99m technetium, is administered as an aerosol . Images are obtained within 3 to 5 h after inhalation . In the present study, we determined the ability of J001 scintigraphy to localize areas of sarcoidosis involvement in 22 patients compared with gallium scanning in ten of them . Nineteen patients underwent bronchoalveolar lavage (BAL) and serum angiotensin-converting enzyme (ACE) assay . J001 scintigraphy was also performed on a control group of six patients with extrathoracic melanoma, in whom J001 scintigraphy was used to evaluate the cutaneous extent of the tumor and the lymph node involvement . In this control group, no fixation appeared in the thoracic area . In the sarcoidosis group, 18 positive results were observed . One stage 0 patient had a mediastinal fixation . Five of the six stage 1 patients had a fixation located in the mediastinum, the lungs, and the wrists . Five of the six stage 2 patients had positive foci located in the mediastinum or the lung areas and in the myocardium in one of them . Six of the nine stage 3 patients had positive J001 scintigraphy occurring in the lungs and/or the mediastinum . One patient had a fixation on the main bronchi . J001 scintigraphy and gallium scanning, performed in ten patients, were positive in seven of them . There were discrepancies between the BAL results and J001 scintigraphy, as well as between the ACE results and J001 scintigraphy . In conclusion, 99mTc-J001 scintigraphy appears to be a sensitive and rapid technique for the imaging of thoracic sarcoidosis at the three stages of the disease.

P N G Med J, 1992 Sep, 35(3), 202 - 4
Gentamicin resistance in fatal Klebsiella septicaemia; Trevett AJ et al.; The occurrence of antibiotic resistance in nosocomial infections is a common problem . This case details a fatality associated with Klebsiella infection acquired as a hospital inpatient . The transfer of gentamicin resistance between gram-negative bacteria could present a considerable problem in Papua New Guinean hospitals.

Biochem Pharmacol, 1992 Aug 18, 44(4), 673 - 80
Effects of tripeptides derived from milk proteins on polymorphonuclear oxidative and phosphoinositide metabolisms; Migliore-Samour D et al.; The tripeptide GLF (glycyl-leucyl-phenylalanine) was isolated from human milk proteins . This peptide increased phagocytosis by human and murine macrophages and protected mice against Klebsiella pneumoniae infection . Specific binding sites on human polymorphonuclear leukocytes (PMNs) have been demonstrated recently . The aim of the present research was to study the action of this peptide on rat and human PMN oxidative burst and to investigate the consequences of cell stimulation on polyphosphoinositide hydrolysis . A biphasic stimulating concentration-dependent effect of GLF on PMN chemiluminescence and superoxide anion generation was demonstrated . One of the peaks of the oxidative response occurred around 10(-9) M, which correlates with the Kd of high affinity receptors of GLF . The other maximum, around 10(-4) M, might be due to the hydrophobic nature of the tripeptide . O2- generation mimicked the phorbol myristate acetate response: after a lag period of 2-5 min, O2- release gradually increased for 10-15 min until a plateau was reached . Furthermore, GLF enhanced phosphoinositide breakdown with maximal IP3 production at 10(-7) M . Various analogs of GLF were synthesized in order to define the relative importance of the different amino acids and their position in the tripeptide molecule: glycyl-phenylalanine-leucine was devoid of biological properties but enhanced the activity of GLF on the metabolic burst at high concentrations; peptides leucyl-leucyl-phenylalanine and leucyl-leucyl-tyrosine, which displaced GLF from its specific membrane receptors, exerted stimulating effects on PMN oxidative and phosphoinositide metabolisms . It is quite conceivable that these short peptides, which may be generated in the newborn during digestion and which are able to stimulate phagocytic cells, are implicated in the defense of the neonate immature organism against infection.

Gene, 1992 Aug 1, 117(1), 125 - 30
Cloning, sequencing and heterologous expression of a Klebsiella pneumoniae gene encoding an FAD-independent acetolactate synthase; Peng HL et al.; The gene encoding the valine-resistant and FAD-independent acetolactate synthase of Klebsiella pneumoniae was isolated and expressed in Escherichia coli . The nucleotide sequence of this gene was determined and it exhibited an open reading frame of 1680 bp in length . In vivo expression of the acetolactate synthase-encoding gene in E . coli revealed a single 60-kDa protein which is consistent with the molecular weight calculated from the deduced amino acid sequence of the gene product . The gene product shares about 20-30% homology with the acetolactate synthases of E . coli, yeast and higher plants.

J Bacteriol, 1992 Aug, 174(15), 4893 - 8
Mechanism of Na(+)-dependent citrate transport in Klebsiella pneumoniae; van der Rest ME et al.; Citrate transport via CitS of Klebsiella pneumoniae has been shown to depend on the presence of Na+ . This transport system has been expressed in Escherichia coli, and uptake of citrate in E . coli membrane vesicles via this uptake system was found to be an electrogenic process, although the pH gradient is the main driving force for citrate uptake (M . E . van der Rest, R . M . Siewe, T . Abee, E . Schwartz, D . Oesterhelt, and W . N . Konings, J . Biol . Chem . 267:8971-8976, 1992) . Analysis of the affinity constants for the different citrate species at different pH values of the medium indicates that H-citrate2- is the transported species . Since the electrical potential across the membrane is a driving force for citrate transport, this indicates that transport occurs in symport with at least three monovalent cations . Citrate efflux is stimulated by Na+ concentrations of up to 5 mM but inhibited by higher Na+ concentrations . Citrate exchange, however, is stimulated by all Na+ concentrations, indicating sequential events in which Na+ binds before citrate for translocation followed by a release of Na+ after release of citrate . CitS has, at pH 6.0 and in the presence of 5 mM citrate on both sides of the membrane, an apparent affinity (K(app)) for Na+ of 200 microM . The Na+/citrate stoichiometry was found to be 1 . It is postulated that H-citrate2- is transported via CitS in symport with one Na+ and at least two H+ ions.

Appl Environ Microbiol, 1992 Aug, 58(8), 2649 - 53
Impact of growth conditions on resistance of Klebsiella pneumoniae to chloramines; Stewart MH et al.; The resistance of Klebsiella pneumoniae to inorganic monochloramine (1.5 mg/liter; 3:1 Cl2:N ratio, pH 8.0) was examined in relation to growth phase, temperature of growth, and growth under decreased nutrient conditions . Growth phase did not impact resistance to chloramines . Mid-exponential and stationary-phase cells, grown in a yeast extract-based medium, had CT99 values and standard deviations of 4.8 +/- 0.1 and 4.6 +/- 0.2 mg.min/liter, respectively . Growth temperature did not alter chloramine resistance at short contact times . CT99 values of cells grown at 15 and 23 degrees C were 4.5 +/- 0.2 and 4.6 +/- 0.2 mg.min/liter, respectively . However, at longer contact times, CT99.99 values of cells grown at 15 and 23 degrees C were 14 and 8 mg.min/liter, respectively, suggesting a small resistant subpopulation for cells grown at the lower temperature . Growth under decreased nutrient conditions resulted in a concomitant increase in resistance to chloramines . When K . pneumoniae was grown in undiluted Ristroph medium and Ristroph medium diluted by 1:100 and 1:1,000, the CT99 values were 4.6 +/- 0.2, 9.6 +/- 0.4, and 24 +/- 7.0 mg.min/liter, respectively . These results indicate that nutrient availability has a greater impact than growth phase or growth temperature in promoting the resistance of K . pneumoniae to inorganic monochloramine.

Epidemiol Infect, 1992 Aug, 109(1), 143 - 7
Vector potential of hospital houseflies with special reference to Klebsiella species; Fotedar R et al.; The vector potential of houseflies (Musca domestica) for Klebsiella spp . was investigated . Klebsiella spp . (mostly Klebsiella pneumoniae) were isolated from 36.7% of hospital flies and 28.1% of infected wounds of patients . Antibiograms of Klebsiella spp . showed that 82.0% of isolates from hospital flies and 96.3% from infected wounds were resistant to four or more commonly used antimicrobials . In contrast, from the control group, only 8.7% klebsiella isolates showed similar antimicrobial resistance pattern (P less than 0.001) . Similar strains of Klebsiella spp . were encountered among patients and hospital houseflies . The results indicate that flies in the hospital environs are potential vectors of hospital resistant strains of Klebsiella spp.

Epidemiol Infect, 1992 Aug, 109(1), 121 - 31
The ability of airborne Klebsiella pneumoniae to colonize mouse lungs; Bolister NJ et al.; A strain of Klebsiella pneumoniae was aerosolized and its survival in air at different relative humidities was studied . Survival was dependent upon relative humidity and aerosols were most stable during storage at a relative humidity of 60% . Mice were exposed to aerosols of K . pneumoniae produced at this humidity and lung samples taken at timed intervals after exposure . Fifteen strains of K . pneumoniae were tested for their ability to colonize mice, but only five were detectable in mouse lungs 7 days after exposure . Three of these strains persisted without an increase in bacterial numbers, regardless of the initial inoculum used . Two strains of K . pneumoniae, designated strains 15 and 16, persisted in a similar manner when used at a low dose; however, when the dose received per lung was increased there was a rapid multiplication of bacteria in the lungs.

Epidemiol Infect, 1992 Aug, 109(1), 113 - 20
Transferable antibiotic resistance among thermotolerant coliforms from rural drinking water in India; Gaur A et al.; A total of 231 thermotolerant coliforms was isolated from rural drinking water from four states of India . Of these, 220 isolates were resistant to ampicillin, chloramphemicol, streptomycin and tetracycline . Multiple (MAR), double and single antibiotic resistances were observed in 31.4, 48.6 and 13.7% of the isolates, respectively . Out of 177 antibiotic-resistant isolates examined for transmissibility, only 15.3% were able to transfer their resistances to Escherichia coli K-12 recipient . The resistances were transferred by 32.5% of MAR, 21.9% of double resistant and 7.6% of single resistant isolates . Ampicillin resistance was transferable in 14.69% strains while resistances for the rest of the antibiotics were transferable in less than 4% strains . MAR strains of E . coli and Klebsiella sp . showed highest levels of R-plasmid transfer.

Int J Food Microbiol, 1992 Aug, 16(4), 349 - 52
Preliminary examinations on the enterotoxigenicity of isolates of Klebsiella pneumoniae from seafoods; Singh BR et al.; One hundred and eighty-five seafood samples, consisting of 96 freshwater fish, 37 marine fish, 13 freshwater prawn, 13 marine prawn and 26 molluscs were screened for presence of Klebsiella . Out of these, 12 isolates of Klebsiella were identified, Four K . pneumoniae var . ozaenae were isolated from marine fish samples and eight K . pneumoniae var . pneumoniae, six from freshwater fish and two from freshwater prawns . All 12 isolates were tested for enterotoxigenicity by the vasopermeability factor test in rabbits, the mouse foot pad test, the latex agglutination test and the coagglutination test . One isolate of K . pneumoniae var pneumoniae, isolated from fresh water prawn was found enterotoxigenic.

Antimicrob Agents Chemother, 1992 Aug, 36(8), 1785 - 7
In vitro activities of streptomycin and 11 oral antimicrobial agents against clinical isolates of Klebsiella rhinoscleromatis; Perkins BA et al.; We tested in vitro the activities of streptomycin and tetracycline--antibiotics that have long been used to treat rhinoscleroma--as well as several newer oral agents by using 23 isolates of the causative organism Klebsiella rhinoscleromatis . All isolates were inhibited by clinically achievable concentrations of trimethoprim-sulfamethoxazole, amoxicillin-clavulanate, chloramphenicol, ciprofloxacin, cephalexin, cefuroxime, and cefpodoxime.

Kansenshogaku Zasshi, 1992 Aug, 66(8), 1053 - 61
{Role of capsular polysaccharide and lipopolysaccharide of Klebsiella pneumoniae in experimental mice pneumonia model}; Tateda K et al.; In this study, role of capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of Klebsiella pneumoniae was investigated in experimental mice pneumonia model . Inoculation with K . pneumoniae mucoid strain DT-S into mice lung induced expansive, voluminous lethal pneumonia characterized with thickening of the alveolar septa caused by infiltration of inflammatory cell and packing of bacteria within alveolar spaces . On the other hand, mice lung inoculated with K . pneumoniae DT-X, which was non-mucoid mutant isolated from DT-S during natural passage, showed infiltration of inflammatory cell into alveolar spaces but there was no death of mice during the course of this pneumonia . Inoculation of CPS 100 micrograms of DT-S strain into mice lung induced lesser extent of accumulation of inflammatory cell than that of LPS 4 micrograms of this strain . Stimulation of alveolar and peritoneal macrophage with CPS, even at a concentration of 100 micrograms/ml, induced weaker Interleukin-1 (IL-1) activity than stimulation with LPS 4 micrograms/ml . These results suggest that since CPS of K . pneumoniae DT-S encapsulate bacteria including LPS, CPS may inhibit chemotaxis of inflammatory cell and IL-1 production of macrophage to be induced by LPS during course of pneumonia . It is speculated that existence of CPS have important role in modulating host response to bacterial LPS, and this effect of CPS may be related with difference of pathological findings of lung and lethality between K . pneumoniae DT-S and DT-X.

Genitourin Med, 1992 Aug, 68(4), 269 - 72
A serological test for granuloma inguinale; Freinkel AL et al.; OBJECTIVES--An indirect immunofluorescence technique applied to paraffin embedded tissue sections of lesions containing Donovan bodies was evaluated as a serological test for the diagnosis of granuloma inguinale . METHODS--Sera from patients with proven granuloma inguinale, other sexually acquired genital ulcerations and blood donors from areas where granuloma inguinale is rarely encountered as well as from disease-endemic regions were tested . Sera were tested either unabsorbed or following absorption with whole Klebsiella pneumoniae bacteria . RESULTS--Using unabsorbed sera at a dilution of 1:160 the test was found to have a sensitivity of 100%, specificity of 98%, positive predictive value (PPV) of 89% and negative predictive value (NPV) of 100% . There proved to be no advantage in preabsorbing sera with K . pneumoniae antigen . CONCLUSIONS--In the absence of culture methods for Calymmatobacterium granulomatis, an indirect immunofluorescence technique may prove valuable for the diagnosis of individual cases of granuloma inguinale and as an epidemiological tool in studies of the disease.

