Pseudomonas

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 degradation of p-CT by JS21 was investigated by respirometry, isolation of intermediates, and assay of enzymes in cell extracts . p-CT was converted to 3-chloro-6-methylcatechol by dioxygenase and dihydrodiol dehydrogenase enzymes . 3-Chloro-6-methylcatechol underwent ortho ring cleavage catalyzed by a catechol 1,2-dioxygenase to form 2-chloro-5-methyl-cis,cis-muconate, which was converted to 2-methyl dienelactone . A dienelactone hydrolase converted 2-methyl dienelactone to 2-methylmaleylacetic acid . Preliminary results indicate that a change in wild-type induction patterns allows JS21 to grow on p-CT.

Proc Natl Acad Sci U S A, 1989 Feb, 86(3), 1008 - 12
Cardiac allograft survival in mice treated with IL-2-PE40; Lorberboum-Galski H et al.; IL-2-PE40 is a chimeric protein composed of human interleukin 2 (IL-2) genetically fused to the amino terminus of a modified form of Pseudomonas exotoxin lacking its cell recognition domain . IL-2-PE40, which is extremely cytotoxic to IL-2 receptor-positive cells, was examined for its ability to prevent graft rejection in mice in which activation of T cells is prominent . We demonstrate that intraperitoneally administered IL-2-PE40 specifically and significantly prolongs the survival of vascularized heart allografts in mice . The chimeric toxin, IL-2-PE40, offers an alternative approach to the treatment of autoimmune diseases and transplant rejection in humans.

J Bacteriol, 1989 Feb, 171(2), 1002 - 9
Cotranscription of genes encoding indoleacetic acid production in Pseudomonas syringae subsp . savastanoi; Palm CJ et al.; Indoleacetic acid (IAA) production by the plant pathogen Pseudomonas syringae subsp . savastanoi is essential for tumor formation on olive and oleander . The bacterium produces IAA from tryptophan in reactions catalyzed by tryptophan monooxygenase and indoleacetamide hydrolase . The genetic determinants are, respectively, iaaM and iaaH . In oleander isolates, the genes encoding the IAA biosynthetic enzymes are located on a plasmid; in olive isolates, the genes occur on the chromosome . The IAA genes from the oleander isolate strain EW2009 are located within a 4-kilobase (kb) segment of the 52-kb plasmid pIAA1 . Escherichia coli strains harboring a recombinant plasmid, pCJP3, which contains this 4-kb fragment, excreted IAA into culture media, and crude cell extracts had both tryptophan monooxygenase and indoleacetamide hydrolase activity . In vitro coupled transcription-translation of pCJP3 demonstrated that this fragment coded for proteins of 62 and 47 kilodaltons which correspond to tryptophan monooxygenase and indoleacetamide hydrolase, respectively . Expression of these genes was dependent upon a vector promoter in pCJP3 . However, in the absence of a vector promoter, E . coli containing recombinant plasmids with additional pIAA1 DNA in front of iaaM had high levels of tryptophan monooxygenase . Northern (RNA) hybridization experiments verified that iaaM and iaaH are cotranscribed as a portion of a ca . 4- to 5-kb transcript in vivo . Southern hybridization experiments with IAA plasmids from different oleander strains of P . syringae subsp . savastanoi revealed that all IAA plasmids contained a region of at least 10 kb of homology, with the IAA genes at one end . Repetitive DNA and a copy of IS51 were found at the end of this region of homology.

J Bacteriol, 1989 Feb, 171(2), 807 - 12
Plasmid-mediated production of the phytotoxin coronatine in Pseudomonas syringae pv . tomato; Bender CL et al.; Pseudomonas syringae pv . tomato PT23.2 produces the chlorosis-inducing phytotoxin coronatine . Thirty-eight chlorosis-defective mutants of PT23.2 were previously generated by using the transposon Tn5 . Five mutants contained Tn5 insertions in the indigenous plasmid pPT23A; the remaining 33 mutants either were missing pPT23A (29 mutants) or contained deletions in this plasmid (4 mutants) . These results suggested that pPT23A was involved in coronatine production in strain PT23.2 . This plasmid was introduced into P . syringae pv . syringae PS61, which does not produce coronatine . A bioassay for coronatine suggested that PS61(pPT23A) transconjugants were able to make this phytotoxin . In a chemical analysis, organic acids were isolated from PT23.2, PS61, and the transconjugant PS61(pPT23A); these were derivatized to their methyl esters and analyzed by gas chromatography . The derivatized organic acids extracted from PT23.2 and PS61(pPT23A) contained peaks that corresponded to coronafacic acid, coronafacoylvaline, and coronatine, but these were absent in the extracts from the wild-type strain PS61 . The identification of these components was confirmed by combined gas chromatography-mass spectrophotometry . Therefore, the acquisition of pPT23A by PS61 resulted in biosynthesis of coronafacic acid, coronafacoylvaline, and coronatine, clearly demonstrating the involvement of pPT23A in coronatine production in P . syringae pv . tomato.

Biochemistry, 1989 Jan 24, 28(2), 580 - 5
The acid-triggered entry pathway of Pseudomonas exotoxin A; Farahbakhsh ZT et al.; In this study we examined the pH requirements and reversibility of early events in the Pseudomonas toxin entry pathway, namely, membrane binding, insertion, and translocation . At pH 7.4, toxin binding to vesicles and insertion into the bilayer are very inefficient . Decreasing the pH greatly increases the efficiencies of these processes . Acid-treated toxin exhibits pH 7.4 binding and insertion levels . This indicates that hydrophobic regions that become exposed upon toxin acidficiation become buried again when the pH is raised to 7.4 . In contrast, the change in toxin conformation that occurs upon membrane binding is irreversible . Returning samples to pH 7.4, incubation with excess toxin, or dilution with buffer up to 1000-fold leads to very little loss of bound toxin . Bound toxin exhibits an extremely high susceptibility to trypsin compared to free toxin (at both pH 4 and pH 7.4) . At pH 4, membrane-associated toxin slowly proceeds to a trypsin-protected state; neutralization halts this process . At low pH, toxin was found to bind and insert into DMPC vesicles very efficiently at temperatures both above and below 23 degrees C, the lipid melting point . With fluid targets, the proportion of bound toxin that was photolabeled from within the bilayer peaked rapidly and then decreased with time . With frozen targets, the efficiency of photolabeling peaked but then remained fairly constant.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochim Biophys Acta, 1989 Jan 23, 1001(1), 60 - 7
Polar lipids and fatty acids of Pseudomonas cepacia; Cox AD et al.; The polar lipids and fatty acids produced by the reference strains for seven different O serogroups of Pseudomonas cepacia have been identified . Similar results were obtained for all strains . Contrary to a previous report, the only significant phospholipids in this species are phosphatidylethanolamine and bis(phosphatidyl)glycerol, which contributed 57-83% and 17-43%, respectively, of the total lipid phosphorus . The former lipid was found as two chromatographically distinct fractions . In the less polar fraction and in bis(phosphatidyl)glycerol, the major fatty acids were hexadecanoic acid, cis-9,10-methylenehexadecanoic acid, cis-octadec-11-enoic acid, and cis-11,12-methyleneoctadecanoic acid . In the more polar fraction of phosphatidylethanolamine, the fatty acid in one position is a 2-hydroxy acid, mainly 2-hydroxyhexadecenoic acid, 2-hydroxyhexadecanoic acid, 2-hydroxyoctadecenoic acid, or 2-hydroxymethyleneoctadecanoic acid . Compared with other phospholipids, this fraction of phosphatidylethanolamine was depleted in cis-9,10-methylenehexadecanoic acid . Each strain also produced two ornithine amide lipids . In the major lipid, 3-hydroxyhexadecanoic acid was amide-bound to the alpha-amino group and was itself probably esterified by a 2-hydroxy acid, mainly 2-hydroxyoctadecenoic acid or the derived cyclopropane acid . In the minor ornithine amide lipid, the ester-bound acids were mainly methyleneoctadecanoic acid and hexadecanoic acid . The unusual lipid profiles of P . cepacia are of chemotaxonomic interest.

Biochemistry, 1989 Jan 10, 28(1), 149 - 59
Kinetic and ultraviolet spectroscopic studies of active-site mutants of delta 5-3-ketosteroid isomerase; Kuliopulos A et al.; delta 5-3-Ketosteroid isomerase (EC 5.3.3.1) of Pseudomonas testosteroni promotes the highly efficient isomerization of delta 5-3-ketosteroids to delta 4-3-ketosteroids by means of a direct and stereospecific transfer of the 4 beta-proton to the 6 beta-position, via an enolic intermediate . An acidic residue responsible for the protonation of the 3-carbonyl function of the steroid and a basic group concerned with the proton transfer have been implicated in the catalytic mechanism . Recent NMR studies with a nitroxide spin-labeled substrate analogue have allowed positioning of the steroid into the 2.5-A X-ray crystal structure of the enzyme {Kuliopulos, A., Westbrook, E.M., Talalay, P., & Mildvan, A.S . (1987) Biochemistry 26, 3927-3937}, thereby corroborating the approximate location of the steroid binding site deduced from a difference Fourier X-ray diffraction map of the 4-(acetoxymercuri)estradiol-isomerase complex {Westbrook, E.M., Piro, O.E., & Sigler, P.B . (1984) J . Biol . Chem . 259, 9096-9103} . The steroid lies in a hydrophobic cavity near Asp-38, Tyr-14, and Tyr-55 . In order to assess the role of these amino acid residues in catalysis, the gene for isomerase was cloned, sequenced, and overexpressed in Escherichia coli {Kuliopulos, A., Shortle, D., & Talalay, P . (1987) Proc . Natl . Acad . Sci . U.S.A . 84, 8893-8897}, and the following mutants were prepared: Asp-38 to asparagine (D38N) and Tyr-14 and Tyr-55 to phenylalanine (Y14F and Y55F, respectively) . The kcat value of the D38N mutant enzyme is 10(5.6)-fold lower than that of the wild-type enzyme, suggesting that Asp-38 functions as the base which abstracts the 4 beta-proton of the steroid in the rate-limiting step . Threefold lower Km values in all mutants indicate tighter binding of the substrate to the more hydrophobic sites . In comparison with the wild-type enzyme, the Y55F mutant shows only a 4-fold decrease in kcat while the Y14F mutant shows a 10(4.7)-fold decrease in kcat, suggesting that Tyr-14 is the general acid . The red shift of the ultraviolet absorption maximum of the competitive inhibitor 19-nortestosterone from 248 to 258-260 nm, which occurs upon binding to the wild-type enzyme {Wang, S.F., Kawahara, F.S., & Talalay, P . (1963) J . Biol . Chem . 238, 576-585}, is mimicked in strong acid . This spectral shift was also observed with the D38N and Y55F mutants, but not on binding of the steroid to the Y14F mutant.(ABSTRACT TRUNCATED AT 400 WORDS)

Eur J Biochem, 1989 Jan 2, 178(3), 751 - 62
Nitrous oxide reductase from Pseudomonas stutzeri . Redox properties and spectroscopic characterization of different forms of the multicopper enzyme; Riester J et al.; The oxidation-reduction and spectroscopic properties of various forms of nitrous oxide reductase from Pseudomonas stutzeri were investigated . The high-activity form I of the enzyme (purple, 8 Cu, Mr 140,000) was reduced by a large variety of cationic, anionic and photochemically generated agents . The blue form III was the only product found in these experiments under anaerobic conditions . Reductive (dithionite) and oxidative (ferricyanide) titrations showed that the conversion of the purple form I to the blue species III was fully reversible in the absence of dioxygen . Two kinetically different phases of the reaction of form I with a stoichiometric amount of dithionite (1e- -equivalent/Cu) were detected: in the fast phase (seconds), the purple chromophore with lamba max at 540 nm disappeared almost completely, whereas in the slower phase (minutes) the blue species with lambda max around 650 nm was generated . Irrespective of the nature of the reductant the blue species did not react even at large excess of reductant . It was reoxidized by ferricyanide, hydrogen peroxide and nitric oxide . A new, catalytically inactive derivative of nitrous oxide reductase (form V, 2 Cu, Mr 140,000) was isolated from a transposon Tn5-induced mutant with defective chromophore biosynthesis . The pink color of the mutant protein faded almost completely after addition of 0.5e- -equivalent/Cu . In this case no blue species was found, similar to earlier observations for the regenerated, catalytically inactive protein . Varying with the sample and the pH, 50-80% of the total copper of form I was in an electron-paramagnetic-resonance-(EPR)-silent state as compared to 47% in the mutant protein . The broad, featureless EPR signal recorded at 9.32 GHz for the blue, reduced form III of nitrous oxide reductase represented approximately 20% of the total copper . For the blue species no resolution enhancement was achieved at 34 GHz . At this frequency both forms I and V showed similar EPR signals with apparent g-values at 2.16 and 1.99 . At 9.32 GHz, form V had an EPR signal with gII at 2.18, AII = 3.55 mT (4 or 5 lines, in contrast to form I) and gI at 2.03 . Above 100 K the splitting of the gII region into seven equidistant lines in the EPR signal of the high-activity form I and the hyperfine structure of the perpendicular transition disappeared . Carbon monoxide and nitric oxide, but not nitrous oxide, had marked effects on the spectroscopic properties of the purple form I . Marked effects were also obtained for the exogenous ligands nitrite, azide, cyanate and thiocyanate.(ABSTRACT TRUNCATED AT 400 WORDS)

J Bacteriol, 1989 Jan, 171(1), 602 - 5
Sequence analysis of the agrA gene encoding beta-agarase from Pseudomonas atlantica; Belas R; The nucleotide sequence of the agrA gene encoding an extracellular beta-agarase of Pseudomonas atlantica was determined . An open reading frame of 1,515 nucleotides which corresponded to agrA was found . The nucleotide sequence predicts a primary translation product of 504 amino acids and Mr 57,486 . Comparison of the deduced amino acid sequences of beta-agarase from P . atlantica and the extracellular beta-agarase from Streptomyces coelicolor A3(2) suggests that these proteins share several domains in common.

J Clin Microbiol, 1989 Jan, 27(1), 217 - 8
Flavimonas oryzihabitans (Pseudomonas oryzihabitans; CDC group Ve-2): an emerging pathogen in peritonitis related to continuous ambulatory peritoneal dialysis?
Bendig JW, Mayes PJ, Eyers DE, Holmes B, Chin TT.
A case of peritonitis caused by Flavimonas oryzihabitans (Pseudomonas oryzihabitans; CDC group VE-2) in a patient on continuous ambulatory peritoneal dialysis is reported . This is the seventh case of infection caused by this organism reported in the English literature and the third reported case of continuous ambulatory peritoneal dialysis-related peritonitis caused by this organism; it is the first case of infection of any kind caused by this organism in England.

J Bacteriol, 1989 Jan, 171(1), 483 - 7
Physiological studies of the regulation of beta-lactamase expression in Pseudomonas maltophilia; Rosta S et al.; The kinetics of beta-lactamase induction in Pseudomonas maltophilia IID1275/873 were investigated . Upon induction with beta-lactam antibiotics, a correlation was seen between the increase in specific beta-lactamase activity and the generation time, as well as the concentration of inducer in the medium . The specific beta-lactamase activity increased slowly within the first 0.5 generation and then more rapidly; it decreased regularly after about 2 generations of growth in the presence of inducer . This decrease could presumably be attributed to the continuous breakdown of inducer by beta-lactamases in the culture medium . In a chemostat culture with continuous supply of fresh inducer-containing medium, the specific beta-lactamase activity could be stabilized at a high level over several generations . Removal of the beta-lactam after a certain induction time showed that a short exposure of the bacteria to inducer caused induction kinetics comparable to those resulting from continuous exposure of the cells to inducer . The two beta-lactamases of P . maltophilia, L1 and L2, were induced simultaneously under various experimental conditions.

Am J Pediatr Hematol Oncol, 1989 Fall, 11(3), 286 - 91
Comparative effects of mezlocillin and carbenicillin on platelet function and thromboxane generation in patients with cancer; Mehta P et al.; Carbenicillin and mezlocillin are widely used for treatment of Pseudomonas infections in patients with cancer . Carbenicillin has been reported to cause platelet dysfunction and bleeding diathesis in some individuals . We evaluated whether carbenicillin causes deterioration of platelet function in patients with cancer and whether mezlocillin causes similar effects on platelets from normal subjects or from patients with cancer . In these in vitro studies, carbenicillin and mezlocillin decreased ADP and epinephrine-induced platelet aggregation and thromboxane A2 generation similarly, but only in concentrations of 3.2 mg/ml or higher . In contrast, carbenicillin was more potent than mezlocillin in decreasing ristocetin-induced platelet aggregation . We also evaluated effects of these antibiotics on platelet function in 19 patients with cancer who developed fever and neutropenia . These patients received either mezlocillin (10 patients) or carbenicillin (nine patients) in combination with nafcillin and gentamycin . Neither carbenicillin nor mezlocillin had any significant effect on platelet aggregation or thromboxane A2 generation . Lack of effects in vivo was due to defective platelet function in these patients prior to any antibiotics . These defects were most probably related to underlying disease and/or prior chemotherapy . Thus, carbenicillin and mezlocillin can both safely be used in patients with cancer who develop fever and neutropenia, and neither seems to have advantage over the other in terms of platelet function.

J Basic Microbiol, 1989, 29(5), 315 - 8
Inhibition of ornithine carbamoyltransferase from Pseudomonas syringae pv . syringae W50 by phaseolotoxin; Jahn O et al.; In contrast to the producer of phaseolotoxin and Orn-P(O)(NH2)-NH-SO3H (PNSOrn), Pseudomonas syringae pv . phaseolicola, which possesses a sensitive and an insensitive type of ornithine carbamoyltransferase (OCT, E.C . 2.1.3.3.), in Pseudomonas syringae pv . syringae W50, an organism which produce neither phaseolotoxin nor PNSOrn, only one type of OCT could be detected . This enzyme is highly sensitive to phaseolotoxin . This result supports our hypothesis that the existence of an insensitive ornithine carbamoyltransferase is an important prerequisite for the synthesis of phaseolotoxin in P . syringae pv . phaseolicola and that this enzyme does not occur generally in P . syringae spec.

J Basic Microbiol, 1989, 29(5), 299 - 303
Detection of an insensitive ornithine-carbamoyltransferase in strains of Pseudomonas syringae pv . phaseolicola with different phytotoxin-generating capacities; Jahn O et al.; Independently of their capacity to produce phytotoxins, strains of Pseudomonas syringae pv . phaseolicola contain two ornithine carbamoyltransferases (OCT, EC 2.1.3.3) which differ in resistance to phaseolotoxin and Orn-P(O) (NH2)-NH-SO3 H (PNSOrn) . At 18 degrees C, the optimal temperature for product formation, the balance of the two types of OCT was shifted in favour of the insensitive type in phaseolotoxin producing strains, and in favour of the sensitive one in strains with little or no toxin production . The results suggest a causal relationship between the existence of an insensitive enzyme and the synthesis of toxins.

J Gen Microbiol, 1989 Jan, 135 ( Pt 1), 37 - 45
Cloning and nucleotide sequence of the isoamylase gene from a strain of Pseudomonas sp; Tognoni A et al.; A strain of Pseudomonas sp., SMP1, isolated from a soil sample collected in the Monterotondo area (Rome), secreted isoamylase activity into the culture medium . The enzyme was purified and optimal reaction and stability conditions were determined by varying pH and temperature . The chemico-physical properties of the enzyme were similar to those of the isoamylase purified in Japan more than 20 years ago from 'Pseudomonas amyloderamosa' strain SB15 . A genomic library of SMP1 was prepared in Escherichia coli using pUC12 as vector . Two isoamylase-producing colonies were identified out of 6300 screened . The hybrid plasmids isolated from the two clones showed common restriction patterns . The chromosomal portion of one of these plasmids (pSM257) was completely sequenced . Comparison between the deduced amino acid sequence of the isoamylase and the published sequences of other amylolytic enzymes showed the presence of conserved domains.

Mikrobiologiia, 1989 Jan-Feb, 58(1), 71 - 5
{Heterogeneity of a Pseudomonas bacteriophage population}; Romashko AM et al.; Twenty-five isolates of virulent bacteriophages for Pseudomonas belonging to the morphological group C (Bradley, 1967) were obtained from different natural habitats . The phages of each isolate were found to differ from one another in at least one of the following characteristics: the sensitivity to an osmotic shock, to heating at 60 degrees C and to UV; the ability to cause lysis of 86 Pseudomonas strains; the reaction of neutralisation with antisera . At the same time, the phages were related by the existence of common antigens . These properties are responsible for the genetic stability of the bacteriophages in their complicated relationship with their host, Pseudomonas bacteria.

Res Microbiol, 1989 Jan, 140(1), 17 - 20
New serotypes of Pseudomonas cepacia; Werneburg B et al.; O and H serotyping of Pseudomonas cepacia has provided a suitable procedure for epidemiological studies . Our previous reports have described 7 O and 5 H antigens . The study of strains from another geographical origin led us to prepare antisera against those which could not be serotyped and thus to determine 2 new O and 2 new H specificities (O:8 and O:9, H:4 and H:8).

