Pseudomonas

Bioorg Khim, 1988 Feb, 14(2), 166 - 71
{Antigenic bacterial polysaccharides . 28 . The structure of the O-specific lipopolysaccharide chain of Pseudomonas syringae pv . atrofaciens K-1025 and Pseudomonas holci 90a (serogroup II)}; Knirel' IuA et al.; Lipopolysaccharides of serologically related strains of Pseudomonas syringae pv . atrofaciens K-1025 and Pseudomonas holci 90a possess the identical O-specific polysaccharide chains, representing a homopolymer of D-rhamnose . On the basis of methylation, partial and complete Smith degradation, and analysis by 1H- and 13C-NMR-spectroscopy, it was concluded that the repeating unit of the polysaccharide is a branched pentasaccharide of the following structure: (formula; see text)

Biochim Biophys Acta, 1988 Jan 4, 952(1), 48 - 55
Inducible oxacillin-hydrolyzing beta-lactamase in a methylotrophic bacterium; Samuelov NS et al.; A novel beta-lactamase (beta-lactam-hydrolase, EC 3.5.2.6) was detected in a culture of Pseudomonas C, an obligatory methylotroph . This is the first beta-lactamase discovered in a methylotrophic organism . The inducible cell-bound enzyme with broad-spectrum activity against penicillins, was purified 77-fold from cell extracts of the methanol-grown bacterium, and its molecular weight was estimated to be 30,000 . As a group, the isoxazolyl penicillins are the favored substrates, while cephalosporins are resistant to hydrolysis and act as mild competitive inhibitors . The activity of this M-OXA beta-lactamase focused as a single band at an acidic pI value (5.5) similar to that of PSE- and TEM-type enzymes, but can be clearly distinguished from other OXA-type beta-lactamases, all of which focus in the alkaline region . The enzyme is coded by a non-transferable gene . Based on the sum of its physical and biochemical properties, the M-OXA beta-lactamase is distinguishable from all previously described beta-lactamases, although immunological studies revealed some cross reactivity with the plasmid mediated OXA-2 enzyme.

Microbios, 1988, 53(215), 119 - 28
Surface characteristics of Pseudomonas cepacia; Eaves DJ et al.; Two major surface characteristics of Pseudomonas cepacia were examined in this study: reactivity with lectins and hydrophobicity . The results indicated that the surfaces of P . cepacia strains are heterogeneous with regard to the distribution of lectin receptors . Only lima bean agglutinin was found to strongly agglutinate all strains . The strains were also heterogeneous with regard to hydrophobicity as determined by adhesion to hexadecane . The degree of hydrophobicity, however, was not significantly altered when selected strains were mixed with either fibronectin or bovine serum albumin . In addition, the strains exhibited no apparent affinities for buccal epithelial cells and gave no evidence for an ability to haemagglutinate human red cells.

Biokhimiia, 1988 Jan, 53(1), 150 - 7
{Isolation and properties of genetic engineered human leukocyte interferons alphaI1 and alphaN}; Eremashvili MR et al.; Using controlled pore glass chromatography and immunoaffinity chromatography on monoclonal antibodies NK-2 immobilized on Sepharose 4B, the electrophoretically homogeneous interferons alpha N and alpha I1 were isolated from the biomass of gene-engineered Pseudomonas sp . strains . In terms of specific activity on human fibroblast diploid cells, interferon alpha I1 does not differ from interferon alpha A, whereas the specific antiviral activity of interferon alpha N is as low as 2.10(7) JU/mg . The procedures for immunometric assay of interferons alpha N and alpha I1 have been elaborated . Various monoclonal antibodies to interferon alpha A and to natural leucocyte interferon were analyzed; among those the antibodies specifically interacting with interferons alpha N and alpha I1 (but not with interferon alpha A) were identified.

J Antibiot (Tokyo), 1988 Jan, 41(1), 7 - 12
MM 42842, a new member of the monobactam family produced by Pseudomonas cocovenenans . II . Production, isolation and properties of MM 42842; Box SJ et al.; A new member of the monobactam family of beta-lactam antibiotics, designated MM 42842, has been detected in a culture of Pseudomonas cocovenenans . The production, isolation and some properties of the antibiotic are described . Structural studies show MM 42842 to be closely related to the previously described antibiotic sulfazecin.

J Antibiot (Tokyo), 1988 Jan, 41(1), 1 - 6
MM 42842, a new member of the monobactam family produced by Pseudomonas cocovenenans . I . Identification of the producing organism; Gwynn MN et al.; A bacterial soil isolate designated 326-32B produces a new member of the monobactam series of antibiotics, MM 42842, and the bulgecins . Identification studies show isolate 326-32B to be a strain of Pseudomonas cocoveneans which is a species previously noted for the production of toxoflavin . A description of P . cocovenenans does not appear to have been previously published and the identify of strain 326-32B was established by means of a direct comparison with the deposited organism P . cocovenenans NCIB 9450 . The properties of strain 326-32B, and P . cocovenenans NCIB 9450 were compared with those of the monobactam and bulgecin producing organisms Pseudomonas acidophila ATCC 31363 and Pseudomonas mesoacidophila ATCC 31433 . The four organisms were found to share certain properties, including the ability to grow at pH 4.0.

Am J Med, 1988 Jan, 84(1), 75 - 81
Indolent epidemic of Pseudomonas cepacia bacteremia and pseudobacteremia in an intensive care unit traced to a contaminated blood gas analyzer; Henderson DK et al.; An epidemic of Pseudomonas cepacia bacteremia and pseudobacteremia occurred in the medical intensive care unit at the Clinical Center of the National Institutes of Health . Sixteen patients in the intensive care unit became colonized or infected with this organism in a 21-month period; whereas P . cepacia had been isolated only 16 times in the preceding 90 months from the entire hospital . Further analysis demonstrated a significant association of the epidemic cases with bloodstream isolation of the organism (p less than 0.001, Fisher's exact test) . Mortality associated with bacteremia caused by P . cepacia was 38 percent . Intensive investigation of the intensive care unit and its surrounding environment eventually demonstrated that a blood gas analyzer in a satellite laboratory adjacent to the intensive care unit was the reservoir for the outbreak . Replacement of the machine resulted in termination of the outbreak, P . cepacia continues to represent an environmental threat to hospitalized patients.

Crit Rev Microbiol, 1988, 15(3), 247 - 68
Genetics of naphthalene catabolism in pseudomonads; Yen KM et al.; In pseudomonads, naphthalene is catabolized in a series of reactions to salicylic acid, which is further degraded via the catechol meta-cleavage, ortho-cleavage, or gentisic acid pathway to Krebs cycle intermediates . The naphthalene catabolic genes have been located on self-transmissible plasmids, in most cases, and implicated to have chromosomal locations in other cases . The best-studied naphthalene catabolic plasmid is NAH7 . It carries two operons, one of which enables the host to utilize naphthalene and the other to utilize salicylate as a carbon and energy source . The product of another NAH7 gene, nahR, is required to turn on both operons in the presence of the inducer, salicylate . Several different naphthalene and salicylate catabolic plasmids have been shown to share sequence homology with NAH7 . These plasmids can undergo structural alterations involving insertions and deletions during conjugations and changes in nutritional conditions . Available evidence suggests that salicylate catabolic plasmids can form from the naphthalene catabolic plasmids by structural alterations of the plasmid DNA . The gene organization and regulation, as well as the genetic instability of the naphthalene catabolic plasmids, are reminiscent of the TOL plasmids and suggest that the naphthalene catabolic plasmids and other catabolic plasmids may have evolved in a short period of time by acquiring and modifying preevolved gene clusters from host chromosomes or other plasmids.

Pediatr Pulmonol, 1988, 4(2), 72 - 7
Derepressed beta-lactamase production as a mediator of high-level beta-lactam resistance in Pseudomonas cepacia; Aronoff SC; A mechanism of high-level resistance to readily and poorly hydrolyzable beta-lactam substrates in Pseudomonas cepacia was identified using a hypersusceptible, beta-lactamase-inducible, non-CF clinical isolate, 75-26, and a drug-resistant mutant derived from this strain . Inoculation of 75-26 onto agar containing 16 micrograms/ml of ceftazidime produced a stable, beta-lactam-resistant mutant at a frequency of 1.7 x 10(-5) . Baseline beta-lactamase production by a representative mutant isolate (75-26z) was almost 40-fold greater than the parent . Both strains produced major beta-lactamase bands with isoelectric points of 6.9, 7.8, 8.1, 8.5, and 9.2 by isoelectric focusing . Compared with the parental strain, multiple satellite bands associated with the major beta-lactamase bands were present in the mutant . Growth of an indicator strain, E . coli ATCC 25922, was inhibited by ceftazidime and piperacillin but was not inhibited after the compounds were preincubated with the beta-lactamase preparation from 75-26z . Preincubation of ceftazidime with beta-lactamase, followed by the addition of a beta-lactamase inhibitor, inhibited the growth of the indicator strain; piperacillin failed to inhibit growth of the indicator strain in a similar experiment . One mechanism of high-level resistance to both poorly and readily hydrolyzable beta-lactam substrates in P . cepacia is derepressed chromosomal beta-lactamase production.

J Basic Microbiol, 1988, 28(9-10), 667 - 72
{Detection and characterization of a levansucrase and a sucrase in Pseudomonas syringae pv . phaseolicola}; Sauerstein J et al.; Pseudomonas syringae pv . phaseolicola, a plant pathogenic pseudomonad, possesses two sucrose-splitting enzymes, a levansucrase and a sucrase . The levansucrase is found both extracellularly and intracellularly, and enzyme synthesis is independent of the carbon source . In addition to levansucrase, cells grown on sucrose contain a sucrase . The two sucrose-splitting enzymes differ in their optimum pH value and optimum temperature as well as in their substrate specificities.

Scand J Gastroenterol Suppl, 1988, 143, 19 - 27
Borderline sweat test: criteria for cystic fibrosis diagnosis; Canciani M et al.; The CF diagnosis can be difficult in subjects with disease consistent symptoms and borderline values of the sweat test . This study aimed to evaluate possibly resolutory criteria . Seventy-one ill subjects with borderline sweat test values (40-70 mEq/l Cl-) aged 0.1-37.7 years (mean, 8.7; SD, 7.3) were studied and compared with 33 age-matched CF patients (Cl- over 75 mEq/l) and 25 healthy subjects . A complex CF score based on 25 variables was set: CF-specific clinical conditions, detailed analysis of sweat test (salt-free diet test included), pancreatic function, Pseudomonas infection, and others . The score ranged from 0 to a maximum of 65 and clearly distinguished the CF patients from the healthy subjects . Therefore the CF score was assumed as the best reference criterion to define the diagnosis in the borderline group: 27 of them (38%) were assigned to the CF condition and 44 (62%) to the healthy one, with a quite clear separation between the 2 subgroups . With respect to these assignments, the most discriminant variable turned out to be the sweat Cl- persisting above 40 mEq/l after 5 days of salt-free diet (mean error, 18.4%) . Neither sweat Cl-/Na+ ratio nor bicarbonate duodenal output showed better discriminant power (mean error, 29% and 25%, respectively) . It is concluded that in borderline situations of sweat test results a definite CF diagnosis can be achieved only by compounding many clinical and laboratory data with a preference for the sweat test after a salt-free diet period . The CF score proposal can be an effective help.

Antonie Van Leeuwenhoek, 1988, 54(6), 567 - 73
Low- and intermediate-copy-number cloning vectors based on the Pseudomonas plasmid pVS1; Itoh Y et al.; The cloning vector pME290 (6.8 kb), which is derived from the Pseudomonas plasmid pVS1 and has about 7 copies, was mutagenized in vitro to provide derivatives with altered copy numbers . Thus, pME292 (about 1-3 copies) and pME294 (about 15-20 copies) were isolated . These vectors were used in the characterization of the P . aeruginosa argF gene encoding ornithine carbamoyltransferase.

Cancer Treat Res, 1988, 37, 161 - 73
Pseudomonas exotoxin--immunotoxins; FitzGerald DJ et al.; Monoclonal antibodies can be coupled with PE to make very potent ITs . Two of these ITs (PE-HB21 and OVB-3-PE) have been shown to have antitumor activity in a nude mouse model of ovarian cancer . PE ITs are at least 10-fold more active than the corresponding RTA IT . Deletion analysis of the structural gene of PE has helped assign specific functions to different portions of the molecule . Current efforts are focused on making ITs with recombinant PE.

Prog Clin Biol Res, 1988, 274, 211 - 26
Phthalate oxygenase, a Rieske iron-sulfur protein from Pseudomonas cepacia; Ballou D et al.; Phthalate oxygenase catalyzes the oxygenation of phthalate to form a cis-dihydrodiol . It is comprised of two proteins: a flavo-iron-sulfur protein with NADH-dependent oxidoreductase activity (POR) and a nonheme iron protein with oxygenase activity (PO) . The latter is a tetramer of 48 kDa units and contains a Rieske {2Fe-2S} center and one mononuclear iron per monomer . The mononuclear iron is likely the site of oxygenation . This system can be isolated in large quantities and is sufficiently stable for detailed mechanistic studies . We briefly describe some of our studies on this system which include kinetics, and visible, magnetic resonance, EXAFS, and ENDOR spectroscopies.