Cesk Epidemiol Mikrobiol Imunol, 1992 Aug, 41(3), 157 - 65
{Plasmid profile and toxicity of Klebsiella pneumoniae strains isolated at a neonatal department}; Kosiarova A et al.; The authors tested the plasmid profile of 11 and the toxicity of 16 strains of K . pneumoniae isolated in a neonatal department . Based on examination of the plasmid profile it may be assumed that very probably a nosocomial spread of the strains was involved . This is suggested by the identical plasmid profile of all strains isolated in the environment and five strains isolated from neonates . On investigation of the enterotoxicity of culture filtrates of the investigated strains the authors did not reveal the presence of a thermolabile or thermostable enterotoxin . In the skin test on rabbits 93.8% of the tested samples were positive . Morphological changes on Vero cells after 48 hours action were produced by four culture filtrates . Only two strains caused haemolysis of agar with 6% sheep and rabbit blood after incubation at 37 degrees C . Another 14 strains caused lightening of the agar with rabbit blood after subsequent incubation at 4 degrees C.

J Bacteriol, 1992 Aug, 174(15), 4913 - 9
Structural analysis of the O-antigen side chain polysaccharides in the lipopolysaccharides of Klebsiella serotypes O2(2a), O2(2a,2b), and O2(2a,2c); Whitfield C et al.; The lipopolysaccharide (LPS) of Klebsiella serotype O2 is antigenically heterogeneous; some strains express multiple antigenic factors . To study this heterogeneity, we determined the structure of the O-antigen polysaccharides in isolates belonging to serotypes O2(2a), O2(2a,2b), and O2(2a,2c), by using composition analysis, methylation analysis, and both 1H and 13C nuclear magnetic resonance spectroscopy . The repeating unit structure of the 2a polysaccharide was identified as the disaccharide {----3)-beta-D-Galf-(1----3)-alpha-D-Galp-(1----} and was identical to D-galactan I, one of two O polysaccharides present in the LPS of Klebsiella pneumoniae serotype O1 (C . Whitfield, J . C . Richards, M . B . Perry, B . R . Clarke, and L . L . MacLean, J . Bacteriol . 173:1420-1431, 1991) . LPS from serotype O2(2a,2b) also contained D-galactan I as the only O polysaccharide, suggesting that the 2b antigen is not an O antigen . The LPS of serotype O2(2a,2c) contained a mixture of two structurally distinct O polysaccharides and provides a second example of this phenomenon in Klebsiella spp . One polymer was identical to D-galactan I, and the other polysaccharide, the 2c antigen, was a polymer with a disaccharide repeating unit structure, {----3)-beta-D-GlcpNAc-(1----5)-beta-D-Galf-(1----} . The 2c structure does not resemble previously reported O polysaccharides from Klebsiella spp . Periodate oxidation confirmed that D-galactan I and the 2c polysaccharide are distinct glycans, rather than representing domains within a single polysaccharide chain . Monoclonal antibodies against the 2c antigen indicated that only LPS molecules with the longest O-polysaccharide chains contained the 2c epitope.

J Formos Med Assoc, 1992 Aug, 91(8), 754 - 9
Adult bacteremic pneumonia: bacteriology and prognostic factors; Chen CW et al.; Bacteremic pneumonia is a highly specified subgroup of pneumonia that is potentially life-threatening . In order to find out the prognostic factors in this subgroup of pneumonia, we conducted a 40-month retrospective analysis of 70 cases in our hospital . The male to female ratio was 54:16 . Forty-one cases were community-acquired bacteremic pneumonia (CABP), and 29 cases were nosocomial bacteremic pneumonia (NBP) . Both CABP and NBP were predominated by gram-negative bacteria . Klebsiella pneumoniae was the most common microorganism isolated in both CABP and NBP . The overall mortality was 62.9% (44/70) . There was no significant difference in the mortality between CABP (61.0%) and NBP (65.5%) . After univariate analysis of all possible prognostic factors, 10 variables were found to have significantly poor prognostic values . They were: 1) the presence of septic shock; 2) the use of ventilatory support; 3) the presence of radiologic spread; 4) treatment in an intensive care unit; 5) male gender; 6) the development of adult respiratory distress syndrome; 7) Klebsiella bacteremic pneumonia in patients with an alcohol habit; 8) patients with ultimately fatal underlying diseases; 9) an initial AaDO2 > 200 mmHg; and 10) an initial arterial pH < 7.25.

Mol Gen Genet, 1992 Aug, 234(2), 201 - 10
Cloning and expression of the genes for xylose isomerase and xylulokinase from Klebsiella pneumoniae 1033 in Escherichia coli K12; Feldmann SD et al.; The genes xylA and xylB were cloned together with their promoter region from the chromosome of Klebsiella pneumoniae var . aerogenes 1033 and the DNA sequence (3225 bp) was determined . The gene xylA encodes the enzyme xylose isomerase (XI or XylA) consisting of 440 amino acids (calculated M(r) of 49,793) . The gene xylB encodes the enzyme xylulokinase (XK or XylB) with a calculated M(r) of 51,783 (483 amino acids) . The two genes successfully complemented xyl mutants of Escherichia coli K12, but no gene dosage effect was detected . E . coli wild-type cells which harbored plasmids with the intact xylAKp 5' upstream region in high copy number (but lacking an active xylB gene on the plasmids) were phenotypically xylose-negative and xylose isomerase and xylulokinase activities were drastically diminished . Deletion of 5' upstream regions of xylA on these plasmids and their substitution by a lac promoter resulted in a xylose-positive phenotype . This also resulted in overproduction of plasmid-encoded xylose isomerase and xylulokinase activities in recombinant E . coli cells.

Biochim Biophys Acta, 1992 Jul 13, 1122(1), 15 - 22
Close amino acid sequence relationship between the new plasmid-mediated extended-spectrum beta-lactamase MEN-1 and chromosomally encoded enzymes of Klebsiella oxytoca; Barthelemy M et al.; Isolated from an Escherichia coli strain MEN-1 is a plasmid-mediated beta-lactamase that confers resistance to methoxy imino third-generation cephalosporins . The protein purified to homogeneity was digested by trypsin, chymotrypsin and endoproteinase Asp-N . Amino acid sequence determinations of the resulting peptides gave rise to the alignment of the 263 residues of the beta-lactamase . From amino acid sequence comparison MEN-1 was found to share more than 72% identity with the chromosomally mediated beta-lactamases of Klebsiella oxytoca . Therefore, MEN-1 is the first transferable extended-spectrum beta-lactamase which is not directly derived from the widespread TEMs or SHV-1 penicillinases with which it presents less than 39% identity.

Carbohydr Res, 1992 Jul 2, 231, 93 - 103
Re-investigation of the structure of the capsular polysaccharide of Klebsiella K15 using bacteriophage degradation and inverse-detected NMR experiments; Parolis H et al.; The structure of the capsular polysaccharide from Klebsiella K15 has been re-investigated, principally by 1D and 2D 1H- and 13C-NMR spectroscopy of the oligosaccharide-alditol obtained by depolymerisation of the polysaccharide with a viral-borne endoglycanase followed by borohydride reduction of the isolated repeating oligosaccharide . The capsular polysaccharide was shown to have the repeating unit: {Formula: see text}

J Bacteriol, 1992 Jul, 174(13), 4324 - 30
Klebsiella aerogenes urease gene cluster: sequence of ureD and demonstration that four accessory genes (ureD, ureE, ureF, and ureG) are involved in nickel metallocenter biosynthesis; Lee MH et al.; The region located immediately upstream from the Klebsiella aerogenes urease structural genes was sequenced and shown to possess an open reading frame capable of encoding a 29.8-kDa peptide . Deletions were generated in this gene, denoted ureD, and in each of the genes (ureE, ureF, and ureG) located immediately downstream of the three structural genes . Transformation of the mutated plasmids into Escherichia coli resulted in high levels of urease expression, but the enzyme was inactive (deletions in ureD, ureF, or ureG) or only partially active (deletions in ureE) . Ureases were purified from the recombinant cells and shown to be identical to control enzyme when analyzed by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis; however, in every case the activity levels correlated to nickel contents as analyzed by atomic absorption analysis . UreD, UreE, UreF, and UreG peptides were tentatively identified by gel electrophoretic comparison of mutant and control cell extracts, by in vivo expression of separately cloned genes, or by in vitro transcription-translation analyses; the assignments were confirmed for UreE and UreG by amino-terminal sequencing . The latter peptides (apparent M(r)s, 23,900 and 28,500) were present at high levels comparable to those of the urease subunits, whereas the amounts of UreF (apparent M(r), 27,000) and UreD (apparent M(r), 29,300) were greatly reduced, perhaps because of the lack of good ribosome binding sites in the regions upstream of these open reading frames . These results demonstrate that all four accessory genes are necessary for the functional incorporation of the urease metallocenter.

FEMS Microbiol Lett, 1992 Jul 1, 73(1-2), 47 - 52
The role of Eh, pH and iron in the bactericidal power of human plasma; Bullen JJ et al.; The bactericidal power of fresh human plasma against Klebsiella pneumoniae and Escherichia coli was extremely sensitive to changes in Eh and pH . At a high Eh (approx . +200 mV) the bacteria were destroyed, but rapid regrowth occurred when the Eh was lowered to approx . -400 mV . Abolition of the bactericidal effect was also produced by adding ferric iron at a high Eh (approx . +200 mV) . Lowering the pH to 6.50 reduced or prevented the bactericidal effect . These results are probably related to the availability of iron for bacterial growth, and could be important for understanding the development of infection in injured or diseased tissue.

Mol Microbiol, 1992 Jul, 6(14), 1943 - 8
NADH formation by Na(+)-coupled reversed electron transfer in Klebsiella pneumoniae; Pfenninger-Li XD et al.; Citrate is fermented by Klebsiella pneumoniae to 2 acetate, 0.5 formate and 1.2 CO2 . The formation of less than 1 formate and greater than 1 CO2 per citrate can be accounted for by the oxidation of formate to CO2 in order to provide reducing equivalents for the assimilation of citrate into cell carbon . A membrane-bound electron transport chain is apparently involved in NADH synthesis by these cells . The electrons from formate oxidation to CO2 are used to reduce ubiquinone to ubiquinol by membrane-bound formate dehydrogenase and ubiquinol further delivers its electrons to NAD+, if this endergonic reaction is powered by delta mu Na+ . The endogenous NADH level of K . pneumoniae cells thus increased in the presence of formate in response to a delta pNa+ greater than -100 mV . NADH formation was completely abolished in the presence of oxygen or after addition of hydroxyquinoline-N-oxide, a specific inhibitor of the Na(+)-translocating NADH:ubiquinone oxidoreductase . The increase of endogenous NADH was dependent on the delta pNa+ applied to the cells . Inverted membrane vesicles of K . pneumoniae catalysed the reduction of NAD+ to NADH with formate as electron donor after application of delta mu Na+ of about 120 mV consisting of delta pNa+ of 60 mV and delta psi of the same magnitude . Neither the delta pNa+ nor the delta psi of this size alone was sufficient to drive the endergonic reaction . Strictly anaerobic conditions were required for NADH formation and hydroxyquinoline-N-oxide completely inactivated the reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbiologica, 1992 Jul, 15(3), 219 - 26
Cefonicid potentiation of human macrophage activity; Tullio V et al.; This study was undertaken to evaluate the in vitro effects of cefonicid on phagocyte functions such as phagocytosis and intracellular killing of phagocytosed bacteria . At concentrations of half the MIC cefonicid caused human macrophages to ingest and kill Klebsiella pneumoniae at a greater rate than did drug-free macrophages . Bacteria pretreated with subinhibitory concentrations of cefonicid became more susceptible to the phagocytic and bactericidal activity of macrophages than untreated microorganisms . Sub-MIC cefonicid pretreatment of macrophages did not reduce phagocytosis and killing, confirming the inability of beta-lactam antibiotics to cross biological membranes.

Antibiot Khimioter, 1992 Jul, 37(7), 31 - 4
{Antibacterial effects of gamma-interferon in experimental Klebsiella infection}; Fil'chakov IV et al.; The results of the experimental study on the effect of the natural and recombinant gamma-interferons (gamma-IFs) of mice on the process of the infection caused by Klebsiella sp . are presented . The infection was reproduced by intraperitoneal contamination of mice with a virulent culture of Klebsiella pneumoniae 5055, line SHK . The gamma-IFs were administered to the animals in a dose of 250 units per mouse on days 1 and 3 after the contamination . Survival of the animals, clearance of the pathogen from the blood and liver and functional activity of the phagocytes in the contaminated mice were investigated . It was shown that both the natural and recombinant gamma-IF stimulated the phagocytic activity and oxidative metabolism of the phagocytes in the contaminated mice . Activation of these functions after the use of the natural gamma-IF correlated with its marked protective effect and accelerated elimination of the pathogen from the host which was not observed after the use of the recombinant gamma-IF.

Med Trop (Mars), 1992 Jul-Sep, 52(3), 291 - 7
{Scleroma and rhinoscleroma}; Claveau AM; Scleroma is a specific granulomatous disease of bacterial origin, chronic evolution, with election in respiratory tracts; nose lesion is practically constant and so justifies the term Rhinoscleroma . Although they are many endemic foci throughout the world and in particular in Africa, it is an uncommon disease, often not recognized for polymorphic . The cases recorded out of already known foci concerned generally migrants; autochtonous cases in France are really infrequent . Within the evolutive lesions diagnosis is made out of the isolation of the responsible germ (Klebsiella rhinoscleromatis) and the presence of the specific Mikulicz' cell . To find out localizations in the same patient, we utilize X Rays and endoscopy . Antibiotic therapy is of long duration but treats the affection and prepares the way of surgery intervention to eliminate cicatricial deformities . Relapses are caused by shortness of therapy and some difficulty to monitor patients . Mortel in the past, scleroma should nowadays, be better recognized to be better treated and cured.

J Antimicrob Chemother, 1992 Jul, 30(1), 3 - 16
Extended spectrum beta-lactamase from Klebsiella oxytoca, not belonging to the TEM or SHV family; Wu SW et al.; In clinical isolates of Klebsiella oxytoca resistance to cefuroxime and aztreonam was mediated by a beta-lactamase, designated KH, (pI 5.25) which could be transferred into Escherichia coli by electroporation, but not by conjugation . The transformants produced two enzymes with pIs 5.25 and 8.4 respectively, and showed resistance to cefuroxime, aztreonam, cefotaxime and ceftazidime . Substrate and inhibition profiles indicated that KH beta-lactamase was different from TEM- or SHV-like enzymes, but similar to chromosomal K1 beta-lactamase . The enzyme profile with pI 8.4 was similar to the enzyme from the recipient and showed elevated activity in transformants . The plasmid profiles of the transformants were different from those of their donors . However, a plasmid fragment of the K . oxytoca isolate KH11 hybridized with a plasmid ranging in size from 4.8 to 7.8 kilobases in all the transformants and most of the donors . Gene probes encoding TEM-1 or SHV-1 did not hybridize with plasmid DNA from the K . oxytoca isolates . Furthermore, a probe of the ampC gene did not hybridize with the plasmid but to DNA fragments of the same size in whole cell DNA preparations from the E . coli XAC recipient and the TKH11 transformants . This indicates that no gross rearrangements in the chromosomal beta-lactamase gene region had occurred in the transformants which could explain the increased expression of the pI 8.4 beta-lactamase.

