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