Microbios, 1989, 57(230), 21 - 6
Observations on cell-wall deficient forms of Pseudomonas maltophilia; Pease PE et al.; Pseudomonas maltophilia cell-wall deficient forms were induced using a medium containing carbenicillin and polyvinyl pyrrolidone . Scanning electron microscopy proved to be a very effective method of demonstrating the various phases . It is suggested that the rapid rate of reversion to bacterial forms could have been due to plasmids.

Proc Natl Sci Counc Repub China B, 1989 Jan, 13(1), 9 - 14
Production of isoamylase by Pseudomonas amyloderamosa mutant strain JD210; Houng JY et al.; Nutritional requirements for the production of isoamylase by Pseudomonas amyloderamosa mutant strain JD210 were investigated . The optimal initial pH for enzyme production in shake-flask cultivation was 5.0 . Maltose and soybean protein hydrolyzate were found to be the best carbon source and nitrogen source, respectively . The enzyme production was drastically inhibited by Zn+2 and Cu+2 . Other metal ions phosphates and surfactants exhibited no significant inhibitory or accelerating effect on enzyme production . According to auxanography and single omission experiments, proline and isoleucine were required for growth . The supplement of 0.1% proline increased enzyme production by around 30% compared with no addition.

Int Arch Allergy Appl Immunol, 1989, 88(3), 304 - 11
Serum antibodies to Pseudomonas pseudoalcaligenes in metal workers exposed to infected metal-working fluids; Mattsby-Baltzer I et al.; Metal workers exposed to aerosol from metal-working fluid were examined with respect to serum antibody to the lipopolysaccharide (LPS) of Pseudomonas pseudoalcaligenes . During 1 year of observation this species grew at high concentrations in the fluid (approximately 10(8) CFU/ml), and the air surrounding the metal-working machines sometimes contained more than 10(5) CFU/m3 . The levels of antibody belonging to the IgG and IgA classes were significantly higher than in blood donors and in workers newly employed . Comparison with employees not working in the machine hall showed a significantly higher IgG, but not IgA, antibody level . For IgM antibody no such difference was found . Among the exposed workers, non-smokers had significantly higher IgG antibody levels than smokers, whereas no such difference was established for IgA and IgM antibodies . In smokers the IgG antibody level seemed to decline in the long-range time, since there was a negative correlation between the time of employment with exposure to metal-working fluids and IgG antibody level . The antibody data indirectly demonstrate that P . pseudoalcaligenes grown in metal-working fluid penetrated the body surface of metal workers to yield an immune response most probably after inhalation of aerosol containing bacteria . In the metal workers this long-term exposure did not lead to any acute or chronic respiratory discomfort.

Pediatr Hematol Oncol, 1989, 6(4), 293 - 305
Congenital dysgranulopoietic neutropenia in two siblings: clinical, ultrastructural, and in vitro bone marrow culture studies; Koren A et al.; Two siblings with congenital neutropenia are reported . The first patient, female, died after Pseudomonas sepsis . The second patient male, suffered from recurrent pyogenic infections, with a more benign course . Bone Marrow (BM) and Peripheral Blood (PB) analysis in the second patient revealed a reduced number of granules and myelin bodies in the PB neutrophils, suggesting a developmental defect of primary and secondary granules . BM promyelocytes were almost normal, but the myelocytes and metamyelocytes showed defective granulogenesis . The BM in vitro granulocyte-macrophage-colony-forming cell (GM-CFC) growth and the PB white blood cells (WBC) granulocyte-macrophage-colony-stimulating factor (GM-CSF) production, which were analyzed in the second patient, showed normal numbers of GM-CFC, with differentiation mostly toward monocytes and a defect in the GM-CSF production capacity . The second patient's PB mononuclear cells or serum did not inhibit normal GM-CFC when added to control BM cells . We suggest that in this specific form of congenital neutropenia, which is probably an autosomal recessive disorder, the abnormal neutrophil granule production and the defective provision of GM-CSF by PB WBC are unique pathognomonic characteristics, possibly associated with the overt neutropenia.

Pediatr Neurol, 1989 Jan-Feb, 5(1), 48 - 52
Melioidosis with multiple cerebral abscesses; Pelekanos JT et al.; Melioidosis from Pseudomonas pseudomallei is common in endemic areas (particularly southeast Asia) and is being recognized with increasing frequency in developed countries . Central nervous system involvement is a rare complication with a high mortality . A patient with multiple cerebral abscesses caused by this organism is presented to demonstrate that successful treatment is possible when a high index of clinical suspicion leads to early diagnosis.

Zh Mikrobiol Epidemiol Immunobiol, 1989 Jan, (1), 21 - 6
{Bacteriocin typing of Pseudomonas cepacia strains isolated from patients and the rhizosphere of plants}; Dodatko TA et al.; The work deals with the bacteriocin typing of 34 P . cepacia strains isolated from different sources with respect to both the capacity of synthesizing bactericins and sensitivity to them . The standard set of strains comprizing 8 P . cepacia bacteriocin-sensitive strains and 6 highly active cepaciacin producer strains was used . 24 P . cepacia strains belonged to 11 different S-types, 20 strains synthetized cepaciacins of new types.

Cornea, 1989, 8(1), 67 - 71
Pseudomonas cepacia keratitis; Levy JH et al.; Pseudomonas cepacia has recently become recognized as a virulent pathogen responsible for nosocomial infections in hosts with altered immunity . It has been implicated in endophthalmitis and conjunctivitis, and is resistant to conventional antipseudomonal therapy . No cases of P . cepacia keratitis have been reported in the literature . We report such a case in association with topical steroid and contact lens use following penetrating keratoplasty . In addition, we developed an experimental model of P . cepacia keratitis in the rabbit . P . cepacia should be considered as a cause of infectious keratitis especially in nosocomial infections in immunocompromised corneas.

Scand J Infect Dis, 1989, 21(6), 697 - 708
Preliminary study on treatment of septic shock patients with antilipopolysaccharide IgG from blood donors; Fomsgaard A et al.; A novel intravenous therapy consisting of polyvalent IgG antibodies to lipopolysaccharide (LPS, endotoxin) obtained from screening of blood donors was used for treatment of patients with profound septic endotoxin shock . Investigation of the anti-LPS IgG pharmacokinetics in the 10 patients revealed time related changes in the plasma concentrations of anti-LPS IgG, endotoxin, tumour necrosis factor (TNF) and the clinical parameters . A decrease in serum concentrations of IgG and IgM antibodies to LPS was observed prior to the immunotherapy as well as in a clinical example of lethal septicemia without anti-LPS immunotherapy . Increasing serum concentrations of anti-LPS IgG during antibody infusion was followed by a decrease in the concentration of endotoxin and TNF . In survivors an IgM and IgG anti-LPS antibody response developed . Using clinical parameters and APACHE II clinical severity scores to measure the clinical condition, a beneficial effect was observed within 24 h corresponding to a decrease in the calculated expected mortality rate from more than 80% to about 50% . Five patients (55%) expired during the study . One patient died in the early septic shock phase . One patient expired due to superimposed hemorrhagic shock . Three immunosuppressed patients died 1-2 weeks after initial recovery, 1 with fungal sepsis and 2 patients due to pseudomonas infection.

Microbiol Immunol, 1989, 33(10), 811 - 20
Inhibition of macromolecular synthesis in cultured macrophages by Pseudomonas pseudomallei exotoxin; Mohamed R et al.; Pseudomonas pseudomallei exotoxin was found to be a potent inhibitor of protein and DNA synthesis in cultured macrophages . Inhibition of DNA synthesis occurred at toxin concentrations as low as 1-2 micrograms/ml and inhibition of 3H-thymidine uptake was almost complete at concentrations of 8 micrograms/ml or more . A close correlation between cell damage and inhibition by DNA synthesis was observed . For protein synthesis, inhibition was obtained at much lower doses (0.06-2.0 micrograms/ml) of the toxin . At similar toxin concentrations, DNA synthesis was marginally affected . Further, it was shown that protein synthesis inhibition occurred almost immediately after incubation, reaching its maximal inhibitory effect of 70% after 6 hr . DNA synthesis, however, was minimally affected by a similar toxin concentration even after 10 hr of incubation . The inhibition of macromolecular synthesis in macrophages by P . pseudomallei exotoxin may be relevant to its modulatory effect on the host defense mechanism.

Microbios, 1989, 60(244-245), 141 - 50
Induction of potassium efflux by cupric ions in Pseudomonas syringae ATCC 12271 and its correlation with cell viability; Cabral JP; In Pseudomonas syringae, Cu2+ induce a significant loss of K+ from the cells . The course of the efflux followed an approximately sigmoidal pattern . The maximum rate of K+ efflux, the time needed to achieve this rate and the maximum amount of K+ released from the cells, were dependent on copper concentration . Pre-treatment with several divalent cations modified markedly the parameters of potassium efflux induced by copper, by increasing the maximum rate of K+ efflux and the amount of K+ released after 4 min of copper treatment, and decreasing the time required to achieve the maximum rate . The addition of copper to cell suspensions resulted in a progressive decrease in the number of viable cells . Pre-treatment with Mg2+ or Ca2+ resulted in a decrease in the lethality of copper ions.

J Basic Microbiol, 1989, 29(7), 441 - 7
Phaseolotoxin production by Pseudomonas syringae pv . phaseolicola: the influence of temperature; Nuske J et al.; Phaseolotoxin (N-sulphodiaminophosphinyl-ornithyl-alanyl-homoarginine) is a phytotoxic secondary metabolite produced by Pseudomonas syringae pv . phaseolicola . The production of the phytotoxin is strongly regulated by temperature . The optimal temperature for phaseolotoxin production is 18 degrees C . Temperatures in the range between 18 degrees C and 30 degrees C inhibit the production of phaseolotoxin in an increasing manner . By temperature shift experiments and the inhibition of protein synthesis by chloramphenicol it was demonstrated, that the synthesis of enzymes related to toxin synthesis and not the regulation of the activity of the responsible enzyme system is affected by higher temperatures.

Gene, 1989, 76(2), 227 - 38
Characterization and nucleotide sequence determination of a repeat element isolated from a 2,4,5-T degrading strain of Pseudomonas cepacia; Tomasek PH et al.; Pseudomonas cepacia strain AC1100, capable of growth on 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), was mutated to the 2,4,5-T- strain PT88 by a ColE1::Tn5 chromosomal insertion . Using cloned DNA from the region flanking the insertion, a 1477-bp sequence (designated RS1100) was identified which was repeated several times on the wild-type chromosome and was also present on AC1100 plasmid DNA . Various chromosomal fragments containing this sequence were cloned and their nucleotide sequence was determined . Examination of RS1100 revealed the presence of 38-39-bp terminal inverted repeats immediately flanked by 8-bp direct repeats . The translated sequence of the single large open reading frame of RS1100 showed structural similarity to the phage Mu transposase and other DNA-binding proteins . Thus the AC1100 repeated sequence has several structural features in common with insertion sequence elements . Three copies of RS1100 were mapped near 2,4,5-t genes encoding degradation of 5-chloro-1,2,4-trihydroxybenzene, an intermediate in 2,4,5-T degradation . Neither RS1100 nor the 2,4,5-t genes hybridized to DNA isolated from Pseudomonas strains, including P . cepacia, suggesting that both gene fragments may be of foreign origin recruited in strain AC1100 . The origin of these two DNA segments as well as the role played by RS1100 in the recruitment of 2,4,5-t genes in AC1100 are presently under investigation.

EMBO J, 1989 Jan, 8(1), 31 - 8
The induction of manganese superoxide dismutase in response to stress in Nicotiana plumbaginifolia; Bowler C et al.; Superoxide dismutases (SODs) are metalloproteins that catalyse the dismutation of superoxide radicals to oxygen and hydrogen peroxide . The enzyme has been found in all aerobic organisms examined, where it plays a major role in the defence against toxic reduced oxygen species which are generated in many biological oxidations . Here we report the complete primary structure of a plant manganese superoxide dismutase (MnSOD), deduced from a cDNA clone of Nicotiana plumbaginifolia . The plant protein is highly homologous to MnSODs from other organisms and also contains an N-terminal leader sequence resembling a transit peptide for mitochondrial targeting . The location of the mature protein within the mitochondria has been demonstrated by subcellular fractionation experiments . We have analysed the expression profile of this MnSOD and found that it is dramatically induced during stress conditions, most notably in tissue culture as a result of sugar metabolism and also as part of the pathogenesis response of the plant, being induced by ethylene, salicylic acid, and Pseudomonas syringae infection . This induction is always accompanied by an increase in cytochrome oxidase activity, which suggests a specific protective role for MnSOD during conditions of increased mitochondrial respiration.

Adv Enzyme Regul, 1989, 28, 323 - 33
Carboxypeptidase G2 enhances trimetrexate cytotoxicity in CCRF-CEM cell lines sensitive and resistant to methotrexate; Romanini A et al.; Carboxypeptidase G2 (CPG2), an enzyme produced by Pseudomonas strain RS-16, degrades folates as well as methotrexate and other folic acid analogs such as aminopterin and dichloromethotrexate, but not the "non-classical" folate antagonist trimetrexate (TMTX) . The possibility of enhancing TMTX cytotoxicity of CPG2 induced depletion of intracellular folates was investigated in human leukemic CCRF-CEM cells and in three methotrexate resistant sublines of these cells with different mechanisms of resistance, CCRF-CEM/E, a subline with increased DHFR; CCRF-CEM/P, a subline defective in polyglutamylation and CCRF-CEM/T, a subline with impaired MTX uptake . The cytotoxic effect was detected using a colorimetric assay with MTT . Dose-effect relationships of single drugs alone and in combination were analyzed by the median effect principle and by the combination indices for the quantitation of synergism or antagonism with the aid of a microcomputer . TMTX alone was very effective on the parent and all the resistant cell lines (CCRF-CEM/E, CCRF-CEM/P, CCRF-CEM/T) with ED50 values in the nanomolar range (1.4, 1.6, 1.5 and 0.7 nM, respectively) following 5 days of exposure . The ED50s of CPG2 for these cell lines were 3.5, 2.6, 26.6 and 7.9 X 10(-5) U/ml, respectively . A synergistic cytotoxic effect of TMTX after simultaneous continuous exposure was observed with CPG2 on CCRF-CEM cells and on the three resistant cell lines.

Nephrol Dial Transplant, 1989, 4(9), 814 - 7
Pseudomonas peritonitis in CAPD patients: characteristics and outcome of treatment; Chan MK et al.; Twenty-five episodes of Pseudomonas peritonitis which occurred over a five-and-a-half-year period were reviewed . Pseudomonas peritonitis accounted for 25/516 (4.8%) of all episodes of peritonitis . Nine of the episodes were first infections in that the patient had not experienced peritonitis before . The rest were repeat peritonitis . There was no difference in any demographic factors between the first episodes and the repeat episodes except exit site infection which was more common among patients who had repeat infections . Overall cure rate of Pseudomonas peritonitis was 20/25 (80%) . Five catheters had to be removed, all in patients who had to be transferred permanently to haemodialysis . In general, ceftazidime in combination with an aminoglycoside was an effective regimen . Oral ofloxacin was not useful despite in vitro sensitivity of the pathogen to the antibiotic.

J Biochem (Tokyo), 1989 Jan, 105(1), 1 - 3
Crystal structure analysis of omega-amino acid:pyruvate aminotransferase with a newly developed Weissenberg camera and an imaging plate using synchrotron radiation; Watanabe N et al.; The three-dimensional structure of omega-amino acid:pyruvate aminotransferase from Pseudomonas sp . F-126, an isologous alpha 4 tetramer containing pyridoxal 5'-phosphate (PLP) as a cofactor, has been determined at 2.0 A resolution . The diffraction data were collected with a newly developed Weissenberg camera with a Fuji Imaging Plate, using synchrotron radiation . The mean figure-of-merit was 0.57 . The subunit is rich in secondary structure and comprises two domains . PLP is located in the large domain . The high homology in the secondary structure between this enzyme and aspartate aminotransferase strongly indicates that these two types of enzymes have evolved from a common ancestor.

Klin Khir, 1989, (3), 48 - 9
{Sensitivity of Pseudomonas bacteria to iodopiron and dimexide determined by electron microscopy}; Litovchenko PP et al.; A structure of the pyocyanic bacterium culture was studied by the method of electron microscopy of native preparations and ultrafine sections of the cells, which contained in the quantity of 10(5)/ml of suspension, and were treated with iodopiron in the concentration of 93.75 mg/ml, 10% dimexide solution and their combination . The increase in damaging effect on bacteria of the combined use of chemopreparations permits to recommend their complex administration in local pyocyanic infection in burned patients.

Proc Natl Acad Sci U S A, 1989 Jan, 86(1), 287 - 91
Chimeric cytotoxin IL2-PE40 delays and mitigates adjuvant-induced arthritis in rats; Case JP et al.; Adjuvant arthritis in rats is a T-cell dependent "autoimmune" disease with close similarities to several forms of human arthritis . Injection of mycobacterial adjuvant leads to T-cell activation and proliferation, processes in which the de novo expression of the interleukin 2 (IL-2) receptor plays a pivotal role . The subsequent massive mononuclear cell infiltration of the joints ultimately results in complete joint destruction . Because activation of the helper/inducer subset of T lymphocytes is critical to the establishment of disease, we reasoned that IL2-PE40, a cytotoxic IL-2-Pseudomonas exotoxin fusion protein that targets the membrane-penetration and ADP-ribosylation domains of the toxin to cells bearing the IL-2 receptor, would be an effective and specific therapy . Adjuvant-injected rats were randomized to treatment with IL2-PE40, phosphate-buffered saline, or either of two control proteins related to IL2-PE40 but lacking either the receptor-binding moiety or an enzymatically active toxin domain and previously demonstrated to lack cytotoxicity in vitro . Intraperitoneal IL2-PE40 given before the establishment of overt clinical disease proved an effective and specific modifier of adjuvant arthritis by clinical, histological, and radiographic criteria . Our data suggest that IL2-PE40 may be effective in those diseases in which activated T-cells play an important role.

Aust J Biotechnol, 1989 Jan, 3(1), 43 - 9
Kinetic properties and contribution to cellulose saccharification of a cloned Pseudomonas beta-glucosidase; Rickard PA et al.; The plasmid pND71, which encodes beta-glucosidase (cellobiase) activity, cloned from the cellulolytic Pseudomonad, PS2-2, was mobilized by conjugation into 10 Pseudomonas strains . The highest specific activity was produced by 17498 (pND71) and the properties of the enzyme produced from this transconjugant were studied . The enzyme was shown to be cell associated, to have a temperature optimum of 37 degrees C, a pH optimum of 7.0 and Km values of 1.33 and 2.94 mM for pNPG and cellobiose respectively . It was competitively inhibited by glucose, with a Ki of 30 mM . Evidence was obtained which suggested that the enzyme was produced constitutively and that synthesis was not repressed by glucose . When culture preparations were used in combination with Trichoderma reesei QM9414 and C30 enzyme preparations to saccharify cellulose, 17498 (pND71) was more effective than preparations of PS2-2 in acting synergistically with T . reesei to solubilize more carbohydrate and produce more glucose.

FEBS Lett, 1988 Dec 19, 242(1), 36 - 40
Cloning, sequencing and expression of the lipase gene from Pseudomonas fragi IFO-12049 in E . coli; Aoyama S et al.; The lipase gene from Pseudomonas fragi IFO-12049 was isolated using the expression library and the primary structure of lipase deduced from the nucleotide sequence was determined . It is composed of 277 amino acid residues and a protein of Mr 29,966, which was close to the value of the lipase expressed in E . coli.

FEBS Lett, 1988 Dec 19, 242(1), 70 - 4
The cupric site in nitrous oxide reductase contains a mixed-valence {Cu(II),Cu(I)} binuclear center: a multifrequency electron paramagnetic resonance investigation; Kroneck PM et al.; Multifrequency electron paramagnetic resonance (EPR) spectra of the Cu(II) site in nitrous oxide reductase (N2OR) from Pseudomonas stutzeri confirm the assignment of the low field g value at 2.18 consistent with the seven line pattern observed at 9.31 GHz, 10 K . S-band spectra at 20 K are better resolved than the X-band spectra recorded at 10 K . The features observed at 2.4, 3.4, 9.31 and 35 GHz are explained by a mixed-valence {Cu(1.5)..Cu(1.5)} S = 1/2 species with the unpaired electron delocalized between two equivalent Cu nuclei . The resemblance of the N2OR S-band spectra to the spectra for the EPR-detectable Cu of cytochrome c oxidase suggests that the S-band spectrum for cytochrome c oxidase measured below 30 K may also contain hyperfine splittings from two approximately equivalent Cu nuclei.

J Immunol, 1988 Dec 15, 141(12), 4224 - 8
IL-2-PE40 is cytotoxic for activated T lymphocytes expressing IL-2 receptors; Ogata M et al.; IL-2-PE40 is a chimeric molecule in which IL-2 is attached to the amino end of modified Pseudomonas exotoxin molecule lacking cell recognition domain . This molecule was extremely toxic for Con A-stimulated spleen cells from mice . Moreover, IL-2-PE40 has suppressive effect against Ag-activated cells; it inhibits the generation of cytotoxic T lymphocyte activity in a MLC . IL-2-PE40 could be a useful agent in IL-2R targeting therapy including immunosuppressive therapy for allograft rejection or some autoimmune diseases.