Microbiol Immunol, 1988, 32(3), 293 - 304
Role of a guanine nucleotide-binding regulatory protein in the hydrolysis of phosphatidylinositol 4,5-bisphosphate in a human T cell line; Hasegawa-Sasaki H et al.; The CD3(T3)/antigen receptor complex appears to function by transducing an antigen signal presented by macrophages into the hydrolysis of phosphatidylinositol 4,5-bisphosphate {PtdIns(4,5)P2} . In order to find out how the CD3/antigen receptor complex regulates the hydrolysis of PtdIns(4,5)P2 to diacylglycerol and inositol trisphosphate, we investigated the possible role played by a guanine nucleotide-binding regulatory protein in PtdIns(4,5)P2 hydrolysis in a human T cell leukemia line, JURKAT . JURKAT cells were made permeable to Al3+, F-, GTP, and a nonhydrolyzable GTP analogue, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), by treatment with pseudomonal cytotoxin . In the presence of AlCl3 NaF stimulated the release of inositol phosphates in the cytotoxin-treated JURKAT cells . NaF plus AlCl3 induced increases in inositol tris-, bis-, and mono-phosphates and decreases in PtdIns(4,5)P2, phosphatidylinositol 4-phosphate, and phosphatidylinositol within 5 min after addition to the cytotoxin-treated cells at 37 C . GTP gamma S stimulated, to some extent, polyphosphoinositide hydrolysis in the cytotoxin-treated JURKAT . The cytotoxin-treated JURKAT cells retained the ability to respond to anti-Leu-4 with polyphosphoinositide hydrolysis . It has been shown that Al3+ in the presence of F- modulates the activity of various guanine nucleotide-binding regulatory proteins . Therefore, the results obtained in this study indicate that a guanine nucleotide-binding regulatory protein regulates the polyphosphoinositide breakdown in JURKAT cells by influencing phosphodiesterase activity.

Arch Microbiol, 1988, 149(6), 492 - 8
Defects in cytochrome cd1-dependent nitrite respiration of transposon Tn5-induced mutants from Pseudomonas stutzeri; Zumft WG et al.; Mutants with defective respiratory nitrite utilization (Nir- phenotype) were obtained by transposon Tn5 insertion into genomic DNA of the ZoBell strain of Pseudomonas stutzeri . Three representative mutants were characterized with respect to their activities of nitrite and nitric oxide reduction, cytochrome cd1 content, and pattern of soluble c-type cytochromes . Mutant strain MK201 overproduced cytochrome c552 about fourfold by comparison with the wild type, but possessed an in vitro functional cytochrome cd1 . Mutant strain MK202 lacked cytochrome cd1 and, simultaneously, had low amounts of cytochrome c552 and the split alpha-peak c-type cytochrome . Strain MK203 synthesized nitrite reductase defective in the heme d1 prosthetic group . Irrespective of these biochemically distinct Nir- phenotypes, all mutants preserved the nitric oxide-reducing capability of the wild type . The mutant characteristics demonstrate that cytochrome cd1 is essential for nitrite respiration of P . stutzeri and establish the presence of a nitric oxide-reducing system distinct from cytochrome cd1 . They also indicate the functional or regulatory interdependence of c-type cytochromes.

J Bacteriol, 1988 Jan, 170(1), 163 - 70
An iron-antagonized fungistatic agent that is not required for iron assimilation from a fluorescent rhizosphere pseudomonad; Gill PR Jr et al.; Fluorescent rhizosphere Pseudomonas sp . strain NZ130 promotes plant growth, and may do so in part because of its production of a growth inhibitory factor that is active against phytopathogenic fungi . Analysis of the inhibitory factor that is active against the phytopathogen Pythium ultimum showed that its activity is antagonized at iron concentrations above 10 microM . The iron-antagonized inhibitor was separated from the fluorescent siderophore of this pseudomonad by gel filtration . Mutants that lacked either the iron-antagonized inhibitor or the fluorescent siderophore were isolated . Results of complementation analysis of these mutants by use of a cosmid library indicated that distinct DNA sequences are required for the production of each factor . Analysis of isogenic mutant strains showed that the genetic requirements for the production of the iron-antagonized inhibitor and the fluorescent siderophore are different, and that only the fluorescent siderophore is required for iron assimilation . Fusions of these same sequences to a beta-galactosidase gene were used to show that the regions required for the production of both the fluorescent siderophore and the iron-antagonized inhibitor were iron-regulated.

Bioorg Khim, 1988 Jan, 14(1), 92 - 9
{Antigenic polysaccharides of bacteria . 27 . Structure of the O-specific polysaccharide chain of lipopolysaccharides from Pseudomonas syringae pv . atrofaciens 2399, phaesolica 120a and Pseudomonas holci 8299 belonging to serotype VI}; Knirel' IuA et al.; Lipopolysaccharides from Pseudomonas syringae pvs atrofaciens 2399 . phaseolicola 120a and Pseudomonas holci 8299, belonging to serogroup VI . possess an identical polysaccharide chain composed of D-rhamnose and D-fucose . On the hasis of methylation, partial acid hydrolysis, 1H- and 13C-NMR data, it was concluded that the backbone of the polysaccharide represents D-rhamnan built up of tetrasaccharide repeating units and alpha-D-fucofuranose residues are attached to the backbone as the monosaccharide branches . The following structure of the repeating unit is established: (Formula: see text).

Bioorg Khim, 1988 Jan, 14(1), 82 - 91
{Antigenic polysaccharides of bacteria . 26 . Structure of O-specific polysaccharides from Pseudomonas cerasi 467 and Pseudomonas syringae pv . syringae strains 218 and P-55 belonging to serogroups II and III}; Knirel' IuA et al.; Serologically active O-specific polysaccharides were obtained on mild acid hydrolysis of lipopolysaccharides from Pseudomonas cerasi 467 and Pseudomonas syringae pv . syringae strains 218 and P-55 . On the basis of 1H- and 13C-NMR analysis, it was concluded that the P . cerasi polysaccharide has the following structure: ----3)-alpha-D-Rhap-(1----3)-alpha-D-Rhap-(1----2)-alpha-D-+ ++Rhap-(1---- which is identical to that of O-specific polysaccharide from P . syringae pv . morsprunorum C28 (Smith A . R . W . et al . Eur . J . Biochem., 1985, V . 149, No 1, p . 73-78) . The polysaccharides from P . syringae pv . syringae strains possess the same backbone but differ by the presence of D-fucose as monosaccharide branches . Methylation and 1H- and 13C-NMR analysis revealed the following structure of these polysaccharides: (Formula: see text) . The degree of substitution of the backbone trisaccharide units by the fucofuranose residues is about 35% for the strain 218 and about 85% for the strain P-55.

Bioorg Khim, 1988 Jan, 14(1), 77 - 81
{Antigenic polysaccharides of bacteria . 25 . Structure of the O-specific polysaccharide chain of Pseudomonas cepacia 673/2 lipopolysaccharide containing L-glycero-D-manno-heptose}; Knirel' IuA et al.; O-Specific polysaccharide, consisting of D-rhamnose and L-glycero-D-manno-heptose (LD-Hep) in a 2 : 1 ratio, was obtained on the mild acid degradation of the Pseudomonas cepacia IMV 673/2 lipopolysaccharide; monosaccharide LD-Hep has not previously been found in O-specific chains of lipopolysaccharides . On the basis of methylation and 13C-NMR data, it was concluded that the polysaccharide is composed of trisaccharide repeating units having the following structure: ----3)-alpha-D-Rha-(1----3)-alpha-D-Rha-(1----2)-alpha-LD-Hep-(1----

Biochim Biophys Acta, 1987 Dec 8, 910(3), 271 - 81
Length, mass, and denaturation of double-stranded RNA molecules compared with DNA; Lang D et al.; The contour lengths of linear, double-stranded (ds) RNAs from mycovirus PcV and Pseudomonas bacteriophage phi 6 have been measured with samples prepared for the electron microscope from 0.05 to 0.5 M NH4Cl solutions . A linear dependence of contour length on the logarithm of ionic strength was found and compared with that of dsDNA (pBR322, linearized and open-circular forms) . Conditions for molecular weight determinations of any natural dsRNA by electron microscopy have been established, and the method has been calibrated with phi 6 dsRNA of known nucleotide sequence . The results imply that dsRNA in 0.20 M NH4Cl solution has a rise per basepair of 0.271 nm, which is shorter than that in the A-conformation (4%) and in the A'-conformation (10%) . The thermal behavior of dsRNA in terms of melting temperature and exhibition of fine structure of melting curves was found to be generally similar to that of dsDNA, as expected from the literature . Folding of dsRNA in ethanolic solution was similar to that of dsDNA . However, in contrast to dsDNA, coiled coils could not be induced by ethanol, which is consistent with dsRNA being stiffer than dsDNA . Concerning dsDNA, the results show that a contraction in rise per basepair by 0.1 nm is coupled with an increase in the winding angle between basepairs by 0.47 degrees, as qualitatively predicted by polyelectrolyte theory.

J Biol Chem, 1987 Dec 5, 262(34), 16484 - 94
Temporal separation of protein toxin translocation from processing events; Hudson TH et al.; Intoxication of Vero cells by ricin, modeccin, diphtheria toxin (DT), and Pseudomonas exotoxin A requires: 1) binding to cell surface receptors; 2) transport to the cytoplasm; and 3) enzymatic inactivation of a component of the protein synthetic machinery . The kinetic profiles of all four toxins consist of a lag followed by the apparent first-order decrease in protein synthesis . Autoradiographic analysis of DT-intoxicated cell populations has demonstrated that two subpopulations of cells exist during the period of decreasing protein synthesis: one population synthesizing at control levels and the other synthesizing little or no protein (Hudson, T . H., and Neville, D . M., Jr . (1985) J . Biol . Chem . 260, 2675-2680) . The present study correlates the autoradiographic data with the rates of protein synthesis decline in cells intoxicated with modeccin, ricin, Pseudomonas exotoxin A, as well DT . In all cases, the first time point which exhibits a decrease in protein synthetic activity also exhibits two subpopulations of cells, one synthesizing protein at control rates and the other synthesizing little or no protein . As the intoxication progresses, cells leave the control population by the rapid cessation of all protein synthesis . These experiments demonstrate that transport of all four toxins to the cytosol is the rate-limiting step during the pseudo first-order decline in protein synthesis . Furthermore, the final step in the transport process (translocation) must result in the release to the cytoplasm of a quantity of toxin sufficient to rapidly inactivate all protein synthesis in that cell . The probability of a translocation event occurring in any cell of the population is established during the lag and remains constant throughout the first-order decrease in protein synthesis . The requirement for acidification during the intoxication by DT, Pseudomonas exotoxin A, or modeccin is restricted to the lag period . Acidification is therefore necessary to establish the probability of translocation, but it is not directly involved in the actual translocation of these toxins . The pseudo first-order passage of DT intoxications through antitoxin and NH4Cl- or monensin-sensitive stages are shown to have the same cellular basis as the pseudo first-order decrease in protein synthesis . A kinetic model is presented which defines the DT intoxication process from one of its earliest events (endocytosis) to its penultimate event (translocation of toxin to the cytosol).(ABSTRACT TRUNCATED AT 400 WORDS)

Tex Heart Inst J, 1987 Dec, 14(4), 401 - 10
Infective endocarditis: diagnosis, treatment, and mortality, as related to surgical timing and infectious organism; Attum AA et al.; To evaluate the timing of surgical treatment in infective endocarditis and to determine the relationship between the risk of mortality and the species of infectious organism, we reviewed a consecutive series of 65 cases involving patients with infective endocarditis who had been treated over a 17-year period . The patients included 41 males and 24 females, who ranged in age from 6 to 85 years (mean, 39.3 years) . Forty-five had native valve endocarditis, 14 had prosthetic valve endocarditis, and six had endocarditis associated with congenital heart defects . Fifty-two patients underwent valve replacement, which was associated with an overall operative mortality of 19% . Those who underwent valve replacement during the early active stage (first 3 weeks) of infection had a higher mortality rate than those who had surgery either during the late active stage (second 3 weeks) of infection or after 6 weeks of antibiotic therapy . S . aureus and Pseudomonas organisms were responsible for the most deaths . On the basis of this study, we recommend that, when cardiovascular function permits, patients who are hemodynamically stable and free of emboli should receive 4 to 6 weeks of antibiotic therapy before undergoing surgical treatment . In contrast, patients with high-risk organisms are more likely to survive if subjected to early surgical intervention . (Texas Heart Institute Journal 1987; 14:401-410)

J Trauma, 1987 Dec, 27(12), 1313 - 22
Treatment of porcine Pseudomonas ARDS with combination drug therapy; Sielaff TD et al.; A combination drug therapy (Poly-5: ibuprofen 12.5 mg/kg, methylprednisolone 30 mg/kg, cimetidine 150 mg, diphenhydramine 10 mg/kg, and ketanserin 0.2 mg/kg) given at 20 and 120 minutes after starting continuous intravenous Pseudomonas (Ps, 5 X 10(8) CFU/20 kg/min) was studied in three groups of swine: saline control (C, n = 9), Ps alone (Ps, n = 8), and Ps plus Poly-5 (n = 5) . PaO2, systemic (SAP) and pulmonary arterial (PAP) pressures, cardiac index (CI), thermal-cardiogreen extravascular lung water (EVLW), pulmonary albumin flux (slope index, SI), and arterial blood serotonin levels (5-HT) were measured . Ps produced significant (p less than 0.05) increases in PAP, EVLW, and SI with decreases in PaO2, CI, and SAP . 5-HT fell significantly compared to baseline . Poly-5 prevented (p less than 0.05) the rise in EVLW and SI and the fall in PaO2 and CI compared to Ps . PAP and SI were maintained at C until 90 and 150 minutes, respectively . SAP fell significantly from C at 30, 60, and 180 minutes . 5-HT was significantly lower than Ps throughout, and significantly lower than baseline at 180 minutes . Combined blockade of arachidonic acid metabolites, histamine, and serotonin receptors prevented hypoxemia, increased pulmonary capillary permeability, and cardiovascular deterioration in this porcine septic ARDS model.

