Science, 1989 Sep 22, 245(4924), 1374 - 7 Bacterial blight of soybean: regulation of a pathogen gene determining host cultivar specificity; Huynh TV et al.; Soybean cultivars resistant to Pseudomonas syringae pathovar glycinea (Psg), the causal agent of bacterial blight, exhibit a hypersensitive (necrosis) reaction (HR) to infection . Psg strains carrying the avrB gene elicit the HR in soybean cultivars carrying the resistance gene Rpg1 . Psg expressing avrB at a high level and capable of eliciting the HR in the absence of de novo bacterial RNA synthesis have been obtained in in vitro culture . Nutritional signals and regions within the Psg hrp gene cluster, an approximately 20-kilobase genomic region also necessary for pathogenicity, control avrB transcription. Presse Med, 1989 Sep 16, 18(28), 1383 - 6 {Chronic bone infections after surgery . Treatment with the new quinolones}; Bricaire F et al.; A prospective open study carried out over 5 years and including 20 patients suffering from chronic bone suppuration following orthopaedic surgery has confirmed the value of the new quinolones (NQ) in these indications . The patients received pefloxacin or ciprofloxacin most often combined with rifampicin or fusidic acid for a mean period of 7 months . Single or multiple organism infections were documented in 14 patients, the majority being Staph . aureus (n = 13) and Pseudomonas (n = 14) . Samples were sterile in 6 cases . Fourteen therapeutic successes and 5 failures were observed . In one patient, improvement was noted but the post-treatment follow-up insufficient to pronounce a cure . Success was obtained in 14 out of 16 patients who had sensitive organisms or sterile samples . The mean post-treatment follow-up (16 months) was satisfactory but insufficient to speak of cure . However, in these patients for whom further surgery, however desirable, is often refused, NQ constitute an improvement which raises hopes of cure or allows further surgery. J Biol Chem, 1989 Sep 15, 264(26), 15157 - 60 Pseudomonas exotoxin: chimeric toxins; Pastan I et al.; Pseudomonas exotoxin binds to and enters cells by receptor-mediated endocytosis . Within the cell it requires exposure to low pH to enable it to translocate to the cell cytoplasm where it inhibits protein synthesis by ADP-ribosylating elongation factor 2 . The toxin has three main structural domains whose functions are: Ia, cell binding; II, translocation; and III, ADP-ribosylation . Key amino acids have been identified within each domain that are required for the function of the toxin . Chimeric toxins were made originally by using chemical cross-linking reagents to couple Pseudomonas exotoxin (or other toxins) to cell-binding proteins . More recently, a variety of Pseudomonas exotoxin-related chimeric toxins have been made by gene fusion technology . These chimeric toxins may be useful clinically for treating various diseases and experimentally for understanding receptor function. Cancer Res, 1989 Sep 15, 49(18), 4990 - 5 Enhanced therapeutic efficacy of an immunotoxin in combination with chemotherapy against an intraperitoneal human tumor xenograft in athymic mice; Pearson JW et al.; A mouse IgG2b anti-pan carcinoma monoclonal antibody, NR-LU-10, was shown to bind homogeneously to ascites xenografts of both ovarian and colon carcinoma . Following linkage to a highly potent holotoxin, Pseudomonas exotoxin A (PE), NR-LU-10 demonstrated high potency and selectivity in vitro (ID50 = 100 pg/ml; elimination of greater than or equal to 4.5 logs of cells) . The conjugate was evaluated for therapeutic efficacy against a human colon tumor (HT-29) transplantable in the peritoneal cavity of nude mice . Beginning 3 days after HT-29 injection, mice received either three or six i.p . injections of 0.5 micrograms of unconjugated NR-LU-10 or immunotoxin conjugate (NR-LU-10/PE) every other day . Mice that received three or six treatments of NR-LU-10 alone had median survival times (MSTs) of 39 and 40 days, respectively, which did not differ significantly from the MST observed for the untreated control groups (MST = 35 days) . In contrast, treatment with three or six injections of 0.5 micrograms NR-LU-10/PE exhibited significantly increased MSTs (P = 0.002) of 50 and 60 days, respectively . Coinjection of unconjugated NR-LU-10 (20 micrograms) and 0.5 micrograms of NR-LU-10/PE blocked the therapeutic effect of the immunotoxin (MST = 33 days) . The therapeutic efficacy of NR-LU-10/PE was further enhanced against HT-29 when administered i.p . during and after cytoreductive chemotherapy . The i.p . administration of 300 mg/lg of cyclophosphamide plus 100 mg/kg of the chemoprotective drug, WR-2721, 10 and 17 days posttumor cell inoculation induced a significant increase in MST from 36 days to 59 days (P = 0.002) . Interestingly, groups of mice that received either two, four, or seven treatments of NR-LU-10/PE following cytoreductive therapy exhibited a further significant increase (P = 0.001) in MSTs of 89, 97, and 105 days, respectively . Therefore, the use of immunotoxin therapy following cytoreductive chemotherapy significantly prolonged survival time of mice bearing the HT-29 colon tumor over that observed with chemotherapy or NR-LU-10/PE alone. FEBS Lett, 1989 Sep 11, 255(1), 27 - 31 The structure of syringomycins A1, E and G; Segre A et al.; By a combination of 1D and 2D 1H- and 13C-NMR, FAB-MS, and chemical and enzymatic reactions carried out at the milligram level, it has been demonstrated that syringomycin E, the major phytotoxic antibiotic produced by Pseudomonas syringae pv . syringae, is a new lipodepsipeptide . Its amino acid sequence is Ser-Ser-Dab-Dab-Arg-Phe-Dhb-4(Cl)Thr-3(OH)Asp with the beta-carboxy group of the C-terminal residue closing a macrocyclic ring on the OH group of the N-terminal Ser, which in turn is N-acylated by 3-hydroxydodecanoic acid . Syringomycins A1 and G, two other metabolites of the same bacterium, differ from syringomycin E only in their fatty acid moieties corresponding, respectively, to 3-hydroxydecanoic and 3-hydroxytetradecanoic acid. Biochemistry, 1989 Sep 5, 28(18), 7233 - 40 X-ray absorption spectroscopy of the {2Fe-2S} Rieske cluster in Pseudomonas cepacia phthalate dioxygenase . Determination of core dimensions and iron ligation; Tsang HT et al.; We have employed X-ray absorption spectroscopy to obtain structural information about the Rieske Fe/S center in the phthalate dioxygenase (PDO) from Pseudomonas cepacia . Native PDO contains a dinuclear Rieske Fe/S center and an additional mononuclear Fe site . In order to study selectively the Fe/S cluster, we measured data for samples in which the mononuclear site was either depleted of metal or reconstituted with Co or Zn . Our results demonstrate that the iron environment in the Rieske cluster is structurally indistinguishable from that found in other Fe/S clusters, thus strongly supporting the suggestion that the unusually high reduction potentials for Rieske clusters are due to electrostatic rather than structural effects . The average Fe-Fe distance is 2.68 (3) A for both oxidized and reduced Rieske clusters . The average Fe-S distance is 2.24 (2) A in the oxidized cluster and 2.28 (2) A in the reduced cluster . Careful analysis of the EXAFS Debye-Waller factors suggests that the bridging and terminal Fe-S distances for the oxidized cluster are 2.20 and 2.31 A, respectively . Taken together with recent ENDOR results, these studies provide a detailed structural model for the Rieske {2Fe-2S} centers. J Med Assoc Thai, 1989 Sep, 72(9), 481 - 6 Splenic abscess at Siriraj Hospital, Thailand; Watanapa P et al.; Splenic abscess is an unusual disease and may be presented either as a localized area of infection in the spleen or as a part of generalized sepsis . Population-based autopsy studies have established the incidence of splenic abscess at between 0.2-0.7 per cent . An eleven-year retrospective study of cases of splenic abscess treated at Siriraj hospital, a total of 9 cases, is presented . Pseudomonas pseudomallei is the most frequent causative agent, found in one-third of the cases, especially if the patient is thalassemic or a resident in the Northeastern part of the country . Thalassemia is also the leading predisposing condition of this malady with the incidence of 33 per cent . There are some differences in the presenting clinical features in Thai patients compared with those reported in the literature . Splenectomy was performed in all but one who died of leukemia preoperatively . The mortality rate of this disease in this series is 11 per cent and we recommend splenectomy under antibiotic coverage as soon as the diagnosis of splenic abscess has been confirmed. Cancer Res, 1989 Sep 1, 49(17), 4791 - 5 Enhancement of the activity of immunotoxins by analogues of verapamil; Pirker R et al.; Verapamil has been shown to enhance immunotoxin activity but only at concentrations that are too high for in vivo use . Therefore, four structural analogues of verapamil (D792, D595, D528, and Sz45) were evaluated for their ability to enhance the in vitro activity of immunotoxins made with either ricin A chain or Pseudomonas exotoxin . The following immunotoxins were used: HB21-PE and 454A12-rRTA which recognize the human transferrin receptor; and 260F9-rRTA which reacts with human ovarian carcinoma and breast carcinoma cells . The activities of the immunotoxins were determined in ovarian carcinoma cells and in KB cells using inhibition of either protein synthesis or colony formation as a measure for the cytotoxicity of the immunotoxins . Each of the four analogues enhanced the activity of ricin A-immunotoxins in a dose-dependent manner . D792 and D595 also increased the activity of IIB21-PE . Low concentrations of either Sz45 or D528 enhanced the activity of HB21-PE, but high concentrations of these two analogues either had less enhancing potency than low concentrations or even decreased the activity of HB21-PE . Specificity of enhancement by the analogues was shown by competition of the activity of the immunotoxins by the corresponding antibody and by inactivity of an irrelevant immunotoxin . The amount of enhancement ranged from 2-fold to greater than 60-fold and was dependent on the cell line and on the experimental conditions . The enhancing ability of the drugs did not correlate with their calcium-antagonistic activity . When compared with verapamil, D792 and D595 had greater enhancing potency with regard to both ricin A-immunotoxins and Pseudomonas exotoxin-immunotoxins . Greater enhancing potency and less in vivo toxicity makes D792 a candidate for use in the enhancement of immunotoxins in vivo. J Med Microbiol, 1989 Sep, 30(1), 17 - 22 Production of an extracellular toxic complex by various strains of Pseudomonas cepacia; Straus DC et al.; Six isolates of Pseudomonas cepacia, representing various serotypes of the organism and possessing similar degrees of virulence in mice, were examined for their production of an extracellular toxic complex (ETC) in vitro . This compound is lethal for mice and produces extensive lung pathology in rats; it is composed of a surface carbohydrate antigen, lipopolysaccharide and protein . All six isolates produced the ETC . The LD50 values for the six ETC preparations ranged from 395 micrograms for strain 61g to 1750 micrograms for strain 90ee . Only two of the six ETC preparations contained ketodeoxyoctonate detectable by the methods used, and these two were the most toxic . Rabbit antiserum to the ETC of a serotype D strain could significantly protect mice only against serotype D strains . Examination of the various phases of growth of P . cepacia showed that there was extracellular release of the ETC beginning in the early logarithmic phase and continuing through the late stationary phase . The presence of the ETC in the supernatant fluids was due to release of this material rather than to cell lysis . In addition, at least one strain of P . cepacia was shown to produce an alginic acid-like compound. J Arthroplasty, 1989 Sep, 4(3), 263 - 9 Reimplantation of infected total hip arthroplasties in the absence of antibiotic cement; Wilson MG et al.; Twenty-two patients with deep infection of the hip were reimplanted and followed for a minimum of 3 years . All reimplantations were done without antibiotic-impregnated cement . Nine were done using cemented and 13 using cementless components . Two patients had recurrent infection . Both of these had primary Pseudomonas infections and had cemented revisions . At 3 or more years, 91% of patients are infection-free as determined by clinical evaluation, erythrocyte sedimentation rate, and, in a few, aspiration . Cemented hips had less pain than cementless hips, although both had equivalent functional scores . The significance of these findings is that reimplantation of infected hips can be successfully accomplished without antibiotic-impregnated cement . Cementless fixation can therefore be used . Clinical results with cementless hips for reimplantation will improve with current designs and techniques. Ann Otol Rhinol Laryngol, 1989 Sep, 98(9), 721 - 5 Use of ceftazidime for malignant external otitis; Kimmelman CP et al.; During the past 2 years we have used ceftazidime (Fortaz), a third-generation cephalosporin, in the treatment of eight patients with progressive necrotizing "malignant" external otitis . Ceftazidime is very active against Pseudomonas species and provides penetration into the CSF . Our results suggest that this medication has several advantages over the previously recommended combinations of aminoglycosides and semisynthetic penicillins, including improved cure rate, lower toxicity, and simpler administration schedules . We review our experience with ceftazidime in the treatment of