J Biol Chem, 1988 Dec 15, 263(35), 18650 - 6
Interleukin 2 (IL2) PE40 is cytotoxic to cells displaying either the p55 or p70 subunit of the IL2 receptor; Lorberboum-Galski H et al.; IL2-PE40 is a chimeric protein composed of human interleukin 2 (IL2) genetically fused to the amino terminus of a modified form of pseudomonas exotoxin (PE) . Internalization of IL2 via the individual p55 and p70 subunits of the IL2 receptor was studied using IL2-PE40 on several mouse and human cell lines expressing either the p55, the p70, or both IL2 receptor subunits . Internalization was assessed by measuring inhibition of protein synthesis caused by the toxin moiety of IL2-PE40 . The results demonstrate that IL2 internalization is mediated by either the p55 receptor subunit or by the p70 subunit but is much more efficient when high affinity receptors composed of both subunits are present . IL2-PE40 is a powerful reagent for studying IL2 receptor interactions and for analyzing pathways of the immune response and its regulation.

J Gen Microbiol, 1988 Dec, 134 ( Pt 12), 3231 - 7
Regulation of autotrophic metabolism in Pseudomonas oxalaticus OX1 wild-type and an isocitrate-lyase-deficient mutant; Meijer WG et al.; In Pseudomonas oxalaticus the activity and synthesis of the Calvin cycle enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) are regulated by inactivation and endproduct repression, respectively . Phosphoenolpyruvate (PEP) has been suggested to function as a signal molecule for the latter control system . During growth of the organism in carbon-source-limited continuous cultures with various ratios of acetate and formate in the feed, the RuBisCO levels varied considerably, but no correlation was observed with the intracellular concentrations of PEP . To study whether the repression exerted by acetate utilization was dependent on the synthesis of glycolytic intermediates from this compound, an acetate-negative mutant defective in isocitrate lyase was isolated and characterized . Clear evidence was obtained that in this mutant acetate is as effective in repressing RuBisCO synthesis as in the wild-type . It therefore appears more likely that acetyl-CoA or a closely related metabolite functions as a signal molecule in the regulation of RuBisCO synthesis.

Proc Natl Acad Sci U S A, 1988 Dec, 85(24), 9738 - 42
Cytotoxic activity of an interleukin 6-Pseudomonas exotoxin fusion protein on human myeloma cells; Siegall CB et al.; A chimeric toxin composed of human interleukin 6 (IL-6) attached to a portion of Pseudomonas exotoxin (PE) devoid of its own cell recognition domain has been produced in Escherichia coli . The fusion protein (IL-6-PE40) is cytotoxic to a human myeloma cell line expressing IL-6 receptors but has no effect on IL-6 receptor-negative cells . The specificity of IL-6-PE40 cytotoxicity was demonstrated through competition with excess IL-6 and neutralization with an antibody to IL-6 . IL-6-PE40 may be useful in the selective elimination of myeloma cells and other cells with high numbers of IL-6 receptors.

J Leukoc Biol, 1988 Dec, 44(6), 529 - 34
Inhibition of human neutrophil and Pseudomonas elastases by the amyloid P-component: a constituent of elastic fibers and amyloid deposits; Vachino G et al.; The amyloid P-component (AP), a ubiquitous component of amyloid fibrils, is also a plasma protein and a connective tissue constituent . Its proximity to elastin, in particular, suggested that AP might serve to protect elastic tissue from hydrolytic enzymes . The inhibition of pancreatic elastase by AP has been reported . In the present study, the effects of AP on human neutrophil elastase and Pseudomonas elastase were investigated, and AP was shown to interfere with the cleavage of soluble elastin . As indicated by Michaelis-Menten analysis, AP is acting as a noncompetitive inhibitor . C-reactive protein, which is structurally similar to AP, had no effect on either elastase . AP was also found to inhibit the degradation of secondary amyloid fibrils by neutrophil elastase when these structures were first partially purified and then reexposed to AP . AP's ability to inhibit elastase was compared with alpha-1 antitrypsin in the presence and absence of oxidizing agents . These substances, which are released by inflammatory cells, are known to abrogate alpha-1 antitrypsin's anti-protease capacity . This contributes to elevated levels of free proteases in the circulation and extravascular spaces during severe inflammation . AP is not susceptible to oxidation and remains a functional inhibitor under these conditions . The potential role of AP as an elastase inhibitor is discussed.

J Bacteriol, 1988 Dec, 170(12), 5479 - 88
Molecular analysis of a pathogenicity locus in Pseudomonas syringae pv . syringae; Mukhopadhyay P et al.; One of the chromosomal regions of Pseudomonas syringae pv . syringae encoding pathogenicity factors had been mapped into a 3.9-kilobase-pair fragment in previous studies . Promoter probe analysis indicated the existence of a promoter near one end of the fragment . DNA sequencing of this fragment revealed the existence of a consensus promoter sequence in the region of the promoter activity and two open reading frames (ORFs) downstream . These ORFs, ORF1 and ORF2, encoded putative polypeptides of 40 and 83 kilodaltons, respectively . All ORF1::Tn5 as well as ORF2::Tn5 mutant strains were nonpathogenic on susceptible host bean plants and were unable to elicit hypersensitive reactions on nonhost tobacco plants . The deduced amino acid sequence of the 83-kilodalton polypeptide contained features characteristic of known integral membrane proteins . Fusion of the lacZ gene to ORF2 led to the expression of a hybrid protein inducible in Escherichia coli . The functions of the putative proteins encoded by ORF1 and ORF2 are unknown at present.

Clin Chest Med, 1988 Dec, 9(4), 669 - 78
Pulmonary host defense: defects that lead to chronic inflammation of the airway; Hornick DB; Current knowledge of pulmonary host defense can help us to understand the unique relationship between CF patients and P . aeruginosa colonization of the lung . Subtle defects in CF host defense, such as those identified in mucociliary clearance and in the CF IgG opsonin, allow P . aeruginosa to persist . Not all the defects are attributable to the host . In several examples, the defects are induced by P . aeruginosa, presumably in an effort to maintain its foothold . The examples of the latter discussed here have included the effect of pseudomonas-derived products on mucociliary action, alpha 1 PI function, and the formation of ineffective IgG opsonins and immune complexes . Overall, P . aeruginosa is the cause of significant morbidity and, eventually, mortality in these patients . As we approach the identification of the genetic defect central to this disease, it is hoped that we will gain more insight into the pathogenesis of the P . aeruginosa lung lesion in CF and develop more effective ways of preventing P . aeruginosa colonization of the CF patients' lungs.

Clin Biochem, 1988 Dec, 21(6), 347 - 52
Determination of human aspartate aminotransferase isoenzymes by their differential sensitivity to proteases; Teranishi H et al.; The effect of various proteases (trypsin, chymotrypsin, subtilisin, protease 401, and thermolysin) on the mitochondrial isoenzyme (m-AST) and cytoplasmic isoenzyme (c-AST) of human and swine aspartate aminotransferase (AST;EC 2.6.1.1) was evaluated . All procedures including the reaction with proteases and the subsequent determination of the AST activity were carried out in an automatic analyzer . The mammalian c-AST was efficiently inactivated by chymotrypsin, subtilisin and protease 401 while m-AST activity decreased very slowly with these proteases . Thermolysin and trypsin showed much less effect on c-AST activity . Especially, chymotrypsin at concentrations of 0.5-1.0 g/L inactivated human c-AST almost completely but showed no detectable inactivating effect on m-AST . Thus chymotrypsin appears to be the most suitable protease for the differential determination of AST isoenzymes in human serum . Further studies on the effects of proteases with AST from other species showed that Escherichia coli AST resembled mammalian m-AST while Pseudomonas AST resembled c-AST.

Mol Gen Genet, 1988 Dec, 215(1), 165 - 72
Deletion mutagenesis of the ice nucleation gene from Pseudomonas syringae S203; Green RL et al.; The ice nucleation gene inaZ, from Pseudomonas syringae S203, was manipulated to produce a series of defined rearrangements in its coding sequence without changing the reading frame . The effects of these mutations on the ice nucleation phenotype were determined in a heterologous host, Escherichia coli K12 . Deletions which disrupted the periodicity of 16 codons, in a repetitive region of inaZ, caused the frequencies of ice nuclei in the bacterial population to be significantly depressed; the nuclei with thresholds at warmer temperatures were most affected . In contrast, when the periodicity was left intact, deletions and duplications in the same region had only slight effects on nucleation activity . Deletions removing part or all of one of the nonrepetitive regions (that encoding the amino-terminal domain of the InaZ protein) did not abolish nucleation activity, but caused it to be limited to cooler threshold temperatures . In contrast, the non-repetitive carboxy-terminal domain of the InaZ protein was shown to be essential for ice nucleation at all temperatures . The differential requirements (for periodicity, and for the amino-terminus) in forming nuclei with different thresholds may be significant for understanding what determines the threshold temperature of an ice nucleus.

Biochem J, 1988 Dec 1, 256(2), 673 - 6
The nucleotide sequence and deduced amino acid sequence of the cytochrome cL gene of Methylobacterium extorquens AM1, a novel class of c-type cytochrome; Nunn DN et al.; The nucleotide sequence and deduced amino acid sequence of the cytochrome cL of Methylobacterium extorquens (Pseudomonas AM1; Methylobacterium AM1) shows that this cytochrome c is completely different, except for its haem-binding site, from all other cytochromes.

J Bacteriol, 1988 Dec, 170(12), 5689 - 97
Genetic organization and regulation of proteins associated with production of syringotoxin by Pseudomonas syringae pv . syringae; Morgan MK et al.; Many strains of Pseudomonas syringae pv . syringae produce one of two low-molecular-weight, peptide-containing phytotoxins, either syringomycin (SR) or syringotoxin (ST) . An analysis of Tn5-induced ST-mutants revealed alterations in the presence of two large proteins (ca . 470 and 435 kilodaltons) . Apparent truncated forms of the 470 (ST1)- or 435 (ST2)-kilodalton proteins were observed in some mutants . Mapping of the Tn5 insertions and size determinations of truncated proteins suggested that both ST1 and ST2 are in the same transcriptional unit, with ST1 being directly upstream of ST2 . When an ST-producing strain of P . syringae pv . syringae was grown under toxin-inducing conditions, ST1 and ST2 were first detected at the end of the exponential phase of growth, which was when the first accumulation of ST was observed . High iron levels were essential for efficient ST production . At concentrations of FeCl3 of between 0.2 and 5 microM, the amount of toxin accumulated was almost directly proportional to the iron concentrations . The amount of ST1 and ST2 present showed a corresponding increase in response to iron concentrations . Our genetic and physiological data implicate ST1 and ST2 in the biosynthesis of syringotoxin.

J Bacteriol, 1988 Dec, 170(12), 5680 - 8
Physical and functional analyses of the syrA and syrB genes involved in syringomycin production by Pseudomonas syringae pv . syringae; Xu GW et al.; The syrA and syrB genes involved in syringomycin production in Pseudomonas syringae pv . syringae B301D were identified from an EcoRI-pLAFR3 cosmid library and then physically and functionally analyzed in relation to plant pathogenicity . Homologous recombination of the genes required for syringomycin production from cosmids pGX183 (syrA) and pGX56 (syrB), respectively, introduced into nontoxigenic (Tox-) Tn5 mutants W4S2545 and W4S770 resulted in the concomitant restoration of toxin production and full virulence . The disease indices of the Tox+ strains obtained by recombination of the cloned, homologous DNA into the corresponding Tn5 mutant were essentially equivalent to that of strain B301D-R and significantly higher than those of W4S2545 and W4S770 . A 12-kilobase (kb) EcoRI fragment from pGX183 was subcloned (i.e., pGX15) and found to contain the sequences necessary for syringomycin production . A map of pGX15 prepared by a combination of restriction endonuclease digestions and Tn5 mutagenesis showed that the syrA sequence was 2.3 to 2.8 kb . Marker exchange of syrA::Tn5 from pGX15 into B301D-R yielded nonpathogenic phenotypes, indicating that syrA is a regulatory gene since it is necessary for both syringomycin production and pathogenicity . The 4.9-kb EcoRI fragment from pGX56 was subcloned (i.e., pGX4) and shown to carry the syrB sequence which was 2.4 to 3.3 kb . Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of protein extracts from B301D-R associated five proteins, ranging from approximately 130,000 to approximately 470,000 in molecular weight, with syringomycin production . The syrA and syrB genes were required for the formation of proteins SR4 (approximately 350,000) and SR5 (approximately 130,000), which are believed to be components of the syringomycin synthetase complex.

Bioorg Khim, 1988 Dec, 14(12), 1678 - 83
{Antigenic polysaccharides of bacteria . 34 . Structure of O-specific polysaccharide chains of lipopolysaccharides from Pseudomonas cepacia strains IMV 4207 (Serotype A) and IMV 598/2}; Knirel' IuA et al.; On the basis of non-destructive analysis by means of 1H and 13C NMR spectroscopy and calculation of specific optical rotation, it was concluded that O-specific polysaccharide of Pseudomonas cepacia strain IMV 4207 (serotype A) has the structure (I): (formula; see text) Two structurally different polysaccharides were found in the ratio of approximately 2.5:1 in P . cepacia strain IMV 598/2 which is serologically related to serotype A in Nakamura classification and serotype 2 in Heidt classification . The minor polysaccharide has the structure (I) whereas the major one possesses the structure (II) which is characteristic of the formerly studied O-specific polysaccharide of P . cepacia strain IMV 4137 belonging to serotype 2: ----4)-beta-D-Galp-(1----2)-alpha-L-Rhap-(1----.

Gene, 1988 Nov 30, 71(2), 267 - 77
Cloning, physical mapping and expression of chromosomal genes specifying degradation of the herbicide 2,4,5-T by Pseudomonas cepacia AC1100; Sangodkar UM et al.; A genomic library of total DNA of Pseudomonas cepacia AC1100 was constructed on a broad-host-range cosmid vector pCP13 in Escherichia coli AC80 . A 25-kb segment was isolated from the library which complemented a Tn5-generated, 2,4,5-trichlorophenoxyacetic acid-negative (2,4,5-T-) mutant, P . cepacia PT88 . This mutation was partially characterized and appeared to be lacking functional enzyme required for metabolism of an intermediate of the 2,4,5-T pathway, recently identified as 5-chloro-1,2,4-trihydroxybenzene {Chapman et al., Abstr . Soc . Environ . Toxicol . Chem . USA 8 (1987) 127} . A simple colorimetric assay was developed to detect the presence of this active enzyme in intact cells and was used to determine the expression of complementing genes . Subcloning experiments showed that a 4-kb BamHI-PstI fragment and a 290-bp PstI-EcoRI fragment, separated by 1.3-kb, were required for complementation . Both fragments are identified to be chromosomal in origin . Hybridization studies using the subcloned fragments revealed that in addition to a Tn5 insertion, mutant PT88 contained an extensive chromosomal deletion accounting for its 2,4,5-T- phenotype . The cloned fragments did not show homology to plasmid DNAs carrying degradative genes for toluene, naphthalene and 3-chlorobenzoate.

Mikrobiologiia, 1988 Nov-Dec, 57(6), 977 - 82
{Rate of endogenous respiration of Pseudomonas cells}; Arzamastsev AA; A method is proposed for determining the rate of endogenous respiration in Pseudomonas cells grown under the conditions of batch cultivation . The method takes into account the dynamic characteristics of the gauge and of the secondary instrument . The rate of Pseudomonas endogenous respiration was shown to be at the beginning of the range established earlier for other bacteria.

Mikrobiologiia, 1988 Nov-Dec, 57(6), 1024 - 30
{Immunochemical characteristics of a lipopolysaccharide from Pseudomonas aurantiaca}; Zdorovenko GM et al.; A lipopolysaccharide was isolated from Pseudomonas aurantiaca IMB 31 by extraction with aqueous phenol and purified by ultracentrifugation . The lipopolysaccharide was confined to the phenol phase . Fucosamine (2-amino-2,6-dideoxygalactose) (36%) and bacillosamine (2,4-diamino-3,4,6-trideoxyglucose) (23%) were identified as hypothetic components of the O-chain in the carbohydrate moiety of the macromolecule using the techniques of paper chromatography, gas-liquid chromatography and ion-exchange chromatography on an amino acid analyser . Rhamnose, glucose, galactose, glucosamine and galactosamine were detected as hypothetical components of the core in the lipopolysaccharide composition, as well as 2-keto-3-deoxyoctonic acid, heptose, alpha-alanine and phosphorus, usual components of the core in Pseudomonas . The following predominant fatty acids were identified in the composition of lipid A using the techniques of gas-liquid chromatography with standard compounds and gas-liquid mass spectrometry: 3-OH C10:0 (14.4%), C12:0 (30.5%), 2-OH C12:0 (14.9%), 3-OH C12:0 (17.4%), C16:0 (9.9%) . The serological relationship between P . aurantiaca strains was studied, and their phylogenetic relationship with P . fluorescens is discussed.

J Clin Microbiol, 1988 Nov, 26(11), 2441 - 3
Peritonitis caused by Pseudomonas mesophilica in a patient undergoing continuous ambulatory peritoneal dialysis; Rutherford PC et al.; We describe a case of recurrent peritonitis caused by Pseudomonas mesophilica in a diabetic man receiving continuous ambulatory peritoneal dialysis . Stagnant water on a bath rail used for support by the patient while showering was implicated as the probable source of these infections . We believe this to be the first report of the isolation of this water-associated bacterium in such infections.

J Biochem (Tokyo), 1988 Nov, 104(5), 681 - 2
Enzymatic transesterification with the lipase from Pseudomonas fragi 22.39 B in a non-aqueous reaction system; Nishio T et al.; The lipase from Pseudomonas fragi 22.39 B catalyzed the transesterification in ester and alcohol mixtures without any other solvent . Activated esters, such as vinyl and phenyl esters, were excellent acyl donors for the reaction, and the activity was enhanced by increasing the carbon number of the fatty acid fraction of the esters . Primary alcohols were esterified faster than secondary ones in this reaction system, while tertiary alcohols such as alpha-terpineol did not react at all . The lipase exhibited stereoselectivity in the esterification of alcohols such as 2-octanol.

Appl Environ Microbiol, 1988 Nov, 54(11), 2683 - 8
Degradation of 2-hydroxybiphenyl and 2,2'-dihydroxybiphenyl by Pseudomonas sp . strain HBP1; Kohler HP et al.; Pseudomonas sp . strain HBP1 was found to grow on 2-hydroxy- and 2,2'-dihydroxy-biphenyl as the sole carbon and energy sources . The first step in the degradation of these compounds was catalyzed by an NADH-dependent monooxygenase . The enzyme inserted a hydroxyl group adjacent to the already existing hydroxyl group to form 2,3-dihydroxybiphenyl when acting on 2-hydroxybiphenyl and to form 2,2',3-trihydroxybiphenyl when acting on 2,2'-dihydroxybiphenyl . To be substrates of the monooxygenase, compounds required a 2-hydroxyphenyl-R structure, with R being a hydrophobic group (e.g., methyl, ethyl, propyl, sec-butyl, phenyl, or 2-hydroxyphenyl) . Several chlorinated hydroxybiphenyls served as pseudosubstrates by effecting consumption of NADH and oxygen without being hydroxylated . Further degradation of 2,3-dihydroxy- and 2,2',3-trihydroxybiphenyl involved meta cleavage, with subsequent formation of benzoate and salicylate, respectively.

J Antimicrob Chemother, 1988 Nov, 22(5), 765 - 70
Imipenem/cilastatin as initial therapy for febrile neutropenic patients; Liang R et al.; Imipenem 2 g daily was administered intravenously to 40 evaluable patients with neutropenia and fever . Twenty-three patients had acute leukaemia and 17 malignant lymphoma . The overall response rate was 70.0% . Of the 14 patients with documented infection, 9 (64.3%) responded . Poorer responses were observed in patients with pneumonia (40%) or pseudomonal infection (50%) . The response rate was significantly higher among patients with increasing neutrophil counts during therapy (P less than 0.02) . Fungal infection was a common cause of treatment failure . Gastrointestinal side effects and skin rashes were occasionally seen . No patient developed central nervous system toxicity . Imipenem is a practical alternative to antibiotic combinations for management of neutropenic infection . However, careful monitoring is essential in the subgroups of patients with pneumonia or pseudomonal infections, who may require modifications of therapy.

Clin Chem, 1988 Nov, 34(11), 2291 - 4
New enzymatic determination of sialic acid in serum; Teshima S et al.; This enzymatic method for determination of sialic acid involves use of neuraminidase (EC 3.2.1.18), N-acetylneuraminate lyase (EC 4.1.3.3), acylglucosamine 2-epimerase (EC 5.1.3.8), N-acetylhexosamine oxidase (from Pseudomonas sp.), and peroxidase (EC 1.11.1.7) . Because the method does not require pyruvic acid in the assay medium, interference by pyruvic acid in serum can be avoided . This simple, accurate assay is little affected by other substances in serum.