Protein Sci, 1992 Jun, 1(6), 722 - 6
A proposed link between nitrogen and carbon metabolism involving protein phosphorylation in bacteria; Reizer J et al.; We demonstrate that certain phosphoryl transfer proteins of the bacterial phosphotransferase system (PTS), the fructose- and mannitol-specific IIA proteins or domains, are homologous to a class of proteins, one of which is known to affect transcription of some of the nitrogen-regulatory sigma 54-dependent operons in Klebsiella pneumoniae . The phosphorylatable histidyl residue in the homologous PTS proteins and the consensus sequence in the vicinity of the active-site histidine are fully conserved in all members that comprise this family of proteins . A phylogenetic tree of the eight protein members of this family was constructed, and a "signature" sequence that can serve for the identification of new protein members of this family is proposed . These observations suggest that PTS-catalyzed protein phosphorylation may provide a regulatory link between carbon and nitrogen assimilation in bacteria.

J Mol Biol, 1992 Jun 5, 225(3), 591 - 607
DNA supercoiling response of the sigma 54-dependent Klebsiella pneumoniae nifL promoter in vitro; Whitehall S et al.; Transcription from the sigma 54-dependent Klebsiella pneumoniae nifL and glnAp2 promoters is activated by the general nitrogen regulatory protein NTRC . Unlike the glnAp2 promoter, which is relatively insensitive to changes in DNA supercoiling, transcription from nifL in vitro in a chloride-based buffer is supercoiling-dependent at physiological salt concentrations . The replacement of chloride with an acetate-based buffer decreases the stringency of the nifL supercoiling response, but open complexes formed on linear nifL promoter DNA under these conditions are unstable and less extensive than those found on supercoiled (form I) DNA . We have introduced mutations in particular elements of the nifL promoter that increase its homology to glnAp2 . At the wild-type nifL promoter, sigma 54-RNA polymerase makes only limited contacts with the promoter in the absence of NTRC . However, a G to T change at -26 (nifL74) allows the formation of a stable closed complex with sigma 54-holoenzyme on both linear and form I templates in the absence of the activator . The combination of C to T mutations at -3 and -1 (nifL18) increases the A+T rich nature of the melted region and stabilizes open complexes formed on linear DNA . Open complex formation as a function of superhelical density was assessed at each promoter . Formation of open complexes at glnAp2 peaks at -0.024 and declines at higher superhelical densities, whereas at the wild-type nifL promoter, open complex formation peaks at -0.067 and is not detectable at superhelical densities less than -0.032 . Both the nifL74 and nifL18 mutations altered the supercoiling response, increasing the ability to form open complexes at low superhelical densities . The presence of the nifL74 and nifL18 mutations in combination further altered the response of the promoter to DNA supercoiling . These observations suggest that the promoter as a whole, and not any one promoter element, mediates the transcriptional response to DNA supercoiling.

J Biochem Biophys Methods, 1992 Jun, 24(3-4), 309 - 20
Stripping interfering sugars from samples using adapted bacteria; Boe IN et al.; Bacteria adapted to individual sugars quickly remove targeted sugars--stripping them--from samples in which unwanted sugars interfere . Adapted bacteria are equivalent to specific reagents for removal of sugars down to bacterial Km values, micromolar to submicromolar concentrations . Bacterial stripping is a simple method, useful when background sugars in micro-to millimolar concentrations (or larger) interfere with analysis of sought-for sugars . Bacteria such as Escherichia coli and Klebsiella are easily adapted to individual sugars such as lactose, fructose, etc., by growing the bacteria on them . Hence one can easily create (and store) many kinds of cells ready to sponge up or strip out unwanted compounds . E . coli specifically remove several sugars from samples containing 100-500 nmol of sugars, using 1-5 mg of adapted cells, and 25 degrees C temperatures . Stripping requires 1-5 min and consists of mixing cells and sample, spinning down the cells, and withdrawal of stripped supernate . A 1-5 min interval is adequate for uptake and stripping, but far too short for cells to metabolize the sugars that were taken up . Hence the cells do not leak metabolites, but act as specific adsorbants without injection of appreciable byproducts into the sample.

Am J Vet Res, 1992 Jun, 53(6), 1059 - 67
Evaluation of sulbactam plus ampicillin for treatment of experimentally induced Klebsiella pneumoniae lung infection in foals; Hoffman AM et al.; Efficacy of sulbactam, a beta-lactamase inhibitor, in combination with ampicillin, was evaluated for treatment of experimentally induced pneumonia caused by beta-lactam-resistant Klebsiella pneumoniae . Infection was experimentally induced in 18 healthy weanling foals that were randomly allocated to 3 treatment groups: sulbactam plus ampicillin (S/A, 3.3 and 6.6 mg/kg of body weight, respectively), ampicillin (6.6 mg/kg), or vehicle only . Foals were treated daily for 7 days; the observer was unaware of treatment status . Compared with ampicillin and vehicle, treatment with S/A resulted in a statistically significant (P less than 0.05) decrease in severity of pneumonia, with regard to bronchoalveolar lavage cytologic findings (decreased total cell and neutrophil numbers, and increased lymphocyte numbers) and extent of macroscopic lesions in lung tissue of the noninoculated regions . Marked trends toward improvement of S/A-treated foals were observed for quantitative results of bacteriologic culture of bronchoalveolar lavage fluid samples (P less than 0.07), macroscopic pathologic features of the whole lung (P less than 0.1), and histopathologic variables (P less than 0.07), compared with ampicillin- and vehicle-treated foals . Treatment effects were not observed for radiographic, hematologic, and blood gas abnormalities that resulted from infection . In conclusion, the combination of sulbactam plus ampicillin was found to have synergistic effects in vivo, to reduce the extent and severity of experimentally induced gram-negative lung infection in foals.

J Am Vet Med Assoc, 1992 Jun 1, 200(11), 1704 - 6
Cholangiohepatitis in a dog; Forrester SD et al.; Cholangiohepatitis was diagnosed in a dog with a 4-day history of anorexia, vomiting, fever, and icterus . Additional findings included signs of depression, dehydration, hepatosplenomegaly, and abdominal discomfort . Exploratory laparotomy was performed, and specimens of liver, spleen, and bile were obtained . Histologic evaluation of liver and spleen revealed acute, suppurative cholangio-hepatitis and splenitis, respectively . Cultures of liver and bile yielded Klebsiella sp . The dog responded to rehydration and intravenous administration of chloramphenicol . Although uncommon, cholangiohepatitis should be suspected in dogs with anorexia, fever, vomiting, icterus, and signs of abdominal discomfort . Definitive diagnosis requires bacterial cultures of liver and bile . Administration of an appropriate antibiotic should resolve clinical signs.

Ann Rheum Dis, 1992 Jun, 51(6), 783 - 9
Absence of autoantibodies to peptides shared by HLA-B27.5 and Klebsiella pneumoniae nitrogenase in serum samples from HLA-B27 positive patients with ankylosing spondylitis and Reiter's syndrome; de Vries DD et al.; Some microorganisms which are pathogenic in humans share amino acid sequences with human proteins (molecular mimicry) . It has been suggested that molecular mimicry might be a reason for autoimmunity as a result of immunological cross reactivity . A homologous sequence of six amino acids has been found in both Klebsiella pneumoniae nitrogenase and the HLA-B27.5 molecule . In addition, (auto)antibodies to a synthetic peptide that contained the HLA-B27.5/klebsiella mimicking epitope have been detected in serum samples from HLA-B27 positive patients with ankylosing spondylitis and Reiter's syndrome . Confirmation of these data is important, because ankylosing spondylitis and Reiter's syndrome have so far been assumed to be 'seronegative' rheumatic diseases . It was, however, not possible to confirm the presence of autoantibodies against the mimicking peptide in serum samples from patients with ankylosing spondylitis and Reiter's syndrome . Serum samples from 81 patients with ankylosing spondylitis, 38 patients with Reiter's syndrome, and 81 healthy blood donors were tested against the 'mimicking peptide' in an enzyme linked immunosorbent assay (ELISA) . Some of the serum samples from patients showed high but non-specific binding to the mimicking peptide . A highly significant correlation between binding to plastic coated with the mimicking peptide, to plastic coated with an irrelevant peptide, and even to non-coated plastic was observed . The nature of the serum component(s) in these patient serum samples (and some control serum samples) responsible for the high non-specific binding to plastic remains unclear . It was also shown that antibodies to the HLA-B27 peptide (containing the mimicking epitope) induced in rabbits do not cross react with the klebsiella peptide and vice versa.

South Med J, 1992 Jun, 85(6), 638 - 41
Pyogenic hepatic abscess and septic pulmonary emboli associated with Klebsiella ozaenae bacteremia; Chowdhury P et al.; We have reported a case of cryptogenic liver abscess with bacteremia and septic pulmonary emboli due to Klebsiella ozaenae . This organism has not previously been reported as a cause of intra-abdominal infection and this is only the eighth reported case of bacteremia . K ozaenae is generally considered an opportunist of low virulence and a respiratory tract colonizer implicated in ozena (atrophic rhinitis) . More information is required to understand the epidemiology and pathogenicity of human disease caused by K ozaenae.

Infect Immun, 1992 Jun, 60(6), 2529 - 35
Mechanisms of Klebsiella pneumoniae resistance to complement-mediated killing; Merino S et al.; The different mechanisms of Klebsiella pneumoniae resistance to complement-mediated killing were investigated by using different strains and isogenic mutants previously characterized for their surface components . We found that strains from serotypes whose K antigen masks the lipopolysaccharide (LPS) molecules (such as serotypes K1, K10, and K16) fail to activate complement, while strains with smooth LPS exposed at the cell surface (with or without K antigen) activate complement but are resistant to complement-mediated killing . The reasons for this resistance are that C3b binds far from the cell membrane and that the lytic final complex C5b-9 (membrane attack complex) is not formed . Isogenic rough mutants (K+ or K-) are serum sensitive because they bind C3b close to the cell membrane and the lytic complex (C5b-9) is formed.

Mol Gen Genet, 1992 Jun, 233(3), 372 - 8
DNA sequence analysis of the lamB gene from Klebsiella pneumoniae: implications for the topology and the pore functions in maltoporin; Werts C et al.; We have determined the sequence of the lamB gene from Klebsiella pneumoniae . It encodes the precursor to the LamB protein, a 429 amino acid polypeptide with maltoporin function . Comparison with the Escherichia coli LamB protein reveals a high degree of homology, with 325 residues strictly identical . The N-terminal third of the protein is the most conserved part of the molecule (1 change in the signal sequence, and 13 changes up to residue 146 of the mature protein) . Differences between the two mature proteins are clustered mainly in six regions comprising residues 145-167, 173-187, 197-226, 237-300, 311-329, and 367-387 (K . pneumoniae LamB sequence) . The most important changes were found in regions predicted by the two-dimensional model of LamB folding to form loops on the cell surface . In vivo maltose and maltodextrin transport properties of E . coli K12 and K . pneumoniae strains were identical . However, none of the E . coli K12 LamB-specific phages was able to plaque onto K . pneumoniae . Native K . pneumoniae LamB protein forms highly stable trimers . The protein could be purified by affinity chromatography on starch-Sepharose as efficiently as the E . coli K12 LamB protein, indicating a conservation of the binding site for dextrins . However, none of the monoclonal antibodies directed against native E . coli K12 LamB protein recognized native purified K . pneumoniae LamB protein . These data indicate that most of the variability occurs within exposed regions of the protein and provide additional support for the proposed model of LamB folding.(ABSTRACT TRUNCATED AT 250 WORDS)

Indian J Biochem Biophys, 1992 Jun, 29(3), 291 - 5
Absorption and fluorescence studies on interaction between cationic dyes and Klebsiella K7 capsular polysaccharide; Mitra A et al.; Interaction of cationic dyes, pinacyanol chloride, acridine orange and phenosafranin, with Klebsiella K7 capsular polysaccharide has been investigated by spectrophotometric and spectrofluorometric measurements . The acidic polysaccharide induce a metachromatic blue shift of the absorption band of pinacyanol chloride from 600 nm to 495 nm, indicating strong metachromasy . Stoichiometry of polyanion and dye cation (1:1.5) in the polymer-dye compound formed by the interaction between pinacyanol chloride dye and K7 polymer indicate that both glucuronic acid and pyruvic acid act as the potential anionic sites for interaction . Both spectrophotometric titration of pinacyanol chloride and spectrofluorometric titration of acridine orange and phenosafranin dyes by the polymer gave quite comparable equivalent weights for the polymer . Dye-polymer interaction studies indicated induction of metachromasy in the cationic dye by the anionic biopolymer, establishing its chromotropic character.

J Antimicrob Chemother, 1992 Jun, 29(6), 629 - 38
Characterization of chromosomally encoded penicillinases in clinical isolates of Klebsiella pneumoniae; Petit A et al.; Four Klebsiella pneumoniae strains, isolated in two geographically distant French hospitals, were found to produce constitutive beta-lactamases with an unusual isoelectric point for this species (8.1) . The four enzymes were chromosomally encoded and related to the Ambler's class A plasmid-mediated SHV-type enzymes . The four enzymes were predominantly penicillinases, with good activity against amino- and ureidopenicillins . They poorly hydrolysed cephalothin, cephaloridine and cefoperazone and did not hydrolyse third-generation cephalosporins and aztreonam . The four enzymes were susceptible to the inhibitory effect of clavulanic acid . Finally, the strains were all found to produce an acetyl-esterase . The acetyl-esterases catalysized hydrolysis of the acetoxy group of cephalothin and cefotaxime although this did not affect their antibacterial activities . These esterases were not susceptible to the inhibitory effect of clavulanic acid and EDTA.