eight patients. Mikrobiologiia, 1989 Sep-Oct, 58(5), 818 - 24 {Lytic activity of Pseudomonas bacteriophages}; Romashko AM et al.; None of the 24 Pseudomonas syringae bacteriophages were found to be identical in the spectrum of lytic action . The phages were subdivided into five groups according to the number of sensitive bacterial strains and their qualitative composition. Mol Plant Microbe Interact, 1989 Sep-Oct, 2(5), 262 - 72 Localization of ice nucleation activity and the iceC gene product in Pseudomonas syringae and Escherichia coli; Lindow SE et al.; Ice nucleation activity and the iceC gene product were quantified in different subcellular fractions of the Pseudomonas syringae source strain and in Escherichia coli containing the cloned iceC gene to determine the activity of this protein in different subcellular locations . Ice nuclei were nearly completely retained during isolation of cell envelopes but exhibited a decrease in the temperature at which they were expressed . Ice nucleation activity was found in Triton X-100 insoluble membrane fragments as well as in slowly sedimenting and high-density membrane fragments . Nearly all ice nucleation activity was associated with the outer membrane because the partitioning of 3-ketodeoxyoctonate (a lipopolysaccharide component) and ice nuclei in cell fractions were similar to and opposite that of NADH oxidase (a cytoplasmic membrane component) . The iceC gene product had an apparent mass of 150,000 Da based on migration in SDS-polyacrylamide gels . This protein was not found in soluble cell components . Nearly all of the iceC gene product, which occurred in low abundance, was associated with the outer membrane of both P . syringae and E . coli . Therefore, the iceC gene product is located at and is maximally active in or on the outer membrane of cells of the source strain and heterologous strains. Biochem Biophys Res Commun, 1989 Aug 30, 163(1), 172 - 6 A rapid solution immunoassay to quantify binding of the human immunodeficiency virus envelope glycoprotein to soluble CD4; McQuade TJ et al.; We developed a particle concentration fluorescent immunoassay to quantify the binding in solution of the human immunodeficiency virus (HIV) external glycoprotein (gp120) to soluble CD4 (sCD4) . The assay is rapid (1 hr), quantitative, and requires as little as 0.1 pmole of gp120 per evaluation . We find that gp120, purified from recombinant baculovirus infected insect cells, is suitable for the assay . Moreover, sCD4s obtained either from recombinant E . coli or mammalian cells, consisting of the N-terminal two domains (about 180 amino acids) as well as linked to the active regions of Pseudomonas exotoxin A, bind gp120 similarly. J Biol Chem, 1989 Aug 25, 264(24), 14256 - 61 Functional analysis of domains II, Ib, and III of Pseudomonas exotoxin; Siegall CB et al.; Pseudomonas exotoxin is composed of three structural domains that are responsible for cell recognition, membrane translocation, and ADP-ribosylation . The substitution of the cell recognition domain (domain Ia) with a growth factor such as transforming growth factor alpha (TGF alpha), creates a cell-specific cytotoxic agent, TGF alpha-PE40, which kills cells bearing epidermal growth factor (EGF) receptors . We have used TGF alpha-PE40 to define the role of sequences in domains II, Ib, and III . Various mutations were made in these domains and mutant forms of TGF alpha-PE40 expressed in Escherichia coli . Mutant proteins were then tested for their ADP-ribosylation, EGF receptor-binding, and cell-killing activities . Additionally, the amino boundary of domain III, which contains the ADP-ribosylation activity, was determined by deletion analysis . Data indicate that (i) the functional amino terminus of domain III is near amino acid 400; (ii) deletion of various regions in domain II or conversion of cysteines 265 and 268 to serines results in a loss of cytotoxicity which ranged from 10-fold to more than 150-fold, indicating that domain II is essential for full expression of cytotoxicity; (iii) deletion of the amino terminus of domain Ib results in a molecule with somewhat increased cytotoxic activity, indicating that domain Ib is not essential for the cytotoxic effect of TGF alpha-PE40; and (iv) TGF alpha-PE40, produced by denaturing and refolding of insoluble material from inclusion bodies, binds better to EGF receptors and is about 10-fold more cytotoxic to cells bearing EGF receptors than is the secreted form of soluble TGF alpha-PE40. Biochem Biophys Res Commun, 1989 Aug 15, 162(3), 1528 - 34 Specificity of endoproteinase Asp-N (Pseudomonas fragi): cleavage at glutamyl residues in two proteins; Ingrosso D et al.; Endoproteinase Asp-N, a metalloprotease from a mutant strain of Pseudomonas fragi, has been reported to specifically cleave on the N-terminal side of aspartyl and cysteic acid residues . We utilized this enzyme to generate fragments for determining the amino acid sequence of the D-aspartyl/L-isoaspartyl methyltransferase isozyme I from human erythrocytes . Surprisingly, we identified cleavage sites for this enzyme at the N-terminal side of several glutamyl residues in addition to the expected cleavage sites at aspartyl residues . The ability of this enzyme to cleave polypeptides at both glutamyl and aspartyl residues was confirmed by mapping additional sites on erythrocyte carbonic anhydrase I . These results indicate that a more appropriate name for this enzyme may be Endoproteinase Asp/Glu-N. Biochem J, 1989 Aug 15, 262(1), 233 - 40 The role of cytochrome c4 in bacterial respiration . Cellular location and selective removal from membranes; Hunter DJ et al.; The cellular location of cytochrome c4 in Pseudomonas stutzeri and Azotobacter vinelandii was investigated by the production of spheroplasts . Soluble cytochrome c4 was found to be located in the periplasm in both organisms . The remaining cytochrome c4 was membrane-bound . The orientation of this membrane-bound cytochrome c4 fraction was investigated by proteolysis of the cytochrome on intact spheroplasts . In P . stutzeri, 78% of the membrane-bound cytochrome c4 could be proteolysed, whilst 82% of the spheroplasts remained intact, suggesting that the membrane-bound cytochrome c4 is on the periplasmic face of the membrane in this organism . Cytochrome c4 was not susceptible to proteolysis on A . vinelandii spheroplasts, in spite of being digestible in the purified state . Cytochrome c5 was shown to have a similar cellular distribution to cytochrome c4 . Selective removal of cytochrome c4 from membranes of P . stutzeri was accomplished by the use of sodium iodide and propan-2-ol, with the retention of most of the ascorbate-TMPD (NNN'N'-tetramethylbenzene-1,4-diamine) oxidase activity associated with the membrane . Sodium iodide removed most of the cytochrome c4 from A . vinelandii membranes with retention of 62% of the ascorbate-TMPD oxidase activity . Cytochrome c4 could be returned to the washed membranes, but with no recovery of this enzyme activity . We conclude that cytochrome c4 is not involved in the ascorbate-TMPD oxidase activity associated with the membranes of these two organisms. Appl Environ Microbiol, 1989 Aug, 55(8), 1860 - 4 Plasmid expression and maintenance during long-term starvation-survival of bacteria in well water; Caldwell BA et al.; Strains of enteric bacteria and pseudomonads containing plasmid R388::Tnl721 (Tpr, Tcr) or pRO101 (Hgr, Tcr) were starved for over 250 days in sterile well water to evaluate effects of starvation-survival on plasmid expression and maintenance . Viable populations dropped to between approximately 0.1 and 1% of the initial populations . Escherichia coli(pRO101) and Pseudomonas cepacia(pRO101) lost both viability and plasmid expression at a lower rate than strains containing R388::Tnl721 . Three patterns of host-plasmid interaction were detected: (i) no apparent loss of plasmid expression, (ii) loss of plasmid expression on initial recovery with subsequent expression upon resuscitation, and (iii) loss of capability to produce functional plasmid resistance. J Infect Dis, 1989 Aug, 160(2), 253 - 60 Enzyme-linked immunosorbent assay for the diagnosis of clinical and subclinical melioidosis; Ashdown LR et al.; An enzyme-linked immunosorbent assay (ELISA) for the detection of specific IgG and IgM antibody to Pseudomonas pseudomallei was developed . The IgG-ELISA was compared with the indirect fluorescence assay for IgG antibody (IgG-IFA) and the indirect hemagglutination (IHA) test in studies with serum specimens from persons from endemic areas for melioidosis and from persons from nonendemic areas of Australia . The sensitivity and specificity of the IgG-ELISA were 90% and 99%, respectively, comparable to those obtained with the IgG-IFA . The IgG-ELISA was more sensitive than the IHA test (74%) and was more suitable than the IgG-IFA as a serologic screening test for melioidosis . The IgM-ELISA was compared with the IgM-IFA as a marker of disease stage in patients with melioidosis . There was good diagnostic agreement between the tests; 92% of patients with active disease gave IgM-ELISA titers greater than or equal to 1:5,120 and 93% of patients with subclinical melioidosis had IgM-ELISA titers less than or equal to 1:1,280 . Of the overlap group of patients with a borderline IgM-ELISA titer of 1:2,560, approximately 33% were clinical cases . An uncommon disease stage consisting of a self-limited, short-term, flu-like, pyrexial illness accompanied by elevated serum IgM-ELISA titers (greater than or equal to 1:5,120) was seen in a small number of patients residing in endemic Australia. J Bacteriol, 1989 Aug, 171(8), 4320 - 5 Transcription of the isoamylase gene (iam) in Pseudomonas amyloderamosa SB-15; Fujita M et al.; S1 nuclease mapping of RNA prepared from Pseudomonas amyloderamosa SB-15 suggested that the iam gene coding for isoamylase (glycogen 6-glucanohydrolase {EC 3.2.1.68}) is transcribed from two promoters . The transcription start site for the upstream promoter (termed P1) was located -182 base pairs from the first nucleotide of the initiation codon of iam, whereas the start site for the downstream promoter (termed P2) was 99 base pairs downstream of the P1 start site . Transcriptions from these promoters were induced by maltose and were not repressed by glucose . The promoter regions contained sequences homologous to the consensus sequence recognized by sigma 54 RNA polymerase of enteric bacteria and found in promoters of other Pseudomonas species . Northern (RNA) hybridization provided evidence that the iam gene is transcribed as monocistronic mRNAs with an approximate size of 2.6 kilobases. J Bacteriol, 1989 Aug, 171(8), 4267 - 71 Mechanism of biosynthesis of unsaturated fatty acids in Pseudomonas sp . strain E-3, a psychrotrophic bacterium; Wada M et al.; Biosynthesis of palmitic, palmitoleic, and cis-vaccenic acids in Pseudomonas sp . strain E-3 was investigated with in vitro and in vivo systems . {1-14C}palmitic acid was aerobically converted to palmitoleate and cis-vaccenate, and the radioactivities on their carboxyl carbons were 100 and 43%, respectively, of the total radioactivity in the fatty acids . Palmitoyl coenzyme A desaturase activity was found in the membrane fraction . {1-14C}stearic acid was converted to octadecenoate and C16 fatty acids . The octadecenoate contained oleate and cis-vaccenate, but only oleate was produced in the presence of cerulenin . {1-14C}lauric acid was aerobically converted to palmitate, palmitoleate, and cis-vaccenate . Under anaerobic conditions, palmitate (62%), palmitoleate (4%), and cis-vaccenate (34%) were produced from {1-14C}acetic acid, while they amounted to 48, 39, and 14%, respectively, under aerobic conditions . In these incorporation experiments, 3 to 19% of the added radioactivity was detected in released 14CO2, indicating that part of the added fatty acids were oxidatively decomposed . Partially purified fatty acid synthetase produced saturated and unsaturated fatty acids with chain lengths of C10 to C18 . These results indicated that both aerobic and anaerobic mechanisms for the synthesis of unsaturated fatty acid are operating in this bacterium. Biopolymers, 1989 Aug, 28(8), 1485 - 9 The osmotic coefficients of the sodium form of some biopolymers; Jang LK et al.; The osmotic coefficients phi p,Na of dilute solutions of the sodium form of some weakly acidic polymers are theoretically predicted in this work . Based on the measured value 0.73 of gamma Na, the activity coefficient of free Na+, of the completely ionized humic acid (sodium salt) in a salt-free solution, the effective interligand distance b is calculated to be 11.34 A by using Manning's counterion condensation theory {Manning, G . S . (1969) J . Chem . Phys . 51(3), 924} . The corresponding values of gamma Na (measured experimentally) and b for the completely ionized exopolymer of Pseudomonas atlantica are 0.624 and 7.57 A when cultivated at a dilution rate D = 0.015 h-1, 0.647 and 8.19 A at D = 0.025 h-1, and 0.613 and 7.29 Aat D = 0.06 h-1 . For alginic acid (in the completely ionized sodium form), gamma Na = 0.40 and b = 4.71 A . The osmotic coefficients phi p,Na for the partially and the completely ionized polymers are then predicted with Manning's theory as well. FEMS Microbiol Lett, 1989 Jul 15, 51(1), 219 - 22 Isolation of 9-hydroxy-delta-tetradecalactone from lipid A of Pseudomonas diminuta and Pseudomonas vesicularis; Arata S et al.; A lipid component was isolated from the fatty acid fraction of acid hydrolysates of lipid A derived from Pseudomonas diminuta JCM 2788 and Pseudomonas vesicularis JCM 1477 lipopolysaccharides . By structural analysis of the lipid and its trimethylsilyl and acetyl derivatives by thin-layer chromatography, gas chromatography-mass spectrometry, mass spectrometry, infrared spectrometry and 13C-NMR, it was identified as 9-hydroxy-delta-tetradecalactone. FEMS Microbiol Lett, 1989 Jul 15, 51(1), 197 - 200 Evidence for the presence of a new NAD+-dependent formate dehydrogenase in Pseudomonas sp . 