Appl Environ Microbiol, 1988 Nov, 54(11), 2756 - 8
Novel method for studying plasmid transfer in undisturbed river epilithon; Bale MJ et al.; A method for in situ mating experiments is described which involved overnight incorporation of donors containing the mercury resistance plasmid pQM1 and recipients into the epilithon on separate river stones . The stones were then joined to begin the mating . Transfer frequencies obtained were between 2.2 x 10(-1) and 2.5 x 10(-6) per recipient and appeared to depend on the donor-to-recipient ratio (489/1 to 0.0047/1) and not on the river temperature (12 to 19 degrees C) . Controls showed that the low density of donors and recipients at the end of the experiment (3.4 x 10(2) to 7.0 x 10(5) cm-2) did not significantly affect the heterotrophic bacterial count (1.43 x 10(6) to 6.39 x 10(6) cm-2) nor the fluorescent-pseudomonad count (2.3 x 10(4) to 9.33 x 10(4) cm-2).

Circulation, 1988 Nov, 78(5 Pt 2), III66 - 72
Staged cardiac transplantation . Total artificial heart or ventricular-assist pump?
Pae WE Jr, Pierce WS, Myers JL, Wisman CB, Campbell DB, Waldhausen JA.
Because of donor organ unavailability, staged cardiac transplantation has been performed in eight patients with The Pennsylvania State University pneumatic total artificial heart (two patients) or with the Pierce-Donachy ventricular-assist pump (six patients) . Of the six patients who received the ventricular-assist pump, four received cardiac allografts after 3, 11, 21, and 31 days of pump support, respectively . Two patients died before transplantation; the causes of death were non-device-related complications . One additional patient died of Pseudomonas sepsis after staged cardiac transplantation . The remaining three patients are alive and have been followed up for as long as 2 years after staged cardiac transplantation . Of the two patients who were supported with the total artificial heart, one underwent staged cardiac transplantation after 11 days of support . Unfortunately, this patient succumbed to fungal sepsis 17 days later . The remaining patient, who received the total artificial heart after rejection of a transplanted heart, expired 379 days later before a suitable donor organ could be located . These experiences indicate that 1) the ventricular-assist pump and total artificial heart can provide reasonably safe effective circulatory support until a patient's overall physiological status is optimal, until a donor organ can be located for transplantation, or both; 2) there is a need for short-, intermediate-, and long-term support system capabilities; and 3) regardless of the patients' underlying pathology (ischemic versus nonischemic cardiomyopathy), in most instances, the simpler external ventricular-assist pump is capable of satisfactory hemodynamic circulatory assistance.

Circulation, 1988 Nov, 78(5 Pt 2), III47 - 57
Reversal of recalcitrant cardiac allograft rejection with methotrexate; Costanzo-Nordin MR et al.; Refractory cardiac transplant rejection is a major therapeutic dilemma . The effectiveness of methotrexate (MTX) in autoimmune diseases prompted us to explore its efficacy in 10 cardiac transplant recipients, aged 20-53 years (39 +/- 13 years; mean +/- SD), with biopsy evidence of drug-refractory cardiac allograft rejection . Nine cardiac transplant recipients were maintained on triple antirejection therapy (cyclosporine, azathioprine, and prednisone), and the remaining recipient was maintained on cyclosporine and prednisone . Rejection episodes treated with MTX occurred 20-422 days (165 +/- 137 days) after transplantation and were the sixth episode of rejection for one recipient, the third for four recipients, the second for four recipients, and the first for one recipient . Before MTX administration, cardiac allograft rejection persisted despite intensified immunosuppression including OKT3 antibody . MTX, given intravenously, orally, or by both routes at a dose of 10-175 mg (85 +/- 62 mg), reversed rejection in nine of 10 recipients (90%) within 7-63 days (26 +/- 18 days) . Elevated pulmonary artery wedge pressures were reduced to normal levels after MTX therapy (15.2 +/- 3.5 before vs . 10.6 +/- 3.0 mm Hg after; p less than 0.05) . Leukopenia occurred in five cardiac transplant recipients after treatment with MTX . Adverse reactions to MTX resolved after MTX therapy was discontinued in all but one recipient . This recipient received one of the larger MTX doses (150 mg) and developed fatal Pseudomonas pneumonia . Twelve moderate rejection episodes recurred in nine recipients, seven episodes of which were successfully re-treated with MTX . Two of these seven recipients have now been rejection-free for 15 months . Of five recurrent episodes of cardiac transplant rejection not treated with MTX, two could not be reversed and were fatal . MTX may be a valuable drug for reversing refractory cardiac allograft rejection . Before MTX therapy gains wider use, however, a clearer understanding of its enhanced ability to suppress the bone marrow is needed.

Antimicrob Agents Chemother, 1988 Nov, 32(11), 1636 - 9
Outer membrane permeability in Pseudomonas cepacia: diminished porin content in a beta-lactam-resistant mutant and in resistant cystic fibrosis isolates; Aronoff SC; Since beta-lactam resistance is a feature of Pseudomonas cepacia isolates causing pulmonary infections in cystic fibrosis (CF), this study was undertaken to determine whether alterations in beta-lactam permeability mediate drug resistance in this species . A beta-lactam-susceptible non-CF isolate (strain 75-26), a resistant mutant derived from 75-26 by selection for cross-resistance to ciprofloxacin and ceftazidime, and two resistant CF isolates of P . cepacia were used . Permeability constants were calculated from the rate of nitrocefin hydrolysis in intact bacterial cells . Qualitative changes in outer membrane proteins were determined electrophoretically . The permeability constants of the mutant and the resistant CF isolates were lower than the value for the reference strain, 75-26 . Whereas the lipopolysaccharide side chains were present in the test and reference strains, the resistant mutant and the CF isolates contained reduced amounts of the 36-kilodalton (kDa) outer membrane protein and failed to express the 27-kDa outer membrane protein . These observations suggest that the 27-kDa outer membrane protein may be a major porin or a major protein component of the porin complex in P . cepacia and that decreased expression of the 36-kDa outer membrane and loss of the 27-kDa porin are associated with high-level beta-lactam resistance in some CF isolates of P . cepacia.

Appl Environ Microbiol, 1988 Nov, 54(11), 2877 - 80
Delivery system for creation of one-step in vivo lac gene fusions in Pseudomonas spp . involved in biological control; O'Sullivan DJ et al.; The suicide plasmid pVA838 carrying the operon fusion transposon Tn5-lac was used as a delivery system to introduce Tn5-lac into Pseudomonas sp . strain M114 . Random, in vivo lac gene fusions were successfully isolated in a one-step conjugation approach with this vector system . Tn5-lac-containing exconjugants were recovered at a frequency of approximately 10(-7) per recipient . However, when the mating temperature was increased from the normal growth temperature (28 degrees C) to 34 degrees C, the frequency was increased to approximately 10(-4) per recipient . A number of in vivo lac gene fusions were isolated and characterized in strain M114, a potentially important bacterium for biological control purposes.

Presse Med, 1988 Oct 26, 17(37), 1910 - 3
{Ceftazidime on animal experimental models}; Carbon C; Experiments on animal models represent an important stage in the development of antibiotics . As regards ceftazidime, they have confirmed that its activity is time-dependent in vivo, that there is no post-antibiotic effect and that combining this cephalosporin with an aminoglycoside is useful in neutropenic animals, notably in case of Pseudomonas infection . The potential usefulness of ceftazidime in the treatment of meningitis has also been demonstrated . However, animal models have limited value when one tries to extrapolate to man the therapeutic results obtained . In particular, they provide no information on optimal dosage, critical concentrations or desirable duration of treatment.

J Biol Chem, 1988 Oct 15, 263(29), 15211 - 6
Nonlinear relationship between concentration and activity of a bacterial ice nucleation protein; Southworth MW et al.; The expression level of an ice nucleation gene (inaZ) was varied in Escherichia coli to observe the relationship between activity and gene product . The ice nucleation activity increased as the 2nd to 3rd power of the membrane concentration of the inaZ gene product, implying that molecules of InaZ protein interact cooperatively in groups of two to three at the rate-limiting step of ice nucleus assembly . The 2nd to 3rd power relationship was independent of the threshold temperature at which ice nucleation was measured and was consistent over a 500-fold range of protein concentration . Such a relationship indicates that the same rate-limiting step must be common to the formation of ice nuclei displaying all the various threshold temperatures within a bacterial population . Observations of Pseudomonas syringae, expressing the inaZ gene at various levels, were consistent with a similar relationship and hence a similar mechanism of ice nucleus assembly in Pseudomonas.

FEBS Lett, 1988 Oct 10, 238(2), 325 - 8
Cloning and high-level expression of a chloroperoxidase gene from Pseudomonas pyrrocinia in Escherichia coli; Wolfframm C et al.; A chloroperoxidase gene from Pseudomonas pyrrocinia was cloned into Escherichia coli using the cosmid vector pJB8 . The gene coding for the chloroperoxidase could be localized to a 1.5 kb fragment of DNA which was subcloned into the high-copy-number plasmid pUC18 . In one subclone increased halogenating activity could be found which was 570-fold greater than in P . pyrrocinia . The halogenating enzyme was identified as the chloroperoxidase by SDS-polyacrylamide gel electrophoresis.

Biokhimiia, 1988 Oct, 53(10), 1718 - 27
{Isolation and properties of monomeric and oligomeric forms of gene-engineered human leukocyte interferon alphaA from Pseudomonas sp.}; Borukhov SI et al.; Using stepwise ion-exchange and gel-permeation high performance liquid chromatography and SDS-PAAG gel electrophoresis, it was demonstrated that the non-reduced gene-engineered interferon alpha A is represented by multiple forms, namely, four monomers, four dimers, two trimers and one tetramer . All the protein forms were obtained in an individual state and characterized in terms of antiviral activity and immunochemical properties . The heterogeneity of the protein is due both to the formation of anomalous intermolecular disulfide bonds and to the existence of reduced S-S bonds . The antiviral activity of the dimers, trimers and tetramers expressed as units per mole of protein is equal to that for the monomeric form, i.e., the interaction of one monomeric subunit of the covalently-linked oligomer is sufficient for the manifestation of the protein antiviral activity . This suggests that the antiviral status of the cell does not depend on the amount internalized interferon molecules of their processing products but is controlled by the cell receptor whose internalization and, possibly, processing stimulate the transcription of genes involved in the triggering of the immune response.

Eur Respir J, 1988 Oct, 1(9), 868 - 9
Pseudomonas thoracic empyema secondary to nosocomial rhinosinusitis; Meyer P et al.; Three cases of Pseudomonas thoracic empyema occurring in nasotracheally intubated patients are reported . Paranasal rhinosinusitis, a well documented complication of prolonged nasotracheal intubation, could be the primary infectious location . Massive respiratory tract colonization leads to extensive necrotizing pulmonary lesions . Failure of diagnosis and treatment of sinus involvement could be responsible for persistent or recurrent pleural empyema . Treatment includes continuous pleural drainage, sinusitis treatment and antibiotics . This complication should be considered in the choice between early tracheostomy and prolonged nasotracheal intubation in Intensive Care Unit (ICU) patients.

Appl Environ Microbiol, 1988 Oct, 54(10), 2478 - 85
New naphthalene-degrading marine Pseudomonas strains; Garcia-Valdes E et al.; Over 100 strains that utilized naphthalene as the only carbon and energy source were isolated from samples of marine sediments taken from a heavily polluted area . The isolates were characterized taxonomically and physiologically . Most of these strains belonged to the genus Pseudomonas, and seven of them did not fit any previous taxonomic description . They differed from type strains in a few biochemical characteristics and in the utilization of aromatic compounds . None had catechol 1,2-dioxygenase activity, and catechol 2,3-dioxygenase was responsible for the aromatic ring cleavage . DNA hybridization demonstrated a close relationship between two isolates and the Pseudomonas stutzeri type strain, and between five isolates and the Pseudomonas testosteroni type strain . On the basis of nutritional and enzymatic characteristics, it was assumed that the seven isolates represent new biovars belonging to the species P . testosteroni and P . stutzeri that are able to degrade aromatic hydrocarbons.

J Laryngol Otol, 1988 Oct, 102(10), 872 - 6
Relapsing malignant otitis externa successfully treated with ciprofloxacin; Morrison GA et al.; Two cases are presented, both patients with advanced relapsing malignant otitis externa . The antibiotic ciprofloxacin has strong anti-pseudomonal activity . It was given orally for six months to both patients, following administration of the traditional parenteral antibiotic courses, and in each case the disease has been extinguished . We therefore recommend that the management of patients shown to have malignant otitis externa should include strict diabetic control, regular local aural toilet, gentamicin ear drops and a six week course of parenteral antipseudomonal antibiotic agents (usually gentamicin and azlocillin) together with metronidazole to cover any anaerobic element in the infection . This regimen should be followed by a six month course of oral ciprofloxacin (750 mg b.d.) . Indium scans should be used to monitor recovery . We believe that this regimen can significantly reduce the morbidity and mortality of patients suffering from malignant otitis externa with cranial nerve involvement.

Can J Microbiol, 1988 Oct, 34(10), 1135 - 41
Degradation of polycyclic aromatic hydrocarbons and aromatic heterocycles by a Pseudomonas species; Foght JM et al.; Enrichment cultures were established with the aromatic fraction of a crude oil and screened for aromatic-degrading pseudomonads, using a sprayed plate technique . One isolate identified as Pseudomonas sp . HL7b was chosen for further study because it oxidized several polycyclic aromatic hydrocarbons and aromatic heterocycles without an apparent lag . Using capillary gas chromatography, spectrophotometry, and radiorespirometry, it was found to be capable of mineralizing and (or) oxidizing a wide range of polycyclic aromatic hydrocarbons, S-, N-, and O-heterocyclic analogues, and alkyl polycyclic aromatic hydrocarbons, but not aliphatic hydrocarbons . The isolate displayed two colonial morphologies which correlated with variation in degradative phenotype and hydrophobicity as measured by polystyrene adherence . Four cryptic plasmids were observed in both colonial types . Pseudomonas sp . HL7b degraded dibenzothiophene co-metabolically by a recognized pathway, but this degradation was constitutive, rather than inducible as reported for other bacteria.

J Bacteriol, 1988 Oct, 170(10), 4924 - 30
Cloning and sequencing of Pseudomonas genes encoding vanillate demethylase; Brunel F et al.; A 2,598-base-pair (bp) SalI-HincII DNA fragment has been cloned which codes for vanillate demethylase, the enzyme responsible for the demethylation of vanillate (3-methoxy-4-hydroxybenzoate) to protocatechuate (3,4-dihydroxybenzoate) . Complementation and insertional inactivation experiments have shown that this fragment carries two genes (vanA and vanB) which are predominantly cotranscribed from a promoter upstream of vanA . Nucleotide sequencing of the SalI-HincII fragment confirmed the genetic data: two open reading frames of 987 and 942 bp were present in the transcribed orientation . These had a very high G + C content in the third base of each codon, which is characteristic of Pseudomonas chromosomal genes . Expression of the genes in Escherichia coli with the T7 RNA polymerase-promoter system gave rise to two polypeptides of 36 and 33 kilodaltons which could be identified by deletion analysis as the products of vanA and vanB, respectively . A search of the protein sequence data bank indicated that the vanB gene product was related to the ferredoxin family.

J Antibiot (Tokyo), 1988 Oct, 41(10), 1351 - 7
Enzymatic formation of glidobactamine: a peptide nucleus of glidobactins A, B and C, new lipopeptide antitumor antibiotics; Numata K et al.; Glidobactin deacylating activity was found in a bacterial strain of Pseudomonas sp . Glidobactamine, a key intermediate for acyl analogues of glidobactin, was isolated from the enzymatic degradation products of glidobactins after treatment using a column of fibrous active gel on which the cells of the Pseudomonas strain were immobilized . The chemical structure of glidobactamine was confirmed as the intact peptide moiety of glidobactins by chemical reformation of glidobactin A from glidobactamine and 2,4-dodecadienoic acid which is the constitutive fatty acid of glidobactin A.

J Bacteriol, 1988 Oct, 170(10), 4846 - 54
Characterization and expression of two avirulence genes cloned from Pseudomonas syringae pv . glycinea; Tamaki S et al.; Two avirulence genes, avrB and avrC, from race 0 of Pseudomonas syringae pv . glycinea, were sequenced and found to encode single protein products of 36 and 39 kilodaltons, respectively . The proteins had neither recognizable signal peptide sequences nor significant stretches of hydrophobic amino acids that might indicate membrane association . Both avrB and avrC had relatively low position 3 and overall G+C contents, which suggests that they may have been recently introduced into P . syringae pv . glycinea . The deduced amino acid sequences of the proteins encoded by avrB and avrC shared 42% identical amino acids . However, when introduced into race 4 of P . syringae pv . glycinea, each gene directed a unique pattern of hypersensitive reactions on several differential soybean cultivars . The avrC protein was overproduced in Escherichia coli cells and deposited as insoluble inclusion bodies in the cell cytoplasm . The avrC protein could be solubilized with urea-octyl glucoside treatment, but neither the solubilized protein nor the intact inclusion bodies elicited a hypersensitive reaction in soybean leaves.

J Bacteriol, 1988 Oct, 170(10), 4501 - 8
Analysis of the Pseudomonas solanacearum polygalacturonase encoded by pglA and its involvement in phytopathogenicity; Schell MA et al.; A major endopolygalacturonase excreted by Pseudomonas solanacearum was purified to greater than 95% homogeneity and shown to have an isoelectric point of 9.0 and a subunit molecular mass of 52 kilodaltons (kDa) . The gene encoding this enzyme (pglA) was isolated from a genomic library of P . solanacearum DNA based on its expression in Escherichia coli and shown to be contained on a 1.8-kilobase DNA fragment . The identity of the pglA gene product and the 52-kDa polygalacturonase was demonstrated by immunoadsorption and isoelectric focusing experiments . The cloned pglA gene was apparently expressed from its own promoter in E . coli and its product was partially secreted into the periplasm . The pglA gene was insertionally inactivated in vitro and used to mutate the chromosomal pglA gene of P . solanacearum by marker exchange mutagenesis . The resulting mutant strain was deficient in production of the 52-kDa polygalacturonase and took twice as long to wilt and kill tomato plants as the wild-type parent in plant bioassay experiments . Complementation in trans with the wild-type cloned pglA gene restored virulence to near wild-type levels . The data indicate that the pglA gene is important, but not absolutely necessary, for pathogenesis.

Proc Natl Acad Sci U S A, 1988 Oct, 85(20), 7521 - 5
Subunit S1 of pertussis toxin: mapping of the regions essential for ADP-ribosyltransferase activity; Pizza M et al.; The toxicity of pertussis toxin is mediated by the ADP-ribosyltransferase activity of subunit S1 . To understand the structure-function relationship of subunit S1 and guide the construction of nontoxic molecules suitable for vaccines, we constructed and expressed in Escherichia coli a series of amino-terminal and carboxyl-terminal deletion mutants as well as a number of molecules containing amino acid substitutions . The shortest peptide still retaining enzymatic activity contains amino acids 2-179 . Within this region we identified three mutants in which amino acid substitutions abolish the enzymatic activity . Mutation of amino acids 8 and 9 or 50 and 53, located within the region of the S1 subunit of pertussis toxin homologous to cholera toxin, causes loss of enzymatic activity . Outside this homology region, substitution of Glu-129 with glycine or aspartic acid also eliminates the enzymatic activity of the S1 subunit . In this respect, Glu-129 resembles the glutamic acid that is crucial for the catalytic activity of diphtheria and Pseudomonas toxins . Once introduced into the Bordetella pertussis chromosome, the above mutations should lead to the synthesis of nontoxic pertussis toxin molecules suitable for vaccine production.

Bioorg Khim, 1988 Oct, 14(10), 1413 - 8
{Antigenic polysaccharides of bacteria . 33 . The structure of O-specific polysaccharide chain of lipopolysaccharide from Pseudomonas cepacia serotype 6}; Knirel' IuA et al.; On mild acid degradation of the Pseudomonas cepacia serotype 6 lipopolysaccharide, the O-specific polysaccharide was obtained, which contains D-mannose and D-galactose residues in the ratio approximately 1:1, as well as O-acetyl groups . On the basis of 1H and 13C NMR analysis, calculation of specific optical rotation, and methylation, it was concluded that the polysaccharide possesses the following structure: (formula; see text) Regularities in glycosidation effects in 13C NMR spectra of 1,3-linked disaccharides containing furanoside residues are discussed.