J Bacteriol, 1987 Dec, 169(12), 5821 - 6
Nucleotide sequences of the genes for two distinct cephalosporin acylases from a Pseudomonas strain; Matsuda A et al.; Two genes, acyI and acyII, for distinct cephalosporin acylases from Pseudomonas sp . strain SE83 (A . Matsuda, K . Matsuyama, K . Yamamoto, S . Ichikawa, and K.I . Komatsu, J . Bacteriol . 169:5815-5820, 1987) were sequenced . Each sequence contained a single open reading frame for two nonidentical subunits, predicting a common precursor . Some homologies at the amino acid level were found between the acyII-encoded enzyme, but not the acyI-encoded one, and other related acylases.

Proc Natl Acad Sci U S A, 1987 Dec, 84(24), 8893 - 7
Isolation and sequencing of the gene encoding delta 5-3-ketosteroid isomerase of Pseudomonas testosteroni: overexpression of the protein; Kuliopulos A et al.; We describe the cloning, sequencing, and overexpression of the steroid isomerase (3-oxosteroid delta 5-delta 4-isomerase, EC 5.3.3.1) gene of Pseudomonas testosteroni . A genomic library of P . testosteroni total DNA constructed from partial EcoRI digests ligated to a lambda gtWES vector was probed with a 23-base oligonucleotide mixture {ATGAAC(T)ACC(A,T)CCG(C,A)GAG(A)CAC(T)ATGAC} corresponding to the NH2-terminal sequence of steroid isomerase . Subclones derived from a recombinant phage containing a 5400-base-pair insert were sequenced and found to contain the expected 375-nucleotide open reading frame flanked at both ends by in-frame TGA termination codons . The DNA sequence agreed with the above 125-amino acid sequence except for codons 22, 24, 33, and 38, all of which encoded Asp rather than Asn, and codon 77, which encoded Glu rather than Gln . A 1370-base-pair fragment was inserted into pUC 19 plasmid vector and used to construct a strain of Escherichia coli JM 101 that overexpressed the isomerase gene in the presence of isopropyl beta-D-thiogalactopyranoside . Cytosolic extracts of this strain contained a major soluble protein that migrated with the steroid delta-isomerase subunit on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . These cytosolic extracts had 10-50% of the specific activity of crystalline isomerase, depending on the method of preparation . The recombinant enzyme was crystallized in both monoclinic (flat plates) and hexagonal (bipyramids) crystal forms, described previously for the enzyme isolated from P . testosteroni . The kinetic properties of the crystalline recombinant enzyme, including specific activity, Km for 5-androstene-3,17-dione (340 microM), and Ki for the competitive inhibitor 19-nortestosterone (11.9 microM), agreed closely with the values reported for the isolated enzyme.

Z Gastroenterol, 1987 Dec, 25(12), 749 - 55
{Role of Pseudomonas maltophilia and "Pseudomonas like bacteria group Va" in the etiology of Crohn disease}; Purrmann J et al.; In 1976, Parent and Mitchell isolated cell-wall defective bacteria of the species Pseudomonas maltophilia and Pseudomonas like bacteria group Va from resected bowel tissue of 8 consecutively operated patients with Crohn's disease . The control group, which included patients with ulcerative colitis, was negative in this respect . The isolated strains were available for our own investigations . New Zealand white rabbits were inoculated with killed bacteria to produce specific antisera . Frozen sections of affected tissue from 6 consecutively operated Crohn patients were incubated with the various antisera and investigated by indirect immunofluorescence . No positive reactions were demonstrable in repeated tests with different dilutions of antisera . Our results do not support the hypothesis that bacteria of the species Pseudomonas maltophilia and Pseudomonas like bacteria group Va are an etiologic factor in Crohn's disease.

Eur J Epidemiol, 1987 Dec, 3(4), 343 - 6
Current status of Pseudomonas cepacia typing systems; Rabkin CS et al.; Pseudomonas cepacia is an important nosocomial pathogen for which measures of isolate relatedness are being developed . Typing systems based on 4 different strain characteristics have been proposed: serologic reactions, biochemical reactions, plasmid profiles, and bacteriocin production and susceptibility . Serology and bacteriocins distinguish many types, but the sensitivity and specificity of these techniques have not been determined . Improved methods for typing P . cepacia are needed to determine the reservoirs and modes of transmission of this emerging nosocomial pathogen.

Cancer, 1987 Dec 1, 60(11), 2596 - 604
Combined antiestrogen and cytotoxic therapy with pseudomonas vaccine immunotherapy for metastatic breast cancer . A prospective, randomized trial; Hortobagyi GN et al.; One hundred thirty-three consecutive, previously untreated patients who had metastatic breast cancer were treated with a combination of 5-fluorouracil, doxorubicin (Adriamycin), and cyclophosphamide (FAC) . They were randomly assigned to receive nonspecific immunotherapy with a heptavalent pseudomonas vaccine . Sixty-five patients were treated with pseudomonas vaccine, whereas 68 did not receive immunotherapy . In addition, all patients with estrogen receptor-positive tumors or tumors with an estrogen receptor status were also treated with tamoxifen . To allow clinical assessment of hormone sensitivity in vivo, tamoxifen was started 6 weeks before chemotherapy except in patients who had life-threatening disease . After the initial 6 weeks of tamoxifen, 3% of patients had achieved a complete remission, 9% a partial remission, while 16% achieved a minor response . The maximum response after tamoxifen and chemotherapy included complete remissions in 20% of patients and partial remissions in 61% of patients for an overall remission rate of 81% . The median response duration was 15 months, and the median survival time, 27 months . There were no differences in remission rate, remission duration, or survival time between the groups treated with or without pseudomonas vaccine . Eleven patients with limited metastatic disease received radiotherapy consolidation to initially involved sites . In these patients the median time from radiotherapy to progression of disease was 33 months, and the median survival time was 46 months . We conclude that nonspecific immunotherapy with pseudomonas vaccine failed to increase remission rate or survival time . Furthermore, the addition of tamoxifen to FAC chemotherapy did not improve the remission rate or duration compared to a recent, historical control group of patients treated with only FAC chemotherapy.

Mol Gen Genet, 1987 Dec, 210(2), 270 - 6
Genetic analysis of a transposon carrying toluene degrading genes on a TOL plasmid pWW0; Tsuda M et al.; Toluene degrading (xyl) genes on a Pseudomonas TOL plasmid pWW0 are located within a 39-kb DNA portion . The 56-kb region including these xyl genes and its 17-kb derivative with a deletion of the internal 39-kb portion transposed to various sites on target replicons such as pACYC184 and R388 in Escherichia coli recA strains . Thus the 56- and 17-kb regions were designated Tn4651 and Tn4652, respectively . Genetic analysis of Tn4652 demonstrated that its transposition occurs by a two-step process, namely, cointegrate formation and its subsequent resolution . The presence in cis of DNA sequences of no more than 150 bp at both ends of Tn4652 was prerequisite for cointegrate formation, and this step was mediated by a trans-acting factor, transposase, which was encoded in a 3.0-kb segment at one end of the transposon . Cointegrate resolution took place site-specifically within a 200-bp fragment, which was situated 10 kb away from the transposase gene . Based on the stability of cointegrates formed by various mini-Tn4652 derivatives, it was shown that the cointegrate resolution requires two trans-acting factors encoded within 1.0- and 1.2-kb fragments that encompass the recombination site involved in the resolution.

J Bacteriol, 1987 Dec, 169(12), 5815 - 20
Cloning and characterization of the genes for two distinct cephalosporin acylases from a Pseudomonas strain; Matsuda A et al.; Pseudomonas sp . strain SE83 converts cephalosporin C and 7 beta-(4-carboxybutanamido)cephalosporanic acid (GL-7ACA) to 7-aminocephalosporanic acid (7ACA) . A DNA library of this strain was constructed in Escherichia coli and screened for the ability to deacylate GL-7ACA to 7ACA . Apparently, two distinct genes, designated acyI and acyII, were cloned on 4.8- and 6.0-kilobase-pair BglII fragments, respectively . The enzymes encoded by the two genes showed different substrate specificities, and the acyII-encoded enzyme was found to yield 7ACA from cephalosporin C by direct deacylation . Expression of the two genes in E . coli was strongly dependent on a promoter of the vector . The coding regions for acyI and acyII were localized on the 2.5- and 2.8-kilobase-pair fragments, respectively, by subcloning experiments, and high expression of both genes was obtained by placing them under the control of the lacUV5 promoter . The acyII-encoded enzyme was purified and shown to be composed of two nonidentical subunits with molecular weights of 26,000 and 57,000 . Maxicell analysis revealed three acyII-specific polypeptides, two of which corresponded to the above subunits . The third polypeptide with a molecular weight of 83,000 was suggested to be the precursor of both subunits.

J Bacteriol, 1987 Dec, 169(12), 5789 - 94
Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv . glycinea; Staskawicz B et al.; A wide-host-range cosmid cloning vector, pLAFR3, was constructed and used to make cosmid libraries of partially digested Sau3A DNA from race 0 and race 1 of Pseudomonas syringae pv . glycinea . Two avirulence genes, avrB0 and avrC, cloned from race 0, elicited the hypersensitivity reaction (HR) on specific cultivars of soybean . Race 4 transconjugants containing avrB0 induced a dark brown necrotic HR within 24 h on the soybean cultivars Harosoy and Norchief, whereas race 4 transconjugants containing avrC induced a light brown necrotic HR within 48 h on the soybean cultivars Acme, Peking, Norchief, and Flambeau . An additional avirulence gene, avrB1, cloned from race 1, appeared to be identical to avrB0 from race 0 . The avrB0 and avrC genes from race 0 were characterized by restriction enzyme mapping, Southern blot analysis, Tn5 transposon mutagenesis, and site-directed gene replacements . The effects of these three genes on the in planta bacterial growth of race 4 transconjugants have also been examined . The identification and cloning of avrB1 provides genetic evidence for a gene-for-gene interaction in the bacterial blight disease of soybean, as avrB1 from race 1 interacts with the soybean disease resistance locus, Rpg1.

J Antibiot (Tokyo), 1987 Nov, 40(11), 1520 - 9
Xylocandin: a new complex of antifungal peptides . II . Structural studies and chemical modifications; Bisacchi GS et al.; Xylocandins A1, A2, B1, B2, C1, C2, D1 and D2 are new antifungal peptides isolated from Pseudomonas cepacia ATCC 39277 . The molecular weights of the xylocandins were determined by fast atom bombardment mass spectrometry (A1 m/z 1,215; A2 1,199; B1 1,229; B2 1,213; C1 1,097; C2 1,081; D1 1,083; D2 1,067) . Each xylocandin is a cyclic peptide containing glycine, serine, asparagine (1-3 residues), beta-hydroxytyrosine, and an unusual amino acid with the formula C18H37NO5 . Additionally A1, A2, D1 and D2 contain 2,4-diaminobutyric acid; A1, B1, C1 and D1 contain erythro-beta-hydroxyasparagine; and A1, A2, B1 and B2 contain xylose . For each xylocandin pair, an erythro-beta-hydroxyasparagine residue in the first component of the pair is thus replaced by an asparagine in the second component, accounting for the 16 dalton mass difference for each pair . Chemical modification of A1 and A2 at the diaminobutyric acid and beta-hydroxytyrosine residues was used to probe structural requirements for activity.