101 cells grown on a molybdenum-containing medium; Karzanov VV et al.; The facultatively methylotrophic bacterium Pseudomonas sp . 101, grown on methanol in presence of molybdate, contains a new formate dehydrogenase (N-FDH) catalyzing NAD+-dependent oxidation of formate . The activity of this N-FDH could also be measured in presence of artificial electron acceptors, ferricyanide and 2,6-dichlorophenol indophenol . This new enzyme is absent in cells grown on a methanol-containing medium with tungstate, where only another two, previously described formate dehydrogenases, which are active only with NAD+ or only with artificial acceptors, respectively, were determined . The N-FDH was partially purified by a combination of ion-exchange and gel-filtration chromatography, and was shown to differ in its properties from the known NAD+-dependent counterpart. Am J Ophthalmol, 1989 Jul 15, 108(1), 64 - 7 Ulcerative keratitis associated with contact lens wear; Koidou-Tsiligianni A et al.; From October 1982 through June 1986, 658 patients developed ulcerative keratitis . In 196 of these patients it was contact lens-related . Fifty-nine patients wore extended-wear contact lenses for cosmetic purposes . On culture, Pseudomonas species was the organism most frequently isolated from the ulcers associated with contact lens wear . No cases of fungal keratitis were found in the contact lens group as compared to 40 cases (17%) in the noncontact lens group . Compared to results of a similar study covering January 1977 through September 1982, current results showed a larger number of patients using extended-wear lenses for cosmetic reasons (59 vs one) and overall younger age. Vopr Med Khim, 1989 Jul-Aug, 35(4), 84 - 9 {Isolation, various physico-chemical and catalytic properties of L-methionine-gamma-lyase from Pseudomonas taetrolens}; Zanin VA et al.; Homogeneous preparation of L-methionine gamma-lyase was isolated from Ps . taetrolens . As shown by gel filtration and gradient polyacrylamide gel electrophoresis molecular mass of the native L-methionine gamma-lyase was 130-135 kDa . Polyacrylamide gel electrophoresis in presence of 0.1% SDS showed that L-methionine gamma-lyase proved to be a tetramer, which consisted of identical subunits with a molecular mass of 34 kDa . Pyridoxal-5'-phosphate was bound to the enzyme in the ratio of four moles of the cofactor per a mole of protein . The absorption spectrum of the enzyme exhibited maximal values at 420 nm, which is specific for a number of pyridoxal phosphate-containing enzymes . L-methionine gamma-lyase from Ps . taetrolens was found to be dissimilar in its physicochemical and catalytic properties to the same enzymes from other sources. Mol Cell Biol, 1989 Jul, 9(7), 2860 - 7 Epidermal growth factor receptor binding is affected by structural determinants in the toxin domain of transforming growth factor-alpha-Pseudomonas exotoxin fusion proteins; Edwards GM et al.; TGF-alpha-PE40 is a hybrid protein composed of transforming growth factor-alpha (TGF-alpha) fused to a 40,000-dalton segment of Pseudomonas exotoxin A (PE40) . This hybrid protein possesses the receptor-binding activity of TGF-alpha and the cell-killing properties of PE40 . These properties enable TGF-alpha-PE40 to bind to and kill tumor cells that possess epidermal growth factor (EGF) receptors . Unexpectedly, TGF-alpha-PE40 binds approximately 100-fold less effectively to EGF receptors than does native TGF-alpha (receptor-binding inhibition IC50 = 540 and 5.5 nM, respectively) . To understand the factors governing receptor binding, deletions and site-specific substitutions were introduced into the PE40 domain of TGF-alpha-PE40 . Removal of the N-terminal 59 or 130 amino acids from the PE40 domain of TGF-alpha-PE40 improved receptor binding (IC50 = 340 and 180 nM, respectively) but decreased cell-killing activity . Substitution of alanines for cysteines at positions 265 and 287 within the PE40 domain dramatically improved receptor binding (IC50 = 37 nM) but also decreased cell-killing activity . Similar substitutions of alanines for cysteines at positions 372 and 379 within the PE40 domain did not significantly affect receptor-binding or cell-killing activities . These studies indicate that the PE40 domain of TGF-alpha-PE40 interferes with EGF receptor binding . The cysteine residues at positions 265 and 287 of PE40 are responsible for a major part of this interference. J Med Assoc Thai, 1989 Jul, 72 Suppl 2, 20 - 2 An epidemic of Pseudomonas cepacia bacteraemia in Ramathibodi Hospital; Wanaying B; An outbreak of P . cepacia bacteraemia involving 16 patients in Ramathibodi hospital from February to May 1988 was reported . The sources of infection were contaminated intravenous succinyl choline and metaraminol used by anaesthetists . Multiple dose preparations of the drugs were used . The drugs were prepared in bulk in an unhygienic room . Contamination occurred during or after the preparation . The outbreak was terminated by the improvement of hygiene in the anaesthetic preparation room, and the discontinuation of multiple-dose drugs. Appl Environ Microbiol, 1989 Jul, 55(7), 1724 - 9 Influence of Pseudomonas syringae culture conditions on initiation of the hypersensitive response of culture tobacco cells; Yucel I et al.; The inhibitor sensitivity and timing of the ionic response of suspension-cultured tobacco cells were used as a bioassay for the Pseudomonas syringae signal that elicits the hypersensitive response in resistant plants . The ionic response of tobacco cell suspensions inoculated with P . syringae pv . syringae 61 and P . syringae pv . pisi grown in rich media was inhibited by rifampin, tetracycline, and streptomycin during a 2- to 2.5-h induction stage . Coculturing the bacteria with tobacco cells for 3 h or more before inoculating fresh tobacco cells specifically abolished the sensitivity of the ionic response to these inhibitors and reduced the response time of the tobacco cells from 3 to 1 h . The apparent activation of the bacteria during coculture was not dependent on the plant cells and could be achieved by incubating the bacteria in a nitrogen-deficient medium containing a metabolizable carbon source . Addition of proteose peptone and Casamino Acids to this medium suppressed activation of the bacteria . The results suggest that the hypersensitive response-eliciting signal forms late in the induction stage, perhaps as a result of the derepression of some of the P . syringae genes functional in elicitation of the hypersensitive response . The nature of the activated state remains elusive but is consistent with the accumulation of protein(s) whose activity indirectly elicits the ionic response. Virology, 1989 Jul, 171(1), 229 - 38 Quantitation of the adsorption and penetration stages of bacteriophage phi 6 infection; Olkkonen VM et al.; The enveloped dsRNA bacteriophage phi 6 uses the pilus of Pseudomonas syringae as its receptor . It enters the host cell by fusion of the virus envelope with the host outer membrane, followed by penetration of the cytoplasmic membrane by the phage nucleocapsid . In this investigation we quantitated the adsorption and penetration of phi 6wt and a host range mutant, phi 6h 1s, to five bacterial strains . Adsorption rate constants were measured for the different phage-host combinations, the constant for phi 6wt with the standard host was 3.3 X 10(10) ml/min . Infections with 14C-labeled phage at different phage/cell ratios were used to measure the numbers of adsorbing and entering virions/sensitive cell . At high phage/cell ratios (200-250) the standard host adsorbed on the average 35-40 wild-type virions/cell, the saturation level being somewhat higher . It was shown that at phage/host cell ratios of 0.1-1 practically every virion produces an infectious center . The average number of entering phage particles per infectious center reached saturation around the phage/cell ratio of 50 and did not exceed 3 for the standard host . The phi 6 preparations used in this study had a specific infectivity of 0.7-0.9. J Bacteriol, 1989 Jul, 171(7), 3767 - 74 Excretion of the egl gene product of Pseudomonas solanacearum; Huang JZ et al.; Pseudomonas solanacearum is an important phytopathogen which excretes a variety of extracellular enzymes . Pulse-chase experiments showed that one of these enzymes, a beta-1,4-endoglucanase (EGL) encoded by the egl gene, is synthesized as a higher-molecular-weight precursor polypeptide (pEGL) which is subsequently excreted into the extracellular medium as a 43-kilodalton mature protein . S1 nuclease transcript mapping and DNA sequence analysis were used to identify the transcription start site and the possible translation start site of egl . Pulse-chase experiments and comparison of the putative NH2-terminal amino acid sequence of pEGL with the actual NH2-terminal amino acid sequence of mature excreted EGL suggested that pEGL has a 45-residue leader sequence preceding the N terminus of EGL which is proteolytically cleaved during export to the extracellular environment . The first 20 residues of the leader sequence resembled a typical lipoprotein signal peptide . The excretion of EGL by P . solanacearum apparently requires a membrane potential since it was blocked by carbonyl cyanide m-chlorophenyl hydrazone. FEMS Microbiol Lett, 1989 Jul 1, 51(1), 85 - 8 The barrier function of the outer membrane of Pseudomonas maltophilia in the diffusion of saccharides and beta-lactam antibiotics; Yamazaki E et al.; This paper reports that the efficiency of solute diffusion through the outer membrane of Pseudomonas maltophilia is roughly 3 to 5% of that of Escherichia coli . This is despite the fact that the outer membrane pore(s) is only a little smaller than that of E . coli . These results suggest that P . maltophilia has a low copy number of porin(s) . The outer membrane of antibiotic resistant clinical isolates showed even less efficient permeability towards saccharides and antibiotics than the laboratory strains. J Gen Microbiol, 1989 Jul, 135 ( Pt 7), 1969 - 78 Degradation of p-toluenesulphonic acid via sidechain oxidation, desulphonation and meta ring cleavage in Pseudomonas (Comamonas) testosteroni T-2; Locher HH et al.; Pseudomonas (Comamonas) testosteroni T-2 completely converted p-toluenesulphonic acid (TS) or p-sulphobenzoic acid (PSB) to cell material, CO2 and sulphate, with growth yields of about 5 g protein (mol C)-1 . PSB and sulphite were excreted as transient intermediates during growth in TS-salts medium . All reactions of a catabolic pathway involving sidechain oxidation and cleavage of the sulphonate moiety as sulphite were measurable in the soluble portion of cell extracts . Degradation of TS and PSB was inducible and apparently involved at least two regulons . TS was converted to p-sulphobenzyl alcohol in a reaction requiring NAD(P)H and 1 mol O2 (mol TS)-1 . This alcohol was in an equilibrium (in the presence of NAD+) with p-sulphobenzaldehyde, which was converted to PSB in an NAD(P)+-dependent reaction . PSB was desulphonated to protocatechuic acid in a reaction requiring NAD(P)H and 1 mol O2 (mol PSB)-1 . Experiments with 18 O2 confirmed involvement of a dioxygenase, because both atoms of this molecular oxygen were recovered in protocatechuate . Protocatechuate was converted to 2-hydroxy-4-carboxymuconate semialdehyde by a 4.5-dioxygenase. Mikrobiol Zh, 1989 Jul-Aug, 51(4), 68 - 74 {The biological activity and physicochemical properties of a new bacteriocin from a strain of Pseudomonas cepacia 5779}; Dodatko TA et al.; Pseudomonas cepacia 5779 bacteriocin (cepaciacin) whose producer was revealed due to application of the special screening system has been studied for its certain biological and physicochemical properties . Possessing a narrow range of action, it inhibits only the P . cepacia strains . Its biosynthesis occurs more intensely on the rich nutrient media, the highest quantities of cepaciacine being revealed at the terminal stage of the produced log growth . UV irradiation or mitomycin C introduction into the medium stimulated biosynthesis of this bacteriocin . Cepaciacin P . cepacia 5779 is a complex consisting of several protein subunits and carbon part . The protein-carbohydrate ratio is 3:1 . The molecular weight of the complex is 1.8 x 10(6) Da . Lipopolysaccharides isolated from the indicator strain being added, cepaciacine loses its activity . This bacteriocin is stable in the narrow range of pH, thermolabile, decomposes under the effect of proteases and is, evidently, a representative of a new type of the bacteriocin-like substances. Klin Padiatr, 1989 Jul-Aug, 201(4), 299 - 303 {Autologous peripheral stem cell transplantation in children}; Emminger W et al.; 19 children between 3 and 23 years underwent 79 leukapheres for collection of blood stem cells . In children suffering from acute lymphoblastic leukemia (ALL), Non Hodgkin's Lymphoma (NHL) and Ewing's Sarcoma (ES) we collected 6.87 x 10(4) CFU-GM/kg (range 2,65-21.7), if collections were started with the first platelet rise . In children with peripheral primitive neuroectodermal tumors (PNET) and neuroblastoma (NBL) we gained only 1.20 x 10(4) CFU-GM/kg (range 0.09-2.24) . 17 children received high dose chemoradiotherapy and peripheral stem cell +/- bone marrow rescue . 9 suffered from solid tumors, 8 from hematopoietic malignancies . 