J Biol Chem, 1988 Sep 25, 263(27), 13725 - 32
Purification and characterization of a novel bacterial non-heme chloroperoxidase from Pseudomonas pyrrocinia; Wiesner W et al.; The first bacterial chloroperoxidase that is capable of catalyzing the chlorination of indole to 7-chloroindole was detected in Pseudomonas pyrrocinia ATCC 15958, a bacterium that produces the antifungal antibiotic pyrrolnitrin (Wiesner, W., van Pee, K.H., and Lingens, F . (1986) FEBS Lett . 209, 321-324) . Here we describe the purification and characterization of this bacterial non-heme chloroperoxidase . The enzyme was purified by DEAE-cellulose chromatography at different pH values, molecular sieve chromatography, and Bio-Gel HTP hydroxylapatite . After the last purification step, chloroperoxidase was homogeneous by polyacrylamide gel electrophoresis and ultracentrifugation . Based on gel filtration and ultracentrifugation results, the molecular weight of the enzyme was 64,000 +/- 3,000 . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band with the mobility of a 32,000 molecular weight species . Therefore, in solution at neutral pH, this chloroperoxidase is a dimer . The enzyme did not exhibit any absorbance in the visible region of the spectrum . The isoelectric point was 4.1 . Chloroperoxidase was specific for I-, Br-, and Cl- and was not inhibited by azide, but was inhibited by cyanide and F- . This procaryotic chloroperoxidase catalyzed the bromination of monochlorodimedone but not its chlorination and has no peroxidase or catalase activity . The pH optimum of the enzyme was between 4.0 and 4.5, and the enzyme was stable between pH 3.5 and 8.5 and showed no loss of activity when incubated at 60 degrees C for 2 h . Chloroperoxidase also chlorinated 4-(2-amino-3-chlorophenyl) pyrrole to yield aminopyrrolnitrin, the immediate precursor of pyrrolnitrin . This suggests very strongly that chloroperoxidase is involved in the biosynthesis of the antibiotic pyrrolnitrin.

Gene, 1988 Sep 15, 69(1), 121 - 9
Nucleotide sequence of the gene for the delta 5-3-ketosteroid isomerase of Pseudomonas testosteroni; Choi KY et al.; The structural gene for the delta 5-3-ketosteroid isomerase of Pseudomonas testosteroni has been sequenced by the dideoxy method . The sequence obtained confirms the amino acid (aa) sequence of Benson et al . {J . Biol . Chem . 246 (1971) 7514-7525} at all but 5 aa residues of the 125-aa polypeptide . Amino acid residues 22, 24, 33, and 38, reported to be asparagines by Benson et al., are found to be encoded by aspartic acid codons . Amino acid residue 77, reported to be a glutamine by Benson et al., is encoded by a glutamic acid codon . The identification of aa 38 as aspartic acid, coupled with its presence in the active site, as indicated by previous affinity and photoaffinity-labeling studies and confirmed independently by x-ray crystallographic studies, strengthens the hypothesis that Asp-38 is the aa responsible for the 4 beta to 6 beta proton transfer which is part of the enzymatic reaction.

Am J Ophthalmol, 1988 Sep 15, 106(3), 279 - 81
Use of collagen corneal shields in the treatment of bacterial keratitis; Sawusch MR et al.; We used an animal model of Pseudomonas keratitis to compare treatment by topical tobramycin with and without the presence of a commercially available collagen corneal shield . Pilot studies showed a significant, 30-fold increase in penetration of tobramycin into the anterior chamber in eyes with a collagen shield in place . Twenty albino rabbit eyes were inoculated with P . aeruginosa to produce stromal keratitis . After 12 hours of topical tobramycin dosing, eyes with a collagen corneal shield in place had a statistically significant (P less than .01) decrease in colony forming unit counts in comparison to treated eyes without a shield and control eyes.

J Biol Chem, 1988 Sep 15, 263(26), 13203 - 7
Mutational analysis of domain I of Pseudomonas exotoxin . Mutations in domain I of Pseudomonas exotoxin which reduce cell binding and animal toxicity; Jinno Y et al.; Pseudomonas exotoxin (PE) is a single polypeptide chain that contains 613 amino acids and is arranged into three structural domains . Domain I is responsible for cell recognition, II for translocation of PE across membranes and III for ADP ribosylation of elongation factor 2 . Treatment of PE with reagents that react with lysine residues has been shown to lead to a reduction in cytotoxic activity apparently due to a modification of domain I (Pirker, R., FitzGerald, D . J . P., Hamilton, T . C., Ozols, R . F., Willingham, M . C., and Pastan, S . (1985) Cancer Res . 45, 751-757) . To determine which lysine residues are important in cell recognition, all 12 lysines in domain I were converted to glutamates by site-directed mutagenesis . Also, two deletion mutants encompassing almost all of domain I (amino acids 4-252) or most of domain I (amino acids 4-224) were studied . The mutant proteins were produced in Escherichia coli, purified, and tested for their cytotoxic activity against Swiss 3T3 cells and in mice . The data indicate that conversion of lysine 57 to glutamate reduces cytotoxic activity towards 3T3 cells 50-100-fold and in mice about 5-fold . Deletion of amino acids 4-224 causes a similar reduction in toxicity towards cells and mice . Deletion of most of the rest of domain I (amino acids 4-252) causes a further reduction in toxicity toward cells and mice indicating this second region between amino acids 225 and 252 of domain I is also important in the toxicity of PE . Competition assays indicated that the ability of PEGlu57 to bind to 3T3 cells was greatly diminished, accounting for its diminished cytotoxic activity.

Surgery, 1988 Sep, 104(3), 494 - 9
Surgical intensive care unit pneumonia; Mock CN et al.; With use of an objective numerical rating system for the assessment of the presence or absence of pneumonia on a chest x-ray film, 81 patients in the surgical intensive care unit with positive sputum cultures were assigned to either colonization (C; 39 patients) or pneumonia (P; 42 patients) groups . Respiratory failures preceding the first positive sputum culture and hepatic and/or renal failure were more frequent in the P group . Escherichia coli and Pseudomonas species, as well as polymicrobial sputa, were more common in the P group . Positive blood or pleural cultures with the same organism found in the sputum were noted in 10 of 11 P patients and only 3 of 10 C patients . Broad-spectrum antibiotic therapy directed at all sputum pathogens decreased mortality in the P group but not in the C group . We conclude that an objective rating system for chest x-ray diagnosis provides a reasonable method for separating patients with pneumonia from those with colonization . We recommend antibiotic therapy directed at all sputum pathogens in patients in surgical intensive care units . For such therapy to be successful, however, diagnostic criteria must be precise and exclude patients with colonized pathogens.

Laryngoscope, 1988 Sep, 98(9), 934 - 9
The evolving treatment of necrotizing external otitis; Kraus DH et al.; Necrotizing external otitis, or malignant external otitis, as initially described by Chandler, is a life-threatening Pseudomonas infection of the external auditory canal and skull base, which occurs most commonly in elderly diabetic patients . Historically, radical surgical intervention was the primary method of treatment . The treatment of choice has shifted during the past 20 years to aggressive systemic antibiotic therapy, with surgery reserved for those patients whose disease is resistant to medical therapy . Using this approach, 19 patients with necrotizing external otitis were treated at the Cleveland Clinic Foundation during the past 8 years . A 90% rate of cure was obtained . The diagnostic approach to patients suspected of having necrotizing external otitis, a classification scheme defining the extent of disease, delivery of systemic antibiotic therapy, indications for surgical intervention, and overall effectiveness of treatment are reviewed.

Appl Environ Microbiol, 1988 Sep, 54(9), 2311 - 7
Possible mechanisms responsible for the reduced intestinal flora in hibernating leopard frogs (Rana pipiens); Banas JA et al.; Mechanisms and factors that normally control the large intestinal flora were investigated to determine whether changes in these parameters could account for the decreased bacterial concentration and facultative nature of the flora found in hibernating frogs . It appeared that low temperatures and limited nutrients were the main factors responsible for the decrease in the bacterial concentration and may also have been responsible for the increase in the proportions of facultative organisms, since no change in the redox potential was seen . The hibernating frogs were extremely sluggish in the removal of India ink particles from the circulatory system by the Kupffer cells of the liver compared with nonhibernating frogs . They were unable to mount an antibody response to bovine serum albumin, but their serum did exhibit killing of Pseudomonas paucimobilis, suggesting opsonization by preformed antibody and complement . The role of these host factors in protecting the hibernating frog against this indigenous flora is discussed.

Jpn J Antibiot, 1988 Sep, 41(9), 1319 - 24
{Clinical evaluation of S 6472 granule preparation (sustained-release cefaclor) in chronic bronchitis}; Tanimoto H et al.; The S 6472 granule preparation (sustained-release cefaclor preparation) was administered to 15 cases of chronic bronchitis for its clinical evaluation; a daily dosage of 750 mg was orally given in 2 divided doses after breakfast and dinner for a duration of 7 to 22 days . Clinical effects were good in 11 cases, fair in 1 case and poor in 3 cases . Among the 13 cases other than 2 unadaptable Pseudomonas infections, good effectiveness was found in 11 cases (efficacy rate: 84.6%) . There appeared to be no side effects or abnormal laboratory test values due to administration of this drug.

Microb Pathog, 1988 Sep, 5(3), 197 - 205
Effect of pyochelin on Pseudomonas cepacia respiratory infections; Sokol PA et al.; Exogenously supplied pyochelin influenced the virulence of Pseudomonas cepacia pyochelin-negative strains in a chronic pulmonary infection model in rats . Groups of rats were inoculated transtracheally with agar beads containing P . cepacia or P . aeruginosa strains, saturated with either pyochelin or PBS . Supplementation of the inocula with pyochelin had no effect on the number of bacteria recovered from the lungs . The availability of pyochelin, however, increased the degree of pathology observed in lungs infected with pyochelin-negative strains of P . cepacia . The area of pathological involvement in the lung was about 2-fold larger, when pyochelin was present . Inclusion of pyochelin in the inoculum had no effect on the degree of pathology observed in lungs infected with a pyochelin-positive P . aeruginosa strain . Pyochelin was shown to stimulate in vitro growth of P . cepacia, but it had no effect on production of lipase or protease, factors which may be involved in P . cepacia virulence . These studies support our hypothesis that pyochelin may be important for dissemination in P . cepacia infections.

Aust Vet J, 1988 Sep, 65(9), 261 - 4
Evaluation of four serological tests for the diagnosis of caprine melioidosis; Thomas AD et al.; A complement fixation (CF) test, 2 indirect haemagglutination (IHA-A; IHA-L) tests which differed in antigen preparation and technique, and a microtitre agglutination (MA) test were compared in the serodiagnosis of melioidosis in goats . One hundred and eighteen experimental serums and 3143 field serums from goats in endemic and non-endemic areas of north Queensland were used in the evaluation . Culture of samples for Pseudomonas pseudomallei from 112 goats provided substantiating evidence of infection . The IHA-A test was the most sensitive, and the CF test the most specific . We advocate the use of the IHA-A as a screening test followed by the CF test for confirmation of active melioidosis . The IHA-A test is the better indicator of past infection.

Appl Environ Microbiol, 1988 Sep, 54(9), 2185 - 91
DNA amplification to enhance detection of genetically engineered bacteria in environmental samples; Steffan RJ et al.; The polymerase chain reaction (PCR) was performed to amplify a 1.0-kilobase (kb) probe-specific region of DNA from the herbicide-degrading bacterium Pseudomonas cepacia AC1100 in order to increase the sensitivity of detecting the organism by dot-blot analysis . The 1.0-kb region was an integral portion of a larger 1.3-kb repeat sequence which is present as 15 to 20 copies on the P . cepacia AC1100 genome . PCR was performed by melting the target DNA, annealing 24-base oligonucleotide primers to unique sequences flanking the 1.0-kb region, and performing extension reactions with DNA polymerase . After extension, the DNA was again melted, and the procedure was repeated for a total of 25 to 30 cycles . After amplification the reaction mixture was transferred to nylon filters and hybridized against radiolabeled 1.0-kb fragment probe DNA . Amplified target DNA was detectable in samples initially containing as little as 0.3 pg of target . The addition of 20 micrograms of nonspecific DNA isolated from sediment samples did not hinder amplification or detection of the target DNA . The detection of 0.3 pg of target DNA was at least a 10(3)-fold increase in the sensitivity of detecting gene sequences compared with dot-blot analysis of nonamplified samples . PCR performed after bacterial DNA was isolated from sediment samples permitted the detection of as few as 100 cells of P . cepacia AC1100 per 100 g of sediment sample against a background of 10(11) diverse nontarget organisms; that is, P . cepacia AC1100 was positively detected at a concentration of 1 cell per g of sediment . This represented a 10(3)-fold increase in sensitivity compared with nonamplified samples.

J Cataract Refract Surg, 1988 Sep, 14(5), 500 - 4
Collagen shield drug delivery: therapeutic concentrations of tobramycin in the rabbit cornea and aqueous humor; Unterman SR et al.; Collagen shields made of porcine collagen were placed in a solution containing tobramycin sulfate (40 or 200 mg/ml) for five minutes, then applied to rabbit eyes . One, four, or eight hours after application, the corneas, aqueous humor samples, and shields were assayed for antibiotic . At all intervals, the concentration of antibiotic in the corneas and aqueous humor samples exceeded the mean inhibitory concentration for tobramycin, as determined for most strains of Pseudomonas . Shields immersed in 200 mg/ml tobramycin produced significantly higher concentrations of antibiotic in the cornea at one hour than subconjunctival injections of tobramycin (20 mg) (P = .0001) . Shields immersed in 40 mg/ml tobramycin produced higher, although not significantly higher, concentrations of antibiotic in the cornea at one hour than subconjunctival injections of tobramycin (20 mg) (P = .318) . Shields immersed in commercially available tobramycin drops or injectable tobramycin solution (40 mg/ml) caused no epithelial damage visible by slitlamp examination . Collagen shields containing antibiotics can serve as a vehicle for drug delivery and may prove superior to current methods for preoperative and postoperative antibiotic prophylaxis and the initial treatment of bacterial keratitis.

Pediatr Infect Dis J, 1988 Sep, 7(9), 634 - 6
Neonatal melioidosis: a report of 5 cases; Lumbiganon P et al.; Melioidosis, caused by Pseudomonas pseudomallei, occurs in tropical areas and is diagnosed mostly in adults . In Khon Kaen, a province of northeast Thailand, five cases of infantile melioidosis were managed at Srinagarind Hospital . The patient's specimens were submitted to microbiologic and serologic examination for P . pseudomallei demonstrated by indirect hemagglutination . Possible modes of transmission such as environment, perinatal exposure and venereal transmission were investigated.

Rev Infect Dis, 1988 Sep-Oct, 10(5), 915 - 21
Experience with trimethoprim-sulfamethoxazole in treatment of infective endocarditis; Street AC et al.; The published experience with trimethoprim-sulfamethoxazole (TMP-SMZ) for treatment of infective endocarditis was reviewed . Among 62 cases, a high proportion had unusual causative organisms: 60% of cases were due to Coxiella burnetii or Pseudomonas species . Only 17% of patients had previously normal cardiac valves . Patients often had complicated courses in which TMP-SMZ was tried only after other treatment regimens had failed, yet a successful outcome was achieved in 61% of cases . Thirty-five patients were treated with antibiotics alone, while the other 27 patients required combined medical and surgical management . TMP-SMZ has a limited role in the management of infective endocarditis; specific guidelines for its use, including proper laboratory control, have been delineated.

Bioorg Khim, 1988 Sep, 14(9), 1208 - 13
{Antigenic polysaccharides of bacteria . 32 . The structure of O-specific polysaccharide chains of Pseudomonas cepacia serotype B and E lipopolysaccharides containing D-fucose}; Knirel' IuA et al.; On the basis of acid hydrolysis, methylation, 1H and 13C NMR analysis, and calculation of specific optical rotation, the following structures were established for O-specific polysaccharides of Pseudomonas cepacia serotypes B and E: ----3)-beta-D-Galf-(1----3)-alpha-D-Fucp-(1----serotype B ----3)-beta-D-GlcpNAc-(1----3)-alpha-D-Fucp-(1----serotype E A characteristic feature of the polysaccharides is the presence of D-fucose, rather rare for bacterial antigens.

J Bacteriol, 1988 Sep, 170(9), 4399 - 401
Induction of the copper resistance operon from Pseudomonas syringae; Mellano MA et al.; Cupric sulfate induced mRNA specific to the copper resistance gene cluster previously cloned from Pseudomonas syringae pv . tomato PT23 . mRNA from each of the four genes of this cluster responded in a similar manner to induction over time and with different concentrations of cupric sulfate . Promoter fusion constructs indicated the presence of a single copper-inducible promoter upstream from the first open reading frame.

J Biol Chem, 1988 Aug 25, 263(24), 12077 - 84
Ketopantoic acid reductase of Pseudomonas maltophilia 845 . Purification, characterization, and role in pantothenate biosynthesis; Shimizu S et al.; Ketopantoic acid reductase (EC 1.1.1.169), an enzyme that catalyzes the formation of D-(-)-pantoic acid from ketopantoic acid, was purified 6,000-fold to apparent homogeneity with a 35% overall recovery from Pseudomonas maltophilia 845 and then crystallized . The relative molecular mass of the native enzyme, as estimated by the sedimentation equilibrium method, is 87,000 +/- 5,000, and the subunit molecular mass is 30,500 . The enzyme shows high specificity for ketopantoic acid as a substrate (Km = 400 microM, Vm = 1,310 units/mg of protein) and NADPH as a coenzyme (Km = 31.8 microM) . Only 2-keto-3-hydroxyisovalerate (Km = 8.55 mM, Vm = 35.8 units/mg) was reduced among a variety of other carbonyl compounds tested . The reaction is reversible (Km for D-(-)-pantoic acid = 52.1 mM), although the reaction equilibrium greatly favors the direction of D-(-)-pantoic acid formation . That the enzyme is responsible for the synthesis of D-(-)-pantoic acid necessary for the biosynthesis of pantothenic acid in P . maltophilia 845 is indicated by the observations that only this enzyme is missing in D-(-)-pantoate (or pantothenate)-requiring mutants derived from P . maltophilia 845 among several enzymes (i.e . ketopantoyl lactone reductase (EC 1.1.1.168) and acetohydroxy acid isomeroreductase (EC 1.1.1.86}, which may be concerned in the formation of D-(-)-pantoic acid, assayed, whereas it is present in substantial amounts in the parent strain and in spontaneous revertants of the mutants.

Eur J Biochem, 1988 Aug 15, 175(3), 633 - 41
Ornithine-containing lipids of some Pseudomonas species; Kawai Y et al.; Ornithine-containing lipids purified by thin-layer chromatography were found to represent 2-15% of the total extractable cellular lipids in two or three strains each of four Pseudomonas species: P . aeruginosa, P . fluorescens, P . stutzeri and P . cepacia . The structures of the ornithine-containing lipids were elucidated by chemical analysis, thin-layer chromatography, gas-liquid chromatography, gas-liquid chromatography/mass spectrometry (electron impact or secondary ion) and infrared absorption spectroscopy . At least six molecular species of ornithine-containing lipids were present in common in all of the preparations of the four Pseudomonas species . The structure which was the most abundantly in P . fluorescens (about 60% of the total amount of the ornithine-containing lipid) was 3-hydroxyhexadecanoic acid amide-linked to ornithine and esterified to hexadecanoic acid . In addition to this structure, 3-hydroxyoctadecenoic acid amide-linked to ornithine and esterified to hexadecanoic acid was a dominant structure in the ornithine-containing lipids of P . aeruginosa, P . stutzeri or P . cepacia . In P . cepacia, another ornithine-containing lipids with a terminal polar fatty acid, 3-hydroxyhexadecanoic acid amide-linked to ornithine and esterified to 2-hydroxynonadecacyclopropanoic acid or 2-hydroxyoctadecenoic acid, was found; its content, which represented 8-11% of the total extractable cellular lipids, was higher than that of the ornithine-containing lipids with a terminal nonpolar fatty acid . These ornithine-containing lipids exhibited hemagglutinating activity . Additionally, it was very interesting that hydroxy fatty acids included in the ornithine-containing lipids were not found in the phospholipids which represented more than 80% of the total extractable cellular lipids.

Virology, 1988 Aug, 165(2), 489 - 98
In vitro replication and transcription of the segmented double-stranded RNA bacteriophage phi 6; Ewen ME et al.; In vitro conditions that support viral-specific replication and transcription have been developed from Pseudomonas phaseolicola cells infected with the segmented double-stranded RNA bacteriophage phi 6 . Transcription activity, previously shown to occur by semiconservative strand displacement, labeled (+) strands of all three genome segments and produced all three corresponding genome length messenger RNAs . Replication activity for each of the three double-stranded RNA segments is observed . Our criteria for replication were formation of genomic length double-stranded RNA products and at least (-) strand synthesis activity . Mn2+ and Sarkosyl together selectively inhibited transcription . Analysis of replication alone suggested that replication templates are the viral (+) messenger RNAs.

J Med Microbiol, 1988 Aug, 26(4), 269 - 80
The importance of extracellular antigens in Pseudomonas cepacia infections; Straus DC et al.; A clinical isolate of Pseudomonas cepacia from a cystic fibrosis patient was examined for its ability to produce extracellular toxic material . The organism was grown to stationary phase in a defined medium and toxic material was isolated by ultrafiltration, ion-exchange chromatography on DEAE-Sephacel and gel-filtration chromatography on Sepharose 4B . It consisted of a surface carbohydrate antigen, lipopolysaccharide and protein, and had an LD50 (when injected intraperitoneally into mice) of 395 +/- 20 micrograms . The toxicity appeared to be associated with the lipopolysaccharide portion of the complex, because boiling for 15 min and exposure to proteolytic enzymes had no effect on toxicity . However, saponification destroyed the toxicity of the compound . Studies employing radial immunodiffusion with the sera of mice infected with this organism demonstrated production of the complex in vivo at levels approaching those sufficient to produce death . When sublethal amounts of this complex were placed in the lungs of specific-pathogen-free rats, the lung pathology observed after 12, 24, 36 and 48 h was extensive . However, antibody generated in rabbits against this material could protect mice against the complex, as well as against challenge by the homologous organism . These data indicate that extracellular toxic material produced by P . cepacia may be responsible for the lethality and lung tissue destruction normally associated with an active pneumonia caused by this organism.