Drug Intell Clin Pharm, 1987 Nov, 21(11), 879 - 81
Increased theophylline concentrations secondary to ciprofloxacin; Rybak MJ et al.; An 82-year-old man with a history of myasthenia gravis and heart failure was admitted to the hospital with respiratory failure . Aminophylline and eventually theophylline therapy were initiated to improve respiratory status . During the hospital stay, the patient developed a resistant pseudomonal pneumonia . After failure with conventional antibiotics, ciprofloxacin was initiated because of favorable sensitivity and the planned avoidance of aminoglycoside therapy . Seventy-two hours after initiation of ciprofloxacin, the patient's theophylline level rose from a steady-state baseline of 9.8 micrograms/ml to 34.7 micrograms/ml . After the theophylline dose was reduced by approximately 67 percent, the patient's theophylline serum concentration returned to baseline (10 micrograms/ml) . Until more data concerning the interaction of theophylline and ciprofloxacin are available, we recommend close monitoring of theophylline serum concentrations in patients receiving concomitant ciprofloxacin.

J Bacteriol, 1987 Nov, 169(11), 4980 - 3
Purification and properties of alpha-pinene oxide lyase from Nocardia sp . strain P18.3; Griffiths ET et al.; alpha-Pinene oxide is an intermediate in the degradation of alpha-pinene by Nocardia sp . strain P18.3 and some Pseudomonas strains . The epoxide is cleaved by a lyase which catalyzes a concerted reaction in which both rings of the bicyclic structure are cleaved with the formation of cis-2-methyl-5-isopropylhexa-2,5-dienal . The enzyme has been purified to homogeneity from Nocardia sp . strain P18.3 . It was induced by growth with alpha-pinene and constituted 6 to 7% of the soluble protein of cell extracts . The apparent molecular weight of the native enzyme was 50,000 by ultracentrifugal analysis . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave two dissimilar subunits with apparent molecular weights of 17,000 and 22,000 . The enzyme was devoid of prosthetic groups, had no cofactor requirement, and had a broad pH activity range, a Km for alpha-pinene oxide of 9 microM, and a turnover number of 15,000 . Inhibitors included sulfhydryl reactive compounds, terpene epoxides, and pinane derivatives with substituent groups at carbon 3 . A mechanism for the concerted reaction has been proposed in which decyclization is initiated by donation of a proton from the catalytic center to the oxygen of the epoxide with consequent destabilization . In vitro the enzyme was inactivated during catalysis, and a reactive cationic intermediate may be responsible for this phenomenon . The enzyme should be classified as a lyase EC 4.99.-.-.

Prikl Biokhim Mikrobiol, 1987 Nov-Dec, 23(6), 747 - 53
{Isolation, purification and stability of a glutamin(asparagin)ase preparation from Pseudomonas boreopolis 526}; Pekhov AA et al.; A technique for purification of glutamine asparaginase from Pseudomonas boreopolis 526 is described which provides a 37% yield of the enzyme homogeneous according to electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate . The effect of pH, freezing, thawing and lyophilic drying on the stability of glutamine asparaginase was studied . The enzyme is rather stable at pH 4.8 and 4 degrees C . Lyophilic drying of the homogeneous enzyme without addition of stabilizers resulted in a decrease of its activity an is accompanied by formation of protein conglomerates with molecular weights of 280,000 and 660,000 D . Freezing and thawing decreased the activity of the nature enzyme by 40-50%.

Biochem Cell Biol, 1987 Nov, 65(11), 968 - 77
Analysis of the lipopolysaccharide of Pseudomonas maltophilia 555; Di Fabio JL et al.; The phenol phase soluble lipopolysaccharide of Pseudomonas maltophilia strain 555, obtained from cells by the hot aqueous phenol method, was of the smooth type . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hydrolysis, methylation, and 13C and 1H nuclear magnetic resonance analyses showed that this lipopolysaccharide has an O-chain polysaccharide composed of a repeating pentasaccharide unit, containing D-rhamnose (D-Rha, one part), 3-acetamido-3,6-dideoxy-D-galactose (D-Fuc3NAc, one part), and 4-acetamido-4,6-dideoxy-D-mannose (D-Rha4NAc, three parts) and having the structure (formula; see text) The serological cross-reactions between P . maltophilia 555 and Brucella species can now be related to the occurrence of N-acyl derivatives of 4-amino-4,6-dideoxy-D-mannopyranosyl residues in the O-chains of their respective lipopolysaccharide components.

Appl Environ Microbiol, 1987 Nov, 53(11), 2617 - 23
Kinetics of p-nitrophenol mineralization by a Pseudomonas sp.: effects of second substrates; Schmidt SK et al.; The kinetics of simultaneous mineralization of p-nitrophenol (PNP) and glucose by Pseudomonas sp . were evaluated by nonlinear regression analysis . Pseudomonas sp . did not mineralize PNP at a concentration of 10 ng/ml but metabolized it at concentrations of 50 ng/ml or higher . The Ks value for PNP mineralization by Pseudomonas sp . was 1.1 micrograms/ml, whereas the Ks values for phenol and glucose mineralization were 0.10 and 0.25 micrograms/ml, respectively . The addition of glucose to the media did not enable Pseudomonas sp . to mineralize 10 ng of PNP per ml but did enhance the degradation of higher concentrations of PNP . This enhanced degradation resulted from the simultaneous use of glucose and PNP and the increased rate of growth of Pseudomonas sp . on glucose . The Monod equation and a dual-substrate model fit these data equally well . The dual-substrate model was used to analyze the data because the theoretical assumptions of the Monod equation were not met . Phenol inhibited PNP mineralization and changed the kinetics of PNP mineralization so that the pattern appeared to reflect growth, when in fact growth was not occurring . Thus, the fitting of models to substrate depletion curves may lead to erroneous interpretations of data if the effects of second substrates on population dynamics are not considered.

Zh Mikrobiol Epidemiol Immunobiol, 1987 Nov, (11), 44 - 7
{Cell-free Pseudomonas vaccine: its immunochemical characteristics and immunogenicity}; Bandman OA et al.; Two variants of cell-free protein vaccine have been prepared from the mixture of 4 P . aeruginosa strains, serovars 02, 06, 07 and 011, and from a single P . aeruginosa strain, serovar 02 . The preparation contains proteins with molecular weight ranging from 20,000 to 100,000 and the admixture of lipopolysaccharide in negligible amounts (not exceeding 0.08% of dry weight) . The vaccine produces no signs of toxicosis in laboratory animals . The vaccine effectively protects mice challenged with P . aeruginosa of different O-serotypes and stimulates the formation of specific protective antibodies in rabbits.

J Dairy Res, 1987 Nov, 54(4), 535 - 43
Production of extracellular lipases and proteinases during prolonged growth of strains of psychrotrophic bacteria in whole milk; Stead D; Three strains of psychrotrophic Pseudomonas spp., each with different lipolytic and proteolytic phenotypes (including a proteinase-deficient mutant), were cultured separately in whole milk at 7 degrees C . Growth rates were the same during logarithmic growth phase, but during early stationary phase the cell densities were related to the activities of lipase and proteinase in the cultures . Only one strain underwent pronounced death phase . Proteinase activity was not detected in the culture of the proteinase-deficient mutant, but in those of the other strains it increased to a plateau, or continued to increase linearly . Lipase activity of the culture of each strain reached a peak in early stationary phase; in late stationary phase activity was highest for the proteinase-deficient mutant strain where degradation of lipase by bacterial proteinase would have been least . The ability of psychrotrophic bacteria both to survive for long periods and to produce high levels of proteinases and lipases on prolonged incubation in milk emphasizes the spoilage potential arising from psychrotrophic bacteria in inadequately cleaned dairy equipment.

J Antimicrob Chemother, 1987 Oct, 20(4), 541 - 6
Therapeutic efficacy of cefpiramide-ciprofloxacin combination in experimental Pseudomonas infections in neutropenic mice; Fu KP et al.; The therapeutic efficacy of cefpiramide and ciprofloxacin alone and in combination was investigated and compared with that of ticarcillin plus tobramycin against pseudomonal infections in mice made neutropenic by administration of cyclosphosphamide . Therapy with cefpiramide plus ciprofloxacin was significantly more effective than that by either antibiotic alone . These results were consistent with in-vitro synergistic effects . At a higher dose of ciprofloxacin (4 mg/kg) plus cefpiramide (50 mg/kg), the combination therapy protected all neutropenic mice from fatal bacteraemia, and was more protective than ticarcillin (200 mg/kg) plus tobramycin (1 mg/kg) . The peak serum concentration of cefpiramide in infected neutropenic mice was 51 mg/l when they were given 50 mg/kg subcutaneously . Ciprofloxacin attained a peak serum concentration of 1.2 mg/l and a serum half-life of 34 min.

J Am Vet Med Assoc, 1987 Oct 1, 191(7), 811 - 5
Pseudomonas mastitis: difficulties in detection and elimination from contaminated wash-water systems; Erskine RJ et al.; Histories of 4 dairy herds with increased incidence of Pseudomonas mastitis associated with contaminated wash hoses in milking parlors are described . Problems of detection and elimination of the organism from contaminated water sources and the inadequacy of iodide germicides in eliminating Pseudomonas are discussed . In problem herds, greater numbers of organisms often are found in the water left standing in the wash hoses between milkings . Thus, flushing of hoses before their use in the milking process may be beneficial in reducing exposure of the cows to the organism.

J Bacteriol, 1987 Oct, 169(10), 4577 - 80
Physical mapping of transposon Tn5 insertions defines a gene cluster functional in nitrous oxide respiration by Pseudomonas stutzeri; Viebrock A et al.; By transposon Tn5 mutagenesis, 19 strains of Pseudomonas stutzeri were acquired that had defects in nitrous oxide respiration (Nos- phenotype) . A physical map of the mutants showed nearly random Tn5 insertions into genomic DNA within a single region ca . 8 kilobases long . Mutants were characterized immunochemically, enzymatically, and chemically . Several functions related to the synthesis and regulation of nitrous oxide reductase were associated with this DNA region, indicating that in P . stutzeri part of the genetic information necessary to respire nitrous oxide is clustered.

J Cell Physiol, 1987 Oct, 133(1), 127 - 34
Mutant KB cells with decreased EGF receptor expression: biochemical characterization; Hwang J et al.; Mutants of the human KB carcinoma cell line resistant to a cytotoxic conjugate of epidermal growth factor and Pseudomonas exotoxin (EGF-PE) express a pleiotropic phenotype, which includes reduced levels of 125I-EGF binding, without altered affinity for EGF (Lyall et al., 1987) . Here, the EGF-toxin (ET) resistant mutants were further characterized with respect to the amount and size of the EGF receptor and the level of EGF receptor RNA . These data indicate that decreased binding of 125I-EGF in the mutants is due to reduced amounts of EGF receptor, which is associated with decreased mRNA levels . Changes in other proteins in the ET mutants were also examined . Five of the six ET mutants had a decrease in a 78,000 Mr- membrane glycoprotein . In addition, an increase in a protein with a Mr- of 40,000 and a pl = 8.0 was found in all the mutants, and an increase in a series of proteins with a Mr- of 36,000 and a pl of 6.3-6.5 was found in some of the mutants . These results confirm the pleiotropic nature of the EGF-PE resistant mutants and show that reduced EGF binding is due to altered expression of the EGF receptor gene in the mutants.

J Biol Chem, 1987 Sep 5, 262(25), 12164 - 8
Degradation of glyphosate by Pseudomonas sp . PG2982 via a sarcosine intermediate; Kishore GM et al.; The bacterium Pseudomonas PG2982 metabolizes glyphosate (N-(phosphonomethyl)glycine) by converting it to glycine, a one-carbon unit, and phosphate . Here we show that this conversion involves the intermediate formation of sarcosine . When cells are incubated with {14C}glyphosate, the 14C can be entrapped in glycine or sarcosine . With added sarcosine, 14C from all three carbons of glyphosate is recovered solely in sarcosine . In experiments with glycine, radioactivity from the carboxymethyl moiety of glyphosate is trapped in glycine as well as serine, whereas radioactivity from the phosphonomethyl carbon is only incorporated into serine . These results are consistent with a pathway involving the conversion of glyphosate to sarcosine by cleavage of its carbon-phosphorus (C-P) bond, followed by the oxidation of sarcosine to glycine and formaldehyde.

Biochim Biophys Acta, 1987 Sep 3, 902(3), 327 - 34
Preparation of fluorescent derivatives of lipases and their use in fluorescence energy transfer studies in hydrocarbon/water interfaces; Roberts GA et al.; Fluorescein isothiocyanate reacted with a chromobacter and pseudomonad lipase to yield mono-substituted, fully active, enzymes . With the carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) in the non-aqueous phase, fluorescence energy transfer was used to follow the lipase and similarly labelled model proteins in and out of the interface in heptane, and heptane/di-O-palmitoyl-rac-glycerol (a substrate analogue), emulsions . Competitive binding, and displacement by other proteins could also be followed.