Indian Pediatr, 1992 Jun, 29(6), 709 - 13
Causes of mortality in children with acute lymphocytic leukemia; Choudhry VP et al.; Fifty five deaths between January, 1982 to September, 1989 in children with acute lymphoblastic leukemia (ALL) were evaluated to determine the cause of mortality . Fifty cases died during remission . Infection alone was responsible for death in 26 of 55 (47.3%) cases while hemorrhage was seen in 7 (12.7%) children . Infection and hemorrhage together were responsible in another 13 cases . Gastrointestinal tract and pulmonary system were the major sites of bleeding . Infections either alone or in combination with other factors were responsible for death in 42 of 55 (76.5%) of children . Septicemia (n = 11), gastrointestinal (n = 15) and pulmonary infections (n = 10) and meningitis in 2 cases were the major sites of infections . Pseudomonas and Klebsiella in 6 cases each accounted for 54.5% of isolatesPIP: New developments in case management presently afford cures to more than 60% of children with acute lymphoblastic leukemia (ALL) . 287 children diagnosed with ALL were admitted to the All India Institute of Medical Sciences over the period January, 1982 - September, 1989, where they began chemotherapy . 50 died during initial or subsequent induction therapy and 5 died during the maintenance phase . All deaths were subsequently reviewed to identify the causes of mortality . Infection alone caused death in 47.3% of cases, hemorrhage was observed among 12.7%, and infection together with hemorrhage killed another 13 children . Septicemia, gastrointestinal, and pulmonary infections in 11, 15, and 10 cases, respectively, and meningitis in 2 cases were major sites or infection . Pseudomonas and Klebsiella in 6 cases each accounted for 54.5% of isolates . The gastrointestinal tract and pulmonary system were major sites of bleeding . While no definite cause of death was found for 5 cases, infections nonetheless either alone or combined with other factors caused 76.5% of deaths . To improve the long-term event free survival of children with ALL, practitioners must be knowledgeable about the potential spectrum of infections, begin treatment early with appropriate antibiotics, and seek to improve the availability of supportive facilities and modern antibiotics .

Indian Pediatr, 1992 Jun, 29(6), 693 - 8
Neonatal gastrointestinal perforations; Merchant RH et al.; Fourteen neonates presented with clinical and radiological evidence of gastrointestinal perforation, of which 13 were treated by operative intervention . No infant was formula fed, had rectal bleeding or radiological evidence of pneumatosis intestinalis . Operative findings revealed localized perforation in ten of which three were gastric, two distal ileal, three cecal and two in sigmoid colon . Gangrene with extensive perforation of jejunum, ascending colon and transverse colon were seen in one each . Blood cultures were positive in four, one grew E . coli and three Klebsiella while peritoneal cultures were positive in six, one grew E . coli and five Klebsiella . Ten neonates survived and are being followed up . Three of the four cases who died had white blood cell count greater than 25 x 109/L and grew Klebsiella on peritoneal culture . Factors predisposing to gastrointestinal perforations in neonates are discussed, emphasis is made on the cautious use of umbilical, gastric and rectal catheters, and the need for early surgical intervention.

Mol Microbiol, 1992 Jun, 6(12), 1625 - 30
Activator-independent formation of a closed complex between sigma 54-holoenzyme and nifH and nifU promoters of Klebsiella pneumoniae; Buck M et al.; The alternative sigma factor sigma 54 is required for transcription of nitrogen fixation genes in Klebsiella pneumoniae and other diazotrophs . The nif genes, and other E sigma 54-dependent genes whose products are necessary for a wide range of processes, are postively regulated . A unifying model that is well supported by studies on nif and other nitrogen-regulated (ntr) genes includes the central tenet that sigma 54 confers upon core RNA polymerase the ability to recognize and bind specific promoter sequences, but not the ability to isomerize to the open complex without assistance from the appropriate activator protein . Direct physical evidence for formation of an activator-independent complex between E sigma 54 and the NifA-dependent K . pneumoniae nifH and nifU promoters has, to date, been lacking . Using purified components we have now demonstrated formation of the closed complex at these promoters, indicating that it is an intermediate along the pathway to open complex formation . The closed complex was not detected when conserved features of the promoter were altered by mutation, nor was its stability increased when integration host factor protein was bound adjacent to the E sigma 54 recognition sequence.

Zhonghua Jie He He Hu Xi Za Zhi, 1992 Jun, 15(3), 161 - 2, 191
{Studies of bronchoalveolar lavage cells during the airborne Klebsiella pneumoniae infection in immunocompromised mice}; Zhu BH; We compared immunocompromised mice with normal mice during the airborne Klebsiella pneumoniae (KP) infection, to study the course and developing mechanisms of KP pneumonia . There are significant difference in the number and peaking time of PMN, the number of lymphocytes and plasma cells in the bronchoalveolar lavage fluids of different mice groups . These results indicate that the interference of immune specific and nonspecific host responses is an important variable in antibiotic efficacy and the existence of an immunomodulating cytokine network was suggested.

Kansenshogaku Zasshi, 1992 Jun, 66(6), 696 - 708
{Study on lymphocyte-response in early stage of respiratory infection--a view point from experimental Klebsiella pneumoniae pneumonia}; Watanaba H; This study was performed to demonstrate the early immuno-responses of lymphocytes in an experimental pneumonia with K . pneumoniae in mature mice (45 week-old) comparing with it in juvenile ones (4 week-old) . Acute mice pneumonia was made by inhalation with K . pneumoniae DT-S strains into lung . Changes in lymphocytes including their subpopulation in bronchoalveolar lavage fluid (BALF), peripheral blood, hilar lymphnodes and lung tissue were observed after the inhalation . In addition, lung tissue and hilar lymphnode were examined immunohistologically . The following results were obtained: 1 . Total lymphocytes in BALF were more rapidly increased in the mature group than in the juvenile one . But there was no significant change in leukocyte count in peripheral blood between both groups . 2 . Such a rapid increase in lymphocytes in BALF in mature group depended on L3T4-Ly6c cells and L3T4-LFA 1 cells . These cells in juvenile group were not accumulated in BALF at initial phase of the infection . But in contrast, they were gradually increased in peripheral blood and in hilar lymphnode . There was significant time-differences in appearance of these cells in BALF between both groups . It might be, that L3T4-Ly6c cells and L3T4-LFA1 cells observed in BALF in mature animals were induced from bronchus-associated lymphoid tissue (BALT) or small lymph tissue in alveolar interstitium, but in juvenile ones were originated in hilar lymphnode . 3 . Changes in Ly2-Ly6c cell and Ly2-LFA1 cell were shown the same tendency changes in L3T4-Ly6c cell and L3T4-LFA1 cell . 4 . Accumulation of B220:Ly5-LFA1 cell in BALF was not observed in significant difference between the mature group and in the juvenile one . 5 . L3T4 cells were markedly accumulation in subcortex area of hilar lymphnode in the juvenile group, but they were only seen scattered in the mature group . 6 . It can be concluded that active T lymphocyte begins to response in situ in the early stage of respiratory infection in the mature host and this finding is a characteristic lymphocyte response, that is never seen in the juvenile group.

Mutat Res, 1992 Jun, 282(2), 73 - 7
Increased mutagenicity of some nitroimidazoles by non-mutagenic nitrotoluene on Klebsiella pneumoniae (fluctuation test); Voogd CE et al.; In the fluctuation test the mutation frequency of Klebsiella pneumoniae by the 5-nitroimidazoles panidazole and dimetridazole was increased by adding the non-mutagenic substances 4-nitrotoluene or toluene-4-sulfonamide . This effect was not found with 1-methyl-5-nitroimidazole, metronidazole, ronidazole, nimorazole and 1-methyl-2-hydroxymethyl-5-nitroimidazole . It is suggested that the molecules of panidazole or dimetridazole form some association, which is destroyed by 4-nitrotoluene or toluene-4-sulfonamide, thus increasing the concentration of mutagenic particles.

J Mol Biol, 1992 May 20, 225(2), 271 - 86
Central domain of the positive control protein NifA and its role in transcriptional activation; Cannon W et al.; The positive control protein NifA of Klebsiella pneumoniae activates transcription by RNA polymerase containing sigma 54 by catalysing open promoter complex formation . We show that the integrity of the putative ATP-binding pocket in the central domain of NifA is necessary for the positive control function of NifA, but is not required for DNA-binding or recognition of NifA by NifL . The inactive mutant NifA proteins are trans dominant to wild-type NifA and are unable to catalyse formation of open promoter complexes irrespective of whether a closed promoter complex at the nifH promoter has preformed . Formation of the closed complex results in a DNA structural distortion adjacent to the DNA region melted in the open promoter complex . This distortion lies at the leading edge of the E sigma 54 footprint . Although unable to catalyse open complex formation, some mutant NifAs altered the chemical reactivity of the distorted base-pair indicating that they retain the ability to recognize the closed promoter complex . The activation phenotype of partially active NifA molecules was sensitive to promoter sequences known to influence closed complex formation, indicating differences in (1) the susceptibility of the closed complexes towards activation and (2) their requirements for NifA during activation.

J Am Vet Med Assoc, 1992 May 15, 200(10), 1521 - 4
Septic cholangitis and peritonitis in a gelding; Clabough DL et al.; An 8-year-old Arabian gelding with septic cholangitis and peritonitis was successfully treated with trimethoprim/sulfadiazine . The gelding was referred for evaluation of signs of abdominal pain, icterus, fever, and weight loss . Peritoneal fluid analysis revealed septic and suppurative peritonitis . Culture of the peritoneal fluid yielded Escherichia coli and Klebsiella pneumoniae, which were sensitive to trimethoprim/sulfadiazine . On the basis of results of hepatic ultrasonography, a diagnosis of septic cholangitis also was made . The horse was treated with 30 mg of trimethoprim/sulfadiazine/kg, PO, q 12 h for approximately 6 weeks . The horse improved steadily, and telephone follow-up with the owner 1 year later disclosed that the horse had complete return to normal condition, appetite, and attitude . On the basis of our findings, aggressive, long-term anti-inflammatory and antibiotic treatment may result in complete return to health and normal athletic function in horses with septic cholangitis and concurrent septic peritonitis.

Infect Immun, 1992 May, 60(5), 1771 - 8
Monoclonal antibody against Klebsiella capsular polysaccharide reduces severity and hematogenic spread of experimental Klebsiella pneumoniae pneumonia; Held TK et al.; Klebsiella pneumoniae is an important nosocomial pathogen causing severe pulmonary infections . The majority of clinical Klebsiella isolates produce a high-molecular-weight capsular polysaccharide (CPS) which is one of the dominant virulence factors . In the present study, we examined the potency of a murine immunoglobulin M monoclonal antibody (MAb) with specificity to Klebsiella type 2 CPS to protect rats against experimental Klebsiella pneumonia . The MAb did not prevent the invasion of virulent bacteria into the interalveolar space . However, the resolution of infection was accelerated in MAb-treated animals . This was demonstrated by (i) less severe weight loss and (ii) markedly reduced inflammatory reactions in the lung . The elimination of bacteria was significantly increased not only in the lungs but also in the livers of antibody-treated rats . This was reflected by reduced levels of circulating, soluble CPS and MAb-bound CPS . A mixture of human MAbs with specificity to CPS of clinically important Klebsiella serotypes may prove to be a useful tool for the prevention or supportive treatment of Klebsiella pneumonia.

J Biol Chem, 1992 May 5, 267(13), 9398 - 408
The dependence on iron availability of allocation of iron to nitrogenase components in Klebsiella pneumoniae and Escherichia coli; Miller AF et al.; Nitrogenase contains approximately 38 iron ions/complete unit . Therefore, we sought to identify steps and genes involved in nitrogenase production that are responsive to iron availability . We have characterized nitrogenase production in Klebsiella pneumoniae grown in a range of different iron concentrations . We find significant accumulation (50-75%) and normal synthesis rates of the structural polypeptides, even under conditions in which the observed nitrogenase activities are only 14-28% of those observed in iron-sufficient conditions . Thus, maturation instead of synthesis of the structural polypeptides is primarily responsible for the iron dependence of nitrogenase activity . We have also used a binary plasmid system in Escherichia coli to investigate the contributions of various nitrogen fixation (nif) genes to the iron dependence of nitrogenase production . At least one of the nif genes DKTYENXUSVW can modulate synthesis of the structural polypeptide NIF H in response to iron availability . We speculate that an iron-deficient complex of the product(s) of at least one of these genes may repress structural polypeptide synthesis in iron-depleted K . pneumoniae . Such a system would compensate for the inactivity of NIF L in iron-depleted cultures and ensure balanced production of the structural polypeptides of nitrogenase in accordance with the iron available for their maturation.

Appl Environ Microbiol, 1992 May, 58(5), 1711 - 8
Excretion of ammonium by a nifL mutant of Azotobacter vinelandii fixing nitrogen; Bali A et al.; A mutation in the gene upstream of nifA in Azotobacter vinelandii was introduced into the chromosome to replace the corresponding wild-type region . The resulting mutant, MV376, produced nitrogenase constitutively in the presence of 15 mM ammonium . When introduced into a nifH-lacZ fusion strain, the mutation permitted beta-galactosidase production in the presence of ammonium . The gene upstream of nifA is therefore designated nifL because of its similarity to the Klebsiella pneumoniae nifL gene in proximity to nifA, in mutant phenotype, and in amino acid sequence of the gene product . The A . vinelandii nifL mutant MV376 excreted significant quantities of ammonium (approximately 10 mM) during diazotrophic growth . In contrast, ammonium excretion during diazotrophy was much lower in a K . pneumoniae nifL deletion mutant (maximum, 0.15 mM) but significantly higher than in NifL+ K . pneumoniae . The expression of the A . vinelandii nifA gene, unlike that of K . pneumoniae, was not repressed by ammonium.

Br J Ind Med, 1992 May, 49(5), 359 - 64
Scintigraphy with J001X, a Klebsiella membrane glycolipid, for the early diagnosis of chronic berylliosis: results from an experimental model; Diot P et al.; A glycolipid (J001X) isolated from the membrane proteoglycans of a non-pathogenic strain of Klebsiella pneumoniae was developed to bind selectively to macrophages . A scintigraphic technique could thus be developed and applied to an experimental model of lung berylliosis . Six baboons were injected intratracheally with a beryllium metal suspension . Three to 24 months later, they were submitted to both an anatomical and a functional respiratory evaluation . Two baboons were explored at the early stage of alveolitis and four baboons at a more advanced stage characterised by a granulomatous disorder . Scintigraphy was performed using J001X labelled with 99mtechnetium administered as an aerosol . In the six baboons, conventional imaging techniques (chest x ray film, computed tomography scan, gallium scintigraphy), failed to show either any lung abnormality or mediastinal lymph nodes consistent with beryllium disease . In the two recently contaminated baboons, J001X scintigraphy showed a well defined parenchymal fixation facing the contaminated lobe . In the four baboons who were at a more advanced stage of berylliosis, J001X fixation was always focused paratracheally without any significant involvement of the lung parenchyma . The subcarinal and laterotracheal lymph nodes seen at necropsy corresponded to J001X scintigraphic fixations . In conclusion, when compared with conventional techniques such as chest x ray film, computed tomography scan, magnetic resonance imaging, and gallium scintigraphy, J001X scintigraphy has proved its ability to detect occult lesions in experimental berylliosis in baboons . By comparison with gallium scintigraphy, scintigraphy with J001X appears to have superior sensitivity and can be performed in four hours.