J Dairy Res, 1988 Aug, 55(3), 373 - 80
Enhanced inactivation of bacterial lipases and proteinases in whole milk by a modified ultra high temperature treatment; Bucky AR et al.; Cultures of Pseudomonas spp . strains P10, P12 and P15 grown in whole milk which contained approximately 1 x 10(8) viable bacteria ml-1 demonstrated near linear increases in the concentration of short-chain free fatty acids and trichloroacetic acid soluble free amino groups at 20 degrees C, following either ultra high temperature (UHT) treatment (140 degrees C for 5 s) or dual heat treatments (140 degrees C followed by either 57, 60 or 65 degrees C) . The dual heat treatments reduced the rates of lipolysis and proteolysis compared to the UHT treatment by up to 25-fold . The dual heat treatment utilizing 60 degrees C for 5 min also effectively limited both lipase and proteinase activities in raw milk culture samples which had contained either 6 x 10(6), 5 x 10(7) or 1 x 10(8) viable bacteria ml-1 . In this system enzyme activities were reduced by up to 10-fold following dual heat treatment compared to UHT treatment alone.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Aug, (8), 16 - 9
{Prospects of the use of chromosomal transformation for the identification of the causative agent melioidosis}; Riapis LA et al.; Prospects for the use of chromosomal transformation for the identification of Pseudomonas pseudomallei and some other medically important Pseudomonas species are considered . The method is simple, reproducible and highly specific . It permits the screening of a great number of cultures and can facilitate further study of the biology of P . pseudomallei and the epidemiology of melioidosis.

Virology, 1988 Aug, 165(2), 482 - 8
Chemical crosslinking of bacteriophage phi 6 nucleocapsid proteins; Hantula J et al.; phi 6 is a lipid-containing dsRNA bacteriophage of Pseudomonas syringae . Its nucleocapsid (NC) has common features with Reoviridae core particles . We report here the crosslinking of phi 6 NC proteins with cleavable 12-A span chemical crosslinker, dithiobis(succinimidyl propionate) . The crosslinked complexes were analyzed in two-dimensional polyacrylamide gels or by using monoclonal antibodies to uncleaved protein complexes in one-dimensional protein gels . The NC surface protein (P8) forms a series of multimeric homopolymers . The phi 6 lytic enzyme, protein P5, is associated with P8 on the NC surface . The interior NC proteins P1 and P4, associated with the virus polymerase activity, are also in contact with the P8 shell . A P1 + P4 complex is also formed . Only one of the NC proteins (P7) did not easily form complexes with the other NC proteins . These results indicate a very closely packed P8 surface lattice with specific contacts to the internal NC proteins.

Cancer Res, 1988 Jul 15, 48(14), 3919 - 23
Enhancement of the activity of immunotoxins made with either ricin A chain or Pseudomonas exotoxin in human ovarian and epidermoid carcinoma cell lines; Pirker R et al.; The present study evaluates whether the in vitro activity of immunotoxins can be enhanced by verapamil or by various antagonists of calmodulin (dansylcadaverine, trifluoperazine, chlorpromazine) . The following immunotoxins made with either Pseudomonas exotoxin (PE), recombinant ricin A chain (rRTA), or ricin A chain (RTA) were used: HB21-PE and 454A12-rRTA that both recognize the human transferrin receptor; and 260F9-rRTA and 454C11-RTA that both react with human ovarian and breast cancer cells . The cytotoxicity of these immunotoxins was determined in human ovarian carcinoma cell lines and KB cells . Verapamil, that was demonstrated previously to enhance the cell-killing activity of PE immunotoxins, enhanced the activity of several ricin A chain immunotoxins, including 454A12-rRTA, 260F9-rRTA, and 454C11-RTA . Comparing 50% inhibitory dose values for inhibition of protein synthesis by 454A12-rRTA, enhancement ranged from 2- to greater than 25-fold, was dependent on the concentration of verapamil, and was greatest at short incubation times . In addition, the cytotoxicity of HB21-PE and of selected RTA immunotoxins was increased up to 30-fold by the addition of various calmodulin antagonists . The enhancing drugs did not decrease the specificity of the immunotoxins.

J Biol Chem, 1988 Jul 5, 263(19), 9333 - 8
Phospholipid requirement for expression of ice nuclei in Pseudomonas syringae and in vitro; Govindarajan AG et al.; Delipidation of partially purified outer membranes of Pseudomonas syringae by various delipidating agents resulted in a significant loss of ice nucleation activity associated with the cell envelopes of this and other ice nucleation active bacteria . Lipopolysaccharide depletion of such membranes caused no reduction in ice nucleation activity . Both phospholipid content and ice nucleation activity of membranes were decreased by a similar fractional amount with time after treatment with phospholipase A2 . A proportional quantitative relationship between loss of ice nucleation activity and lipid removal with increasing concentrations of sodium cholate and sodium dodecyl sulfate (SDS) was also observed . Significant linear relationships between the amount of lipid removed by phospholipase A2, sodium cholate, and SDS and the loss of ice nucleation activity in P . syringae outer membranes were observed . However, the slopes of these linear relationships for membranes treated with phospholipase A2 (m = 0.80), SDS (m = 0.94), and sodium cholate (m = 0.53) differed . The lower slope value for cholate-treated membranes indicated a partial substitution of sodium cholate for the phospholipids removed . The ice nucleation activity of delipidated outer membranes was restored by reconstitution with various phospholipids in a cholate dialysis procedure . Lipid classes differed in their ability to restore ice nucleation activity to sodium cholate-treated outer membranes . These results suggest that a hydrophobic environment provided either by lipids or certain detergent micelles is required for proper assembly and structural organization of an oligomeric ice protein complex enabling its expression as an ice nucleus.

J Biol Chem, 1988 Jul 5, 263(19), 9470 - 5
Activity of immunotoxins constructed with modified Pseudomonas exotoxin A lacking the cell recognition domain; Kondo T et al.; Pseudomonas exotoxin (PE) contains three domains whose functions are cell recognition, membrane translocation, and ADP ribosylation of elongation factor 2 . PE40 is a form of PE which is missing the cell recognition domain . To study the properties of PE40, it was expressed in Escherichia coli using a vector which contains a T7 phage promoter, an OmpA signal sequence, and that portion of the PE gene encoding PE40 . Upon induction with isopropyl-1-thio-beta-D-galactopyranoside, large amounts of PE40 were secreted, and highly purified PE40 was prepared from the culture medium . PE40 was chemically coupled to different monoclonal antibodies, and protein synthesis inhibition activities of these immunotoxins was assessed on various cell lines . These activities were compared with the activities of the corresponding immunotoxins made with native PE . These data indicate that PE40 may be useful in the construction of certain immunotoxins.

J Thorac Cardiovasc Surg, 1988 Jul, 96(1), 157 - 61
An outbreak of Pseudomonas cepacia bacteremia associated with a contaminated intra-aortic balloon pump; Rutala WA et al.; In January 1983, symptomatic Pseudomonas cepacia bacteremia developed in two patients in the cardiothoracic intensive care unit within 3 days after cardiac operation and insertion of an intra-aortic balloon pump . An epidemiologic and microbiologic investigation revealed that both patients required intra-aortic balloon pumping for circulatory support and that the water reservoir of the intra-aortic balloon pump (SMEC, Inc., Cookeville, Tenn.) contained more than 10(5) Pseudomonas cepacia per milliliter . This organism was also recovered from the purge button and on-off switch of the pump and from the hands of a health care worker who manipulated the water reservoir of the intra-aortic balloon pump . Agarose gel electrophoresis of lysates of Pseudomonas cepacia with rapid methods of deoxyribonucleic acid preparation revealed three identical plasmids of the Pseudomonas cepacia from the water reservoir of the intra-aortic balloon pump and from the infected patients . Transmission from the worker's hands to patients presumably occurred by inoculation of the intravascular lines during management . No additional cases of Pseudomonas cepacia bacteremia were observed after the unit was replaced with a nonwater reservior intra-aortic balloon pump . This report substantiates the ability of Pseudomonas cepacia to multiply in water and to cause epidemic bacteremia, identifies the water reservoir of the SMEC intra-aortic balloon pump as a previously unrecognized hazard for the patient requiring intra-aortic balloon pumping, and documents the value of plasmid analysis in elucidating the mode of transmission of nosocomial Pseudomonas cepacia infections.

Rev Infect Dis, 1988 Jul-Aug, 10(4), 765 - 9
Outer membrane permeability and beta-lactamase content in Pseudomonas maltophilia clinical isolates and laboratory mutants; Mett H et al.; Low outer-membrane permeability appears to be responsible for the generally high degree of antibiotic resistance of Pseudomonas maltophilia . Constitutive overproduction of beta-lactamases affects the sensitivity of these bacteria only to those beta-lactam antibiotics that are hydrolyzed by strain-specific beta-lactamases and that do not efficiently induce these enzymes in inducible strains.

J Gen Microbiol, 1988 Jul, 134 ( Pt 7), 1949 - 60
Genetic diversity and relationships of two pathovars of Pseudomonas syringae; Denny TP et al.; To determine genetic relationships within and between two pathovars of Pseudomonas syringae, strains typical of P . syringae pv . tomato (P . s . tomato) and selected strains of P . syringae pv . syringae (P . s . syringae) were characterized by three methods . DNA-DNA hybridization experiments showed that strains of P . s . tomato and P . s . syringae were, respectively, 86-100% and 37-47% homologous to DNA from a P . s . tomato reference strain when tested under stringent conditions . An analysis of electrophoretic variation in enzymes encoded by 26 loci placed 17 P . s . tomato strains studied in a group of four electrophoretic types, and these strains had a mean genetic diversity per locus of 0.076 . Six P . s . syringae strains formed a second group of six electrophoretic types, which had a higher mean genetic diversity per locus of 0.479 . The mean genetic distance separating P . s . tomato from P . s . syringae (D = 0.94) was unexpectedly large for strains of a single species . An analysis of restriction fragment length polymorphisms (RFLPs) with three cloned hybridization probes demonstrated that each of the P . s . tomato and P . s . syringae strains was unique . A method was developed to quantify the RFLP difference between pairs of strains, and cluster analysis revealed relationships among P . s . tomato, but not among P . s . syringae, that were similar to those based on enzyme polymorphisms . Implications of these findings for bacterial systematics and epidemiology are discussed.

Mol Gen Genet, 1988 Jul, 213(1), 72 - 7
Identification and characterization of Tn4653, a transposon covering the toluene transposon Tn4651 on TOL plasmid pWW0; Tsuda M et al.; A Pseudomonas TOL plasmid pWW0 possesses toluene degradative pathway (xyl) genes . Unstable maintenance of a pWW0 derivative in Escherichia coli allowed us to identify two transposable elements each carrying all the xyl genes . One element corresponded to a 56 kb transposon, Tn4651, which we had previously characterized . The other element newly identified in this study was 70 kb long, and this element, designated Tn4653, completely included Tn4651 . Genetic analysis of Tn4653 demonstrated that its transposition involves two steps, i.e . cointegrate formation and its subsequent resolution . The former step required a trans-acting factor, transposase, which was encoded in a 3.0 kb fragment at one end of Tn4653, and the latter step was inferred to be mediated by the factors necessary for resolution of the Tn4651-mediated cointegrate . The transposase functions were not interchangeable between the two transposons.

Rev Infect Dis, 1988 Jul-Aug, 10(4), 806 - 17
Molecular cloning and expression of the imipenem-hydrolyzing beta-lactamase gene from Pseudomonas maltophilia in Escherichia coli; Dufresne J et al.; The L-1 penicillinase structural gene, blaS, from Pseudomonas maltophilia was cloned into the vector pACYC184 . The pMON01 recombinant plasmid selected by ampicillin resistance carried a 2.6-kilobase (kb) Sau3A fragment of P . maltophilia DNA and was confirmed to express L-1 beta-lactamase by comparative isoelectric focusing . A detailed physical map was constructed, and the blaS structural gene was localized with a 17-mer oligonucleotide mixed probe encoding the L-1 NH2-terminal amino acid sequence . Induction studies confirmed constitutive expression . Isolation of a complete beta-lactamase operon was attempted by construction of a P . maltophilia genomic library into phage lambda 2001 . A recombinant phage was selected by DNA hybridization; the 13.4-kb DNA insert was physically mapped and subcloned into the high-copy-number plasmid pACYC184 and into the low-copy-number vector pLG338 . The expression of the cloned blaS L-1 structural gene and levels of beta-lactamase synthesis were studied in Escherichia coli . The protein synthesized was found to be similar to the L-1 beta-lactamase of the prototype P . maltophilia, although expression levels were gene dosage dependent for beta-lactamase synthesis.

Mikrobiologiia, 1988 Jul-Aug, 57(4), 629 - 33
{Analysis of DNA-DNA homologies in obligate methylotrophic bacteria}; Doronina NV et al.; The genotypic affinity of 19 bacterial strains obligately dependent on methanol or methylamine as carbon and energy sources was studied by techniques of molecular DNA hybridization . The high homology level (35-88%) between motile strain Methylophilus methanolovorus V-1447D and nonmotile strain Methylobacillus sp . VSB-792 as well as other motile strains (Pseudomonas methanolica ATCC 21704, Methylomonas methanolica NRRL 5458, Pseudomonas sp . W6, strain A3) indicates that all of them belong to one genus . Rather high level of homology (62-63%) was found between Methylobacillus glycogenes ATCC 29475 and Pseudomonas insueta ATCC 21276 and strain G-10 . The motile strain Methylophilus methylotrophus NCIB 10515 has a low homology (below 20%) to other of the studied obligate methylobacteria . Therefore, at least two genetically different genera of obligate methylobacteria can be distinguished, namely Methylophilus and Methylobacillus, the latter being represented by both motile and nonmotile forms.

Appl Environ Microbiol, 1988 Jul, 54(7), 1886 - 8
Phosphate starvation induces uptake of glyphosate by Pseudomonas sp . strain PG2982; Fitzgibbon J et al.; Pseudomonas sp . strain PG2982 has the ability to use the phosphonate herbicide, glyphosate, as a sole phosphorus source (J . K . Moore, H . D . Braymer, and A . D . Larson, Appl . Environ . Microbiol . 46:316-320, 1983) . Glyphosate uptake is maximal in the late log phase of growth and is induced by phosphate starvation . Uptake is inhibited by phosphate and arsenate, but not by the amino acids glycine and sarcosine . The Km and Vmax for glyphosate uptake were calculated to be 23 microM and 0.97 nmol/mg (dry weight) per min, respectively . A phosphate transport system with a broad substrate specificity may be responsible for glyphosate uptake.

J Clin Microbiol, 1988 Jun, 26(6), 1241 - 3
Postoperative infant septicemia caused by Pseudomonas luteola (CDC group Ve-1) and Pseudomonas oryzihabitans (CDC group Ve-2); Freney J et al.; Pseudomonas luteola (CDC group Ve-1) and Pseudomonas oryzihabitans (CDC group Ve-2) were both isolated from the same blood culture of a 5-month-old infant, 8 days after open-heart surgery . He quickly responded to appropriate antibiotics . Carbon substrate assimilation tests and fatty acid analysis clearly differentiated these two rarely pathogenic organisms.

J Bacteriol, 1988 Jun, 170(6), 2879 - 83
Nucleotide sequence and organization of copper resistance genes from Pseudomonas syringae pv . tomato; Mellano MA et al.; The nucleotide sequence of a 4.5-kilobase copper resistance determinant from Pseudomonas syringae pv . tomato revealed four open reading frames (ORFs) in the same orientation . Deletion and site-specific mutational analyses indicated that the first two ORFs were essential for copper resistance; the last two ORFs were required for full resistance, but low-level resistance could be conferred in their absence . Five highly conserved, direct 24-base repeats were found near the beginning of the second ORF, and a similar, but less conserved, repeated region was found in the middle of the first ORF.

J Cell Physiol, 1988 Jun, 135(3), 502 - 8
Epidermal growth factor-dependent growth of human KB cells in a defined medium and altered growth factor requirements of KB mutants resistant to EGF-Pseudomonas exotoxin conjugates; Amano F et al.; A serum-free culture system was established for human KB carcinoma (HeLa) cells that consisted of a chemically defined medium and several growth factors including epidermal growth factor (EGF), insulin, transferrin, hydrocortisone, and ethanolamine . EGF and insulin showed the greatest effects on the growth rate of KB cells . Insulin-like growth factor I (IGF-I) at the same concentration as insulin stimulated cell growth less than insulin . Transferrin, hydrocortisone, or ethanolamine had no growth-stimulatory effects alone but were stimulatory when combined with EGF and/or insulin . Transforming growth factor-beta inhibited growth and triiodothyronine stimulated growth . The growth factor requirements were established for several KB mutants with low EGF receptor levels that had been selected for resistance to a conjugate of EGF with Pseudomonas exotoxin (EGF-PE) . Three of five KB mutants did not respond to EGF; two other mutants responded to a lesser extent than the parental KB cells . Four mutants had a reduced response to insulin and responded to T3; one mutant (ET-30) responded to neither . These results indicate that KB cells selected for EGF-PE resistance have lost their growth response to EGF and illustrate the usefulness of serum-free medium for studying the growth factor requirements of mutants with altered receptor levels.

South Med J, 1988 Jun, 81(6), 796 - 8
Pseudomonas paucimobilis empyema after cardiac transplantation; Cover TL et al.; Empyema caused by P paucimobilis and mouth flora occurred in a 56-year-old man two months after orthotopic cardiac transplantation . Successful treatment was accomplished with chest tube drainage and four weeks of intravenous cefazolin and clindamycin.

J Cell Physiol, 1988 Jun, 135(3), 527 - 32
Genetic characterization of human KB cell lines resistant to epidermal growth factor: Pseudomonas exotoxin conjugates; Amano F et al.; Pleiotropic human KB cell mutants, selected for resistance to a conjugate of epidermal growth factor with Pseudomonas exotoxin (PE-EGF), were characterized genetically . These mutants have a pleiotropic phenotype, which includes reduced number of EGF receptors and reduced growth rate . Hybrid cells between HeLa D98 and four out of five of these resistant cell lines were more resistant to PE-EGF than hybrids formed between HeLa D98 and parental KB cells . This result indicates that the phenotype of PE-EGF resistance is incompletely dominant in four out of five cases and recessive in one out of five variants . In three separate experiments, transfection of DNA from two of the dominant resistant cell lines resulted in transformation of wild-type KB cells to PE-EGF resistance, confirming the dominant nature of these mutations, which affect levels of EGF receptor in KB cells.

Postgrad Med J, 1988 Jun, 64(752), 426 - 30
Serious complications following treatment of chronic idiopathic thrombocytopenic purpura; Wanachiwanawin W et al.; Six patients had serious complications as consequences of treatment of idiopathic thrombocytopenic purpura . Five had splenectomy-related complications, one of them developed fatal intra-abdominal bleeding . Three patients acquired operation-related serious infection, two of them died . Serious neutropenia after vinblastine-loaded platelets occurred in one patient leading to pseudomonas septicaemia and panophthalmitis with permanent vision loss of left eye . Recurrence thrombocytopenia occurred in every case during serious complications . Early detection by awareness of the possibility of serious complications can reduce morbidity and mortality occurring after therapy of idiopathic thrombocytopenic purpura.

J Biochem (Tokyo), 1988 Jun, 103(6), 1045 - 9
L-alanine: 4,5-dioxovalerate aminotransferase from Pseudomonas riboflavina: purification and inactivation by methylglyoxal; Rhee H et al.; L-Alanine:4,5-dioxovalerate aminotransferase, which catalyzes transamination between L-alanine and 4,5-dioxovalerate to yield delta-aminolevulinate and pyruvate, has been purified from Pseudomonas riboflavina IFO 3140 . The enzyme had a molecular weight of 190,000 and consisted of four identical subunits . It was crystallized as pale yellow needles . The enzyme used L-alanine (relative activity 100), beta-alanine (39), and L-ornithine (14) as amino donors . gamma-aminobutyrate (55) and epsilon-aminocaproate (34) were also effective as amino donors . The reaction proceeded according to a ping-pong mechanism and the Km values for L-alanine and 4,5-dioxovalerate were 1.7 and 0.75 mM, respectively . The activity of the enzyme is strongly inhibited by pyruvate, hemin, and methylglyoxal . Methylglyoxal interacted with the enzyme and brought about a complete inactivation.

Microbiologia, 1988 Jun, 4(2), 125 - 8
Heat stability of the extracellular lipase from a Pseudomonas strain isolated from refrigerated raw milk; Garcia-Collia P et al.; A study on the thermal stability of the extracellular lipase of a Pseudomonas strain has been made . The D-values at 140 degrees C ranged from 0.54 to 0.85 min, depending on the heating medium . The average Z-value and activation energy were 36.2 degrees C and 8.18 x 10(4) J . mol-1, respectively . The enzyme showed maximum activity at pH 8.5 and 37 degrees C.