Appl Environ Microbiol, 1987 Sep, 53(9), 2129 - 32
Isolation of a Pseudomonas stutzeri strain that degrades o-xylene; Baggi G et al.; A Pseudomonas stutzeri strain capable of growing on o-xylene was isolated from enrichment cultures . The organism grew on 2,3- and 3,4-dimethylphenol but not on 2-methylbenzyl alcohol, o-tolualdehyde, or o-toluate . P . stutzeri was not able to utilize m-xylene, p-xylene, or 1,2,4-trimethylbenzene, but growth was observed in the presence of the corresponding alcohols and acids . From the Pseudomonas cultures supplied with o-xylene, 2,3-dimethylphenol was isolated and identified . When resting P . stutzeri cells were incubated with 2,3-dimethylphenol, the reaction mixture turned greenish yellow and showed spectral properties identical to those of the 3,4-dimethylcatechol meta ring fission product . Catechol 2,3-oxygenase was induced by growth on o-xylene or on 2,3- or 3,4-dimethylphenol . The suggested hypothesis is that the first metabolic steps of growth on o-xylene involve the direct oxygenation of the aromatic nucleus, followed by meta pathway reactions.

J Gen Microbiol, 1987 Sep, 133 ( Pt 9), 2487 - 95
Catabolism of arginine, citrulline and ornithine by Pseudomonas and related bacteria; Stalon V et al.; The distribution of the arginine succinyltransferase pathway was examined in representative strains of Pseudomonas and related bacteria able to use arginine as the sole carbon and nitrogen source for growth . The arginine succinyltransferase pathway was induced in arginine-grown cells . The accumulation of succinylornithine following in vivo inhibition of succinylornithine transaminase activity by aminooxyacetic acid showed that this pathway is responsible for the dissimilation of the carbon skeleton of arginine . Catabolism of citrulline as a carbon source was restricted to relatively few of the organisms tested . In P . putida, P . cepacia and P . indigofera, ornithine was the main product of citrulline degradation . In most strains which possessed the arginine succinyltransferase pathway, the first step of ornithine utilization as a carbon source was the conversion of ornithine into succinylornithine through an ornithine succinyltransferase . However P . cepacia and P . putida used ornithine by a pathway which proceeded via proline as an intermediate and involved an ornithine cyclase activity.

J Hosp Infect, 1987 Sep, 10(2), 179 - 86
Pseudomonas peritonitis in continuous ambulatory peritoneal dialysis: laboratory predictors of treatment failure; Craddock CF et al.; Five episodes of pseudomonas peritonitis complicating continuous ambulatory peritoneal dialysis (CAPD) which were not cured by intraperitoneal antibiotics were studied to assess causes for treatment failure . The activity of gentamicin and ceftazidime against these strains was decreased in the presence of sterile used dialysate compared with nutrient broth . Likewise, kinetic studies showed that in dialysate therapeutically used concentrations of antibiotics failed to kill the isolates over 24 h . All five pseudomonas strains were adherent to silicone rubber Tenckoff catheter segments . An in vitro model of CAPD peritonitis demonstrated that persistence of viable adherent bacteria, after exposure to therapeutic concentrations of gentamicin and ceftazidime, contributes to the failure of antibiotics to cure pseudomonas CAPD peritonitis.

J Biochem (Tokyo), 1987 Sep, 102(3), 481 - 6
Evidence against proton pump activity by cytochrome c oxidase of Pseudomonas AM1; Sone N et al.; Proteoliposomes reconstituted from purified cytochrome c oxidase of Pseudomonas AM1 and from a heptyl beta-D-thioglucoside-extract of its membranes showed respiratory control but did not show H+ pumping upon a pulse with reduced cytochrome c . The stoichiometries of respiration-dependent H+ translocation in the resting cells respiring ascorbate via N,N,N',N'-tetramethyl-p-phenylenediamine were measured by the oxygen-pulse and initial rate methods . The apparent H+/O ratio of about 2 was due to 2H+ release from the hydrogen-donating substrate . These results strongly suggested that Pseudomonas AM1 does not pump H+ intrinsically, although the enzyme catalyzes electron transfer across the membranes.

J Biol Chem, 1987 Aug 25, 262(24), 11857 - 63
Purification to homogeneity and initial physical characterization of secondary amine monooxygenase; Alberta JA et al.; Secondary amine monooxygenase from Pseudomonas aminovorans grown on trimethylamine has been purified 265-fold to apparent homogeneity . The purified enzyme exhibits a specific activity of 14.7 mumol of NADPH oxidized per min per mg of protein, a native molecular weight of 210,000, and nondisulfide-linked subunits of molecular weight 42,000, 36,000, and 24,000, each of which is required for activity . The enzyme is extremely labile during purification; rapid handling and the presence of 5% ethanol are essential to enzyme stability . Storage at 77 K in the presence of NADH (1 mM) also confers considerable stability to the purified enzyme . The heme prosthetic group in the enzyme has been identified as protoporphyrin IX . The quantification of prosthetic group components reveals the presence of 1.6 mol of flavin as FMN, 2.0 mol of heme iron, 4.0 mol of acid-soluble (nonheme) iron, and 3.6 mol of free sulfide/210,000 molecular weight enzyme . Ferric and ferrous-CO secondary amine monooxygenase exhibit electronic absorption spectra that are very similar to those of analogous myoglobin derivatives and, therefore, quite distinct from parallel forms of cytochrome P-450, the most extensively studied heme iron-containing monooxygenase . Like myoglobin and, again, in contrast to P-450, this enzyme forms a very stable dioxygen complex . In fact, it is this oxygen-bound form of the enzyme that is obtained from the purification procedure . Once again, the absorption spectrum of oxygenated secondary amine monooxygenase is nearly identical to that of oxymyoglobin . The spectroscopic similarities between secondary amine monooxygenase and myoglobin suggest the presence of an endogenous histidine fifth ligand to the heme iron of the enzymes.

Eur J Biochem, 1987 Aug 3, 166(3), 575 - 9
NAD(P)+-independent aldehyde dehydrogenase from Pseudomonas testosteroni . A novel type of molybdenum-containing hydroxylase; Poels PA et al.; Aldehyde dehydrogenase from Pseudomonas testosteroni was purified to homogeneity . The enzyme has a pH optimum of 8.2, uses a wide range of aldehydes as substrates and cationic dyes (Wurster's blue, phenazine methosulphate and thionine), but not anionic dyes (ferricyanide and 2.6-dichloroindophenol), NAD(P)+ or O2, as electron acceptors . Haem c and pyrroloquinoline quinone appeared to be absent but the common cofactors of molybdenum hydroxylases were present . Xanthine was not a substrate and allopurinol was not an inhibitor . Alcohols were inhibitors only when turnover of the enzyme occurred in aldehyde conversion . The enzyme has a relative molecular mass of 186,000, consists of two subunits of equal size (Mr 92,000), and 1 enzyme molecule contains 1 FAD, 1 molybdopterin cofactor, 4 Fe and 4 S . It is a novel type of NAD(P)+-independent aldehyde dehydrogenase since its catalytic and physicochemical properties are quite different from those reported for already known aldehyde-converting enzymes like haemoprotein aldehyde dehydrogenase (EC 1.2.99.3), quino-protein alcohol dehydrogenases (EC 1.1.99.8) and molybdenum hydroxylases.

Can J Microbiol, 1987 Aug, 33(8), 658 - 62
alpha-Santonin 1,2-reductase and its role in the formation of dihydrosantonin and lumisantonin by Pseudomonas cichorii S; Naik U et al.; 1,2-Dihydrosantonin is the first stable product in the degradative pathway of alpha-santonin by Pseudomonas cichorii S . Its formation is catalyzed by an oxidoreductase, which is NADH or NADPH dependent and has an apparent Km value of 66.66 microM for santonin and 44.33 microM for NADH . The enzyme activity is stable at pH 6.0, 7.0, and 8.0, and is not affected by EDTA and divalent metal ions . It is postulated that the enzymic reduction of santonin occurs via formation of a transient zwitterionic intermediate, which undergoes nonenzymatic 1,4-sigmatropic rearrangement to yield lumisantonin during the solvent extraction process . Lumisantonin is, thus, not a true metabolic intermediate but an artifact.

Biochem J, 1987 Aug 1, 245(3), 899 - 901
Pseudomonas toxin binds triton X-114 at low pH; Sandvig K et al.; Pseudomonas toxin was found to bind Triton X-114 at pH values below pH 5.0, whereas much less binding was observed at higher pH values . The toxin bound Triton X-114 at a higher pH value in the presence of 0.14 M-NaCl, -KCl or -NaNO3 than at low salt concentrations . The results suggest that low pH in an intracellular compartment facilitates the transport of Pseudomonas toxin across the membrane and into the cytosol by inducing a conformational change in the molecule.

Med J Aust, 1987 Jul 20, 147(2), 95 - 6
Subacute pulmonary melioidosis in a temperate climate; Wilson JW et al.; Previous reports of cases of melioidosis that were seen in nonendemic areas of Australia describe recrudescences of latent infection . We describe the case of a patient who presented in the cooler climate of Melbourne with a probable primary, subacute pulmonary infection with Pseudomonas pseudomallei . This case illustrates several points that bear consideration in the management of atypical pneumonia and, more specifically, pulmonary melioidosis . Historical and occupational clues are easily missed or unrecognized, while a persistent growth of gentamicin-resistant Pseudomonas species should arouse suspicion . Septicaemic melioidosis carries a poor prognosis, and treatment should be early and aggressive; use of the newer, third-generation cephalosporin agents should be considered . Given active support in a well-equipped intensive care unit, together with appropriate antibiotic therapy, patients may eventually be cured of this infection, but a high mortality rate is still encountered.

J Biol Chem, 1987 Jul 5, 262(19), 9340 - 6
Purification and some properties of component B of the 4-chlorophenylacetate 3,4-dioxygenase from Pseudomonas species strain CBS 3; Schweizer D et al.; 4-Chlorophenylacetate 3,4-dioxygenase system from Pseudomonas sp . CBS 3 consists of two protein components, a red-brown iron-sulfur protein (component A) which functions as dioxygenase and an orange-colored reductase (component B) . Component B was purified by a five-step procedure . Criterion of purity was sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which also showed that the protein consists of one polypeptide species with a molecular weight of 35,000 . With gel chromatography on Sephadex G-100 also, a molecular weight of 35,000 was found for the native enzyme, suggesting a monomeric structure for the reductase enzyme . The isoelectric point was determined as pH 4.8 . The visible absorption spectrum of the homogeneous protein exhibits maxima at 336, 394, and 458 nm . One mol of FMN, 2.1 mol of iron, and 1.7 mol of acid-labile sulfide were found in one mol of component B . The EPR-spectrum of the reduced form of the enzyme (with NADH) showed two types of signals . The signal at g values of 2.03, 1.94, and 1.90 was assigned to an iron-sulfur cluster of the {2Fe-2S}-type . The properties of the other signal type at g = 2.004 are characteristic of flavosemiquinone radical which does not show coupling to an other paramagnetic center . The apparent Km values for 2,6-dichlorophenolindophenol, cytochrome c, and NADH were calculated from Lineweaver-Burk plots as 6.3, 2.3, and 32 microM, respectively . Inhibitors of the reductase are metal-chelating reagents and sulfhydryl-inhibiting compounds . Component B reduces several redox compounds: 2,6-dichlorophenolindophenol, potassium hexacyanoferrat III, cytochrome c, methylene blue, and nitro blue tetrazolium.

Appl Environ Microbiol, 1987 Jul, 53(7), 1626 - 31
Widespread occurrence of bacterial thiol methyltransferases and the biogenic emission of methylated sulfur gases; Drotar A et al.; A majority of heterotrophic bacteria isolated from soil, water, sediment, vegetation, and marine algae cultures methylated sulfide, producing methanethiol . This was demonstrated with intact cells by measuring the emission of methanethiol with a sulfur-selective chemiluminescence detector, and in cell extracts by detection of sulfide-dependent thiol methyltransferase activity . Extracts of two Pseudomonas isolates were fractionated by gel-filtration and ion-exchange chromatography, and with sulfide as the substrate a single peak of thiol methyltransferase activity was seen in each case . Extracts of several bacterial strains also contained thiol methyltransferase activity with organic thiols as substrates . Thus, S-adenosylmethionine-dependent thiol methyltransferase activities are widespread in bacteria and may contribute to biogenic emissions of methylated sulfur gases and to the production of methyl thioethers.