J Infect Dis, 1992 May, 165(5), 898 - 903
Dietary fish-oil supplementation in experimental gram-negative infection and in cerebral malaria in mice; Blok WL et al.; Dietary fish-oil supplementation interferes with eicosanoid production and appears to decrease production of interleukin-1 (IL-1) and tumor necrosis factor (TNF) . The effect of fish oil was investigated in an intramuscular Klebsiella pneumoniae infection in Swiss mice and in cerebral malaria induced by Plasmodium berghei in C57B1/6 mice . After a low inoculum of K . pneumoniae, 90% of fish oil-fed mice survived; survival in control mice fed equal amounts of corn or palm oil or normal chow was 30%, 40%, and 0, respectively . Cerebral malaria occurred in only 23% of fish oil-fed mice; in the controls, cerebral malaria developed in 61%, 81%, and 78%, respectively . Contrary to what was expected, lipopolysaccharide-induced ex vivo production of IL-1 alpha and TNF alpha by peritoneal cells was significantly enhanced in fish oil-fed mice compared with controls . Indomethacin treatment did not alter the outcome in these two infections, thus arguing against reduced prostaglandin synthesis as an explanation for the increase in resistance to infection.

Hepatology, 1992 May, 15(5), 799 - 803
The prevalence and prognostic significance of spontaneous bacterial peritonitis in severe acute hepatitis with ascites; Chu CM et al.; The prevalence and prognostic significance of spontaneous bacterial peritonitis were prospectively studied in a series of 82 acute hepatitis patients decompensated with ascites . The in-hospital prevalence of spontaneous bacterial peritonitis was 31.7% (26 of 82 patients) . Twenty cases were culture positive, including one with multiple isolates, and six cases were culture negative . E . coli and Klebsiella pneumoniae were the most common pathogens, accounting for 71.4% (15 of 21) of the total isolates, whereas only 9.5% were gram-positive organisms . No significant difference in the age, sex, cause of acute hepatitis, liver biochemistry, prothrombin time and ascites fluid concentration of total protein was noted between patients with spontaneous bacterial peritonitis and those without spontaneous bacterial peritonitis, except that bacteremia was recognized significantly more frequently in the former (57.7% or 15 of 26 patients) than in the latter (25.0% or 14 of 56 patients, p less than 0.005) . In addition, patients with spontaneous bacterial peritonitis, when compared with those without spontaneous bacterial peritonitis, were more likely to have kidney failure (57.7% vs . 30.4%, p less than 0.05) and had a significantly higher mortality rate (73.1% vs . 39.3%, p less than 0.01) . Among patients without spontaneous bacterial peritonitis, the prevalence of kidney failure and gastrointestinal hemorrhage and the mortality rate in patients with bacteremia (57.1%, 64.3% and 71.4%, respectively) were significantly higher than in those without bacteremia (21.4%, 19.0% and 28.6%, respectively; p less than 0.05, p less than 0.01 and p less than 0.01, respectively) . In conclusion, 31.7% of severe acute hepatitis patients with ascites were recognized as having spontaneous bacterial peritonitis; the other 17.1% had bacteremia.(ABSTRACT TRUNCATED AT 250 WORDS)

Mikrobiol Zh, 1992 May-Jun, 54(3), 14 - 8
{The surface structures and biological properties of Klebsiella strains}; Martynenko LD et al.; The electron-microscopic method has been used to study the surface structures and biological properties (adhesiveness, antibiotic-resistance, biochemical properties) in 67 strains of Klebsiella isolated from one-year-old children with intestinal infections (56) and from healthy ones (11) . It is found that bacterial cells in 58.2% of strains have fimbriae of the 3d type, in 20.9%--of the 1st type . No correlation has been revealed between the type of fimbriae and the studied properties.

Philos Trans R Soc Lond B Biol Sci, 1992 Apr 29, 336(1276), 73 - 81; discussion 81-2
Nitrogenase of Klebsiella pneumoniae: an MgATP hydrolysing energy transduction system with similarities to actomyosin and p21 ras; Thorneley RN; The mechanism of ATP hydrolysis by nitrogenase shows some similarity to that proposed for actomyosin and for GTP hydrolysis by p21 ras . All three systems involve the formation of an active complex from two component proteins, nucleotide-induced changes in protein conformation, energy transduction that in the case of nitrogenase involves a decrease in redox potential of metal centres, and a slow dissociation of the protein complex . Metal ion activation (Mg2+ or Ca2+) and in-line displacement of ADP by H2O without enzyme phosphorylation are also common features . At 5 degrees C, stopped-flow calorimetry shows that the kinetic and thermodynamic parameters for endothermic, reversible on-enzyme cleavage of MgATP by nitrogenase and myosin subfragment 1 are remarkably similar . {18O4}Pi-water exchange studies also show that ATP cleavage on nitrogenase and myosin are reversible.

Biochim Biophys Acta, 1992 Apr 14, 1138(4), 318 - 26
Enhanced localization of liposomes with prolonged blood circulation time in infected lung tissue; Bakker-Woudenberg IA et al.; In an experimental model of unilateral pneumonia caused by Klebsiella pneumoniae in rats we investigated whether intravenous administration of liposomes with prolonged blood circulation time resulted in significant localization of liposomes in infected lung tissue . Liposomes (100 nm) composed of hydrogenated phosphatidylinositol:hydrogenated phosphatidylcholine:cholesterol (molar ratio, 1:10:5) radiolabeled with gallium-67-deferoxamine showed relatively long blood circulation time . The degree of localization of these long circulating liposomes in the infected left lung was significantly higher compared to that of localization of 110 nm egg phosphatidylglycerol:egg phosphatidylcholine:cholesterol (molar ratio, 1:10:5) liposomes which exhibited relatively short blood circulation time . At 16 h after administration of the long circulating liposomes (when 10% of the injected dose was still present in the bloodstream) localization of liposomes in the infected left lung was increased up to 10-fold compared to the left lung of uninfected rats, and appeared to be highly correlated with the intensity of the infection . In the uninfected right lung the localization of long circulating liposomes was not increased . The degree of localization of liposomes in the infected tissue is dependent on the residence time of liposomes in the blood compartment . The extent of localization of long circulating liposomes in infected tissue appeared to be dependent on the liposomal dose administered.

Ann Med, 1992 Apr, 24(2), 129 - 36
Liver transplantation restores low serum levels of very low density and high density lipoproteins in end-stage primary biliary cirrhosis; Nikkila K; Amounts of serum lipids were determined in 10 women suffering from end-stage primary biliary cirrhosis (PBC) in order to evaluate the concentrations of and changes in very low density (VLDL), high density (HDL) lipoprotein cholesterol and apoprotein A-I before and after liver transplantation . The concentrations of some preoperative serum total lipids, especially HDL cholesterol and VLDL triglyceride, and apoprotein A-I, were significantly lower, but those of serum total triglycerides and cholestanol (a metabolite of cholesterol) were higher in the PBC patients than in 12 controls matched for age, sex, and body mass index . The relation between the serum concentrations of HDL cholesterol and cholestanol was markedly different before transplantation in the PBC group and in the control group . Liver transplantation was followed by a significant and rapid increase in serum apoprotein A-I and HDL cholesterol concentrations, which were affected by CMV and Klebsiella infections and acute rejections and immunosuppressive treatments . As a result the patients serum values did not quite return to normal during the three month follow up . The concentrations of serum VLDL cholesterol, triglycerides and cholestanol returned to normal during the follow up, and the changes in cholestanol were inversely related to the increases in amounts of HDL cholesterol . Thus, the restoration of low HDL concentrations after liver transplantation suggests that the liver plays a key role in HDL metabolism and indicates that concentrations of serum HDL cholesterol may be useful in monitoring the recovery of liver function.

Can J Ophthalmol, 1992 Apr, 27(3), 143 - 7
Endogenous Klebsiella pneumoniae endophthalmitis in diabetic patients; Liao HR et al.; We describe 12 patients with endogenous endophthalmitis caused by Klebsiella pneumoniae seen between 1983 and 1988 . Three patients had bilateral involvement . Eleven patients had diabetes mellitus, either newly diagnosed or poorly controlled . In six cases the infection was associated with a liver abscess . In all eyes the outcome was light perception or worse; nine eyes had no light perception . Six eyes were enucleated or eviscerated.

Int J Syst Bacteriol, 1992 Apr, 42(2), 252 - 6
Identification of Klebsiella pneumoniae by DNA hybridization and fatty acid analysis; Spierings G et al.; On the basis of the idea that DNA sequences encoding cell surface-exposed regions of outer membrane proteins are genus or species specific, two oligonucleotide probes which were based on the PhoE protein of Klebsiella pneumoniae were evaluated . In slot blot hybridizations and in polymerase chain reactions, no cross-hybridizations were observed with non-Klebsiella strains . When the probes were tested on 75 different K-antigen reference Klebsiella strains, 16 strains were not recognized although they did produce PhoE protein under phosphate starvation . To determine whether these 16 strains belong to (a) different species, the reference strains were also tested for the ability to produce indole and to grow at 10 degrees C and their whole-cell fatty acid patterns were analyzed by gas chromatography . A strong correlation was observed among (i) reaction with the probes, (ii) the inability to produce indole, (iii) the inability to grow at 10 degrees C, and (iv) the presence of the hydroxylated fatty acid C14:0-2OH . From these results we conclude that the two oligonucleotides are specific for the species K . pneumoniae . Furthermore, analysis of fatty acid patterns appears to be a useful tool to distinguish K . pneumoniae from other Klebsiella species.

Acta Anaesthesiol Scand, 1992 Apr, 36(3), 289 - 91
An outbreak of Klebsiella oxytoca septicemias associated with the use of invasive blood pressure monitoring equipment; Ransjo U et al.; Seven cases of septicemia with piperacillin-resistant Klebsiella oxytoca (PRKO) occurred at 2-monthly intervals in a thoracic surgery intensive care unit . All PRKO isolates were serotyped, and phenotyped with a biochemical typing system . Only one patient in the unit was found to be colonized in stool or respiratory tract with PRKO, and this strain was different from the septicemia strains in phenotype though not in serotype . Environmental cultures, from humidifiers, oxygenators, pressure transducers, etc . were negative . PRKO of the epidemic phenotype was recovered from several non-patient transducer domes . The outbreak ended when transducer heads were disinfected and the use of non-patient domes was abolished.

Radiat Res, 1992 Apr, 130(1), 61 - 4
Short and long courses of ofloxacin therapy of Klebsiella pneumoniae sepsis following irradiation; Brook I et al.; Exposure to whole-body irradiation is associated with fatal gram-negative sepsis . The optimal length of therapy of such infection is not established . The effect of short and long courses of oral therapy with the quinolone ofloxacin for orally acquired Klebsiella pneumoniae infection was tested in B6D2F1 mice exposed to 8.0 Gy of bilateral radiation from 60Co . A dose of 10(8) organisms was given orally 4 days after irradiation, and therapy was started 1 day later . Cultures of the ileum 7 days after irradiation showed the recovery of K . pneumoniae in 7 of 10 untreated mice and in 3 of 20 treated with ofloxacin . However, 14 days after irradiation K . pneumoniae was isolated in 5 of 6 untreated mice, in 7 of 9 that received the short course of therapy, and in one of those that received the long course of therapy (P less than 0.05) . At Day 7, K . pneumoniae was isolated from the livers of 6 of 10 untreated mice, and from none of those receiving ofloxacin (P less than 0.05) . At 14 days, K . pneumoniae was isolated in 4 of 6 untreated animals, in 4 of 9 that received the short course of therapy, and in none of the mice that received the long course of therapy (P less than 0.05) . Only 3 of 20 (15%) untreated mice survived for 30 days as compared to 11 of 20 (55%) mice treated for 7 days with ofloxacin and 18 of 20 (90%) mice treated for 21 days with ofloxacin (P less than 0.05) . These survival data illustrate the efficacy of a 21-day course over a 7-day course of ofloxacin therapy for orally acquired K . pneumoniae infection in irradiated hosts.

J Med Microbiol, 1992 Apr, 36(4), 250 - 4
Klebsiella capsular type K7 in relation to toxicity, susceptibility to phagocytosis and resistance to serum; Podschun R et al.; Klebsiella strains possessing capsule type K7 are found predominantly in respiratory secretions . To investigate the importance of this K antigen in virulence, 13 K7 strains were compared with K2 capsulate isolates which are generally regarded as highly virulent . The toxicity of the strains was determined in a mouse peritonitis model . Generally, K7 isolates were significantly less toxic for mice than K2 strains . In the absence of serum, neither capsule type showed much stimulation of leucocytes, measured as the chemiluminescence (CL) response of human polymorphonuclear leucocytes (PMNL) . However, in the presence of normal human serum, CL values with K7 strains increased considerably, whereas the CL response to K2 isolates was unaffected . Correspondingly, intracellular killing by PMNL was observed with K7 strains only, whereas K2 isolates proved to be relatively resistant to phagocytic destruction . No correlation was found between capsule type K7 and serum resistance . These data suggest that, in contrast to K2, capsule type K7 may not be a critical factor in the virulence of K7-capsulate Klebsiella strains nor does it seem to act as an antiphagocytic barrier.

J Bacteriol, 1992 Apr, 174(8), 2485 - 92
A monoamine-regulated Klebsiella aerogenes operon containing the monoamine oxidase structural gene (maoA) and the maoC gene; Sugino H et al.; The Klebsiella aerogenes gene maoA, which is involved in the synthesis of monoamine oxidase, was induced by tyramine and the related compounds, subjected to catabolite and ammonium ion repression, and cloned . The nucleotide sequence of the region involved in monoamine oxidase synthesis was determined . Two open reading frames, the maoA gene and a hitherto unknown gene (maoC), were found . These are located between a potential promoter sequence and a transcriptional terminator sequence . A region of the Escherichia coli chromosome that was highly homologous to the Klebsiella maoA gene was found . The potential maoA gene is located at 30.9 min on the E . coli chromosome . Analysis of the amino acid sequences of the first 11 amino acids from the N terminus of the purified monoamine oxidase agrees with those deduced from the nucleotide sequence of the maoA gene . The leader peptide extends over 30 amino acids and has the characteristics of a signal sequence . Primer extension and S1 nuclease mapping of transcripts generated in vivo suggests that the tyramine-induced mRNA starts at a site 62 bases upstream from the ATG initiation codon of the maoC gene . In the putative promoter region, a high degree of similarity to the consensus sequence for the binding site of cyclic AMP receptor protein was found . Thus, the mao region is composed of two cistrons, and the mao operon is regulated by monoamine compounds, glucose, and ammonium ions.