Orthop Rev, 1988 Jun, 17(6), 601 - 4
Pseudomonas osteomyelitis of the metatarsal sesamoid . A case report; Browne T et al.; Osteomyelitis of the metatarsal sesamoid is rare . Few cases of Pseudomonas osteomyelitis have been reported . This report includes a current review of the literature and case report . The difficulty in diagnosis and suggested treatment are discussed.

Antimicrob Agents Chemother, 1988 Jun, 32(6), 819 - 26
Cloning and expression of the imipenem-hydrolyzing beta-lactamase operon from Pseudomonas maltophilia in Escherichia coli; Dufresne J et al.; The L-1 penicillinase structural gene, blaS, from Pseudomonas maltophilia has been cloned into the vector pACYC184 . The pMON01 recombinant plasmid selected by ampicillin resistance carried a 2.6-kilobase Sau3A fragment of P . maltophilia DNA and was confirmed to express L-1 beta-lactamase by comparative isoelectric focusing . A detailed physical map was constructed, and the blaS structural gene was localized with a 17-mer oligonucleotide mixed probe encoding the L-1 N-terminal amino acid sequence . Induction studies confirmed constitutive expression . Isolation of a complete beta-lactamase operon was attempted by construction of a P . maltophilia genomic library into phage lambda 2001 . A recombinant phage was selected by DNA hybridization, and the 13.4-kilobase DNA insert was physically mapped and subcloned into plasmid vectors . Expression and L-1 beta-lactamase synthesis were studied in Escherichia coli.

J Bacteriol, 1988 Jun, 170(6), 2584 - 91
Isolation and characterization of pathogenicity genes of Pseudomonas syringae pv . tabaci; Salch YP et al.; Pseudomonas syringae pv . tabaci BR2 produces tabtoxin and causes wildfire disease on tobacco and bean plants . Approximately 2,700 Tn5 insertion mutants of a plasmid-free strain, PTBR 2.024, were generated by using suicide plasmid pGS9 . Of these Tn5 mutants, 8 were no longer pathogenic on tobacco plants and 10 showed reduced symptoms . All of the eight nonpathogenic mutants caused typical wildfire disease symptoms on bean plants . Two of the nonpathogenic mutants failed to produce tabtoxin . The eight nonpathogenic mutants have Tn5 insertions into different EcoRI and SalI restriction fragments . The EcoRI fragments containing Tn5 from the eight nonpathogenic mutants were cloned into vector pTZ18R or pLAFR3 . A genomic library of the parent strain was constructed in the broad-host-range cosmid pLAFR3 . Three different cosmid clones that hybridized to the cloned Tn5-containing fragment from one of the nonpathogenic mutants, PTBR 4.000, were isolated from the genomic library . These clones contained six contiguous EcoRI fragments (a total of 57 kilobases {kb}) . A 7.2-kb EcoRI fragment common to all three restored pathogenicity to mutant PTBR 4.000 . None of the six EcoRI fragments hybridized to Tn5-containing fragments from the other seven mutants . The 7.2-kb fragment was conserved in P . syringae pv . tabaci and P . syringae pv . angulata, but not in other pathovars or strains . Because the mutants retained pathogenicity on bean plants and because of the conservation of the 7.2-kb EcoRI fragment only in pathovars of tobacco, we suggest that genes on the fragment might be related to host specificity.

Biochemistry, 1988 May 31, 27(11), 3990 - 6
Cloning and nucleotide sequence of the 2,3-dihydroxybiphenyl dioxygenase gene from the PCB-degrading strain of Pseudomonas paucimobilis Q1; Taira K et al.; The bphC gene encoding 2,3-dihydroxybiphenyl dioxygenase was cloned from biphenyl-degrading and chlorinated biphenyl-degrading Pseudomonas paucimobilis Q1, and its complete nucleotide sequence was determined . The DNA-derived protein sequence provides the primary structure of 298 amino acids . Polyclonal antibodies raised against this protein from P . paucimobilis Q1 failed to cross-react with the previously isolated 2,3-dihydroxybiphenyl dioxygenase from Pseudomonas pseudoalcaligenes KF707 {Furukawa, K., & Arimura, N . (1987) J . Bacteriol . 169, 924-927 . Furukawa, K., Arimura, N., & Miyazaki, T . (1987) J . Bacteriol . 169, 427-429}, despite the close similarities of these proteins in terms of their native as well as subunit molecular weights, cofactor, and enzymatic activities . The sequence homology of the 2,3-dihydroxybiphenyl dioxygenase from the two different sources is examined.

Biochemistry, 1988 May 3, 27(9), 3398 - 403
Localization of the active site of diphtheria toxin; Zhao JM et al.; Information about the location of the active site of diphtheria toxin was derived from proteolysis studies and an analysis of its sequence . It was found that a specific trypsin cleavage within whole diphtheria toxin occurs at Lys-39 . Therefore, Lys-39 appears to be a surface residue . Furthermore, protection from proteolysis could be obtained upon binding of either the substrate beta-nicotinamide adenine dinucleotide (oxidized form) (NAD+) or a competing ligand, adenylyl(3'-5')uridine 3'-phosphate (ApUp) . The protection by ApUp, which binds to the toxin very tightly, required only stoichiometric levels . The most likely explanation of these results is that both NAD+ binding and ApUp binding block trypsin either through a steric mechanism or through a local conformational change, suggesting Lys-39 may be near the active site . Further evidence supporting this conclusion comes from comparison of the previously determined sequences of diphtheria toxin and of Pseudomonas exotoxin A, a protein that catalyzes an identical reaction . We find a significant degree of homology between the N-terminal halves of the catalytic domains of these two proteins, which apparently represents active-site residues, and that Lys-39 is in the center of the homologous sequence . Furthermore, the location of the amino acid that is the homologue of Lys-39 within the crystal structure of Pseudomonas exotoxin A is also in agreement with a location in or near the active site . Other unusual features in the sequences of diphtheria toxin and Pseudomonas exotoxin A are also described, and on the basis of the experiments presented, a possible function for ApUp is considered.

Biochemistry, 1988 May 3, 27(9), 3277 - 85
Studies of electron-transfer properties of salicylate hydroxylase from Pseudomonas cepacia and effects of salicylate and benzoate binding; Einarsdottir GH et al.; The pH dependence of the redox behavior of salicylate hydroxylase from Pseudomonas cepacia as well as the effects of salicylate, benzoate, and chloride binding is described . At pH 7.6 in 0.02 M potassium phosphate buffer E1(0')(EFl ox/EFl.-) is -0.150 V and E2(0')(EFl.-/EFl red H-) is -0.040 V versus the standard hydrogen electrode (SHE) . A maximum of 5% of FAD anion semiquinone is thermodynamically stabilized under these conditions . However, in coulometric and dithionite titrations more semiquinone is kinetically formed, indicating slow transfer of the second electron . The potential/pH dependence is consistent with a two-electron, one-proton transfer . Upon salicylate binding the midpoint potential is shifted 0.020 V negative from -0.094 to -0.114 V vs SHE at pH 7.6 . A maximum of 7% of the neutral semiquinone is stabilized both in potentiometric and coulometric titrations . This small potential shift indicates that the substrate is bound nearly to the same extent to all three oxidation states of the enzyme . It is clear that the substrate binding does not make the reduction of the flavin thermodynamically more favorable . In contrast to salicylate, the potential shift caused by the effector, benzoate, is much more significant . (A maximum potential shift of -0.07 V is calculated.) Benzoate binds most tightly to the oxidized form and is least tightly bound to the two-electron-reduced form of the enzyme . For the reduction of the free enzyme the transfer of the second electron or the transfer of the proton is rate limiting, as is shown by the kinetic formation of the anionic semiquinone.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1988 May, 54(5), 1199 - 202
Enzymatic dehalogenation of chlorinated nitroaromatic compounds; Thiele J et al.; 4-Chlorobenzoate dehalogenase from Pseudomonas sp . strain CBS3 converted 4-chloro-3,5-dinitrobenzoate to 3,5-dinitro-4-hydroxybenzoate and 1-chloro-2,4-dinitrobenzene to 2,4-dinitrophenol . The activities were 0.13 mU/mg of protein for 4-chloro-3,5-dinitrobenzoate and 0.16 mU/mg of protein for 1-chloro-2,4-dinitrobenzene compared with 0.5 mU/mg of protein for 4-chlorobenzoate.

J Clin Microbiol, 1988 May, 26(5), 979 - 84
Production of lipase by clinical isolates of Pseudomonas cepacia; Lonon MK et al.; Ten clinical isolates of Pseudomonas cepacia from the sputum of cystic fibrosis patients were examined for the ability to produce lipase . Lipase substrates used included egg yolk agar, four different polyoxyethylene sorbitans (Tweens), and p-nitrophenylphosphorylcholine, a chromogenic substrate used to assay for phospholipase C . Lipase activity was detected in the filtrates of organisms grown to the exponential phase in either tryptose minimal medium or chemically defined medium . Lipase activity increased in the filtrates if the cultures were allowed to proceed into the stationary phase . None of the isolates produced phospholipase C . Lipase activity on Tween 20 ranged from 41.6 X 10(-3) to 640.0 X 10(-3) U/micrograms of protein . The activity was similar or slightly lower when Tween 40, 60, or 80 was used as the substrate . There was no correlation between lipase activity on Tween and that demonstrated on egg yolk agar . Lipase activity increased as pH increased from 7.0 to 9.0 . Boiling for 5 min resulted in 66% loss of enzyme activity . The remaining activity continued to decrease with increasing boiling time . The enzyme was purified by gel filtration on Sephadex G-200, and the resultant preparation, when subjected to polyacrylamide gel electrophoresis, resulted in a single protein band (molecular weight, approximately 25,000) from which lipase activity could be eluted . The purified lipase was not cytotoxic to HeLa cells, nor was it toxic when injected intravenously into mice.

Am J Med, 1988 May, 84(5), 965 - 7
Melioidosis: a reminder; Morrison RE et al.; Recrudescent pulmonary melioidosis developed in two patients 12 and 16 years after their last travels to an endemic area . In one, a clinically silent prostatic abscess may have been the focus; and in both, the diagnosis was difficult to make even when the laboratory was notified of the possibility of infection with Pseudomonas pseudomallei . Recrudescent melioidosis should be considered in febrile patients who have been in endemic areas regardless of the interval from last exposure to the development of disease.

J Bacteriol, 1988 May, 170(5), 2412 - 3
Abundant expression of Pseudomonas genes for chlorocatechol metabolism; Ngai KL et al.; The respective specific activities of catechol 1,2-oxygenase II (catechol 1,2-dioxygenase; EC 1.13.11.1) and muconate cycloisomerase II (chloromuconate cycloisomerase; EC 5.5.1.7) in crude extracts of chlorobenzoate-grown Pseudomonas cells corresponded to about 16 and 11% of the soluble cell protein . High levels of protein synthesis appeared to compensate for a loss in catalytic activity that accompanied evolutionary acquisition of broad substrate specificity required for the enzymes to accommodate halogenated substrates.

Obstet Gynecol, 1988 May, 71(5), 798 - 800
Pleuroamniotic shunting for decompression of fetal pleural effusions; Blott M et al.; Pleuroamniotic shunting was performed at 22-35 weeks' gestation in 11 fetuses with pleural effusions . Eight of the infants, born two to 16 weeks after shunting, had no evidence of pulmonary hypoplasia . Three died in the neonatal period; one because of pseudomonas septicemia, one because of pulmonary hypoplasia caused by an associated diaphragmatic hernia, and the third because of a major cardiac defect . Pleural effusions and their prenatal decompression offer an experimental human model for the study of the effect of intrathoracic compression on pulmonary development.

Jpn J Cancer Res, 1988 May, 79(5), 626 - 31
Tumor necrosis factor-inducing activities of lipid A preparations from Pseudomonas diminuta and Pseudomonas vesicularis; Arata S et al.; Tumor necrosis factor (TNF)-inducing activities of lipid A preparations from P . diminuta and P . vesicularis, which contain mainly 2 mol of 2,3-diamino-2,3-dideoxy-D-glucose and 1 mol of nonglycosidic phosphate as the backbone component and have partly different fatty acid compositions, were examined . TNF was induced by injecting various lipid A fractions into mice that had previously been sensitized with Mycobacterium bovis BCG vaccine . A major component of lipid A of both strains, referred to as A3 fraction, exhibited stronger TNF-inducing activity than A2 fraction having incomplete acyl residues . The removal of ester-linked fatty acyl groups by mild hydrazinolysis of the P . diminuta lipid A results in a marked decrease of the activity . These results suggest that the structure of the hydrophobic part, including the amide-linked acyloxyacyl group(s), of the lipid A molecule play an important role in inducing TNF in the sera of mice.

J Bacteriol, 1988 May, 170(5), 2367 - 73
Role of indoleacetic acid-lysine synthetase in regulation of indoleacetic acid pool size and virulence of Pseudomonas syringae subsp . savastanoi; Glass NL et al.; The phytopathogen Pseudomonas syringae subsp . savastanoi incites the production of galls on olive and oleander plants . Gall formation is dependent upon the bacterial synthesis of the phytohormone indole-3-acetic acid (IAA) . Strains isolated from oleander galls are capable of further metabolizing IAA to an amino acid conjugate, 3-indoleacetyl-epsilon-L-lysine (IAA-lysine); bacterial olive gall isolates lack this activity . In this study, the cloned gene for IAA-lysine synthetase (iaaL+) was introduced into strains isolated from olive and oleander galls to determine its effect on the regulation of IAA pool size and virulence . IAA-lysine was synthesized by isolates from olive galls when iaaL+ was introduced by conjugation, but the amount of IAA which accumulated in culture by the transconjugant was reduced by one-third . When the iaaL+ locus of an oleander gall isolate was inactivated by Tn5 mutagenesis, the resulting mutant did not convert IAA to IAA-lysine; however, it accumulated fivefold more IAA in culture than the wild type did . When inoculated into oleander plants, the iaaL mutant did not cause typical gall symptoms, nor did it replicate within host tissue similarly to the wild type.

J Bacteriol, 1988 May, 170(5), 2306 - 11
Cloning and sequencing of a plasmid-borne gene (opd) encoding a phosphotriesterase; McDaniel CS et al.; Plasmid pCMS1 was isolated from Pseudomonas diminuta MG, a strain which constitutively hydrolyzes a broad spectrum of organophosphorus compounds . The native plasmid was restricted with PstI, and individual DNA fragments were subcloned into pBR322 . A recombinant plasmid transformed into Escherichia coli possessed weak hydrolytic activity, and Southern blotting with the native plasmid DNA verified that the DNA sequence originated from pCMS1 . When the cloned 1.3-kilobase fragment was placed behind the lacZ' promoter of M13mp10 and retransformed into E . coli, clear-plaque isolates with correctly sized inserts exhibited isopropyl-beta-D-thiogalactopyranoside-inducible whole-cell activity . Sequence determination of the M13 constructions identified an open reading frame of 975 bases preceded by a putative ribosome-binding site appropriately positioned upstream of the first ATG codon in the open reading frame . An intragenic fusion of the opd gene with the lacZ gene produced a hybrid polypeptide which was purified by beta-galactosidase immunoaffinity chromatography and used to confirm the open reading frame of opd . The gene product, an organophosphorus phosphotriesterase, would have a molecular weight of 35,418 if the presumed start site is correct . Eighty to ninety percent of the enzymatic activity was associated with the pseudomonad membrane fractions . When dissociated by treatment with 0.1% Triton and 1 M NaCl, the enzymatic activity was associated with a molecular weight of approximately 65,000, suggesting that the active enzyme was dimeric.

Schweiz Med Wochenschr, 1988 Apr 16, 118(15), 558 - 64
{Septic melioidosis following a visit to India}; Thurnheer U et al.; A case is reported of lethal septicemic melioidosis due to Pseudomonas pseudomallei in a 40-year-old woman who had been in India . The epidemiology, clinical findings and management of this unusual disease are discussed . The diagnostic value of serological tests and in vitro sensitivity of Pseudomonas pseudomallei to various antibiotics are outlined . Melioidosis should be considered in the differential diagnosis of any febrile condition in a person returning from a tropical country.

FEBS Lett, 1988 Apr 11, 231(1), 102 - 6
Pyruvate carboxylase from Pseudomonas citronellolis: shape of the enzyme, and localization of its prosthetic biotin group by electron microscopic affinity labeling; Fuchs J et al.; Pseudomonas citronellolis is known to contain a pyruvate carboxylase with an alpha 4 beta 4 composition . All the other pyruvate carboxylases investigated so far are made up of four seemingly identical subunits . Nevertheless, this exceptional pyruvate carboxylase exhibits a size and overall shape similar to other pyruvate carboxylases . Electron microscopic affinity labeling with avidin revealed that the prosthetic biotin groups (one per alpha beta unit, i.e . four per enzyme particle) are located close to the inter-unit junctions of pairs of alpha beta units making up the enzyme . This position of the prosthetic biotin groups is very similar to the location of the biotin in the other carboxylases.

Thorax, 1988 Apr, 43(4), 318 - 22
Deposition of carbenicillin aerosols in cystic fibrosis: effects of nebuliser system and breathing pattern; Newman SP et al.; Antibiotic aerosol treatment is successful in treating Pseudomonas infection in some patients with cystic fibrosis, but the amount of drug reaching the lungs is unknown . The deposition patterns of carbenicillin aerosols delivered from two commercially available nebuliser systems (the Turret nebuliser plus Maxi compressor and the Inspiron nebuliser plus Traveller compressor) have been compared in six patients with cystic fibrosis during tidal breathing . The aerosol mass median diameters were 3.2 and 7.3 microns . In addition, the aerosol from the Turret-Maxi nebuliser system was inhaled by a combination of tidal and deep breathing . After two minutes' breathing via a mouthpiece the mean (SEM) deposition in the lungs was 15.60 (1.5) mg carbenicillin with the Turret nebuliser plus Maxi compressor, but only 6.54 (1.09) mg with the Inspiron nebuliser plus Traveller compressor; the distribution pattern within the lung was significantly more peripheral with the former nebuliser system . These differences may be ascribed partly to the smaller droplet size from the Turret system and partly to the higher nebulisation rate from the more powerful Maxi compressor . Tidal plus deep breathing produced a further small but non-significant increase in lung aerosol deposition . A seventh patient, who failed to complete the trial, had little aerosol deposited in his lungs because he inhaled through his nose . These results emphasise the importance of correct selection of nebuliser equipment for antibiotic aerosol treatment.

J Lipid Res, 1988 Apr, 29(4), 459 - 68
Biotransformation of ursodeoxycholic acid by Pseudomonas sp NCIB 10590; Owen RW et al.; The metabolism of ursodeoxycholic acid by Pseudomonas sp NCIB 10590 has been studied in phosphate-buffered mineral salts . The organism completely metabolized ursodeoxycholic acid in 24 hr, and time-course experiments revealed that maximum product formation occurred at 14 hr . The major products detected and identified at 14 hr were 7 beta-hydroxychol-4-en-3-one-24-oic acid, 7 beta-hydroxy-3-oxo-pregna-1,4-diene-20-carboxylic acid, and 7 beta-hydroxyandrosta-1,4-diene-3,17-dione . Several minor intermediates were isolated and evidence is given for the following structures: 7 beta-hydroxy-5 beta-cholan-3-oxo-24-oic acid, 7 beta-hydroxyandrost-4-en-3,17-dione, 7 beta,17 beta-dihydroxyandrosta-1,4-diene-3-one, 3-hydroxy-1,3,5(10)-9,10-seco-androstatriene-3,17-dione-7 beta-ol, and 22 alpha-hydroxymethylpregna-1,4-diene-3-one-7 beta-ol.

Biull Eksp Biol Med, 1988 Apr, 105(4), 426 - 9
{The nature of functional groups in the active center of antitumor glutamin-(asparagin-)ase}; Lebedeva ZI et al.; The effect of two reagents on glutamin (asparagin) ase from Pseudomonas aurantiaca-548 has been studied . 2,3-butanedione which modified arginine residues was ineffective for the inactivation of the enzyme . The enzyme was completely inactivated in the presence of N-ethyl-5-phenylisoxazolium-3'-sulfonate (Woodward's reagent K) . The effects of pH, reagent concentration, competitive inhibitors and their analogues on the rate or degree of enzyme inactivation were studied . The experimental results suggest that the carboxyl groups localized at the active site of glutamin (asparagin) ase are probably essential for the substrate binding.