Br J Urol, 1987 Jul, 60(1), 51 - 3
The longer-term results of undiversion; Galloway NT et al.; Thirty patients have been undiverted over the past 7 years . All but six were originally diverted for neuropathic problems of incontinence and/or upper tract compromise . Eighteen were undiverted because of deteriorating renal function . Only two were reconstructed for purely social reasons . Twenty-eight patients have stable renal function, though three are enuretic and two have persistent Pseudomonas infection . Ureteric "failure" remains the most difficult problem.

Mol Biol (Mosk), 1987 Jul-Aug, 21(4), 1050 - 5
{Regulation of expression of the htpR gene in Escherichia coli}; Kiselev VI et al.; The hybrid operon htpR: ATP was obtained, in which the expression of the aminoglycoside phosphotransferase gene is controlled by the promoter of the htpR gene of Escherichia coli . It was shown that the transcription of the htpR gene is not induced during the "heat shock" . The basal level of the htpR gene transcription in htpR mutants was found to be two times higher than that in the strains of the "wild type" . These findings suggested that the expression of the htpR gene is autoregulated . Hybridization experiments demonstrated that the genome of Pseudomonas bacteria contains the htpR-like gene.

Proc Natl Acad Sci U S A, 1987 Jul, 84(13), 4538 - 42
Activity of a recombinant fusion protein between transforming growth factor type alpha and Pseudomonas toxin; Chaudhary VK et al.; The transforming growth factor type alpha gene has been fused to a modified Pseudomonas toxin gene from which the cell-recognition domain has been deleted . The chimeric gene has been expressed in Escherichia coli, and the chimeric protein, PE40-TGF-alpha, has been highly purified . PE40-TGF-alpha kills cells expressing epidermal growth factor receptors and has little activity against cells with few receptors . This chimeric protein might be useful in treating cancers that contain high numbers of epidermal growth factor receptors.

Biochemistry, 1987 Jun 30, 26(13), 3927 - 37
Positioning of a spin-labeled substrate analogue into the structure of delta 5-3-ketosteroid isomerase by combined kinetic, magnetic resonance, and X-ray diffraction methods; Kuliopulos A et al.; We have shown by kinetic and magnetic resonance measurements that a spin-labeled substrate analogue, spiro{doxyl-2,3'-5' alpha-androstan}-17'beta-ol, binds at the substrate site of crystalline delta 5-3-ketosteroid isomerase (steroid delta-isomerase; EC 5.3.3.1) of Pseudomonas testosteroni . The spin-labeled steroid is a linear competitive inhibitor with a Ki value (25 +/- 5 microM) that is consistent with dissociation constants obtained by direct binding measurements based on changes in the electron paramagnetic resonance spectrum of the nitroxide, longitudinal relaxation rates of water protons, and longitudinal and transverse relaxation rates of carbon-bound protons of the isomerase . These binding studies yield a stoichiometry for the nitroxide of 1 per subunit of the enzyme . Measurements of the longitudinal relaxation rates of water protons indicate that the 3-doxyl portion of the spin-label is highly immobilized yet is exposed to solvent . Paramagnetic effects of the nitroxide on T1 defined distances to several previously assigned {Benisek, W . F., & Ogez, J . R . (1982) Biochemistry 21, 5816-5825} and newly assigned protons of the enzyme . These distances were then used to locate (with an accuracy of +/- 2 A) the nitroxide moiety at a unique position in a partially refined 2.5-A resolution X-ray structure of native isomerase . Three of five additional proton resonance peaks, attributed to ring-shielded methyl groups, could be assigned to specific residues on the basis of distances from the spin-label in the X-ray structure . The remaining portion of the spin-labeled steroid was then docked into the X-ray structure in a hydrophobic cavity of the enzyme . This position of the steroid is consistent with the steroid binding site previously proposed {Westbrook, E . M., Piro, O . E., & Sigler, P . B . (1984) J . Biol . Chem . 259, 9096-9103} . However, the rotational orientation of this steroid about its long axis could not be unambiguously established . If we assume that steroid substrates and the spin-labeled inhibitor bind to the same site, but with reversal of the 3- and 17-positions, then the phenolic hydroxyl of Tyr-55 is optimally positioned to function as the general acid that protonates the 3-keto group of the substrate, facilitated by the negative end of the dipole of a 10-residue alpha-helix, the only helix in the molecule.(ABSTRACT TRUNCATED AT 400 WORDS)

Biochem J, 1987 Jun 15, 244(3), 565 - 70
An NAD+-dependent alanine dehydrogenase from a methylotrophic bacterium; Bellion E et al.; A study was made of the NAD+-dependent alanine dehydrogenase (EC 1.4.1.1) elaborated by the methylotrophic bacterium Pseudomonas sp . strain MA when growing on succinate and NH4Cl . This enzyme was purified 400-fold and was found to be highly specific for NH3 and NAD+; however, hydroxypyruvate and bromopyruvate, but not alpha-oxoglutarate or glyoxylate, could replace pyruvate to a limited extent . The Mr of the native enzyme was shown to be 217,000, and electrophoresis in SDS/polyacrylamide gels revealed a minimum Mr of 53,000, suggesting a four-subunit structure . The enzyme, which has a pH optimum of 9.0, operated almost exclusively in the aminating direction in vitro . It was induced by NH3 or by alanine, and was repressed by growth on methylamine or glutamate . It is suggested that this enzyme has two roles in this organism, namely in NH3 assimilation and in alanine catabolism.

Dtsch Med Wochenschr, 1987 Jun 12, 112(24), 963 - 6
{Progressive multifocal leukoencephalopathy . Late complication in chronic lymphatic leukemia}; Hofeler H et al.; In a 44 year old patient with chronic lymphatic leukemia and secondary antibody deficiency syndrome, a disorder of articulation and a left hemiparesis developed during a thrombocytopenic phase . Computer tomography of the cranium, CSF diagnostics and the electroencephalogram did not provide any indication for the cause of the rapidly progressive cerebral symptoms . In the NMR tomogram, a diffusedly increasing intensity in the T2-weighted tomograms were shown around the central region in the right brain . The patient died of a Pseudomonas septicemia . At autopsy, a typical finding of progressive multifocal leukoencephalopathy was found in the areas altered in the NMR tomography . Papova-like virions could be demonstrated in glial cells by electron microscopy.

J Clin Microbiol, 1987 Jun, 25(6), 1113 - 4
Recurrent Pseudomonas luteola (CDC group Ve-1) peritonitis in a patient undergoing continuous ambulatory peritoneal dialysis; Connor BJ et al.; Recurrent Pseudomonas luteola (CDC group Ve-1) peritonitis occurred in a patient undergoing continuous ambulatory peritoneal dialysis . Catheter removal was required for cure despite therapy based on antibiotic susceptibilities . This is the third report in the English literature of severe P . luteola infection and the first report of peritonitis caused by this organism.

J Med Microbiol, 1987 Jun, 23(4), 353 - 7
A competitive immunosorbent assay for detection of Pseudomonas pseudomallei exotoxin; Ismail G et al.; The development of monoclonal antibody and enzyme-linked immunosorbent assay (ELISA) techniques has made possible the detection of specific antigens at extremely low concentrations . Diagnosis of recalcitrant diseases such as melioidosis depends upon either early isolation and identification of the causative organism or the identification of a specific marker antigen, Pseudomonas pseudomallei exotoxin, in serum; the latter is better because it allows more rapid and simple diagnosis . A method of detecting exotoxin concentrations of greater than 16 ng/ml by an ELISA based on a monoclonal antitoxin is here described; it is significantly more sensitive than the mouse lethality test (lower threshold 30 micrograms/ml) currently in use and an in-vitro cytotoxicity test (lower threshold 10 micrograms/ml) that we have developed and describe here.

J Bacteriol, 1987 Jun, 169(6), 2906 - 10
Molecular cloning and biological characterization of the recA gene from Pseudomonas syringae; Hickman MJ et al.; We have identified a recombinant plasmid, pCUV8, from a cosmid library of Pseudomonas syringae genomic DNA which contains a functional analog of the Escherichia coli recA gene . The plasmid was initially identified by its ability to restore UV resistance to E . coli HB101 . Quantitative analysis demonstrated that it restored both recombination proficiency and UV resistance to an E . coli recA deletion mutant . By these criteria, pCUV8 appears to contain the P . syringae recA gene . Several pathogenic and epiphytic strains of P . syringae, but not E . coli, showed sequence homology to pCUV8 under normal stringency.

Ann Ophthalmol, 1987 Jun, 19(6), 204 - 6
Bilateral simultaneous Pseudomonas keratitis with myopic extended-wear contact lenses; Shivitz IA; Bacterial keratitis is reported more frequently in the literature with the use of extended-wear contact lenses . This report describes a case of bilateral simultaneous pseudomonas keratitis in an elderly patient wearing extended-wear contact lenses for correction of myopia . The lenses were not properly disinfected prior to the development of the corneal ulcers . When bacterial keratitis is suspected with the use of extended-wear contact lenses, prompt diagnosis and treatment are necessary . Appropriate microbiologic testing of the contact lenses, lens cases, and lens solutions should also be undertaken.

J Bacteriol, 1987 Jun, 169(6), 2440 - 8
Cloning and characterization of Saccharomyces cerevisiae genes that confer L-methionine sulfoximine and tabtoxin resistance; Marek ET et al.; Pseudomonas tabaci produces a toxin, tabtoxin, that causes wildfire disease in tobacco . The primary target of tabtoxin is presumed to be glutamine synthetase . Some effects of tabtoxin in tobacco can be mimicked by methionine sulfoximine (MSO), a compound that is known to inactivate glutamine synthetase . To understand how organisms can be made resistant to tabtoxin and MSO, we used Saccharomyces cerevisiae . We demonstrate that yeast strains carrying the glutamine synthetase gene, GLN1, on a multicopy plasmid overproduced glutamine synthetase and showed increased drug resistance . These and other data indicate that glutamine synthetase is the primary target of tabtoxin and MSO in S . cerevisiae . We also isolated three S . cerevisiae DNA inserts of 2.1, 2.3, and 2.8 kilobases that conferred tabtoxin and MSO resistance when the inserts were present on a multicopy plasmid . These plasmids conferred resistance to MSO by blocking intracellular transport of the drug . Transport appeared to occur by one or more methionine permeases . Resistance to tabtoxin could also occur by blockage of intracellular transport, but the drug was transported by some permease other than a methionine permease . These drug resistance plasmids did not block transport of citrulline, indicating that they did not affect the general amino acid permease.

Plasmid, 1987 May, 17(3), 240 - 7
Conservation of plasmids among plant-pathogenic Pseudomonas syringae isolates of diverse origins; von Bodman SB et al.; Thirty isolates of Pseudomonas syringae pv . tabaci, pv . angulata (pathogens on tobacco), pv . coronafaciens, and pv . striafaciens (pathogens on oats) were examined for plasmid DNAs . The strains were obtained from plants throughout the world, some over 50 years ago . Of the 22 tobacco pathogens, 16 contain predominantly one type of plasmid, the pJP27.00 type . The remaining six tobacco-specific strains do not harbor detectable plasmids . The oat pathogens contain one, two, or three plasmids . DNA homology studies indicate that the plasmid DNAs are highly conserved . More importantly, the plasmids harbored by strains isolated from one host plant are conserved most stringently; e.g., the plasmids from the tobacco pathogens are, with one exception, indistinguishable by restriction endonuclease digestion and Southern hybridization . There is also extensive homology among plasmids indigenous to the oat-specific P . syringae pv . coronafaciens and pv . striafaciens strains.

J Appl Bacteriol, 1987 May, 62(5), 403 - 12
Volatile compounds produced by meat pseudomonads and relate reference strains during growth on beef stored in air at chill temperatures; Edwards RA et al.; Volatile compounds produced by 31 strains of pseudomonads and by reference strains of Pseudomonas fragi and Ps . fluorescens biotype 1 during growth on beef stored at 6 degrees C in air were analysed by gas chromatography-mass spectrometry of headspace gases . Compounds of major sensory significance were ethyl and methyl esters of C2-C8 fatty acids and sulphur-containing compounds which included methane- and isopropanethiols and their related sulphides and thioesters but not hydrogen sulphide . Ester production was mainly associated with growth of some, but not all, Ps . fragi and related meat strains but sulphur-containing compounds were produced by all but a single meat strain . A minority of other meat strains produced greater amounts of methyl ketones, secondary alcohols and unsaturated hydrocarbons believed to be of lipid origin.

J Bacteriol, 1987 May, 169(5), 1993 - 6
Metabolism of aromatic compounds by Caulobacter crescentus; Chatterjee DK et al.; Cultures of Caulobacter crescentus were found to grow on a variety of aromatic compounds . Degradation of benzoate, p-hydroxybenzoate, and phenol was found to occur via beta-ketoadipate . The induction of degradative enzymes such as benzoate 1,2-dioxygenase, the ring cleavage enzyme catechol 1,2-dioxygenase, and cis, cis-muconate lactonizing enzyme appeared similar to the control mechanism present in Pseudomonas spp . Both benzoate 1,2-dioxygenase and catechol 1,2-dioxygenase had stringent specificities, as revealed by their action toward substituted benzoates and substituted catechols, respectively.