9 were transfused with peripheral stem cells only, 8 received bone marrow in addition . Time to reach 0.5 x 10(9)/l granulocytes was very short-median 31 days (12-65), in 4 children receiving more than 5 x 10(4) CFU-GM/kg 12 to 13 days, only . On January 31st, 1989 6/17 children are alive in complete remission after a median observation time of 14.5 months (3-26) after autologous stem cell transfusion, one child is alive in "no remission", 7 died with relapse, 3 died because of infections (2 x aspergillosis, 1 x pseudomonas septicemia) . The collection of blood derived stem cells by leukaphereses was well tolerated even in very small children and easily repeatable . With optimal timing high stem cell numbers were obtainable, resulting in a very short duration of posttransplant granulocytopenia. J Gen Microbiol, 1989 Jul, 135 ( Pt 7), 1989 - 96 The genetics of bile acid degradation in Pseudomonas spp.: location and cloning of catabolic genes; Leppik RA; Four Pseudomonas spp . capable of utilizing bile acids as sole carbon source were examined for the presence of plasmids . One plasmid was found in Pseudomonas sp . RAL8, but no plasmids could be detected in the other three strains . Mitomycin C curing of RAL8 did not affect the ability of the strain to grow on bile acids . This suggested that the genetic information for bile acid catabolism in all four strains was chromosomally located . To isolate bile acid catabolic genes . DNA from RAL8 was partially digested with Sau3A, then the DNA fragments cloned into the broad-host-range cosmid vector pMMB33 . The resulting gene bank was screened by plate-mating with two stable RAL8 mutants . Four of the gene bank clones were found to give a positive complementation with one or both mutants . Examination of the plasmids in the four clones revealed that they were unstable, but detailed mapping enabled a 52 kb restriction map to be derived . Further complementation work showed that two of the bile acid catabolic genes are located close together on the map, and may be contiguous. J Gen Microbiol, 1989 Jul, 135 ( Pt 7), 1979 - 88 Steroid catechol degradation: disecoandrostane intermediates accumulated by Pseudomonas transposon mutant strains; Leppik RA; Eleven transposon mutant strains affected in bile acid catabolism were each found to form yellow, muconic-like intermediates from bile acids . To characterize these unstable intermediates, media from the growth of one of these mutants with deoxycholic acid was treated with ammonia, then the crude product was methylated with diazomethane . Four compounds were subsequently isolated; spectral evidence suggested that they were methyl 12 alpha-hydroxy-3-oxo-23,24-dinorchola-1,4-dien-22-oate, methyl 4-aza-12 beta-hydroxy-9(10)-secoandrosta-1,3,5-triene-9,17-dione-3-carboxyl ate, 4-aza-9 alpha, 12 beta-dihydroxy-9(10)-secoandrosta-1,3,5-trien-17-one-3- methyl carboxylate and 4 alpha-{3'-propionic acid}-5-amino-7 beta-hydroxy-7 alpha beta-methyl- 3a alpha, 4,7,7a-tetrahydro-1-indanone-delta-lactam . It is proposed that the mutants are blocked in the utilization of such muconic-like compounds as the 3,12 beta-dihydroxy-5,9,17-trioxo-4(5),9(10)- disecoandrostal (10),2-dien-4-oic acid formed from deoxycholic acid . A further mutant was examined, which converted deoxycholic acid to 12 alpha-hydroxyandrosta-1,4-dien-3,17-dione, but accumulated yellow products from steroids which lacked a 12 alpha-hydroxy function, such as chenodeoxycholic acid . The products from the latter acid were treated as above; spectral evidence suggested that the two compounds isolated were methyl 4-aza-7-hydroxy-9(10)-secoandrosta-1,3,5- triene-9,17-dione-3-carboxylate and 4 alpha-{1'alpha-hydroxy-3'-propionic acid}-5-amino-7a beta-methyl-3a alpha,4,7,7a-tetrahydro-1-indanone-delta-lactam. Am J Clin Pathol, 1989 Jul, 92(1), 96 - 100 Cytomegalovirus infection involving the skin in immunocompromised hosts . A clinicopathologic study; Lee JY; Cytomegalovirus (CMV) infection involving the skin in three transplant patients is presented . Patient 1, whose infection apparently was localized only to a cutaneous wound induced by extravasated ionotropic solution, survived . Mixed CMV and Candida infections developed in patient 2 in the cutaneous ulcer . He died of disseminated herpes simplex virus infection in two weeks . Patient 3 had CMV pneumonia and purpuric maculopapular eruption . He died of Pseudomonas sepsis 17 weeks later . Eighteen cases with CMV skin lesions are reported in the English literature . The clinical findings and the outcome of the current and the reported cases are analyzed . All patients were immunocompromised . CMV infection, when detected in the skin, appears to be associated with grave prognosis . Seventeen of 20 patients whose final outcome was recorded died within six months after the onset of CMV skin lesions . The outcome of one case is unknown . The mortality was 85% . The fatal cases had either concurrent disseminated CMV infection or mixed cutaneous or systemic infections . When the infection is localized in the skin wounds, the prognosis seems fairly good . All three such patients survived. Eur Respir J Suppl, 1989 Jul, 7, 663s - 665s Nutrition for the respiratory insufficient patient; Mohsenin V et al.; Malnutrition is fairly common in patients with chronic obstructive pulmonary disease, the more severe the airway obstruction the more severe the nutritional status . The consequences of nutritional depletion on respiratory and immune systems are ventilatory compromise and susceptibility to infection . Diaphragm muscle mass and thickness is decreased in patients with COPD . This results in decreased maximum voluntary ventilation and diminished inspiratory pressure . Malnutrition is one of the causes of failure to wean in patients with respiratory failure . Malnutrition also has profound effects on cell-mediated immune response and humoral immunity with reduced levels of secretory IgA . In patients with COPD, colonization of respiratory tract bears a direct relationship with parameters of nutritional status . Patients with significant nutritional impairment have more tracheal cell bacteria adhered to and the tracheas were more frequently colonized by Pseudomonas species . The improvement of nutrition in these patients resulted in less bacterial cell binding to tracheal epithelial cells. Cancer Res, 1989 Jul 1, 49(13), 3562 - 7 Chemoimmunotoxin therapy against a human colon tumor (HT-29) xenografted into nude mice; Pearson JW et al.; The efficacy of intracavitary chemoimmunotoxin therapy for cancer treatment was evaluated using the human colon carcinoma (HT-29) which had been xenografted i.p . into nude mice . Mice bearing HT-29 were treated with an immunotoxin consisting of the monoclonal antibody OVB3 coupled to Pseudomonas exotoxin (OVB3-PE), with cyclophosphamide (Cy), or with both OVB3-PE plus Cy . Mice given injections i.p . of 3 x 10(6) HT-29 ascites cells developed a localized disease that presented as both malignant ascites and solid tumor confined to the peritoneal cavity . All mice died within 30 to 40 days . Mice that received either three or six injections of OVB3-PE at a dose of 0.5 micrograms every other day beginning 3 days post-tumor inoculation exhibited significantly increased median survival times (MSTs) (P = 0.002) of 62 and 68 days, respectively, as compared to a MST of 33 days for the controls . OVB3 alone or an irrelevant monoclonal antibody conjugated to PE exhibited no antitumor activity . The therapeutic effects of the immunotoxin could be blocked by giving a large amount of unconjugated OVB3 at the same time . Treatment of mice with Cy alone at the maximal tolerated dose (250 mg/kg) on Days 10 and 17 after tumor inoculation increased the MST from 33 days to 54 days . The maximum tolerated dose could be increased to 300 mg/kg per injection if the Cy treatment was preceded by 100 mg/kg of S-2-(3-aminopropylamino)ethyl phosphorothioic acid (WR-2721), a sulfhydryl compound that selectively protects normal tissue against the toxicity of radiation and alkylating agents . Cy plus WR-2721 treatment on Days 10 and 17 increased the MST from 35 to 61 days (P = 0.002) . Interestingly, groups of mice that received either two, four, or seven treatments of OVB3-PE following Cy plus WR-2721 therapy exhibited a further increase (P less than 0.002) in MSTs to 81, 87, and 96 days, respectively . Thus, the combination of cytoreductive chemotherapy with the OVB3-PE was significantly more effective for the intracavitary treatment of established HT-29 colon cancer xenografts than either chemotherapy or immunotoxin therapy alone. Korean J Intern Med, 1989 Jul, 4(2), 174 - 7 Hypersensitivity pneumonitis by a cool-mist vaporizer: a detailed microbiologic and immunologic study; Chung JC et al.; A patient with hypersensitivity pneumonitis caused by a contaminated cool-mist vaporizer was evaluated . A detailed microbiologic and immunologic study was done, and a Pseudomonas species was isolated as the possible causative organism by inhalational provocative and serologic tests. Biochem J, 1989 Jun 15, 260(3), 857 - 62 The second subunit of methanol dehydrogenase of Methylobacterium extorquens AM1; Nunn DN et al.; The nucleotide and deduced amino acid sequence of a novel small (beta) subunit of methanol dehydrogenase of Methylobacterium extorquens AM1 (previously Pseudomonas AM1) has been determined . Work with the whole protein has shown that is has an alpha 2 beta 2 configuration. FEBS Lett, 1989 Jun 5, 249(2), 348 - 52 Detection and localization of a new enzyme catalyzing the beta-aryl ether cleavage in the soil bacterium (Pseudomonas paucimobilis SYK-6); Masai E et al.; Cleavage of the arylglycerol-beta-aryl ether linkage is the most important process in the biological degradation of lignin . We determined the activity of the enzyme cleaving the beta-aryl ether linkage in membranes of Pseudomonas paucimobilis SYK-6 . This enzyme was tightly associated with the cellular membrane and catalyzed the unique and reductive cleavage of compound II but not cleavage of compound I . This enzymatic activity was stimulated by addition of NADH . On the basis of this evidence, we present a model of the specific cellular assimilation of beta-aryl ether by P . paucimobilis SYK-6. J Antimicrob Chemother, 1989 Jun, 23(6), 831 - 5 Ciprofloxacin, imipenem and rifampicin: in-vitro synergy of two and three drug combinations against Pseudomonas cepacia; Kumar A et al.; The in-vitro activity of ciprofloxacin, imipenem and rifampicin, singly, and in two and three drug combinations was evaluated against 16 isolates of Pseudomonas cepacia . All 16 isolates were resistant to rifampicin; nine isolates were susceptible to ciprofloxacin; six were susceptible and seven had intermediate susceptibility to imipenem . The imipenem and rifampicin combination was synergistic for one of 16, imipenem and ciprofloxacin synergistic for seven of 16 and the three antibiotic combination was synergistic for 12 of 16 isolates . The three antibiotic combination demonstrated synergism with two isolates which were resistant to all three drugs . Combinations of two and three antibiotics resulted in enhanced killing of P . cepacia in this in-vitro study. Burns, 1989 Jun, 15(3), 167 - 70 Pilot study into the IgG1 and IgG2 subclass response to polyvalent Pseudomonas vaccine in burned adults; Frame JD et al.; The IgG1 and IgG2 subclass response to thermal injury has been measured in a group of eight burned adults who received a single intramuscular injection of 0.5 ml of polyvalent pseudomonas vaccine (PPV), within hours of burn injury . The response in three of these patients is compared with the response in three matched patients who did not receive the vaccine . This single dose regimen of PPV did not appear to stimulate the early production of IgG1 or IgG2 and if subclass deficiency contributes to the risk of sepsis or toxin-mediated disease, as previous workers have established (Schur et al., 1970; Oxelius et al., 1981), then there is no apparent benefit in active immunization to reduce the risk in the early postburn period. Proc Natl Acad Sci U S A, 1989 Jun, 86(11), 4215 - 9 Cytotoxic activity of a recombinant fusion protein between interleukin 4 and Pseudomonas exotoxin; Ogata M et al.; A recombinant chimeric toxin in which the cell binding domain of Pseudomonas exotoxin (PE) was replaced by murine interleukin 4 (IL-4) was produced in Escherichia coli . This chimeric protein, IL-4-PE40, was cytotoxic to murine IL-4 receptor-bearing cell lines but had little effect on human cell lines lacking receptors capable of binding murine IL-4 . A mutant form of IL-4-PE40 (termed IL-4-PE40 asp553) with very low ADP-ribosylating activity displayed mitogenic activity similar to that of IL-4 rather than cytotoxic activity . Because the cytotoxic effects of IL-4-PE40 were blocked by excess IL-4 or by neutralizing antibody to IL-4 (11B11), we conclude that the cytotoxic effect of IL-4-PE40 is specifically mediated through IL-4 receptors . IL-4-PE40 could be a useful reagent for specific elimination of cells bearing IL-4 receptors. J Urol, 1989 Jun, 141(6), 1463 - 6 Effects of anti-lipid A human monoclonal antibody on lipopolysaccharide-induced toxicity to the kidney; Tune BM et al.; Studies were done to evaluate the effects of the human monoclonal anti-lipid A IgM antibody A6(H4C5) on several components of the hemodynamic and renal toxicity of the cell wall lipopolysaccharide of E . coli 0111:B4 . Antibody (0.25 to four mg./kg . BW) was administered 0.5 hour before, or premixed for one hour with, lipopolysaccharide (0.05 mg./kg., a 14 to 18% lethal dose), and the following measurements made over 0.5 to 3.5 hours of study: systemic arterial blood pressure, renal plasma flow, and glomerular filtration . The proximal tubular cell cytotoxicity of 90 mg./kg . of the cephalosporin cephaloridine