Eur J Biochem, 1992 Apr 1, 205(1), 417 - 24
Cloning and expression of an Arabidopsis nitrilase which can convert indole-3-acetonitrile to the plant hormone, indole-3-acetic acid; Bartling D et al.; From an Arabidopsis thaliana cDNA expression library, a cDNA clone was isolated, characterized and sequenced which, at the amino acid level, resembled the Klebsiella ozaenae bromoxynil nitrilase encoded by the bxn gene . The cDNA contained a long open reading frame, starting from two possible neighbouring ATG codons and capable of encoding 340 or 346 amino acids with calculated molecular masses of 37526 Da or 38176 Da, respectively . The sequence similarity between the deduced polypeptides from the Arabidopsis cDNA and bxn was clustered in three domains, one at the C-terminus, one in the center and one near the N-terminus of the two proteins, suggesting important functional elements in these parts of the proteins . The cDNA was cloned into different vectors under the control of the lacZ promotor and was functionally expressed by induction with isopropyl-beta-D-thiogalactoside . Using a combination of high-performance liquid chromatography, monoclonal-antibody based enzyme-linked immunosorbent assay and mass spectroscopy, it was shown that the isolated cDNA clone encodes an enzymatically active nitrilase which is able to convert indole-3-acetonitrile to the plant growth hormone, indole-3-acetic-acid.

J Bacteriol, 1992 Apr, 174(7), 2344 - 51
Cloning and nucleotide sequence of a negative regulator gene for Klebsiella aerogenes arylsulfatase synthesis and identification of the gene as folA; Azakami H et al.; A negative regulator gene for synthesis of arylsulfatase in Klebsiella aerogenes was cloned . Deletion analysis showed that the regulator gene was located within a 1.6-kb cloned segment . Transfer of the plasmid, which contains the cloned fragment, into constitutive atsR mutant strains of K . aerogenes resulted in complementation of atsR; the synthesis of arylsulfatase was repressed in the presence of inorganic sulfate or cysteine, and this repression was relieved, in each case, by the addition of tyramine . The nucleotide sequence of the 1.6-kb fragment was determined . From the amino acid sequence deduced from the DNA sequence, we found two open reading frames . One of them lacked the N-terminal region but was highly homologous to the gene which codes for diadenosine tetraphosphatase (apaH) in Escherichia coli . The other open reading frame was located counterclockwise to the apaH-like gene . This gene was highly homologous to the gene which codes for dihydrofolate reductase (folA) in E . coli . We detected 30 times more activity of dihydrofolate reductase in the K . aerogenes strains carrying the plasmid, which contains the arylsulfatase regulator gene, than in the strains without plasmid . Further deletion analysis showed that the K . aerogenes folA gene is consistent with the essential region required for the repression of arylsulfatase synthesis . Transfer of a plasmid containing the E . coli folA gene into atsR mutant cells of K . aerogenes resulted in repression of the arylsulfatase synthesis . Thus, we conclude that the folA gene codes a negative regulator for the ats operon.

Eur J Clin Microbiol Infect Dis, 1992 Apr, 11(4), 349 - 52
Isolation of Klebsiella terrigena from clinical specimens; Podschun R et al.; In a three-year survey conducted from 1988 to 1990 Klebsiella isolates from human clinical specimens were subjected to additional tests to identify any Klebsiella terrigena strains . Ten strains of Klebsiella terrigena (0.4%) were found among 2355 indole-negative Klebsiella isolates . Most of the isolates were recovered from the respiratory tract . In the API20EC system almost exclusively biotypes no . 1777771 and 1777671 were observed . Serotyping revealed capsule types K2, K5 and K18 in two strains each . In antibiotic susceptibility tests the strains were shown to be comparable in sensitivity to Klebsiella pneumoniae.

Mol Microbiol, 1992 Apr, 6(8), 1035 - 47
Mapping and characterization of the promoter elements of the regulatory nif genes rpoN, nifA1 and nifA2 in Rhodobacter capsulatus; Preker P et al.; The promoter elements responsible for the expression of the regulatory nif genes rpoN, nifA1 and nifA2 of Rhodobacter capsulatus were mapped by exonuclease-III-mediated deletions and by primer extension analysis . The rpoN promoter maps 600 bp upstream of rpoN and has the characteristic features of a -24/-12 promoter . The upstream activator sequence (UAS) displays two mismatches with the NIFA consensus sequence and is located 37 bp upstream of a perfect -24/-12 promoter element . The spacing and/or the helical phasing of these two promotor elements was found to be important for promoter function . In addition, an UAS half-site may contribute to optimal promoter function . The rpoN UAS can partially substitute for the UAS of the nifE promoter . An open reading frame with homology to Klebsiella pneumoniae NIFU was identified between the rpoN promoter and rpoN and termed nifU2 since another nifU-like gene (nifU1) is located in a conventional nifUSVW operon in nif region A . Thus, rpoN, encoding an alternative sigma factor for RNA polymerase, is cotranscribed with a nifU analogous gene from an rpoN-dependent promoter . Mapping of the promoter elements involved in the expression of nifA copy 1 and copy 2 identified a novel promoter type . A conserved distal promoter element is likely to represent the binding site of NTRC in R . capsulatus . The DNA region preceding the mapped 5' ends of the nifA transcripts displays much less homology . The distance between the distal and proximal elements is about 100 bp.

Rev Latinoam Microbiol, 1992 Apr-Jun, 34(2), 77 - 81
{Characterization of a substance which protects Klebsiella pneumoniae from the bactericidal effect of normal human serum}; Rodas-Suarez O et al.; Chemical composition of a substance produced by Klebsiella pneumoniae which is accumulated in chemically defined medium was determined . This substance (the protective factor) protects the bacterium from being killed by normal human serum . Protective factor was treated with DNase, lysozyme and an alkaline protease . The two former enzymes did not affect the protective factor . On the other hand, when alkaline protease was used, the protective activity was totally lost . Results showed that the protective factor is a protein.

Infect Immun, 1992 Apr, 60(4), 1577 - 88
Adherence to respiratory epithelia by recombinant Escherichia coli expressing Klebsiella pneumoniae type 3 fimbrial gene products; Hornick DB et al.; We examined the role of Klebsiella fimbrial types 1 and 3 in mediating adherence to human buccal and tracheal cells and to lung tissue sections . We found that clinical isolates of Klebsiella pneumoniae producing type 3 fimbriae and Escherichia coli HB101 containing a recombinant plasmid encoding expression of Klebsiella type 3 fimbriae (pFK10) demonstrated increased adherence to tracheal cells, trypsinized buccal cells, and lung tissue sections, in contrast to nonfimbriate and to type 1 fimbriate bacteria . Adherence by type 3 fimbriate bacteria was inhibited by purified type 3 fimbriae and Fab fragments derived from type 3 fimbrial-specific polyclonal immunoglobulin G . Type 3 fimbriae mediated attachment to the basolateral surface of tracheal cells and to the basal epithelial cells and the basement membrane regions of bronchial epithelia . Using an E . coli transformant (pDC17/pFK52), which expresses nonadherent P fimbrial filaments, along with the type 3 fimbrial adhesin (MrkD), we demonstrated that type 3 fimbrial attachment to respiratory cells was attributable to the MrkD adhesin subunit . Subsequent experiments demonstrated that the epithelial target of the type 3 fimbrial adhesin was most likely a peptide molecule rather than a carbohydrate . The results of this study demonstrate that, in vitro, the Klebsiella type 3 fimbrial adhesin mediates adherence to human respiratory tissue.

FEBS Lett, 1992 Mar 23, 300(1), 67 - 70
The sodium ion pumping oxaloacetate decarboxylase of Klebsiella pneumoniae . Metal ion content, inhibitors and proteolytic degradation studies; Dimroth P et al.; Oxaloacetate decarboxylase of Klebsiella pneumoniae was shown to contain between 0.6 and 1.0 mol zinc per mol enzyme in different preparations . The decarboxylase activity was completely abolished after 15 min incubation with 1 mM Hg(NO3)2 in phosphate buffer, while the activity decreased only 20% if the incubation was performed in MES/Tris buffer . Treatment of the isolated subunits with Hg(NO3)2 indicated that the binding site for Hg2+ ions is on the alpha subunit . Other inhibitors of the decarboxylase are KSCN and diethylstilbestrol . Inactivation of the enzyme with 2% 1-butanol was significantly reduced by 100 mM NaCl . Sodium ions also protected the isolated beta + gamma subunits from a digestion with trypsin.

Mil Med, 1992 Mar, 157(3), 130 - 6
Therapies for radiation injuries: research perspectives; Brook I et al.; Exposure to radiation damages the immune, hematopoietic, and gastrointestinal components of the host defense system . This may lead to serious endogenous or exogenous infections . When radiation injury is combined with other physical trauma, e.g., burn or wound, the resulting damage to these systems is synergistic, and treatment for infection requires multiple approaches . This paper reviews successful single and combined therapeutic modalities for infections in irradiated mice and irradiated mice inflicted with trauma that are currently conducted at the Armed Forces Radiobiology Research Institute . The models of endogenous and exogenous infection and combined injury are described . The management of wounds infected with bacteria, exogenous systemic infection due to gram-negative enteric bacteria, and the chemoprophylaxis of enteric-derived systemic infection with quinolones is described . Infections can be treated successfully with proper antimicrobial therapy . In gamma- and neutron-irradiated mice, the immunomodulator trehalose dimycolate (TDM) was effective in treating endogenous infection . TDM with the antimicrobial ceftriaxone was effective in treating exogenous infection due to Klebsiella pneumoniae . Improvement in managing infection in irradiated and injured hosts will require further research using these diagnostic and therapeutic modalities . Accurate biological dosimetry is critical in determining if victims are at risk of developing infection . We found that radiation induced changes in plasma diamine oxidase activity; monitoring these changes was a useful indicator of the severity of radiation injury.

J Gen Microbiol, 1992 Mar, 138 ( Pt 3), 423 - 8
Anaerobic 2-ketogluconate metabolism of Klebsiella pneumoniae NCTC 418 grown in chemostat culture: involvement of the pentose phosphate pathway; Simons JA et al.; Under anaerobic 2-ketogluconate-limited growth conditions (D = 0.1 h-1), Klebsiella pneumoniae NCTC 418 was found to convert this carbon source to biomass, acetate, formate, CO2, ethanol and succinate . The observed fermentation pattern is in agreement with the simultaneous functioning of the pentose phosphate pathway and the Entner-Doudoroff pathway in 2-ketogluconate catabolism . When cultured at pH 8.0 apparent YATP values were lower than those found at culture pH 6.5 . This difference can be explained by assuming that at high culture pH values approximately 0.5 mol ATP was invested in the uptake of 1 mol 2-ketogluconate . Sudden relief of 2-ketogluconate-limited conditions led to lowering of the intracellular NADPH/NADP ratio and (possibly as a result of this) to inhibition of biosynthesis . Whereas production of ethanol stopped, lactate was produced at high rate . This product was formed, at least partly, via the methylglyoxal bypass.

Mikrobiol Zh, 1992 Mar-Apr, 54(2), 75 - 80
{The microbiological and pathomorphological characteristics of Klebsiella infection}; Sel'nikova OP et al.; Biological properties of 350 strains of Klebsiella pneumoniae isolated in newborns during the outbreak of intrahospital infection have been studied . Experimental infection with isolated strains of a similar plasmid profile is simulated, interrelation between the presence of plasmids and ability of the strain to induce generalized Klebsiella infection is shown . Pathological processes in the newborns with generalized infection and in animals with the reproduced experimental Klebsiella infection are shown to be similar in principle.

Infection, 1992 Mar-Apr, 20(2), 66 - 72
Salicylate or bismuth salts enhance opsonophagocytosis of Klebsiella pneumoniae; Domenico P et al.; After treatment of encapsulated Klebsiella pneumoniae with salicylate or bismuth compounds, phagocytic uptake by human peripheral white blood cells or rat alveolar macrophages was assessed . Without salicylate pretreatment of bacteria, a 30-60% net increase in viable bacteria resulted in phagocytic assays after a 1 hour incubation . With salicylate pretreatment, dose-related decreases in bacterial counts were seen, achieving a maximal reduction of 60% with 240 microM salicylate pretreatment . Bacterial variants producing less capsule were more serum sensitive and more readily phagocytosed . Micrographs of Giemsa-stained cells revealed phagocytic uptake of multiple bacteria after salicylate pretreatment, but virtually no uptake of untreated bacteria . Opsonization with polyclonal antiserum decreased bacterial cell counts by 20% without and by 90% with salicylate pretreatment of bacteria . Pretreatment of bacteria with bismuth salts also enhanced opsonophagocytosis of encapsulated bacteria . Thus, agents known to reduce capsule expression in K . pneumoniae also enhance phagocytic uptake of bacteria.

Res Microbiol, 1992 Mar-Apr, 143(3), 347 - 51
Putative nickel-binding sites of microbial proteins; Wu LF; Nickel is biologically important because of its catalytic role in the mechanisms of action of metalloenzymes, and also because of its toxic cellular effects . There exist at least 3 groups of nickel-binding proteins in microorganisms: nickel-specific transporters, accessory proteins involved in nickel incorporation and nickel-containing enzymes . The differences in their physiological functions determine the nature of the ligands and the structures of the nickel-binding sites . The homology among the accessory proteins HypB, ORF4 and UreG suggests that the mechanism of nickel incorporation into hydrogenases in Escherichia coli is the same as or similar to that into hydrogenases of Rhodobacter capsulatus and into urease of Klebsiella aerogenes.

Zh Mikrobiol Epidemiol Immunobiol, 1992 Mar, (3), 30 - 3
{Immunobiological properties and therapeutic effectiveness of preparations from Klebsiella bacteriophages}; Bogovazova GG et al.; The purified preparations of Klebsiella bacteriophages, viz . the monovalent preparation of K . pneumoniae bacteriophage and the polyvalent bacteriophage preparation for the treatment of infections caused by K . ozaenae, K . rhinoscleromatis scleromatis and K . pneumoniae sensu lato, have been obtained . The bacteriophage preparations have proved to be nontoxic and safe for laboratory animals after the intraperitoneal injection of these preparations followed by the pathomorphological study of the internal organs of the animals . The clinical study of the newly developed bacteriophage preparations in the course of the treatment of purulent inflammatory diseases in 109 patients has revealed that the preparations are not reactogenic and exhibit sufficient effectiveness in the therapy of ozena, rhinoscleroma and Klebsiella infections with different localization of the infectious process.