J Bacteriol, 1987 May, 169(5), 1954 - 9
Self-protection of Pseudomonas syringae pv . "tabaci" from its toxin, tabtoxinine-beta-lactam; Knight TJ et al.; An extracellular toxin, tabtoxinine-beta-lactam (T beta L), is produced by Pseudomonas syringae pv . "tabaci." This toxin irreversibly inhibits its target, glutamine synthetase; yet P . syringae pv . "tabaci" retains significant amounts of glutamine synthetase activity during toxin production in culture . As part of our investigation of the self-protection of P . syringae pv . "tabaci," we compared the effects of T beta L on Tox+ (T beta L-producing, insensitive to T beta L) and Tox- (T beta L nonproducing, sensitive to T beta L) strains . The extent of protection afforded to the Tox- strain when induced to adenylylate glutamine synthetase was tested . We concluded that an additional protection mechanism was required . A detoxification activity was found in the Tox+ strain which opens the beta-lactam ring of T beta L to produce the inactive, open-chain form, tabtoxinine . Whole cells of the Tox+ strain incubated for 24 h with {14C}T beta L (0.276 mumol/3 X 10(10) cells) contained {14C}tabtoxinine (0.056 mumol), and the medium contained T beta L (0.226 mumol) . Extracts of spheroplasts of the Tox+ stain also converted T beta L to tabtoxinine, whereas extracts of the Tox- strain did not alter T beta L . The conversion was time dependent and stoichiometric and was destroyed by boiling for 30 min or by the addition of 5 mM EDTA . Penicillin, a possible substrate and competitive inhibitor of this lactamase activity, inhibited the conversion of T beta L to tabtoxinine . Periplasmic fluid did not catalyze the conversion of T beta L.

Pathol Biol (Paris), 1987 May, 35(5), 616 - 9
{Pseudomonas cepacia septicemias: therapeutic difficulties}; Beytout J et al.; From Summer 1983 to Summer 1986, 34 cases of septicemia due to Pseudomonas cepacia could be detected in several intensive care units in the university hospital in Clermont-Ferrand (France) . Intravascular catheters can be involved in the inoculation of this bacterial agent: a previous respiratory tract infection or a drained abscess can be the portal of entry of the bacteremia . Three patients died from the septicemia and the overall prognosis of the intensive care patients looks significatively worsened . The removing of the catheters and drains, the opening of an infected collection were useful but not sufficient to overcome . The choice of a good antibiotic was not easy; only ceftazidime, minocycline and cotrimoxazole have a fair activity in vitro . We only assessed the good results of ceftazidime . Pseudomonas cepacia is also resistant for many antiseptics . The large use of disinfecting procedures in intensive care units promotes the diffusion of this bacteria.

Biochem Int, 1987 May, 14(5), 871 - 8
Physiological roles of two enzymes with fumarase activity in two Pseudomonads; Oda Y et al.; Effects of Fe2+ ions on the levels of two enzymes (fumarase and mesaconase) with fumarase activity in two Pseudomonads grown under various nutritional conditions were investigated . Fe2+ ions decreased fumarase but increased mesaconase . A high level of mesaconase was found in Ps . arvilla which was unable to metabolize itaconate . The level of mesaconase in the itaconate-grown cells of Ps . fluorescens was almost the same as that in the glucose-grown cells . This suggests that mesaconase is not an enzyme involved in the metabolism of C5-branched-chain dicarboxylates but presumably, taking the place of fumarase, plays a role in the operation of the tricarboxylic acid cycle in the cells grown in the medium containing Fe2+ ions more than 10 nmol/ml.

Antibiot Med Biotekhnol, 1987 May, 32(5), 348 - 53
{Genetic determinants of resistance to antibiotics in Pseudomonas bacteria}; Anisimova LA et al.; Seven hundred and eighty nine drug resistant strains of Pseudomonas isolated in hospitals, from antibiotic production sewage and mine soils were studied in experiments on colony hybridization with the use of 32P-labeled plasmid DNA fragments containing various antibiotic resistance genetic determinants . The genes controlling production of aminoglycoside-3'-phosphotransferase, aminoglycoside-3'-adenilyl transferase, type I dihydropteroate synthetase (DHPS I) and DHPS II were detected in the strains of all the ecological niches . More than 90 per cent of the clinical strains hybridized with the samples containing these genes, though in the soil strains the main mechanisms of streptomycin and sulfanilamides resistance were probably controlled by the other nonhomologous genetic determinants studied . The gene controlling production of aminoglycoside-3'-phosphotransferase II was detected in the strains isolated both in the hospitals (49 per cent) and from the sewage (45 per cent), while the gene of aminoglycoside-3'-phosphotransferase I was detected only in the population of the clinical strains (52 per cent) . The majority of the tetracycline resistant strains isolated in the hospitals and from the sewage hybridized with the samples containing the classes A and C tetracycline resistance genetic determinants . In the soil strains the genetic determinants of the main tetracycline resistance mechanism(s) were not homologous to those of classes A, B and C . Resistance of the strains to trimetoprine was as a rule determined by the genes of type II dihydrofolate reductase.

J Bacteriol, 1987 May, 169(5), 2207 - 14
Outer membrane protein mediating iron uptake via pyoverdinpss, the fluorescent siderophore produced by Pseudomonas syringae pv . syringae; Cody YS et al.; In an iron-limited environment Pseudomonas syringae pv . syringae B301D produces a yellow-green fluorescent siderophore called pyoverdinpss which functions in high-affinity iron transport . Two-dimensional electrophoretic comparisons of the outer membrane proteins of strain B301D identified nine proteins which were expressed at low (50 nM) but not at high (10 microM) iron concentrations . Except for the minor protein 8e, the iron-regulated proteins exhibited high molecular weights ranging from approximately 74,000 to 80,000 . A mutant of strain B301D incapable of iron uptake (Iu-) from ferric pyoverdinpss lacked the 74,000-molecular-weight protein 4a, which was the major iron-regulated outer membrane protein . In contrast, a nonfluorescent mutant (Flu-) unable to synthesize pyoverdinpss showed no quantitative or qualitative difference in its outer membrane profile from that of the wild-type strain . In plant pathogenicity tests the Iu- and Flu- strains caused typical brown necrotic and sunken lesions in immature sweet cherry fruit which were indistinguishable from those of the wild-type strain . Thus, excretion of pyoverdinpss and subsequent Fe(III) uptake do not have a determinative role in the pathogenicity or virulence of P . syringae pv . syringae.

Biochim Biophys Acta, 1987 Apr 30, 912(3), 357 - 64
Structural elements of bactopterin from Pseudomonas carboxydoflava carbon monoxide dehydrogenase; Kruger B et al.; Bactopterin is a novel pterin occurring in bacterial molybdoenzymes as the organic portion of the molybdenum cofactor . Its structure is investigated here . The compound contains a single pterin ring and carries a side chain at carbon atom 6 of the pterin nucleus as indicated by the formation of pterin-6-carboxylic acid upon alkaline permanganate oxidation . Studies with phosphate-cleaving enzymes revealed the presence of two monophosphoric acid monoesters . The affinity of reduced bactopterin for thiol-Sepharose points to the presence of thiol(s) in active bactopterin.

Biochemistry, 1987 Apr 21, 26(8), 2263 - 9
Affinity alkylation of 3-oxo-delta 5-steroid isomerase by steroidal 3 beta-oxiranes: identification of the modified amino acid by reduction with hydroxyborohydride; Bounds PL et al.; The steroidal 3 beta-oxirane (3S)-spiro{5 alpha-androstane-3,2'-oxiran}-17 beta-ol (1 beta) is an active site directed irreversible inhibitor of the 3-oxo-delta 5-steroid isomerase from Pseudomonas testosteroni . Two steroid-bound peptides (TPS1 and TPS2) were isolated by high-performance liquid chromatography (HPLC) from the trypsin digest of enzyme inactivated with 1 beta . The modified tryptic peptides (residues 14-45 of the enzyme) were further digested with chymotrypsin, each giving rise to a single steroid-containing product (CPS1 and CPS2, respectively) derived from residues 31 to 45 of the enzyme . The modified chymotryptic peptides were isolated by HPLC, and the peptide-steroid ester linkage was reduced with sodium hydroxyborohydride . Amino acid analysis of the reduced peptides gave ca . 0.5 residue of homoserine and one less residue of aspartic acid than the corresponding unreduced peptides . Sequence analysis of both reduced chymotryptic peptides revealed that homoserine was located at position 8 in the peptide sequence, corresponding to residue 38 of the enzyme . The finding that the steroidal 3 beta-oxirane, like the 17 beta-oxiranes, inactivates the isomerase via esterification of aspartic acid-38 is strong evidence that this enzyme binds steroids in at least two orientations.

Biochemistry, 1987 Apr 21, 26(8), 2270 - 9
Inhibition of 3(17)beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni by steroidal A ring fused pyrazoles; Levy MA et al.; Several 2,3- and 3,4-steroidal fused pyrazoles have been investigated as potential inhibitors of NAD(P)H-dependent steroid oxidoreductases . These compounds are proven to be potent, specific inhibitors for 3(17) beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni with Ki values of 6-100 nM . In contrast, the activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase from Streptomyces hydrogenans, steroid 5 alpha-reductase from rat prostate, and 3 alpha-hydroxysteroid dehydrogenase from rat liver were unaffected by micromolar concentrations of these compounds . Product and dead-end inhibition studies indicate an ordered association to the beta-dehydrogenase with the cofactor binding prior to substrate or inhibitor . From the results of double inhibition experiments, it is proposed that inhibition occurs through formation of an enzyme-NAD+-inhibitor ternate . On the basis of pH profiles of Vm/Km, Vm, and 1/Ki and of absorbance difference spectra, a hypothetical mechanism of inhibition by the steroidal pyrazoles, drawn by analogy from the inhibition of liver alcohol dehydrogenase by alkylpyrazoles {Theorell, H., & Yonetani, T . (1963) Biochem . Z . 338, 537-553; Andersson, P., Kvassman, J . K., Lindstrom, A., Olden, B., & Pettersson, G . (1981) Eur . J . Biochem . 113, 549-554}, is reconsidered . The pH studies and enzyme modification experiments by diethyl pyrocarbonate suggest the involvement of histidine in binding of the inhibitor . A modified proposal for the structure of the enzyme-NAD+-steroidal pyrazole complex is proposed.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1987 Apr 15, 262(11), 4994 - 9
The biosynthesis of tabtoxinine-beta-lactam . Use of specifically 13C-labeled glucose and 13C NMR spectroscopy to identify its biosynthetic precursors; Unkefer CJ et al.; Tabtoxinine-beta-lactam, an irreversible inhibitor of glutamine synthetase is produced by several pathovars of Pseudomonas syringae . We have examined tabtoxinine-beta-lactam biosynthesis, an important and poorly characterized step in pathogenesis caused by this organism . We have identified the biosynthetic precursors of tabtoxinine-beta-lactam by incorporating 13C from specifically 13C-labeled D-glucose precursors and determining the labeling pattern using 13C NMR spectroscopy . Tabtoxinine-beta-lactam is generated by combining a 4-carbon fragment, a 2-carbon fragment, and a single carbon . The 4-carbon fragment arises from aspartic acid, and the 2-carbon unit is donated from carbons 2 and 3 of pyruvate . The 6-carbon backbone of tabtoxinine-beta-lactam arises from the condensation of fragments from aspartate and pyruvate, probably using reactions analogous to the initial steps in the pathway of lysine biosynthesis.

Chest, 1987 Apr, 91(4), 527 - 32
Colonization of the respiratory tract with Pseudomonas cepacia in cystic fibrosis . Risk factors and outcomes; Tablan OC et al.; Between 1981 and 1983, some 85 patients with cystic fibrosis at Rainbow Babies and Childrens Hospital, Cleveland, developed colonization or infection of the respiratory tract with Pseudomonas cepacia . Twenty-nine (34 percent) of the colonized patients died; four were female patients with fulminant bacteremia with P cepacia prior to death . Case-control studies showed that increasing severity of underlying cystic fibrosis, increasing age, having a sibling with cystic fibrosis who was colonized with P cepacia, and previous hospitalizations were associated with increased risk of colonization . In patients with mild cystic fibrosis, no differences in clinical outcome were seen during the period of study; however, patients colonized with P cepacia who had moderate or advanced cystic fibrosis were hospitalized longer and died sooner after colonization, compared with control subjects with similar severity of cystic fibrosis . The excess mortality associated with such colonization varied in magnitude and trend according to the patient's sex and severity of underlying cystic fibrosis, reflecting the combined influence of colonization with P cepacia, sex, and severity of cystic fibrosis on the mortality of the patients . The source and mode of transmission of P cepacia were not determined, but the data suggest a possible nosocomial source . The results of this investigation showed that colonization with P cepacia most often affected patients with moderate or advanced cystic fibrosis and was associated with an adverse clinical outcome in these patients.