was also quantified in similarly treated animals sacrificed 48 hours later . While one mg./kg . of antibody prevented the reduction by the lipopolysaccharide of renal plasma flow, it did not prevent the nephrotoxic synergy with cephaloridine, and four times the antibody dose did not prevent lipopolysaccharide-induced hypotension or reduced glomerular filtration . These amounts of this antibody protect leukopenic rabbits against the lethality of the slow onset bacteremic model of Pseudomonas conjunctivitis . It is suggested that the incompleteness of protection in this study may be the result of the sensitivity of the assay methods used and/or the acute endotoxemia produced in these animals. J Bacteriol, 1989 Jun, 171(6), 2994 - 3001 Cloning, sequencing, and overexpression of mvaA, which encodes Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl coenzyme A reductase; Beach MJ et al.; We have cloned, determined the primary structure of, and overexpressed in Escherichia coli the gene mvaA, which is the 1,287-base structural gene for the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase {EC 1.1.1.88} of Pseudomonas mevalonii . The amino acid composition of HMG-CoA reductase agreed with that predicted from the nucleotide sequence of mvaA, and DNA-derived sequences were identical to all experimentally determined peptide sequences . Overexpression of mvaA in E . coli yielded quantities of HMG-CoA reductase over 1,500-fold higher than those present in control cultures . Comparison of the primary structure of the P . mevalonii enzyme with the DNA-derived primary structure for a mammalian HMG-CoA reductase revealed two regions of similarity suggestive of functional relatedness . An open reading frame, ORF1, lies on the 3' side of mvaA, and a potential ribosome-binding site for ORF1 overlaps the termination codon of mvaA. J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1469 - 77 13C-NMR studies of acetate and methanol metabolism by methylotrophic Pseudomonas strains; Narbad A et al.; The metabolism of {2-13C}acetate by Pseudomonas M27(Icl-) and Pseudomonas MA(Icl+) was studied in vivo using 13C-NMR spectroscopy . The flux of 13C-label into bicarbonate, glutamate and citrate was observed in both organisms . In addition 13C-labelled alpha, alpha-trehalose was synthesized as a major metabolite by Pseudomonas M27 but not by Pseudomonas MA . The presence of this disaccharide in cell extracts of Pseudomonas AM1(Icl-) grown with {13C}methanol was also observed . The data from analysis of the trehalose multiplet signal observed in the spectra of Pseudomonas M27 cell extracts were consistent with the absence of the glyoxylate cycle in this methylotroph. Anasth Intensivther Notfallmed, 1989 Jun, 24(3), 153 - 61 {Use of pseudomonas immunoglobulin . Indications and results}; Werdan K et al.; In comparison with polyvalent immunoglobulins, Pseudomonas immunoglobulin (Psomaglobin) is enriched several times in antibodies to Ps . lipopolysaccharide antigens and exotoxin A as well as lipid A . The resulting protective action which is superior to polyvalent immunoglobulins in infections with Pseudomonas, was demonstrated both in cell culture (protection against cytotoxicity of Ps . exotoxin A in heart muscle cells) and in animal models of sepsis . In patients suffering from Ps . pneumonia and Ps.-sepsis clinical improvement is seen after application of this immunoglobulin and also in quantifiable by scoring systems, the unequivocal proof of lowering lethality by using this specific immunoglobulin in Pseudomonas infection is to be shown however. J Antimicrob Chemother, 1989 Jun, 23(6), 885 - 90 Serum and sputum concentrations of netilmicin in combination with acylureidopenicillin and cephalosporins in clinical treatment of pulmonary exacerbations in cystic fibrosis; Hjelte L et al.; The pharmacokinetics of netilmicin was studied in 14 patients with cystic fibrosis, aged 4-21 years (mean 16 years) during treatment of pulmonary exacerbations of pseudomonas infection . The patients received 24 courses of netilmicin (10 mg/kg/day) in combination with azlocillin (600 mg/kg/day), cefsulodin (200 mg/kg/day) or ceftazidime (150 mg/kg/day) for 9-14 days . Seven patients received two or three courses of different combinations . Serum and sputum concentrations of netilmicin were determined on day 2 and 6 . Mean (+/- S.E.M.) trough serum values were 1.4 +/- 0.2 mg/l (same on day 2 and 6), peak values at 10 min 13.6 +/- 1.0 and 13.7 +/- 0.9 mg/l, and serum concentration at 1 h 7.5 +/- 0.6 and 7.5 +/- 0.5 mg/l, on days 2 and 6 respectively . The half-life was about 1 h . The pharmacokinetics did not differ on day 2 and 6 . Sputum concentrations increased up to 2-3 h after administration, mean (+/- S.E.M.) peak values being 2.6 +/- 0.6 and 1.5 +/- 0.4 mg/l at day 2 and 6, respectively . The study shows that the pharmacokinetics of netilmicin was not influenced by different combinations with beta-lactams . All patients improved clinically, but pseudomonas growth was only reduced in nine courses . In one case transient resistance to netilmicin developed during the treatment . The clinical efficacy and tolerance were good and similar to those seen with combinations with other aminoglycosides. Nature, 1989 Jun 1, 339(6223), 394 - 7 A recombinant immunotoxin consisting of two antibody variable domains fused to Pseudomonas exotoxin; Chaudhary VK et al.; Antibodies and growth factors have been chemically coupled to different toxins to produce cytotoxic molecules that selectively kill cells bearing appropriate antigens or receptors . Antibody-toxin conjugates (immunotoxins) produced using conventional chemical coupling techniques have several undesirable characteristics . The smallest binding unit of an antibody is an Fv fragment which consists of a light and heavy chain variable domain . Recently, active single chain Fv fragments of antibodies have been produced in Escherichia coli by attaching the light and heavy chain variable domains together with a peptide linker . Here we describe the construction and expression in E . coli of a single chain antibody toxin fusion protein, anti-Tac(Fv)-PE40, in which the variable regions of anti-Tac, a monoclonal antibody to the p55 subunit of the human interleukin-2 receptor, are joined in peptide linkage to PE40, a modified form of Pseudomonas exotoxin lacking its binding domain . Anti-Tac(Fv)-PE40 was very cytotoxic to two interleukin-2 receptor-bearing human cell lines but was not cytotoxic to receptor-negative cells. J Clin Pathol, 1989 Jun, 42(6), 645 - 8 Identification of Pseudomonas pseudomallei in clinical practice: use of simple screening tests and API 20NE; Dance DA et al.; The API 20NE kit and a simple screening system involving Gram's stain, the oxidase reaction, colistin and gentamicin resistance, and colonial characteristics on a differential agar medium, were used to test 400 strains of Pseudomonas pseudomallei . The API kit identified 390 (97.5%) strains correctly on first testing and all but one of the remainder on second testing . Only one strain was initially misidentified (as Ps cepacia) . The screening system was 100% accurate in identifying Ps pseudomallei . In non-endemic areas the API 20NE kit may be used to identify sporadic imported strains of Ps pseudomallei . Such kits may also help to delineate the geographical distribution of melioidosis . In endemic areas the screening tests described offer a cheap, simple, and accurate means of presumptively identifying Ps pseudomallei from clinical specimens. Biochemistry, 1989 May 30, 28(11), 4861 - 71 Electron-nuclear double resonance spectroscopy of 15N-enriched phthalate dioxygenase from Pseudomonas cepacia proves that two histidines are coordinated to the {2Fe-2S} Rieske-type clusters; Gurbiel RJ et al.; We have performed ENDOR spectroscopy at microwave frequencies of 9 and 35 GHz at 2 K on the reduced Rieske-type {2Fe-2S} cluster of phthalate dioxygenase (PDO) from Pseudomonas cepacia . Four samples have been examined: (1) 14N (natural abundance); (2) uniformly 15N labeled; (3) {15N}histidine in a 14N background; (4) {14N}histidine in a 15N background . These studies establish unambiguously that two of the ligands to the Rieske {2Fe-2S} center are nitrogens from histidine residues . This contrasts with classical ferredoxin-type {2Fe-2S} centers in which all ligation is by sulfur of cysteine residues . Analysis of the polycrystalline ENDOR patterns has permitted us to determine for each nitrogen ligand the principal values of the hyperfine tensor and its orientation with respect to the g tensor, as well as the 14N quadrupole coupling tensor . The combination of these results with earlier Mossbauer and resonance Raman studies supports a model for the reduced cluster with both histidyl ligands bound to the ferrous ion of the spin-coupled {Fe2+ (S = 2), Fe3+ (S = 5/2)} pair . The analyses of 15N hyperfine and 14N quadrupole coupling tensors indicate that the geometry of ligation at Fe2+ is approximately tetrahedral, with the (Fe)2(N)2 plane corresponding to the g1-g3 plane, and that the planes of the histidyl imidazoles lie near that plane, although they could not both lie in the plane . The bonding parameters of the coordinated nitrogens are fully consistent with those of an spn hybrid on a histidyl nitrogen coordinated to Fe . Differences in 14N ENDOR line width provide evidence for different mobilities of the two imidazoles when the protein is in fluid solution . We conclude that the structure deduced here for the PDO cluster is generally applicable to the full class of Rieske-type centers. Biochemistry, 1989 May 30, 28(11), 4650 - 5 Structure-activity relationships in the hydrolysis of substrates by the phosphotriesterase from Pseudomonas diminuta; Donarski WJ et al.; The mechanism and substrate specificity of the phosphotriesterase from Pseudomonas diminuta have been examined . The enzyme hydrolyzes a large number of phosphotriester substrates in addition to paraoxon (diethyl p-nitrophenyl phosphate) and its thiophosphate analogue, parathion . The two ethyl groups in paraoxon can be changed to propyl and butyl groups, but the maximal velocity and Km values decrease substantially . The enzyme will not hydrolyze phosphomonoesters or -diesters . There is a linear correlation between enzymatic activity and the pKa of the phenolic leaving group for 16 paraoxon analogues . The beta value in the corresponding Bronsted plot is -0.8 . No effect on either Vmax or Vmax/Km is observed when sucrose is used to increase the relative solvent viscosity by 3-fold . These results are consistent with rate-limiting phosphorus-oxygen bond cleavage . A plot of log V versus pH for the hydrolysis of paraoxon shows one enzymatic group that must be unprotonated for activity with a pKa of 6.1 . The deuterium isotope effect by D2O on Vmax and Vmax/Km is 2.4 and 1.2, respectively, and the proton inventory is linear, which indicates that only one proton is "in flight" during the transition state . The inhibition patterns by the products are consistent with a random kinetic mechanism. Biochemistry, 1989 May 16, 28(10), 4403 - 9 Differences between the manganese- and the iron-containing superoxide dismutases of Escherichia coli detected through sedimentation equilibrium, hydrodynamic, and spectroscopic studies; Beyer WF Jr et al.; The genome of Escherichia coli codes for two superoxide dismutases that may contain either iron (FeSOD) or manganese (MnSOD) at the active site . The crystal structures of MnSODs from two bacterial sources (but not E . coli) have been completed, and structural comparisons with the crystal structure of the FeSOD from either E . coli or Pseudomonas ovalis have been made . Despite the low degree (less than 50%) of sequence homology between the E . coli enzymes, the two proteins are suggested to be structurally homologous . Nonetheless, these enzymes exhibit absolute metal cofactor specificity in conferring enzymatic activity to the inactive apoenzyme . This observation is surprising considering the identity of the active site ligands and the similarities in their geometry and surrounding environment . Using analytical ultracentrifugation, we have determined that the solution properties of these two proteins are different . Thus dialysis of FeSOD but not of MnSOD against phosphate buffer in the presence or absence of EDTA caused dissociation of the homodimer . This dissociation appeared to be related to the loss of iron from native FeSOD . Thus, apoFeSOD but not apoMnSOD existed predominantly as a monomer at protein concentrations below 150 micrograms/mL . ApoMnSOD showed no evidence for dissociation under these conditions . Fluorescence data suggest that the tryptophan environments for the two enzymes are also different . The results of these physical measurements lead us to propose that subtle differences, perhaps at the subunit contact faces, exist in the structures of these crystallographically similar proteins. Rinsho Ketsueki, 1989 May, 30(5), 644 - 9 {Empirical antibiotic therapy in febrile neutropenic patients with acute leukemia}; Yabe H et al.; One hundred and ninety-five episodes of fever during the neutropenic phase of chemotherapy in 49 patients with acute leukemia from 1984 to 1987 were analyzed with the following results: 1) Febrile episodes occurred in 80 percent of the neutropenic (less than 500/microliters) phase lasting more than 7 days after chemotherapy . 2) Febrile episodes consisted of 44 (22%) of established septicemia and 111 (57%) of suspected septicemia . 3) The pathogens causing septicemia were 8 GPC, 38 GNB (22 Pseudomonas species) and 6 fungi . Fungemia was confirmed on an average of 4.8 days after the onset of fever . The mortality of septic events was 10 out of 17 episodes (59%) when treated with antibiotics alone, while 8 out 27 (30%) with the combination of antibiotics plus antifungal drugs . 