Sarcoidosis, 1992 Mar, 9(1), 49 - 53
B-lymphocyte response in peripheral blood of patients with pulmonary sarcoidosis; Barth J et al.; Sarcoidosis is a granulomatous disorder which can be characterized by various immunologic abnormalities including lymphocyte dysfunctions . The purpose of this study was to investigate the B-lymphocyte reactivity in the peripheral blood of 29 various patients with pulmonary sarcoidosis . A significant decrease in the production of IgG, IgM and IgA in supernatants of cultivated sarcoidosis B-cells both after stimulation with a T-cell dependent polyclonal B-cell activator (pokeweed mitogen) and with a T-cell independent polyclonal B-lymphocyte activator (Klebsiella pneumoniae) was seen, which suggested a disturbance in the B-cell differentiation of sarcoidosis patients . Those patients which were known to have a clinically active disease or a high intensity alveolitis in the bronchoalveolar lavage fluid showed a greater reduction in the B-cell differentiation response . Further evaluation of Ig-G subclasses displayed a significant decrease in the secretion of IgG1, IgG3 and IgG4 . In contrast to the differentiation, the proliferation response of sarcoidotic peripheral blood lymphocytes to different mitogens did not differ from healthy controls.

J Biol Chem, 1992 Feb 25, 267(6), 3667 - 73
Ionic interactions in the nitrogenase complex . Properties of Fe-protein containing substitutions for Arg-100; Wolle D et al.; A series of Azotobacter vinelandii strains have been constructed in which the nitrogenase Fe-protein (Av2) was altered by substitutions for Arg-100 . This invariant residue is a likely partner in a salt bridge with the MoFe-protein and, in some species, is the site of reversible regulation by ADP-ribosylation (Pope, M . R., Murrell, S . A., and Ludden, P . W . (1985) Proc . Natl . Acad . Sci . U . S . A . 82, 3173-3177) . Although we find that arginine is the optimum amino acid, other residues in this position could support diazotrophic growth . These results were surprising because Klebsiella pneumoniae Fe-protein substituted by His-100 had been reported to be inactive (Lowery, R . G., Chang, C . L., Davis, L . C., McKenna, M.-C., Stevens, P . J., and Ludden, P . W . (1989) Biochemistry 28, 1206-1212) . Two altered Fe-proteins (Av2-R100Y, the tyrosyl form, and Av2-R100H, the histidyl form) were isolated and, in contrast to this earlier report, we found that both had some activity in acetylene reduction . However, both altered proteins exhibited a decreased maximum velocity (35 and 3% of wild type, respectively) and were strongly inhibited by excess MoFe-protein . These adverse activity parameters were also manifest in the increased sensitivity of the altered proteins to inhibition by salts . Indeed, the salt sensitivity of Av2-R100H is so significant that its activity is masked in the normal assay and is easily missed . In addition, for Av2-R100H, substrate reduction is substantially uncoupled from MgATP hydrolysis . These results suggest that substitutions for Arg-100 may decrease the affinity of the Fe-protein for the MoFe-protein prior to electron transfer but increase affinity after electron transfer . Hence, the role of Arg-100 may be to provide the optimum balance in stabilities of these two complexes for maximum efficiency in substrate reduction.

Presse Med, 1992 Feb 8, 21(5), 203 - 6
{Bacteriological safety in enteral nutrition in intensive care units . Comparison of 4 types of pumps}; Lecso-Bornet M et al.; Enteral nutrition may appear as a risk factor for infection in intensive care patients if bacterial growth is not prevented . The authors have tested in the microbiology laboratory, 4 types of pumps (3 cooling machines: SEC Nutridose/Climadose, Peters NPR 3000, Vial "Alaska" and 1 flow regulator Sherwood Kangaroo 330) using food contaminated with a low inoculum of Klebsiella pneumoniae . Bacterial growth was investigated periodically in the food container and in the feeding tube during the testing of the pumps . Results showed that external refrigeration is the best cooling method . But after 8 hours of working, the best results were obtained with the flow regulator associated with a container cooled by an ice pocket . These in vitro results can be extended in vivo by taking care of changing the sterile tubings for each new dose of food.

Biochemistry, 1992 Feb 4, 31(4), 1216 - 24
Posttranslational modification of Klebsiella pneumoniae flavodoxin by covalent attachment of coenzyme A, shown by 31P NMR and electrospray mass spectrometry, prevents electron transfer from the nifJ protein to nitrogenase . A possible new regulatory mechanism for biological nitrogen fixation; Thorneley RN et al.; A strain of Escherichia coli (71-18) that produces ca . 15% of its soluble cytoplasmic protein as a flavodoxin, the Klebsiella pneumoniae nifF gene product, has been constructed . The flavodoxin was purified using FPLC and resolved into two forms, designated KpFldI and KpFldII, which were shown to have identical N-terminal amino acid sequences (30 residues) in agreement with that predicted by the K . pneumoniae nifF DNA sequence . 31P NMR, electrospray mass spectrometry, UV-visible spectra, and thiol group estimations showed that the single cysteine residue (position 68) of KpFldI is posttranslationally modified in KpFldII by the covalent, mixed disulfide, attachment of coenzyme A . KpFldII was inactive as an electron carrier between the K . pneumoniae nifJ product (a pyruvate-flavodoxin oxidoreductase) and K . pneumoniae nifH product (the Fe-protein of nitrogenase) . This novel posttranslational modification of a flavodoxin is discussed in terms of the regulation of nitrogenase activity in vivo in response to the level of dissolved O2 and the carbon status of diazotrophic cultures.

J Bacteriol, 1992 Feb, 174(3), 1063 - 7
Involvement of rcsB in Klebsiella K2 capsule synthesis in Escherichia coli K-12; Wacharotayankun R et al.; Escherichia coli K-12 harboring a part of the structural genes for the Klebsiella K2 capsular polysaccharide (cpsK*) expresses a large amount of K2 capsular polysaccharide as a thick capsule in the presence of plasmids carrying rmpA and rcsB . We have previously shown that expression of the Klebsiella K2 capsule in E . coli HB101 harboring cpsK* depends on the presence of rmpA, a regulatory gene from a large plasmid of Klebsiella pneumoniae Chedid (O1:K2) . E . coli K-12 JM109, however, produces only a small amount of K2 capsular polysaccharide, even in the presence of plasmids carrying rmpA as well as the cpsK* structural genes . Introduction of the rcsB gene, a positive regulator of colanic acid capsule synthesis in E . coli K-12 which was cloned from HB101 on a plasmid, into JM109 cells carrying cpsK* and rmpA, results in the expression of a thick K2 capsule . By Northern (RNA) hybridization analysis, rcsB has been found to enhance transcription of a long strand of mRNA (longer than 14 kb) from cpsK* . These E . coli transformants which produce a thick K2 capsule also express colanic acid production at high levels . Therefore, rcsB can act as a positive regulator of Klebsiella K2 capsule production and two capsular polysaccharides can be expressed in E . coli simultaneously . With a somewhat different strain background, we have found that both of the colanic acid regulators, rcsA and rcsB, contribute to the basal level of Klebsiella K2 capsule expression but that the presence of multicopy rcsB in either an rcsB or an rcsA mutant of E . coli is sufficient to increase the expression of K2 capsular polysaccharide . These results suggest further parallels between the regulation of colanic acid synthesis in E . coli and the regulation of Klebsiella K2 capsule synthesis.

Arch Biochem Biophys, 1992 Feb 1, 292(2), 583 - 8
Transport of 4-hydroxyphenylacetic acid in Klebsiella pneumoniae; Allende JL et al.; Klebsiella pneumoniae M5a1 has been shown to possess an inducible transport system for 4-hydroxyphenylacetate (4-HPA) . This transport system has a Kt of 16.3 microM and a maximal velocity of 31.2 nmol/min (milligrams dry weight) . The transport system has been inhibited by inhibitors of energy metabolism with a concomitant decrease in cellular ATP concentrations, and the 4-HPA binding activity has been detected in the crude shock extracts . All these observations indicate that 4-HPA uptake is an active transport which involves a periplasmic binding protein and it seems to be energized by phosphate bond energy.

Immun Infekt, 1992 Feb, 20(1), 3 - 6
{Outbreak of a nosocomial infection of SHV2-beta-lactamase-containing Klebsiella pneumonia strains in an operative intensive care unit}; Ritter E et al.; Resistant strains of Klebsiella pneumoniae were found in increasing frequency as a cause of nosocomial infection in an intensive care unit between July and October 1990 . The isolated strains had an almost identical biochemical profile, showed a similar pattern of antibiotic resistance, and produced type SHV2-broad-spectrum betalactamase . Thus, it was assumed that the isolates were copies of identical strains, causing an outbreak of nosocomial infections . The bacteria were resistant to third-generation cephalosporins, such as cefotiam, cefotaxime and ceftriaxone, and also to aminoglycosides and acylaminopenicillins . Approximately half of the strains were resistant to ceftazidim and aztreonam . The bacteria were sensitive in vitro to ciprofloxacin, imipenem, latamoxef and cefotetan . During three months, 10% (11) of all patients became infected; four of these patients (36%) died from septicemia . After conventional hygiene programs had failed to stop the outbreak, the intensive care unit was closed and disinfected, a measure, which effectively interrupted the infection.

Mol Microbiol, 1992 Feb, 6(3), 389 - 94
Role of lipid modification on a starch-debranching enzyme, Klebsiella pullulanase: comparison of properties of lipid-modified and unmodified pullulanases; Yamashita M et al.; Klebsiella pullulanase is a lipoprotein synthesized as a precursor with a signal peptide, which is processed by lipoprotein signal peptidase . To clarify the role of lipid modification of pullulanase, we purified lipid-modified wild-type and the unmodified (mutant) pullulanases and compared their properties . The Km and Vmax values of both pullulanases for pullulan were the same . The optimal pH and temperature, the stabilities over pH and temperature ranges, the specificity of substrates, and the patterns of inhibition of the lipid-modified and unmodified pullulanases were also the same . However, we found that the wild-type pullulanase formed trimers whereas the unmodified enzyme did not, and that the migrations of the two enzymes on sodium dodecyl sulphate/electrophoresis were different when the samples were applied on the gel without heating . The results presented in this paper and in previous work show that the correct processing and translocation of pullulanase in K . aerogenes require modification of lipid . However, the enzymatic properties and physical stabilities of pullulanase were not affected by the lipid modification.

Zh Mikrobiol Epidemiol Immunobiol, 1992 Feb, (2), 62 - 4
{Interferon type I in protective body reactions in an experimental Klebsiella infection}; Fil'chakov IV et al.; The study on mice with experimental generalized Klebsiella infection, carried out with the use of microbiologic, immunologic and pathomorphologic methods, revealed that the intraperitoneal injection of type I interferon into the animals prevented their death and led to the rapid elimination of the infective agent from their body, enhanced the phagocytic and metabolic activity of polymorphonuclear lymphocytes of their peritoneal exudate, decreased the manifestation of microcirculatory and dystrophic changes in the parenchyma of their internal organs.

Zh Mikrobiol Epidemiol Immunobiol, 1992 Feb, (2), 12 - 4
{The resistance plasmid of Klebsiella pneumoniae that controls its cytotoxic activity}; Kalnin KV et al.; R-plasmid (40 MD) isolated from K . pneumoniae hospital strain makes Escherichia coli strain J62 capable of inducing a cytotoxic effect which can be detected in Hep-2 cell culture . In contrast to the initial E . coli strain J62 producing no changes in the monolayer, E . coli J62 cells containing P-plasmid induced pronounced cytotoxic changes and a sharp increase in the number of nonviable Hep-2 cells by hour 24 of interaction.

Mol Microbiol, 1992 Jan, 6(1), 95 - 105
Pullulanase secretion in Escherichia coli K-12 requires a cytoplasmic protein and a putative polytopic cytoplasmic membrane protein; Possot O et al.; The previously uncharacterized third and fourth genes (pulE and pulF) of the pullulanase secretion gene operon of Klebsiella oxytoca strain UNF5023 are, respectively, predicted to encode a 55 kDa polypeptide with a putative nucleotide-binding site, and a highly hydrophobic 44 kDa polypeptide that probably spans the cytoplasmic membrane several times . Expression of pulE in minicells or under the control of a strong bacteriophage T7 promoter resulted in the production of a c . 58 kDa cytoplasmic protein . A representative PulE-beta-galactosidase hybrid protein created by Tnlac mutagenesis was also found mainly in the cytoplasm . These results are in line with the predicted absence from PulE of a region of sufficient hydrophobicity to function as a signal sequence . The PulF polypeptide could not be detected either in minicells or when the gene was transcribed from the T7 promoter, but the acquirement of three pulF-lacZ gene fusions that encoded hybrid proteins with relatively high levels of beta-galactosidase activity indicates that this gene can be transcribed and translated . Gene disruption experiments indicated that both pulE and pulF are required for pullulanase secretion in Escherichia coli K-12 . Both proteins exhibit considerable homology throughout their entire lengths with other proteins involved in protein secretion, pilin assembly, conjugation and transformation competence in a variety of bacteria . In addition, PulE protein has consensus sequences found in a wide variety of nucleotide-binding proteins . This study completes the initial characterization of the pullulanase secretion gene operon, which comprises 13 genes that are all essential for the transport of pullulanase across the outer membrane.

J Cataract Refract Surg, 1992 Jan, 18(1), 106 - 10
Postoperative inflammation following cataract extraction caused by bacterial contamination of the cleaning bath detergent; Kreisler KR et al.; Five of 16 patients who had uncomplicated cataract extraction with intraocular lens (IOL) implantation within a 2.5 day period experienced increased anterior segment inflammation on their first postoperative day . Four of these five patients had phacoemulsification and one a planned extracapsular cataract extraction . All had posterior chamber IOL implantation in the capsular bag . In all patients, this anterior segment inflammation cleared with topical steroids over two to three weeks with no evident residual ocular effects . A careful search for the possible cause of the inflammation showed that the ultrasonic cleaning bath and Weck liquid detergent used to clean the instruments contained Klebsiella pneumoniae bacteria . Further investigation demonstrated the presence of a heat-stable endotoxin produced by the bacteria . We postulate that endotoxin remaining on the instruments after cleaning and sterilization caused this postoperative anterior segment inflammation . To the best of our knowledge, these are the first reported cases caused by contaminated liquid detergent.