J Biochem (Tokyo), 1988 Apr, 103(4), 714 - 21
Interaction of 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase with Reactive Blue 2 and related dyes; Maruyama K; Steady-state kinetic analyses suggest that Pseudomonas ochraceae 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase (4-carboxy-2-hydroxy-cis,cis-muconate-6-semialdehyde: NADP+ oxidoreductase {EC 1.2.1.45}) functions through an ordered BiBi mechanism . The enzyme binds one NADP+ molecule per subunit with a Kd of 4.8 +/- 0.8 microM . The enzyme is adsorbed to a Blue Sepharose CL-6B column and can be eluted therefrom with reagents having high affinity for the enzyme such as NADP+, NAD+, ATP, and Reactive Blue 2 . Equilibrium dialysis and difference spectral titration show the binding of four molecules of Reactive Blue 2 per enzyme subunit . Two of these dye molecules show high-affinity binding with a Kd of 0.03 +/- 0.02 microM . The resulting 1: 2 enzyme-dye complex can be isolated by gel filtration on Bio-Gel P-6 . The kinetic, spectroscopic, and chromatographic properties of the complex indicate that the dye-binding sites are different from the coenzyme binding site . The other two dye molecules, in contrast, bind loosely with a Kd of 0.8 +/- 0.5 microM to a site overlapping the coenzyme binding site . This is confirmed by the following findings: NADP+ effectively abolishes the difference spectrum associated with the enzyme-dye binding, and the slope of the double reciprocal plot showing the competitive inhibition of the dye (Ki = 0.20 +/- 0.02 microM) with respect to NADP+ linearly depends on the square of the dye concentration . Essentially similar results are also obtained with methoxy Reactive Blue 2 and Reactive Blue 4.(ABSTRACT TRUNCATED AT 250 WORDS)

J Bacteriol, 1988 Apr, 170(4), 1907 - 12
Variation in the ability of Pseudomonas sp . strain B13 cultures to utilize meta-chlorobenzoate is associated with tandem amplification and deamplification of DNA; Rangnekar VM; Single-colony isolates of Pseudomonas sp . strain B13 were examined for their ability to utilize benzoate (Ben) and meta-chlorobenzoate (3CB) as the sole carbon source . Scoring of B13 cultures by the replica-plating technique revealed that under nonselective conditions, B13 spontaneously formed four different types of colonies: 3CB+ Ben+, 3CB+ Ben-, 3CB- Ben-, 3CB- Ben+ . Successive testing of each of the four colony types showed that each produced the same four different types of single-colony isolates . Colonies of each class had characteristic phenotypic properties with respect to the accumulation of Ben or 3CB pathway intermediate products . The physical abundance of a 4.3-kilobase DNA encoding the first three enzymes of the chlorocatechol pathway correlated with the 3CB+ phenotype . Increased abundance of the 4.3-kilobase DNA fragment was the result of tandem amplification.

J Bacteriol, 1988 Apr, 170(4), 1445 - 51
Cloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity; Roberts DP et al.; The egl gene of Pseudomonas solanacearum was cloned on a cosmid and expressed in Escherichia coli . Restriction endonuclease mapping, transposon mutagenesis, and subclone analysis showed that the egl gene was located on a 2.7-kilobase XhoI-SalI P . solanacearum DNA fragment . Immunoabsorption experiments and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis showed that the egl gene encodes the 43-kilodalton endoglucanase that is the major excreted endoglucanase of P . solanacearum . In E . coli, the egl gene appeared to be expressed from its own promoter, but its product was restricted to the cytoplasm . The cloned egl gene was mutagenized with Tn5 and used to specifically mutate the chromosomal egl gene of P . solanacearum by site-directed mutagenesis . The resultant mutant was identical to the wild-type strain in production of extracellular polysaccharide and extracellular polygalacturonase as well as several other excreted proteins but produced at least 200-fold less endoglucanase . This mutant strain was significantly less virulent on tomato than the wild-type strain in plant bioassay experiments . Virulence of the endoglucanase-deficient strain was restored to near wild-type levels by complementation in trans with the cloned egl gene, indicating that the egl gene is important but not absolutely required for pathogenesis.

J Biol Chem, 1988 Mar 15, 263(8), 3840 - 4
The post-translational trimethylation of diphthamide studied in vitro; Moehring JM et al.; The amino acid diphthamide is a complex post-translational derivative of histidine that exists in eukaryotic and Archaebacterial elongation factor 2 (EF-2) . Diphtheria toxin and Pseudomonas exotoxin A catalyze the transfer of an ADP-ribose residue from NAD to diphthamide, causing the inactivation of EF-2 . We have used cytosolic extracts of mutant CHO-K1 cells to study the biosynthesis of diphthamide in vitro . We have identified chromatographically a precursor form of diphthamide that exists in one complementation group of mutant cells and have documented the addition of 3 methyl residues from S-adenosylmethionine to this precursor . We have identified the presence of methyltransferase capable of carrying out this reaction in vitro in cells of 15 diverse eukaryotic species.

Biochemistry, 1988 Mar 8, 27(5), 1591 - 7
Mechanism and stereochemical course at phosphorus of the reaction catalyzed by a bacterial phosphotriesterase; Lewis VE et al.; The reaction mechanism for the phosphotriesterase from Pseudomonas diminuta has been examined . When paraoxon (diethyl 4-nitrophenyl phosphate) is hydrolyzed by this enzyme in oxygen-18-labeled water, the oxygen-18 label is found exclusively in the diethyl phosphate product . The absolute configurations for the (+) and (-) enantiomers of O-ethyl phenylphosphonothioic acid have been determined by X-ray diffraction structural determination of the individual crystalline 1-phenylethylamine salts . The (+) enantiomer of the free acid corresponds to the RP configuration . The RP enantiomer of O-ethyl phenylphosphonothioic acid has been converted to the SP enantiomer of EPN {O-ethyl O-(4-nitrophenyl) phenylphosphonothioate} . (SP)-EPN is hydrolyzed by the phosphotriesterase to the SP enantiomer of O-ethyl phenylphosphonothioic acid . The enzymatic reaction therefore proceeds with inversion of configuration . These results have been interpreted as an indication of a single in-line displacement by an activated water molecule directly at the phosphorus center of the phosphotriester substrate . (RP)-EPN is not hydrolyzed by the enzyme at an appreciable rate.

J Clin Microbiol, 1988 Mar, 26(3), 607 - 8
Enzymatic characterization of Pseudomonas cepacia by API ZYM profile; Poh CL et al.; The enzymatic activities of 53 strains of Pseudomonas cepacia were determined by using the API ZYM system . Strong alkaline phosphatase, acid phosphatase, butyrate esterase, caprylate esterase, myristate lipase, leucine arylamidase, and phosphoamidase activities were consistently detected in all strains . Weak activities were observed for valine arylamidase, beta-glucosidase, and N-acetyl-beta-glucosaminidase . No activities could be demonstrated for cystine arylamidase, trypsin, chymotrypsin, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-glucosidase, alpha-mannosidase, and alpha-fucosidase . Enzymatic activities of pseudomonads may provide useful information about their pathogenesis and information for identification of Pseudomonas species.

Cancer Res, 1988 Mar 1, 48(5), 1307 - 11
Potentiation by a biscoclaurine alkaloid, cepharanthine, of the toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin in HeLa cells; Shiraishi N et al.; Cepharanthine, a biscoclaurine alkaloid, causes an 8-fold enhancement of the cytotoxic effect of a conjugate of epidermal growth factor (EGF) coupled with Pseudomonas exotoxin in HeLa cells . Cepharanthine also potentiates the effect of Pseudomonas exotoxin . Cepharanthine does not affect the binding and uptake of 125I-EGF by HeLa cells, but it delays the release of radioactivity associated with 125I-EGF into the medium . Analysis by colloidal silica gradients using cell homogenates suggests that 125I-EGF accumulates in the lysosomes of cells treated with cepharanthine and that {3H}cepharanthine accumulates in lysosomes . The pH in HeLa cell lysosomes is 5.2, and cepharanthine does not significantly increase the pH . Electron microscopy shows an increased number of electron-dense bodies and dilated Golgi apparatus after cepharanthine treatment . Cepharanthine appears to accumulate in lysosomes, and it may delay degradation of EGF-Pseudomonas exotoxin in the cells as does 125I-EGF.

J Trauma, 1988 Mar, 28(3), 362 - 7
Plasma proteolytic activity following burns; Neely AN et al.; Because a number of metabolic events which are triggered by proteolysis in the bloodstream are activated following trauma, net proteolytic activity (P.A.) in the plasma of 37 pediatric burned patients was measured . The P.A . assay involved incubating plasma with a radioiodinated protein substrate and counting the isotopic activity of the hydrolyzed fragments in the acid-soluble fraction . In patients, plasma P.A . increased in direct proportion to the extent of burn injury . To examine additional trends in plasma P.A . suggested by the patient P.A . data, we measured plasma P.A . in a burned rat model: circulating P.A . was significantly elevated at 6 hours and until at least 2 weeks postburn; infection with Pseudomonas and use of the proteolytic debriding agent, Travase, each further elevated this activity; the plasma P.A . was not directly derived from the burn site . We postulate that this elevated circulating P.A . triggers some of the pathologic as well as some physiologic sequelae which follow burn trauma.

Proc Natl Acad Sci U S A, 1988 Mar, 85(6), 1922 - 6
Cytotoxic activity of an interleukin 2-Pseudomonas exotoxin chimeric protein produced in Escherichia coli; Lorberboum-Galski H et al.; A cDNA clone for human interleukin 2 (IL-2) has been fused to the 5' end of a modified Pseudomonas exotoxin (PE) gene that lacks the sequences encoding the cell recognition domain . The chimeric protein IL-2-PE40 was produced in Escherichia coli . It was extremely toxic to IL-2 receptor-positive cells but had no measurable effect on cells lacking the IL-2 receptor . IL-2-PE40 might be a useful cytotoxic agent in the treatment of diseases involving IL-2 receptor-positive cells and in the treatment of allograft rejection.

Bioorg Khim, 1988 Mar, 14(3), 352 - 8
{Antigenic polysaccharides of bacteria . 31 . The structure of the O-specific polysaccharide chain of Pseudomonas aurantiaca IMB 31 lipopolysaccharide}; Knirel' IuA et al.; The O-specific polysaccharide chain of the Pseudomonas aurantiaca IMV 31 lipopolysaccharide contains N-acetyl-L-fucosamine (FucNAc) and di-N-acetyl-D-bacillosamine (2,4-diacetamido-2,4,6-trideoxyglucose, Bac(NAc)2) in the ratio 2:1 . On the basis of methylation, solvolysis with anhydrous hydrogen fluoride, and computer-assisted analysis of 13C-NMR spectrum, it was concluded that the trisaccharide repeating unit of the polysaccharide possesses the following structure: structure: ----3)-beta-D-Bac(NAc)2-(1----3)-alpha-L-FucNAc-(1----3)-alpha-L- FucNAc-(1----.

J Biol Chem, 1988 Feb 15, 263(5), 2270 - 9
Estrogen preferentially stimulates lactosaminoglycan-containing oligosaccharide synthesis in mouse uteri; Dutt A et al.; The effects of steroid hormones on the synthesis of lactosaminoglycan (LAG)-containing oligosaccharides by mouse uteri are reported . The uterine LAG-containing oligosaccharides were degraded partially by Pseudomonas endo-beta-galactosidase, releasing an oligosaccharide of the apparent structure: Gal beta----N-acetylglucosaminyl(----N-acetylgalactosaminyl)beta 1,3----galactose . A larger fraction of the LAG-containing oligosaccharides bound to pokeweed mitogen than to Datura stramonium lectin, suggesting the presence of highly branched structures . LAG-containing oligosaccharides were resistant to sequential digestion with Pronase, nitrous acid, hyaluronidase, and chondroitinase ABC . These polysaccharides exhibited a Gal:GlcNAc:GalNAc ratio of approximately 1.0:1.0:0.3 and were not fucosylated . The ion-exchange behavior of the LAG-containing oligosaccharides before and after mild acid hydrolysis indicated the presence of sialic acid residues . The LAG-containing glycopeptides were highly resistant to beta-elimination but were released quantitatively by hydrazinolysis, demonstrating an N-linkage to protein . Binding to pokeweed mitogen was markedly enhanced following release of these oligosaccharides from peptides by hydrazinolysis, suggesting that peptide-bound oligosaccharides were partially inaccessible to the lectin . Molecular exclusion chromatography of the oligosaccharides released by hydrazinolysis revealed a broad distribution ranging from Mr 4,000 to 15,000 with a median Mr of approximately 8,000 . We extended the above observations by determining how the steroid hormones 17-beta-estradiol (E2) and progesterone affected synthesis of the LAG-containing oligosaccharides in ovariectomized mice . Generally, E2 and a number of E2 agonists stimulated glycoconjugate synthesis; however, chronic E2 treatment or combined treatment with E2 plus progesterone caused the synthesis of most glycosaminoglycans to return to basal levels . In contrast, E2 either alone or in combination with progesterone stimulated synthesis of LAG-containing oligosaccharides in preference not only to glycosaminoglycans but also to other classes of N-linked oligosaccharides . This effect was apparent during both priming and nidatory E2 treatments . Collectively, these data provide the first demonstration of LAG-containing oligosaccharides in uteri and for the hormonally regulated synthesis of lactosaminoglycans . In addition, this is the first demonstration of the ability of steroid hormones to induce the synthesis of certain types of N-linked oligosaccharides in preference to others in the same tissue.

Biokhimiia, 1988 Feb, 53(2), 332 - 40
{Analysis of oligomeric forms of recombinant human leukocyte interferons}; Borukhov SI et al.; Using SDS-PAAG electrophoresis, gel-permeation HPLC and immunoblotting, it was demonstrated that homogeneous preparations of human leukocyte interferons (alpha-INF)-A, -N and -I1 obtained from the biomass of the corresponding producer strains (Pseudomonas sp.) contained several oligomeric forms produced by way of S-S intermolecular cross-linkage and making up to 10-15%, 4-7% and 2-5% of the total monomeric form content in the protein preparations . Immunologic testing with the use of MAB NK-2 and {125I}NK-2 showed that the oligomeric forms of alpha-INF-A, -N and -I1 were present in the protein preparations at all purification stages and seemed to be formed at early steps of interferon synthesis in the cell . The effects of limited proteolysis as well as of acid, alkaline and thermodenaturation on the aggregation and oligomerization of alpha-INF-A were studied . SDS-PAAG electrophoresis performed in the absence of the reducing agents showed that upon denaturation of 10% TCA, the amount of the oligomeric forms in the preparations of homogeneous and especially partly proteolytic INF was significantly increased . The causes and the putative mechanisms of aggregation and oligomerization of INF are discussed.

Appl Environ Microbiol, 1988 Feb, 54(2), 594 - 5
Novel biotransformations of 4-chlorobiphenyl by a Pseudomonas sp; Barton MR et al.; A bacterium, tentatively identified as a representative of the genus Pseudomonas (strain MB86), was isolated from soil contaminated by wood-preservation chemicals by using 4-chlorobenzoate as an enrichment substrate . The pseudomonad was able to grow on 4-chlorobenzoic acid and 4-chlorobiphenyl as sole carbon and energy sources . Spent culture medium from 4-chlorobiphenyl-grown cells contained 4-chlorobenzoic acid, 4'-chloroacetophenone, 2-hydroxy,2-{4'-chlorophenyl} ethane, and 2-oxo,2-{4'-chlorophenyl} ethanol as metabolites . 4'-Chloroacetophenone was produced in large amounts, possibly as a dead-end metabolite.

Appl Environ Microbiol, 1988 Feb, 54(2), 343 - 7
Predictive model of conjugative plasmid transfer in the rhizosphere and phyllosphere; Knudsen GR et al.; A computer simulation model was used to predict the dynamics of survival and conjugation of Pseudomonas cepacia (carrying the transmissible recombinant plasmid R388:Tn1721) with a nonrecombinant recipient strain in simple rhizosphere and phyllosphere microcosms . Plasmid transfer rates were derived for a mass action model, and donor and recipient survival were modeled as exponential growth and decay processes or both . Rate parameters were derived from laboratory studies in which donor and recipient strains were incubated in test tubes with a peat-vermiculite solution or on excised radish or bean leaves in petri dishes . The model predicted donor, recipient, and transconjugant populations in hourly time steps . It was tested in a microcosm planted with radish seeds and inoculated with donor and recipient strains and on leaf surfaces of radish and bean plants also growing in microcosms . Bacteria were periodically enumerated on selective media over 7 to 14 days . When donor and recipient populations were 10(6) to 10(8) CFU/g (wet weight) of plant or soil, transconjugant populations of about 10(1) to 10(4) were observed after 1 day . An initial rapid increase and a subsequent decline in numbers of transconjugants in the rhizosphere and on leaf surfaces were correctly predicted.

Exp Cell Res, 1988 Feb, 174(2), 397 - 410
Enhancement of cytotoxicity of modeccin by nigericin in modeccin-resistant mutant cell lines; Ghosh PC et al.; We have isolated a Chinese hamster ovary cell mutant (DMPR-2) simultaneously resistant to diphtheria toxin and modeccin . In addition to the increased resistance to these two toxins used in the selection, this mutant is more resistant to Pseudomonas toxin and hypersensitive to ricin than the parental cell line . In contrast to the wild-type cells in which nigericin protects cells from modeccin, the cytotoxicity of modeccin in the DMPR-2 mutant is enhanced by nigericin . We have also studied the effects of nigericin and NH4Cl on the cytotoxicity of modeccin in a modeccin-resistant mutant of HeLa cells (ModRI) . The cytotoxicity of modeccin is enhanced by nigericin in ModRI mutant cells, in contrast to the protection of modeccin cytotoxicity by nigericin in the parental HeLa cells . Our results suggest that modeccin can reach the cytosol of mammalian cells by two distinct routes; the major route requires endosomal acidification and the minor route is activated by nigericin.

Arch Pathol Lab Med, 1988 Feb, 112(2), 166 - 72
Pseudomonas cepacia-associated pneumonia in cystic fibrosis . Relation of clinical features to histopathologic patterns of pneumonia; Tomashefski JF et al.; We studied lungs at autopsy from 40 patients with cystic fibrosis (CF) to determine the structural and clinicopathologic features of pneumonia associated with Pseudomonas cepacia respiratory tract colonization . Three clinical groups were identified: group A included 11 patients exhibiting a fulminant course following P cepacia colonization; group B included 20 patients who declined slowly following colonization; and group C included nine patients without P cepacia colonization . Acute pneumonia occurred in all groups but was most extensive and necrotizing in group A . Chronic lobular pneumonia involved all groups equally, whereas interstitial pneumonia predominated in group B . Diffuse alveolar damage occurred infrequently in all groups . Combinations of structural patterns were frequently seen . We conclude that, although there is great overlap in the structural appearance of pneumonia among patients with CF with different bacterial colonization histories, the evidence suggests that P cepacia is a cause of necrotizing pneumonia in some patients . Factors that predispose to this fulminant lung infection are poorly understood.

J Bacteriol, 1988 Feb, 170(2), 617 - 22
Molecular cloning of genes that specify virulence in Pseudomonas solanacearum; Xu PL et al.; The suicide plasmid pSUP2021 was used to introduce Tn5 into the Pseudomonas solanacearum wild-type strain K60 . We isolated eight avirulent mutants after screening 6,000 kanamycin-resistant transconjugants by inoculating eggplant (Solanum melongena L . cv . Black Beauty) and tobacco (Nicotiana tabacum L . cv . Bottom Special) seedlings . The Tn5-containing EcoRI fragments from the eight mutants were unique, suggesting that numerous genes specify virulence in this species . These EcoRI fragments were cloned into pBR322 or pUC12, and one of the clones, pKD810, was transformed into K60 . All of the kanamycin-resistant, ampicillin-sensitive transformants were avirulent . Three randomly selected avirulent transformants were shown to carry the Tn5-containing fragment in place of the wild-type fragment and to exhibit the same hybridization pattern as the original KD810 mutant did . With pKD810 as a probe, we identified cosmids carrying the wild-type virulence genes by using a genomic library of K60 prepared in pLAFR3 . Two of the homologous cosmids, pL810A and pL810C, when introduced into KD810 by transformation, restored virulence and normal growth of this mutant in tobacco . Altogether, these data indicate that the gene(s) interrupted by Tn5 insertion in KD810 is essential for the virulence of P . solanacearum . Further characterization of this gene is now being completed by subcloning, transposon mutagenesis, and complementation analysis.

Bioorg Khim, 1988 Feb, 14(2), 180 - 6
{Antigenic bacterial polysaccharides . 30 . The structure of the polysaccharide chain of lipopolysaccharide of Pseudomonas syringae pv . syringae 281 (serogroup I)}; Knirel' IuA et al.; Anomeric methyl 3-O-(D-mannopyranosyl- and L-rhamnopyranosyl)-beta-D-talopyranosides were synthesised by the stereoselective 1,2-cis- and 1,2-trans manno- and rhamnosylation of methyl 2,4,6-tri-O-acetyl-beta-D-talopyranoside, which has been prepared from methyl beta-D-galactopyranoside by a synthetic scheme including conversion of the C2 configuration . From 13C-NMR spectra of the disaccharides obtained the spectral alpha- and beta-effects of O3-glycosylation of talopyranose were determined.

Bioorg Khim, 1988 Feb, 14(2), 172 - 9
{Antigenic bacterial polysaccharides . 29 . The structure of the polysaccharide chain of Pseudomonas holci 8300 (serogroup I) lipopolysaccharide}; Knirel' IuA et al.; The Pseudomonas holci 8300 lipopolysaccharide has an O-specific polysaccharide chain, containing L-rhamnose and 3-acetamido-3-deoxy-D-fucose residues in the ratio 4:1 . On the basis of methylation, Smith degradation, and 1H- and 13C-NMR spectroscopy data, it was concluded that the polysaccharide is built up of pentasaccharide units of A and B types in the ratio approximately 2.5:1 . In some stretches of the polysaccharide, minor B units form rather long chains, and in the others they alternate with predominant A units . (formula; see text)

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