4) The mortality of suspected sepsis was only 2 out of 111 episodes . Eighty-three (75%) of these 111 episodes responded to antibiotics alone, while 26 (23%) cases needed antibiotics plus antifungal drugs . Our results suggest that in febrile neutropenic patient empiric broad-spectrum antibiotic therapy should be initiated which is especially effective for Pseudomonas species, but if fever persists despite more than 4 or 5 days of antibiotic therapy, additional antifungal therapy should be considered. J Infect Dis, 1989 May, 159(5), 890 - 9 Melioidosis: a major cause of community-acquired septicemia in northeastern Thailand; Chaowagul W et al.; In a prospective study of all patients with Pseudomonas pseudomallei infections admitted to a large provincial hospital in northeastern Thailand, 63 cases of septicemic melioidosis and 206 patients with other community-acquired septicemias were documented during a 1-y period . Apart from P . pseudomallei, the spectrum of bacteria isolated from blood cultures and the overall mortality (32%) were similar to those previously reported elsewhere . Death from septicemia was associated with failure to develop a leukocytosis or pyrexia over 38 degrees C, azotemia, hypoglycemia, and jaundice . Septicemic melioidosis presented mainly in the rainy season, occurred predominantly in rice farmers or their families, and was significantly associated with preexisting diabetes mellitus or renal failure (P = .03) . Blood-borne pneumonia and visceral abscesses were common and the mortality was high (68%; P less than .001) . The response to appropriate treatment was slow (median fever clearance time 5.5 d) and the median duration of hospital stay was 4 w . Septicemic melioidosis is a major cause of morbidity and mortality in northeast Thailand. J Bacteriol, 1989 May, 171(5), 2756 - 61 Mutational changes in physiochemical cell surface properties of plant-growth-stimulating Pseudomonas spp . do not influence the attachment properties of the cells; de Weger LA et al.; Bacteriophage-resistant mutant strains of the root-colonizing Pseudomonas strains WCS358 and WCS374 lack the O-antigenic side chain of the lipopolysaccharide, as was shown by the loss of the typical lipopolysaccharide ladder pattern after analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . These strains differed from their parent strains in cell surface hydrophobicity and in cell surface charge . The observed variation in these physicochemical characteristics could be explained by the differences in sugar composition . The mutant strains had no altered properties of adherence to sterile potato roots compared with their parental strains, nor were differences observed in the firm adhesion to hydrophilic, lipophilic, negatively charged, or positively charged artificial surfaces . These results show that neither physicochemical cell surface properties nor the presence of the O-antigenic side chain plays a major role in the firm adhesion of these bacterial cells to solid surfaces, including potato roots. Mycoses, 1989 May, 32(5), 219 - 23 Subcutaneous granuloma caused by Phialophora richardsiae: case report and review of the literature; Gueho E et al.; A phaeomycotic cyst in a 47-year-old man, caused by Phialophora richardsiae, was treated successfully by excision . A critical review of the literature indicates that the pathological course of P . richardsiae infections usually follows a similar pattern to that of the present case, viz . generation of a well-defined and limited nodule following traumatic subcutaneous introduction of the fungus . The nodule typically accumulates a viscous yellow fluid . The infection remains localized, often being encapsulated by a fibrous layer; adenopathy is not observed . Concomitant bacterial infections are also common; in the present case Pseudomonas stutzeri was identified . Neither bacterial nor fungal infection recurred after surgery. Antibiot Khimioter, 1989 May, 34(5), 378 - 82 {Combined chemotherapy of experimental infection in neutropenia}; Viadro MM et al.; A significant decrease in resistance to infections caused by gramnegative pathogens was observed in mice with neutropenia induced by cytostatics . Efficacy of schemes for combined chemotherapy with beta-lactams, aminoglycosides and a novel peptide antibiotic was studied on model infections in mice with neutropenia . In the neutropenic mice with sepsis caused by Pseudomonas the peptide antibiotic administered parenterally in a single dose of 50 micrograms/kg provided high therapeutic activity . In combination with azlocillin, cefotaxime and amikacin the peptide antibiotic has a synergistic therapeutic action. J Gen Microbiol, 1989 May, 135 ( Pt 5), 1083 - 92 Molecular analysis of a plasmid-encoded phenol hydroxylase from Pseudomonas CF600; Shingler V et al.; Pseudomonas strain CF600 is able to utilize phenol and 3,4-dimethylphenol as sole carbon and energy source . We demonstrate that growth on these substrates is by virtue of plasmid-encoded phenol hydroxylase and a meta-cleavage pathway . Screening of a genomic bank, with DNA from the previously cloned catechol 2,3-dioxygenase gene of the TOL plasmid pWW0, was used in the identification of a clone which could complement a phenol-hydroxylase-deficient transposon insertion mutant . Deletion mapping and polypeptide production analysis identified a 1.2 kb region of DNA encoding a 39.5 kDa polypeptide which mediated this complementation . Enzyme activities and growth properties of Pseudomonas strains harbouring this fragment on a broad-host-range expression vector indicate that phenol hydroxylase is a multicomponent enzyme containing the 39.5 kDa polypeptide as one component. J Bacteriol, 1989 May, 171(5), 2740 - 7 Cloning and sequencing of two tandem genes involved in degradation of 2,3-dihydroxybiphenyl to benzoic acid in the polychlorinated biphenyl-degrading soil bacterium Pseudomonas sp . strain KKS102; Kimbara K et al.; Two genes involved in the degradation of biphenyl were isolated from a gene library of a polychlorinated biphenyl-degrading soil bacterium, Pseudomonas sp . strain KKS102, by using a broad-host-range cosmid vector, pKS13 . When a 3.2-kilobase (kb) PstI fragment of a 29-kb cosmid DNA insert was subcloned into pUC18 at the PstI site downstream of the lacZ promoter, Escherichia coli cells carrying this recombinant plasmid expressed 2,3-dihydroxybiphenyl dioxygenase activity . Nucleotide sequencing of the 3.2-kb PstI fragment revealed that there were two open reading frames (ORFI {882 base pairs} and ORFII {834 base pairs}, in this gene order) . Results of analysis of Tn5 insertion mutants and unidirectional deletion mutants suggested that the ORFI coded for 2,3-dihydroxybiphenyl dioxygenase . When the sequence of ORFI was compared with that of bphC of Pseudomonas pseudoalcaligenes KF707 (K . Furukawa, N . Arima, and T . Miyazaki, J . Bacteriol . 169:427-429, 1987), the homology was 68%, with both strains having the same Shine-Dalgarno sequence . The result of gas chromatography-mass spectrometry analysis of the metabolic product suggested that the ORFII had meta cleavage compound hydrolase activity to produce benzoic acid . DNA sequencing suggested that these two genes were contained in one operon. Biochem Biophys Res Commun, 1989 Apr 14, 160(1), 243 - 9 Detection of essential arginine in bacterial peptidyl dipeptidase-4: arginine is not the anion binding site; Lanzillo JJ et al.; Peptidyl dipeptidase-4 from Pseudomonas maltophilia was modified with the arginine reagents p-hydroxyphenylglyoxal and 2,3-butanedione . The enzyme was inactivated in a pseudo-first-order manner by p-hydroxyphenylglyoxal with a half-time of 72 min . Inactivation by 2,3-butanedione was biphasic with a rapid phase followed by a slower inactivation to less than 10% activity within 24h . The competitive inhibitor thiorphan protected against inactivation by phydroxyphenylglyoxal and by 2,3-butanedione also but to a lesser degree . Inhibitory anions chloride and phosphate did not protect against inactivation by either reagent . These data support the conclusion that an active site arginine is essential for substrate hydrolysis . Furthermore, arginine is not the binding site for the inhibitors chloride and phosphate. J Biol Chem, 1989 Apr 5, 264(10), 5442 - 51 The Pseudomonas oleovorans alkBAC operon encodes two structurally related rubredoxins and an aldehyde dehydrogenase; Kok M et al.; The Pseudomonas oleovorans alkBAC operon encodes seven proteins, of which at least three are involved in alkane hydroxylase (alkBA) and alkanol dehydrogenase (alkC) activities . We have determined the nucleotide sequence of the 2.5-kilobase pair alkA region and analyzed the role of its translation products in alkane oxidation . The alkA region contains three coding sequences, encoding two related rubredoxins (alkF and alkG) of 14- and 18-kDa molecular mass and a 52-kDa aldehyde dehydrogenase (alkH) . Deletion analysis indicated that neither the 14-kDa alkF gene product (rubredoxin 1) nor the amino-terminal part of the 18-kDa alkG gene product (rubredoxin 2) is required for alkane hydroxylase activity in vivo . The product of the alkH cistron restores growth of a P . oleovorans aldehyde dehydrogenase mutant on aliphatic alcohols and aldehydes . Its amino acid sequence shows considerable homology to previously characterized aldehyde dehydrogenases from mammalian and fungal origin . The nucleotide composition of the alk genes (47% G + C) differs considerably from the G + C content of the P . oleovorans genome suggesting that the alk regulon may originate from an unrelated organism. J Biol Chem, 1989 Apr 5, 264(10), 5435 - 41 The Pseudomonas oleovorans alkane hydroxylase gene . Sequence and expression; Kok M et al.; We have identified and sequenced the Pseudomonas OCT plasmid-encoded alkane hydroxylase gene (alkB) and its promoter . The transcription initiation site of the alkBAC mRNA was determined by nuclease S1 mapping . A putative interaction site with RNA-polymerase was identified based on homology of the alk promoter with other Pseudomonas promoters . The alkB gene encodes a 401-amino acid polypeptide which, despite an unusual codon composition, can be expressed at high levels in Escherichia coli and Pseudomonas . The amino-terminal sequence of the purified cytoplasmic membrane alkane hydroxylase was determined and was found to be in agreement with the nucleotide sequence . The translation product of the alkB gene contains nine hydrophobic sequences of which eight are sufficiently long to be membrane-spanning segments . The amino-terminal sequence resembles that of several bacterial integral membrane proteins and is not cleaved off following translation. J Infect Dis, 1989 Apr, 159(4), 654 - 60 Acute suppurative parotitis caused by Pseudomonas pseudomallei in children; Dance DA et al.; During a prospective clinical study of melioidosis in northeast Thailand, suppurative parotitis was observed as a characteristic presentation in children . Parotitis constituted 6.3% of all culture-positive melioidosis and 38% of melioidosis in children . Nine cases are described . None had apparent predisposition to infection, although two patients developed rising mumps virus antibody titers, suggesting a possible relation between these conditions . Complications included abscess formation (nine), spontaneous rupture into the auditory canal (five), facial nerve palsy (two), and septicemia and osteomyelitis with septic arthritis (one each) . All children initially responded to surgical drainage and appropriate antibiotic therapy . Pseudomonas pseudomallei parotitis should be considered in children from endemic areas with fever and facial swelling . It has a good prognosis with appropriate treatment . It may also prove to be a sensitive clinical indicator of the presence of melioidosis within a particular geographic area. Can J Microbiol, 1989 Apr, 35(4), 439 - 43 Isolation and preliminary characterization of a 2-chlorobenzoate degrading Pseudomonas; Sylvestre M et al.; Pseudomonas sp . strain B-300, which is able to utilize 2-chlorobenzoic acid, was isolated from a soil sample by enrichment culture . This strain was shown to grow on 2-chlorobenzoic acid and to completely degrade the substrate with concomitant chlorine ion release . Concentrations of 2-chlorobenzoic acid higher than 0.5% (w/v) were toxic to the cells . Our study also suggested that in the presence of glucose, 2-chlorobenzoic acid is converted to catechol or chlorocatechol; these are in turn transformed to muconic and chloromuconic acid, respectively, suggesting a repression by glucose of some of the degradation pathway enzymes . A similar scheme was already described for 3-chlorobenzoate degradation by pAC25 plasmid. Appl Environ Microbiol, 1989 Apr, 55(4), 946 - 52 Bacterial metabolism of hydroxylated biphenyls; Higson FK et al.; Isolates able to grow on 3- or 4-hydroxybiphenyl (HB) as the sole carbon source were obtained by enrichment culture . The 3-HB degrader Pseudomonas sp . strain FH12 used an NADPH-dependent monooxygenase restricted to 3- and 3,3'-HBs to introduce an ortho-hydroxyl . The 4-HB degrader Pseudomonas sp . strain FH23 used either a mono- or dioxygenase to generate a 2,3-diphenolic substitution pattern which allowed meta-fission of the aromatic ring . By using 3-chlorocatechol to inhibit catechol dioxygenase activity, it was found that 2- and 3-HBs were converted by FH23 to 2,3-HB, whereas biphenyl and 4-HB were attacked by dioxygenation . 