Am Rev Respir Dis, 1992 Jan, 145(1), 31 - 5
Parenteral followed by oral ofloxacin for nosocomial pneumonia and community-acquired pneumonia requiring hospitalization; Gentry LO et al.; We report a multicentric, open trial of intravenous followed by oral ofloxacin, 400 mg every 12 h, as therapy for 100 cases of nosocomial pneumonia and community-acquired pneumonia requiring hospitalization . The typical subject was 57 yr old, and underlying diseases, such as chronic obstructive pulmonary diseases (COPD), diabetes mellitus, and congestive heart failure, were common . For 10 subjects previous therapy had failed . There were 118 pathogens isolated in blood or sputum; S . pneumoniae was the most common (42), followed by H . influenzae (13), Klebsiella spp . (11), and S . aureus (10) . Ofloxacin was administered for an average of 5.7 days intravenously followed by 6.9 days orally . Response to therapy was judged to be cure in 71 subjects, improvement in 24, and failure in 5 . Among the more seriously ill subjects, ofloxacin therapy was successful for four of five immunocompromised subjects, for 12 of 12 subjects with nosocomial pneumonia, three of whom were on the ventilator, and for nine of 10 subjects with community-acquired pneumonia and bacteremia, including seven of eight cases due to S . pneumoniae . Univariate risk factor analysis revealed underlying COPD and/or tachypnea upon admission to be associated with failure of ofloxacin therapy, with bacteremia suggestive of failure . Conversely, ofloxacin was equally effective in cases in whom previous therapy failed and in cases of nosocomial pneumonia, multilobar pneumonia, and/or pneumonia due to S . pneumoniae . Results for P . aeruginosa were inconclusive . Intravenous followed by oral ofloxacin was highly effective in many difficult cases of pneumonia.

Infect Immun, 1992 Jan, 60(1), 95 - 101
Free versus liposome-encapsulated muramyl tripeptide phosphatidylethanolamide in treatment of experimental Klebsiella pneumoniae infection; Melissen PM et al.; The effect of free and liposome-encapsulated muramyl tripeptide phosphatidylethanolamide (MTPPE) on resistance to Klebsiella pneumoniae infection in mice was investigated . It was shown that administration of MTPPE, at 24 h before bacterial inoculation, led to a dose-dependent antibacterial resistance in terms of increased clearance of bacteria from the blood and bacterial killing in various organs . The lowest effective dose of MTPPE was 50 micrograms per mouse . Administration of liposome-encapsulated MTPPE was also effective at a dose of 25 micrograms per mouse . The time of administration of both free and liposome-encapsulated MTPPE, with respect to the appearance of bacteria in the blood, was very important and indicated that repeated administration is necessary to obtain protection for a prolonged period . In view of the toxicity of MTPPE, it was an important observation that repeated administration of MTPPE in the liposome-encapsulated form also produced antibacterial resistance . Administration of free and liposome-encapsulated MTPPE resulted in increased numbers of granulocytes, monocytes, and lymphocytes in the blood of uninfected mice and prevented leukopenia in infected mice.

Nihon Kyobu Shikkan Gakkai Zasshi, 1992 Jan, 30(1), 35 - 44
{Experimental study on development and lung injury in pneumonia under neutropenic condition}; Takeda H; This study was performed to demonstrate the characteristic findings in pneumonia under neutropenic condition . The results were as follows . 1) After nebulizing Klebsiella pneumonia DT-S strains, the survival rate in the neutropenic mouse group rapidly decreased compared with that of the control group . 2) In the neutropenic state, a rapid decline of survival rate was demonstrated during the initial stage of infection . Bacteremia and endotoxemia developed earlier than in the healthy control group . 3) In neutropenic pneumonia, the neutrophil cell count in BALF was significantly lower than that in usual pneumonia . Consequently, a large number of bacteria grew in the alveolar spaces during the early period after inhalation . Neither inflammatory cell infiltration nor thickening of the alveolar wall was present; however, structural destruction of alveolar and vascular walls was found . 4) These pathological changes were also seen following intratracheal inoculation of some extracellular enzymes of Klebsiella pneumoniae DT-S into rabbit lung . However, no destruction of alveolar walls was observed after nebulizing the killed Klebsiella pneumoniae DT-S . It was suggested that the significant amount of enzymes produced by the bacteria growing in the alveoli caused tissue damage, especially in the neutropenic state . 5) Thus the typical inflammatory changes that occurred in the usual state, were not seen in neutropenic pneumonia, and a large number of bacteria grew in the alveoli . Marked tissue damage was caused by extracellular enzymes produced by bacteria . Consequently, bacteria entered the blood stream and bacteremia and endotoxemia developed . These processes observed in neutropenic pneumonia readily progressed to bacteremia of endotoxemia, which are more severe in this state.

Appl Biochem Biotechnol, 1992 Spring, 34-35, 149 - 59
Enhancement of 1,3-propanediol production by cofermentation in Escherichia coli expressing Klebsiella pneumoniae dha regulon genes; Tong IT et al.; 1,3-Propanediol (1,3-PD) is an intermediate in chemical and polymer synthesis . We have previously expressed the genes of a biochemical pathway responsible for 1,3-PD production, the dha regulon of Klebsiella pneumoniae, in Escherichia coli . An analysis of the maximum theoretical yield of 1,3-PD from glycerol indicates that the yield can be improved by the cofermentation of sugars, provided that kinetic constraints are overcome . The yield of 1,3-PD from glycerol was improved from 0.46 mol/mol with glycerol alone to 0.63 mol/mol with glucose cofermentation and 0.55 mol/mol with xylose cofermentation . The engineered E . coli also provides a model system for the study of metabolic pathway engineering.

Autoimmunity, 1992, 12(1), 47 - 52
Experimental autoimmune adrenalitis: a murine model for Addison's disease; Fujii Y et al.; Experimental autoimmune adrenalitis was produced in mice by immunizing 8 times or more at intervals of 30 days with syngeneic adrenal extract mixed with Klebsiella O3 lipopolysaccharide (KO3 LPS) as a potent adjuvant . The cortex regions of the adrenal glands after the 8th injection were definitely infiltrated with polymorphonuclear leukocytes (PMN) . The main infiltrates in the lesions after the 9th injection were replaced by mononuclear cells, such as small lymphocytes and macrophages, and further by fibrous connective tissues . There were no histological changes in the medullary regions . The repeated immunization developed the delayed type hypersensitivity to adrenal extract and production of anti-adrenocortical autoantibody in those immunized mice . Moreover, the adrenalitis could be produced in normal mice by transfer of spleen cells from hyperimmunized mice, suggesting the critical role of the cell-mediated immunity . This experimental model might be useful to study immunological phenomena in the pathogenesis of Addison's disease.

Antimicrob Agents Chemother, 1992 Jan, 36(1), 167 - 71
Synergistic effects of romurtide and cefmenoxime against experimental Klebsiella pneumonia in mice; Tatara O et al.; We investigated the synergistic effects of romurtide (MDP-Lys {L18}) and cefmenoxime (CMX) in the treatment of experimental Klebsiella pneumonia in mice . Mice were infected with 1 x 10(4) CFU of Klebsiella pneumoniae by inhalation of aerosol bacterial suspension . About 90% of untreated animals died within a week; however, the mortality rate of animals treated with CMX alone at a dose of 40 mg/kg/day was 60% at 7 days after the infection . When one or two doses of L18 were administered before or after the infection concomitantly with CMX, a remarkable improvement in the survival rate was observed . There was no significant improvement in the survival rate of animals treated with L18 alone before or after infection . Histopathological sections of the lungs of mice treated with CMX and L18 showed slower progression of infection than those of mice treated with CMX alone . Significant differences were also found in quantitative cultures of viable bacteria in the lungs 1 to 4 days after the infection . Although viable bacterial counts in the lungs of the control and CMX-treated groups showed a rapid increase 24 to 48 h after the infection, they remained lower than the initial counts (x 10(4)) in the lungs of mice treated with combination regimens . From these results, it can be concluded that L18 is a useful biological response modifier in the treatment of acute pulmonary bacterial infections.

Mikrobiol Zh, 1992 Jan-Feb, 54(1), 3 - 9
{An evaluation of the biochemical typing of Klebsiella cultures}; Pokhil SI et al.; The paper deals with the results of analysis of 219 strains of Klebsiella using 12 tests--fermentation of adonite, asparaginic acid, sodium citrate, dulcite, d-tartrate, glutamic acid, inosite, L-proline, sodium malonate; reactions with methyl red, Foges-Proscauer, with 5-ASA . A scheme for determining fermentovars of Klebsiella is suggested which includes the tests--fermentation of adonite, dulcite, d-tartrate, glutamic acid; color reaction with 5-ASA . The groups of Klebsiella different in origin are characterized by nonhomogeneous distribution of different fermentovars . The suggested method of biochemical labelling may be one of the basic ones in complex typing of Klebsiella.

Can J Microbiol, 1992 Jan, 38(1), 28 - 33
Effect of the method of preparing monochloramine upon inactivation of MS2 coliphage, Escherichia coli, and Klebsiella pneumoniae; Berman D et al.; Monochloramine prepared in situ by first adding chlorine to a suspension of microorganisms, followed by subsequent addition of ammonia, inactivated the MS2 coliphage more rapidly than did exposure of phage to monochloramine prepared either by adding chlorine to ammonia or by adding chlorine and ammonia simultaneously . The rapid viral inactivation was apparently due to the exposure of MS2 to free chlorine before the addition of ammonia . The average 99% CT value of MS2 when exposed to free chlorine was 1.3 and 1.1 at 5 and 15 degrees C, respectively . The average 99% CT values of MS2 briefly exposed to the combined action of free chlorine followed by the addition of ammonia to form monochloramine in situ were 19.3 and 1.5 at 5 and 15 degrees C, respectively . No 99% CT values were calculated for the inactivation of MS2 with preformed monochloramine because less than 1 log (90%) of inactivation occurred during a 4-h contact time . Inactivation of MS2 by monochloramine was more rapid at 15 than at 5 degrees C and when the chlorine to nitrogen weight ratio was 5:1 compared with 3:1 . Monochloramine was a more efficient inactivating agent for the coliforms Escherichia coli and Klebsiella pneumoniae than it was for the MS2 coliphage.

Pathol Biol (Paris), 1992 Jan, 40(1), 31 - 5
{Identification of a new penicillinase in a Klebsiella pneumoniae strain of which a mutant also produces an esterase hydrolyzing cephalothin and cefotaxime}; Ben Yaghlane-Bouslama H et al.; Klebsiella pneumoniae strain L 164 produces a penicillinase whose isoelectric point is 8.1, an unusual figure for this bacterial species . This strain exhibits resistance to conventional penicillins and a synergistic effect is seen with clavulanic acid . In contrast, susceptibility to cephalosporins is marked, as shown by the low minimum inhibitory concentrations (MICs) . This phenotype is characteristic of strains with no acquired resistance . A first mutant with MICs for cephalothin and cefotaxime 8-fold to 16-fold those of the initial strain was obtained spontaneously . This mutant's MICs for the other beta-lactams were not substantially changed . In addition to the same penicillinase as the one produced by the parent strain, this mutant produced an acetyl-esterase capable of hydrolyzing the cephalosporins with an acetoxyl side-chain, i.e., cefalothin and cefotaxime, to deacetylated derivates which retain substantial antibacterial activity . Another mutant selected on an amoxicillin gradient produced ten times more penicillinase than the parent strain but no esterase . This second mutant exhibited very high MICs for penicillins and first and second generation cephalosporins . The MIC for cefotaxime was comparable to that seen with the esterase-producing mutant . Among the antimicrobials tested, only third generation cephalosporins and cefoxitin showed adequate activity.

Ann Rheum Dis, 1992 Jan, 51(1), 83 - 6
Immune complexes in ankylosing spondylitis; MacLean IL et al.; Immune complexes have been reported in ankylosing spondylitis (AS) and may implicate infectious agents . Serum samples from 49 patients with AS were assayed for immune complexes by polyethylene glycol precipitation, followed by radial immunodiffusion and pepsinogen binding immunoassay . Both methods showed increases in IgA containing immune complexes, which correlated with serum IgA and with IgA rheumatoid factor concentrations, but did not show increases in other immune complex components . Increased immune complexes were associated with peripheral joint synovitis, but showed no correlation with other clinical or laboratory indices of disease activity . Immune complexes from nine AS serum samples and one AS synovial fluid were electrophoretically separated then probed with anti-Klebsiella pneumoniae, but AS specific antigens were not identified . This study did not suggest a major role for immune complexes in AS without peripheral disease, nor provide serological evidence for the involvement of klebsiella antigens.

Appl Environ Microbiol, 1992 Jan, 58(1), 27 - 31
Metabolism of benzonitrile and butyronitrile by Klebsiella pneumoniae; Nawaz MS et al.; A strain of Klebsiella pneumoniae that used aliphatic nitriles as the sole source of nitrogen was adapted to benzonitrile as the sole source of carbon and nitrogen . Gas chromatographic and mass spectral analyses of culture filtrates indicated that K . pneumoniae metabolized 8.4 mM benzonitrile to 4.0 mM benzoic acid and 2.7 mM ammonia . In addition, butyronitrile was metabolized to butyramide and ammonia . The isolate also degraded mixtures of benzonitrile and aliphatic nitriles . Cell extracts contained nitrile hydratase and amidase activities . The enzyme activities were higher with butyronitrile and butyramide than with benzonitrile and benzamide, and amidase activities were twofold higher than nitrile hydratase activities . K . pneumoniae appears promising for the bioremediation of sites contaminated with aliphatic and aromatic nitriles.

Ukr Biokhim Zh, 1992 Jan-Feb, 64(1), 110 - 3
{Isolation and various properties of alpha-aminocaprolactam hydrolase from Klebsiella aerogenes}; Kliment'eva TA et al.; L-alpha-aminocaprolactam hydrolase possessing the L lysine amidase activity was isolated from Klebsiella aerogenes and purified . The procedure of enzymes purification included cell destruction on USDN-I, fractionation by ammonium sulfate, gel chromatography on G-200 . The preparation of the purified enzyme possessed specific activity of 50 mumol of lysin per 1 mg of protein per hour . Km was 2.6 mM in case of phosphate buffer (ph 7.2) for I-alpha-aminocaprolactam . Besides L-alpha-aminocaprolactam the enzyme hydrolyses lysine amide, leucine amide tryptophanamide . Magnesium ions are necessary for manifestation of catalytic activity of the enzyme.

J Hyg Epidemiol Microbiol Immunol, 1992, 36(2), 129 - 39
Epidemiological analysis of Klebsiella infections at a neonate department; Kolarova M et al.; At the Department of Obstetrics and Gynaecology of a Brno hospital, 133 Klebsiella strains were isolated from 23 mothers and 58 neonates during a period of increased incidence of Klebsiella and 73 strains were examined in the laboratory . The control group was comprised by 78 Klebsiella strains isolated at three other Brno hospitals . Four prevailing Klebsiella types were identified using bacteriocinotyping and phagotyping . Different bacteriocinotypes and phagotypes were found to prevail at certain intervals both at the examined clinic and in control patients from Brno hospitals . The findings suggest circulation of certain Klebsiella types and their introduction to hospitals . Most infants (37 out of 45, i.e . 82%) were colonized by Klebsiella, their mothers remaining without colonization . The factors influencing the onset and transmission of nosocomial infections in infants are discussed.

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