4-HB was metabolized to 2,3,4'-trihydroxybiphenyl . Neither organism attacked chlorinated HBs . The degradation of 3- and 4-HBs by these strains is therefore analogous to the metabolism of biphenyl, 2-HB, and naphthalene in the requirement for 2,3-catechol formation. Mol Gen Mikrobiol Virusol, 1989 Apr, (4), 14 - 8 {The use of the plasmid pTH10 for isolating the donor strains of Pseudomonas mallei}; Ageeva NP et al.; The plasmid pTH10 was transfered by conjugation into the Pseudomonas mallei strains . An attempt to construct the donor strains using the widely known technique employing the homology between the plasmid and chromosome due to the transposon Tn1 carried by the plasmid was unsuccessful . Among the clones resistant to bacteriophage PRD1 the variants were selected with the supposed integration of the plasmid into the chromosome . The latter clones required the ability to transfer the auxotrophic chromosomal markers in conjugation after the repeated conjugational transfer of the plasmid pTH10 into them. Singapore Med J, 1989 Apr, 30(2), 205 - 7 Pseudomonas pseudomallei pneumonia with septicemia--case report; Wei SS et al.; A case of Pseudomonas pseudomallei pneumonia with septicemia is described . The onset was insidious with paucity of systemic symptoms except fever . Diabetes mellitus and alcoholism were associated problems . Initially blood cultures were negative but subsequently P . pseudomallei was isolated . The outcome was fatal . Unless diagnosed early and treated appropriately, patients often succumb to septicemic shock. Antibiot Khimioter, 1989 Apr, 34(4), 251 - 4 {Direct screening of antibiotics-siderophores of bacterial origin}; Smirnov VV et al.; Antifungal activity of 275 strains belonging to 15 species of Pseudomonas was studied with using media containing no iron or supplemented with 100 micrograms/ml of FeCl3 . 33 per cent of the cultures showed lower activity against phytopathogenic fungi in the presence of iron . Addition of this element did not influence the antifungal activity of phenazin and floroglucin derivatives isolated from Pseudomonas cultures . However, its addition markedly lowered the antifungal effect of some crude antibiotics and fluorescent pigments . A scheme for screening siderophore antibiotics with using Pseudomonas cultures is described. Kansenshogaku Zasshi, 1989 Apr, 63(4), 400 - 9 {Detection of inducible beta-lactamase in sputum--clinical studies on Pseudomonas respiratory infection}; Nakahama C et al.; To investigate the clinical incidence of inducible beta-lactamase, we measured the beta-lactamase activity in the sputum of 5 patients with chronic respiratory tract infection due to P . aeruginosa, by using the spectrophotometric method . During the piperacillin (PIPC) therapy given twice a day with a single dose of 2-3 g, sputum samples were collected every 2 hours for 3 days, and on the second day, two grams of Cefmetazole (CMZ) was added to PIPC therapy . The antibiotics concentration of each collected sputum samples were also measured by HPLC . In one out of 5 patients, no beta-lactamase activity in sputum was detected throughout the 3 days . However in three out of 5 patients, after the addition of CMZ to PIPC, the beta-lactamase activity significantly increased 2-3 times (max: 0.03 units/ml) that on PIPC alone, and gradually decreased on the 3rd day when PIPC was given alone . Then the peak concentration of PIPC with the addition of CMZ decreased to 38-73%, compared with that of PIPC alone . These findings were supported by the fact that CMZ showed a high in vitro inducer activity against the isolates from the sputum . In the remaining one patient, high beta-lactamase activity (mean: 0.16 units/ml) and no antibiotics concentration was detected to be constant throughout the 3 days, and it was confirmed for the reason that one of the isolates constitutively produced large amounts of beta-lactamase . These results suggest that inducible and constitutive beta-lactamase would clinically cause undesirable effects in the treatment by some beta-lactams and have a possibility of indirect pathogenesis. Biochimie, 1989 Apr, 71(4), 551 - 7 Kinetic isotope effect and the presteady-state kinetics of the reaction catalyzed by the bacterial formate dehydrogenase; Tishkov VI et al.; The primary kinetic isotope effect of the reaction catalyzed by NAD+-dependent formate dehydrogenase (EC 1.2.1.2.) from the methylotrophic bacterium Pseudomonas sp . 101 has been studied . Analysis of the ratios HVm/DVm and H(Vm/KM)/D(Vm/KM) in the pH range 6.1-7.9 showed that the transfer of hydride ion in ternary enzyme-substrate complex is a limiting step of the reaction, and the formate binding to the binary complex (formate dehydrogenase + NAD+) reached equilibrium when the pH of the medium was increased . An approach has been developed to determine the elementary constants of substrate association (kon) and dissociation (koff) at the stages of the binary--ternary enzyme-substrate complexes for the random equilibrium 2-substrate kinetic mechanism . The kon and koff values obtained for the bacterial formate dehydrogenase by using the proposed approach for NAD+ were (4.8 +/- 0.8)*10(5)M-1s-1 and (90 +/- 10) s-1, and for formate (2.0 +/- 1.0)*10(4) M-1s-1 and (60 +/- 20) s-1, respectively. Antibiot Khimioter, 1989 Apr, 34(4), 282 - 6 {Effectiveness of tobramycin and immunologic preparations in experimental Pseudomonas infection}; Minukhin VV et al.; Certain indices of immunity were studied in mice with burn sepsis due to P . aeruginosa during their treatment with tobramycin (Tb) alone or in combination with immunological drugs . The most significant stimulation of the phagocytic function of peritoneal macrophages was observed when Tb was used in combination with polyvalent corpuscular vaccine of P . aeruginosa . When Tb was used alone or in combination with hyperimmune plasma of P . aeruginosa there was observed close correlation between the phagocytic index and the levels of cyclic adenosine-3',5'-monophosphate in them . Therapy of P . aeruginosa infection with the antibiotic and immunological drugs resulted in much higher levels of agglutinine antibodies in blood serum of the mice than the therapy with Tb alone. J Cell Physiol, 1989 Apr, 139(1), 51 - 7 An epidermal growth factor-ricin A chain (EGF-RTA)-resistant mutant and an epidermal growth factor-Pseudomonas endotoxin (EGF-PE)-resistant mutant have distinct phenotypes; Banker DE et al.; H2Oe12 is a mutant HeLa cell line selected for resistance to the toxicity of a chimeric protein conjugate composed of epidermal growth factor (EGF) and the toxic A chain of ricin (RTA) . ET-28 is a mutant KB cell line selected for resistance to the toxicity of a chimeric protein conjugate composed of EGF and Pseudomonas exotoxin (PE) . In this report we describe the presence or absence, in these mutants, of cross-resistance to the two toxic conjugates and the effects of ammonium chloride, leupeptin, and adenovirus cotreatments on toxin efficacies . ET-28 cells, the EGF-PE-resistant cells, are resistant to both EGF-PE and EGF-RTA . In contrast, H2Oe12 cells, the EGF-RTA-resistant cells, are as sensitive to EGF-PE toxicity as are their parent HeLa cells . Ammonium chloride cotreatment substantially reduces the resistance of H2Oe12 cells to EGF-RTA but has little or no effect on the resistance of ET-28 cells to either EGF-RTA or EGF-PE . Leupeptin has no effect on the toxicity of either chimeric conjugate on any of the four cell lines, effect on the toxicity of either chimeric conjugate on any of the four cell lines, despite its demonstrated ability to inhibit cellular degradation of EGF . In contrast, adenovirus cotreatment enhances the toxicity of EGF-RTA and EGF-PE on all cells tested, and completely nullifies the relative resistance of H2Oe12 and ET-28 cells to these toxic conjugates . H2Oe12 and ET-28 cells appear to be altered in distinct, possibly endosomal, functions. Cancer, 1989 Mar 15, 63(6), 1084 - 91 Activity of pirarubicin (4'-0-tetrahydropyranyladriamycin) in malignant mesothelioma; Sridhar KS et al.; Eight patients with diffuse malignant mesothelioma of the pleura or peritoneum, previously untreated with chemotherapy, were treated with a new anthracycline 4'-0-tetrahydropyranyladriamycin (pirarubicin) . Pirarubicin was given intravenously at the rate of 5 mg per minute, at doses ranging from 35 to 70 mg/m2 once every 21 days . On clinical evaluation, one patient had complete response lasting 4 months . On second-look laparotomy residual tumor was found and she was labelled a partial responder and changed to alternate chemotherapy . Another patient had a partial response of recurrent chest wall tumors lasting 11 months . A third patient had a partial response lasting 4+ months of a pleural-based tumor and resolution of pleural effusion . After the fifth course of chemotherapy, he developed severe granulocytopenia, pseudomonas sepsis, shock, and renal failure . Despite recovery of blood counts to normal within 3 days, renal failure proved fatal . Autopsy revealed only fibrosis and no gross or microscopic evidence of malignant mesothelioma . A fourth patient had improvement in evaluable disease lasting about 4 months; and the remaining four had stable disease for at least 2 months each . The authors conclude that, whenever feasible, noninvasive clinical assessment of tumor response should be supplemented by surgical-pathologic evaluation . Pirarubicin is active in malignant mesothelioma . This is the first report documenting complete tumor eradication after chemotherapy in an adult with malignant mesothelioma. Appl Environ Microbiol, 1989 Mar, 55(3), 767 - 70 Isolation and identification of Pseudomonas spp . from Schirmacher Oasis, Antarctica; Shivaji S et al.; Ten cultures of Pseudomonas spp . were established from soil samples collected in and around a lake in Antarctica . Based on their morphology, biochemical and physiological characteristics, and moles percent G + C of their DNA, they were identified as P . fluorescens, P . putida, and P . syringae . This is the first report on the identification of Pseudomonas spp . from continental Antarctica. J Bacteriol, 1989 Mar, 171(3), 1760 - 2 Structure of an acidic exopolysaccharide of Pseudomonas marginalis HT041B; Osman SF et al.; The exopolysaccharide of Pseudomonas marginalis HT041B has been characterized as a 1,3-linked galactoglucan in which galactose and glucose are in the alpha- and beta-anomeric configurations, respectively . The polysaccharide is substituted with pyruvate at the 4 and 6 positions of galactose and with succinic acid at either the 2 or 4 position of glucose . This polysaccharide has been given the trivial name marginalan. Mikrobiol Zh, 1989 Mar-Apr, 51(2), 32 - 8 {Immunologic and structural studies of lipopolysaccharides from Pseudomonas cepacia}; Soldatkina A et al.; O-serotyping of 30 Pseudomonas cepacia strains isolated from the soil and rhizosphere of different plant species in the territory of the USSR has been performed using 15 O-typing antisera according to the Heidt and Nakamura schemes . It is suggested to introduce two new O-serogroups (serogroups K and L) into the available P . cepacia classification scheme . They are most often met among the P . cepacia strains in different geographical areas of the USSR simultaneously with serogroups 2 (G) and 1 (D) . To elucidate the molecular principles of serological inhomogeneity of the species the immunochemical studies of lipopolysaccharides of a number of P . cepacia strains have been conducted and the structure has been determined for repeating links of O-specific polysaccharides of P . cepacia strains attributed to 4 Nakamura serogroups, 3 Heidt serogroups, to serogroups K and L, as well as for certain strains from the collection of the Institute of Microbiology and Virology of the Ukr . SSR Academy of Sciences. J Bacteriol, 1989 Mar, 171(3), 1333 - 9 Cloning and nucleotide sequence of the gene (amyP) for maltotetraose-forming amylase from Pseudomonas stutzeri MO-19; Fujita M et al.; The gene (amyP) coding for maltotetraose-forming amylase (exo-maltotetraohydrolase) of Pseudomonas stutzeri MO-19 was cloned . Its nucleotide sequence contained an open reading frame coding for a precursor (547 amino acid residues) of secreted amylase . The precursor had a signal peptide of 21 amino acid residues at its amino terminus . An extract of Escherichia coli carrying the cloned amyP had amylolytic activity with the same mode of action as the extracellular exo-maltotetraohydrolase obtained from P . stutzeri MO-19 . A region in the primary structure of this amylase showed homology with those of other amylases of both procaryotic and eucaryotic origins . The minimum 5' noncoding region necessary for the expression of amyP in E . coli was determined, and the sequence of this region was compared with those of Pseudomonas promoters. Biochem Int, 1989 Mar, 18(3), 573 - 80 Purification and some properties of a 2Fe ferredoxin in Pseudomonas ovalis; Ohmori D et al.; A {2Fe-2S} ferredoxin was found in Pseudomonas ovalis which was grown in a medium supplemented with glucose and ammonium sulfate . The molecular weight of the 2Fe ferredoxin was estimated to be 13,000 . It contained 2.2 gramatoms of non-heme iron and 2.3 gramatoms of acid-labile sulfur per mole protein . The absorption and circular dichroism spectra were characteristic of those of {2Fe-2S} type ferredoxins, especially adrenodoxin and putidaredoxin . The electron paramagnetic resonance spectrum of the reduced protein showed an axial symmetry (g = 2.020, g = 1.939) . The amino acid composition was determined. J Bacteriol, 1989 Mar, 171(3), 1763 - 6 Nucleotide sequence of IS492, a novel insertion sequence causing variation in extracellular polysaccharide production in the marine bacterium Pseudomonas atlantica; Bartlett DH et al.; The complete nucleotide sequence of insertion element IS492, which causes reversible inactivation of extracellular polysaccharide production in the marine bacterium Pseudomonas atlantica, is presented . Insertion of IS492 results in the EPS- phenotype, and excision results in restoration of EPS+ . DNA sequencing of the site of insertion in the eps locus showed that insertion of IS492 generates a 5-base-pair repeat and that its excision is precise . IS492 is 1,202 nucleotides in length and contains one large open reading frame encoding a protein of 318 amino acids, a candidate for transposition function . No similarity between IS492 and other transposable elements has been found . Unlike the situation with other insertion sequences, no direct or inverted repeats exist at the termini of IS492. Kansenshogaku Zasshi, 1989 Mar, 63(3), 195 - 202 {Investigations on the etiology of sepsis by using experimental mouse model with leukocytopenia . 1 . Influence of antibiotics}; Yamaguchi K et al.; Investigation on the etiology of septicemia occurring among the immunocompromised patients was performed by using experimental model of the mouse with leukocytopenia . The ddY conventional mice of 4 weeks of age were inoculated with cyclophosphamide (CPM) intraperitoneally 3 to 5 times every other day with a dose of 3 mg/mouse once to make an agranulocytic status . Then, intraperitoneal administrations of various antibiotic regimens consisting of ampicillin (ABPC) alone, ABPC + ceftazidime (CAZ), ABPC + CAZ + cloxacillin (MCIPC), ABPC + CAZ + MCIPC + minocycline (MINO) and saline as a control to these immunosuppressed mice were begun once every day for 10 days after the second inoculation of CPM . The mortality rate of the mice given saline as a control was very high with a frequency of 43.3% and there were significant differences between the saline group and another antibiotic groups other than ABPC (p less than 0.01) . On the other hand, the mortality rate of the group given APBC showed the highest rate of 70% and it was significantly higher than that of the saline control group (p less than 0.05) . The main cause of most of the dead mice was septicemia due to P . aeruginosa and which were isolated from the feces of all these mice and serotype of the strains isolated from the heart blood and feces in the same host corresponded to each other . Moreover, intestinal bacterial flora in mice treated by saline and ABPC which highly showed Pseudomonas sepsis, was occupied dominantly by P . aeruginosa, although P . aeruginosa was not detectable from the experimental environments.(ABSTRACT TRUNCATED AT 250 WORDS) J Trauma, 1989 Mar, 29(3), 284 - 91 Ibuprofen plus prostaglandin E1 in a septic porcine model of adult respiratory distress syndrome; Davies EA et al.; Prostaglandin manipulation has been shown to improve pulmonary dysfunction in animal models of acute respiratory distress syndrome . Using our previously reported porcine model of Pseudomonas-induced respiratory failure, we examined the therapeutic effects of a vasodilating prostaglandin, PGE1, and a reversible cyclooxygenase inhibitor, ibuprofen . Forty-two animals were randomized to seven groups: I--ibuprofen; II--PGE1; III--ibuprofen + PGE1; IV--Pseudomonas + ibuprofen; V--Pseudomonas + PGE1; VI--Pseudomonas + ibuprofen + PGE1; VII--Pseudomonas . Ibuprofen significantly improved pulmonary vasoconstriction, pulmonary hypertension, and hypoxemia, as well as increased survival slightly . PGE1 had no effect on pulmonary dysfunction, but prevented the rise in systemic vascular resistance that occurred in untreated, infected animals and animals treated with ibuprofen alone . Combination therapy improved stroke volume index, a measure of nonpulmonary organ function. J Bacteriol, 1989 Mar, 171(3), 1725 - 32 Cloning and expression in Escherichia coli of Pseudomonas strain LB400 genes encoding polychlorinated biphenyl degradation; Mondello FJ; Pseudomonas strain LB400 is able to degrade an unusually wide variety of polychlorinated biphenyls (PCBs) . A genomic library of LB400 was constructed by using the broad-host-range cosmid pMMB34 and introduced into Escherichia coli . Approximately 1,600 recombinant clones were tested, and 5 that expressed 2,3-dihydroxybiphenyl dioxygenase activity were found . This enzyme is encoded by the bphC gene of the 2,3-dioxygenase pathway for PCB-biphenyl metabolism . Two recombinant plasmids encoding the ability to transform PCBs to chlorobenzoic acids were identified, and one of these, pGEM410, was chosen for further study . The PCB-degrading genes (bphA, -B, -C, and -D) were localized by subcloning experiments to a 12.4-kilobase region of pGEM410 . The ability of recombinant strains to degrade PCBs was compared with that of the wild type . In resting-cell assays, PCB degradation by E . coli strain FM4560 (containing a pGEM410 derivative) approached that of LB400 and was significantly greater than degradation by the original recombinant strain . High levels of PCB metabolism by FM4560 did not depend on the growth of the organism on biphenyl, as it did for PCB metabolism by LB400 . When cells were grown with succinate as the carbon source, PCB degradation by FM4560 was markedly superior to that by LB400. Biochim Biophys Acta, 1989 Feb 28, 973(2), 302 - 7 Thermodynamic efficiency of bacterial growth calculated from growth yield of Pseudomonas oxalaticus OX1 in the chemostat; Rutgers M et al.; In order to determine the thermodynamic efficiency of bacterial growth, Pseudomonas oxalaticus OX1 was grown in carbon-limited continuous cultures . 11 different carbon sources, ranging from oxalate (most oxidised component) to ethanol (most reduced component), were used as limiting substrate in these experiments . From the experimental yield values (expressed as C-mol dry weight produced per C-mol carbon substrate consumed) the thermodynamic efficiencies were calculated . On substrates more reduced than biomass (such as ethanol and glycerol) the thermodynamic efficiency of growth of P . oxalaticus was negative but it reached a maximum of 23 +/- 3% with substrates with a degree of reduction of 3 (citrate) and lower . The actual concentrations of the components involved were incorporated into the calculations but this affected the overall thermodynamic efficiency only to a small extent . This result strengthens the conclusion of Westerhoff et al . (Westerhoff, H.V., Hellingwerf, K.J . and Van Dam, K . (1983) Proc . Natl . Acad . Sci . 80, 305-309) that bacteria have been optimised towards a theoretical thermodynamic efficiency of 24%, corresponding with maximisation of growth rate at optimal efficiency, with highly oxidised substrates. Biochem J, 1989 Feb 15, 258(1), 193 - 8 The structural basis for neutrophil inactivation of C1 inhibitor; Pemberton PA et al.; Limited proteolysis of C1 inhibitor (C1-INH) by neutrophil elastase, Pseudomonas elastase and snake venoms resulted in initial cleavage within the molecule's N-terminus followed by further cleavage within the molecule's C-terminally placed reactive centre . N-Terminal proteolysis occurred at peptide bonds 14-15, 36-37 and 40-41 . This had no effect on either the inhibitory activity or the heat-stability of C1-INH . Proteolysis within the reactive centre occurred at peptide bonds 439-440, 440-441, 441-442 and 442-443 . Cleavage at any one of these sites inactivated C1-INH and conferred enhanced heat-stability upon a previously heat-labile molecule . Released neutrophil proteinases also cleaved and inactivated C1-INH, suggesting that they may physiologically regulate C1-INH during inflammatory episodes. J Biol Chem, 1989 Feb 5, 264(4), 2379 - 84 Structure and function relationship of Pseudomonas exotoxin A . An immunochemical study; Hwang J et al.; We have raised antisera against Pseudomonas exotoxin A (PE) and domains Ia and III to study the structure-function relationships of PE . Anti-PE antibody (AbPE) was shown to abolish the ADP-ribosylation activity of PE . However, neither antidomain Ia antibody nor antidomain III antibody inhibited the ADP-ribosylation activity of PE . This suggests that the inhibition of ADP-ribosylation by AbPE results from the binding of AbPE to the region between domains Ia and III . Since the binding of AbPE to PE did not inhibit NAD hydrolysis in the absence of elongation factor 2, the inhibitory effect of AbPE on ADP-ribosylation may be due to steric hindrance rather than a direct action on the catalytic function . Thus, the interface between domain Ia and III may be the site of entry of elongation factor 2 during ADP-ribosylation . The antibodies were also used to study both the inhibitory effects of PE on protein synthesis and its cytotoxic activity . Either AbPE or antidomain Ia antibody, but not antidomain III antibody, was able to reverse the inhibition of protein synthesis by PE and to block its cytotoxicity . In addition, rabbits immunized with domain Ia acquired tolerance against 100 micrograms of PE injected subcutaneously . These results suggest that domain Ia is the cell-binding domain of PE and may be used for vaccination against PE-mediated diseases. J Clin Microbiol, 1989 Feb, 27(2), 270 - 3 Recovery of Pseudomonas gladioli from respiratory tract specimens of patients with cystic fibrosis; Christenson JC et al.; Pseudomonas gladioli was isolated from 11 patients with cystic fibrosis . It resembled Pseudomonas cepacia on the selective and differential medium OFPBL, producing yellow colonies after 48 to 72 h of incubation . Isolates were characterized biochemically, by DNA hybridization, and by cellular fatty acid analysis . A review of the clinical status of selected patients colonized by P . gladioli did not reveal any apparent association of this organism with infectious complications of cystic fibrosis . Thus, the clinical implications may differ depending on which of these two closely related species is reported by laboratories . Determination of the fatty acid profile of isolates by gas chromatography may be a useful adjunct to biochemical characterization as a means of identification . In contrast to P . cepacia, most isolates of P . gladioli contained 3-OH C10:0 fatty acid under the growth conditions used. J Bacteriol, 1989 Feb, 171(2), 1223 - 4 Spermidine synthesis by Pseudomonas sp . strain Kim, previously reported to lack this polyamine; Rosano CL et al.; Pseudomonas sp . strain Kim has previously been reported to be the only known naturally occurring organism lacking spermidine . We now show that it synthesizes this polyamine . The apparent lack of intracellular levels of spermidine results from an efficient conversion of spermidine to putrescine and hydroxyputrescine. Kyobu Geka, 1989 Feb, 42(2), 141 - 4 {Pseudomonas cepacia endocarditis successfully treated by surgery}; Saitoh Y et al.; The patient was a 3-year-old female with coarctation of the aorta complicated by ventricular septal defect and mitral regurgitation . She underwent surgery for coarctation of the aorta at 7 months of age . We performed direct closure using a pledget for ventricular septal defect and valvoplasty with annuloplasty for mitral regurgitation . Infective endocarditis due to pseudomonas cepacia developed 3 months after the surgery, and echocardiography revealed vegetation in the ventricular septum and anterior leaflet of the mitral valve . After treatment with antibiotics, the second open heart surgery involving removal of the pledget used in the previous operation, reclosure of the ventricular septal defect, and mitral valve replacement was performed . The patient is healthy without recurrence of infective endocarditis 2 years and 2 months after the surgery. DICP, 1989 Feb, 23(2), 151 - 2 Precipitation of benzodiazepine withdrawal following sudden discontinuation of midazolam; Finley PR et al.; Midazolam hydrochloride is an ultra-short acting benzodiazepine recently approved by the Food and Drug Administration for anesthesia induction and preoperative sedation . Frequently, midazolam is also used as an injection or infusion for the treatment of agitation in ventilator-dependent patients . A 53-year-old man underwent a gastrojejunostomy and was later intubated following the development of pseudomonal pneumonia . Midazolam was initiated in an effort to resolve his agitation and the patient continued to receive frequent bolus injections, averaging 22 mg/d over 21 days . Approximately eight hours after midazolam was abruptly discontinued, the patient became increasingly anxious and developed somatic complaints felt to be consistent with benzodiazepine withdrawal syndrome . Symptoms rapidly abated upon the reintroduction of midazolam and the drug was ultimately tapered over a period of four days and discontinued without further incident . Implications derived from the association of long-term midazolam therapy with benzodiazepine withdrawal syndrome are discussed. Appl Environ Microbiol, 1989 Feb, 55(2), 372 - 9 Degradation of p-chlorotoluene by a mutant of Pseudomonas sp . strain JS6; Haigler BE et al.; Pseudomonas sp . strain JS6 grows on chlorobenzene, p-dichlorobenzene, or toluene as a sole source of carbon and energy . It does not grow on p-chlorotoluene (p-CT) . Growth on glucose in the presence of p-CT resulted in the accumulation of 4-chloro-2,3-dihydroxy-1-methylbenzene (3-chloro-6-methylcatechol), 4-chloro-2,3-dihydroxy-1-methylcyclohexa-4,6-diene (p-CT dihydrodiol), and 2-methyl-4-carboxymethylenebut-2-en-4-olide (2-methyl dienelactone) . Strain JS21, a spontaneous mutant capable of growth on p-CT, was isolated from cultures of strain JS6 after extended exposure to p-CT . In addition to growing on p-CT, JS21 grew on all of the substrates that supported growth of the parent strain, including p-dichlorobenzene, chlorobenzene, benzene, toluene, benzoate, p-hydroxybenzoate, phenol, and ethylbenzene . The pathway for