Can J Microbiol, 1987 Apr, 33(4), 286 - 9
Antibiotic-resistant Pseudomonas in bottled drinking water; Hernandez Duquino H et al.; Eight different bottled drinking waters were tested weekly over an 8-month period to determine the diversity of their Pseudomonas population and their sensitivity to eight antibiotics used in treating Pseudomonas infections . Nine species of Pseudomonas were recovered, with P . stutzeri (24%) and P . diminuta (18.8%) being the most common isolates . Sensitivity patterns of environmental and clinical isolates were shown to differ to some degree . Statistical analyses indicated a significant effect of specific antibiotic on the size of the inhibition zone, a significant difference between species and size of inhibition zone, and a strong species-antibiotic interaction . Distribution of species within the brands of water was also significantly different in 68% of the paired comparisons.

Appl Environ Microbiol, 1987 Apr, 53(4), 895 - 7
A new selective medium for isolating Pseudomonas spp . from water; Krueger CL et al.; A new medium, pseudomonas selective isolation agar, was developed to isolate Pseudomonas spp . from water . It consists of 350 micrograms of nitrofurantoin per ml and 2 micrograms of crystal violet per ml in a nutrient agar base . It is more selective for Pseudomonas spp . than are available commercial media . Its ingredients are inexpensive and readily available, and it is easy to prepare.

J Clin Microbiol, 1987 Apr, 25(4), 744 - 5
Pseudomonas sp . group Ve-2 bacterial peritonitis in a patient on continuous ambulatory peritoneal dialysis; Amber IJ et al.; Pseudomonas sp . group Ve-2 peritonitis occurred in a patient on continuous ambulatory peritoneal dialysis who had recently completed intraperitoneal cephalosporin therapy for culture-negative peritonitis . This is the second reported case of peritonitis in this population of patients due to this unusual organism, which is usually resistant to most cephalosporin antibiotics.

J Bacteriol, 1987 Apr, 169(4), 1441 - 6
Lipopolysaccharides of Pseudomonas spp . that stimulate plant growth: composition and use for strain identification; de Weger LA et al.; The outer membrane proteins of a series of fluorescent, root-colonizing, plant-growth-stimulating Pseudomonas spp . having been characterized (L . A . de Weger et al., J . Bacteriol . 165:585-594, 1986), the lipopolysaccharides (LPSs) of these strains were examined . The chemical composition of the LPSs of the three best-studied plant-growth-stimulating Pseudomonas strains WCS358, WCS361, and WCS374 and of P . aeruginosa PAO1 as a reference strain was determined and appeared to differ from strain to strain . The 2,6-dideoxy-2-aminosugar quinovasamine was the most abundant compound in the LPS of strain WCS358 . Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified LPS and of proteinase K-treated cell envelopes revealed ladderlike patterns for most of these strains . These patterns were not substantially influenced by differences in culture conditions . Analysis of proteinase K-treated cell envelopes of 24 root-colonizing Pseudomonas spp . revealed a unique band pattern for each strain, suggesting a great variety in the LPS structures present in these root colonizers . Therefore, electrophoretic analysis of LPS can be used for characterization and identification of the fluorescent root-colonizing Pseudomonas strains.

Bone Marrow Transplant, 1987 Apr, 1(4), 365 - 71
Treatment of marrow graft recipients with thymopentin; Witherspoon RP et al.; Four adult patients with acute non-lymphocytic leukemia were given marrow grafts from HLA-identical siblings following 120 mg/kg cyclophosphamide and 10-12 Gy total body irradiation . All received intermittent intravenous methotrexate as prophylaxis against graft-versus-disease (GVHD) . In an attempt to accelerate immune recovery and prevent GVHD, each patient received thymopentin (TP5) for 100 days after grafting . No adverse effects were seen with TP5 administration . All four patients developed acute GVHD (one grade I, one grade II, and two grade III) . Two patients died of late infections: one at 6 months from Pneumocystis carinii pneumonia and one at 11 months from disseminate Pseudomonas, Candida and cytomegalovirus infection . Two patients survive more than 3.9 years after transplantation with Karnofsky scores of 100% . One required treatment for chronic GVHD and recovered . Delayed-type hypersensitivity, antibody production to specific antigen in vivo, and results of in vitro immunologic studies were not altered by TP5 treatment . We conclude that while the administration of TP5 in these patients as described was not harmful, it did not prevent opportunistic infection, improve immunologic reconstitution or lower the incidences of acute or chronic GVHD from that of our previous experiences without thymopentin.

Ann Ophthalmol . 1987 Apr;19(4):151, 154.
Surgical therapy of a Pseudomonas corneal ulcer in a diabetic; Knopf HL; A 30-year-old white woman with diabetes was seen with a Pseudomonas corneal ulcer . The ulcer progressed despite appropriate antibiotics, and the patient was treated with surgical debridement . Pseudomonas infections in a compromised host are discussed.

Proc Natl Acad Sci U S A, 1987 Apr, 84(8), 2474 - 8
Pseudomonas exotoxin coupled to a monoclonal antibody against ovarian cancer inhibits the growth of human ovarian cancer cells in a mouse model; Willingham MC et al.; In an effort to devise an alternative treatment for human ovarian cancer, we have isolated a monoclonal antibody (OVB-3) that reacts with all ovarian cancers tested (10/10) but few normal tissues . An immunotoxin produced by coupling OVB-3 to Pseudomonas exotoxin kills ovarian cancer cells in tissue culture and prolongs the life of animals bearing human ovarian cancers . These data suggest that this immunotoxin should be evaluated as a treatment for ovarian cancer in women.

J Infect Dis, 1987 Apr, 155(4), 783 - 8
Comparative efficacy of ciprofloxacin, azlocillin, and tobramycin alone and in combination in experimental Pseudomonas sepsis; Johnson M et al.; Ciprofloxacin, azlocillin, and tobramycin, used alone and in combination, were studied in a model of lethal pseudomonas sepsis in rats . Ciprofloxacin alone at all doses studied and tobramycin alone at the highest dose reduced mortality significantly compared with saline controls . In contrast, survival was not improved with either azlocillin alone at all doses studied or lower doses of tobramycin alone . The combination of ciprofloxacin and azlocillin was synergistic in rats with systemic pseudomonas infection . Ciprofloxacin was rapidly absorbed, and effective levels persisted for 1-4 hr relative to the dose administered . These data suggest that ciprofloxacin may be a potent agent, either alone or in combination with azlocillin, in the treatment of severe pseudomonas infections.

Biochem J, 1987 Apr 1, 243(1), 15 - 21
Pseudomonas mutant strains that accumulate androstane and seco-androstane intermediates from bile acids; Leppik RA et al.; Transposon mutant strains which were affected in bile acid catabolism were isolated from four Pseudomonas spp . Two of the mutant groups isolated were found to accumulate 12 alpha-hydroxyandrosta-1,4-diene-3,17-dione as the major product from deoxycholic acid . Strains in one of these two groups were able to grow on steroids such as chenodeoxycholic acid, which lacks a 12 alpha-hydroxy function, whereas the one member of the second group could not . With chenodeoxycholic acid, this latter strain accumulated a yellow muconic-like derivative, tentatively identified as 3,7-dihydroxy-5,9,17-trioxo-4(5),9(10)-disecoandrosta-1(10)2 -dien-4-oic acid . Members of two further mutant groups accumulated either 12 beta-hydroxyandrosta-1,4-diene-3,17-dione or 3,12 beta-dihydroxy-9(10)-secoandrosta-1,3,5(10)-triene-9,17-dione as the major product from deoxycholic acid . The relationship between the catabolism of m- and p-cresol, 3-ethylphenol and the bile acids was also examined.

J Bacteriol, 1987 Apr, 169(4), 1777 - 9
IS2 activates the ilvA gene of Pseudomonas cepacia in Escherichia coli; Barsomian G et al.; The ilvA gene of Pseudomonas cepacia was expressed poorly in Escherichia coli . Insertion of IS2 upstream of the cloned gene dramatically increased its transcription, resulting in an 85-fold increase in threonine dehydratase (deaminase) specific activity . The results confirm earlier reports that IS2 promotes efficient expression of foreign genes in E . coli.

Biochim Biophys Acta, 1987 Mar 18, 912(1), 82 - 6
Amino acid sequences of the two heme c-binding sites of Pseudomonas cytochrome-c peroxidase; Ronnberg M; The amino acid sequences of the two heme c-containing tryptic peptides of Pseudomonas cytochrome-c peroxidase have been determined . The tryptic peptides were isolated from two cyanogen bromide fragments of the protein . Both heme-binding sites have the Cys-X-Y-Cys-His structure characteristic of c-type cytochromes . The sequences of the two peptides show distinct homology with each other, suggesting the occurrence of gene doubling during evolution of the protein molecule . The function of the heme c moieties in the catalytic cycle of the enzyme is discussed on the basis of their homology with the proximal histidine region of peroxidase (horseradish peroxidase and yeast cytochrome-c peroxidase) and cytochromes (horse cytochrome c and Pseudomonas cytochrome c-551).

Biochem Pharmacol, 1987 Mar 15, 36(6), 861 - 8
Enhancement by a synthetic isoprenoid of the toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin; Akiyama S et al.; A newly synthesized isoprenoid, N-solanesyl-N,N'-bis(3,4-dimethoxybenzyl)ethylenediamine, has a verapamil-like structure but no calcium channel blocking activity . The isoprenoid enhanced the cytotoxic effect of a conjugate of epidermal growth factor coupled with Pseudomonas exotoxin in human KB cells . By using iodinated epidermal growth factor ({125I}EGF), the effect of the isoprenoid on intracellular transport of EGF was examined . The isoprenoid did not affect the binding and uptake of {125I}EGF by KB cells . The release of radioactivity associated with {125I}EGF into medium was slow in the presence of the isoprenoid . Density gradient fractionation studies using cell homogenates suggest that {125I}EGF accumulates in an undegraded form in lysosomes when cells are treated with the isoprenoid . The pH value in lysosomes of KB cells was 5.5, and SDB did not affect significantly the pH value at the concentrations used to potentiate the cytotoxicity of chimeric toxins . Electron microscopy showed an increased number of electron-dense bodies in KB cells grown for 24 hr with 17-51 micrograms/ml isoprenoid . The potentiating action of chimeric toxins by the isoprenoid is discussed in relation to the altered lysosomal function in treated cells.

Arch Microbiol, 1987 Mar, 147(2), 174 - 8
Characterization of two ornithine carbamoyltransferases from Pseudomonas syringae pv . phaseolicola, the producer of phaseolotoxin; Jahn O et al.; Two ornithine carbamoyltransferases (OCT 1 and OCT 2) were isolated from Pseudomonas syringae pv . phaseolicola and purified by precipitation with ammonium sulfate, heat denaturation, chromatography on DEAE-Sephadex A-50 and Sephadex G-200 . Molecular weights of both enzymes: 110,000; optimal activity: pH 8.5 to 9.5 (OCT 1), pH 8.4 (OCT 2); apparent Km for ornithine: 7 X 10(-4) (both enzymes); apparent Km for carbamoyl-phosphate: 7 X 10(-4) (OCT 1), 2.8 X 10(-3) (OCT 2) . Both enzymes possess only an anabolic function . OCT 1 is highly inhibited by low concentrations of phaseolotoxin and Orn-P(O)(NH2)-NH-SO3H, OCT 2 is insensitive to both compounds . The inhibition of OCT 1 is reversible.

Southeast Asian J Trop Med Public Health, 1987 Mar, 18(1), 94 - 6
In-vitro susceptibility of Pseudomonas pseudomallei isolated in Malaysia to some new cephalosporins and a quinolone; Cheong YM et al.; The current drugs recommended for treatment of melioidosis are tetracycline, chloramphenicol and cotrimoxazole . Unfortunately these drugs are not the drug of choice in an acutely ill patient with septicaemia prior to the availability of laboratory results . With the discovery of the new cephalosporins which have a broad spectrum of activity clinicians are using them either alone or in combination with other antibiotics in such critical situations . Hence, an in-vitro study was carried out on the susceptibility of 41 strains of P . pseudomallei isolated in Malaysia, to these new cephalosporins and a new quinolone . The results showed that all the cephalosporins tested had some activity on the strains tested, with ceftazidime being the most active drug . Pefloxacin had very poor activity . However, further clinical studies are required to determine the duration, dosage and in-vivo activity of the antibiotics.

J Heart Transplant, 1987 Mar-Apr, 6(2), 96 - 9
Early, aggressive open lung biopsy in heart transplant recipients; Miller R et al.; Fourteen open lung biopsies were done in 13 patients out of a total of 63 heart transplant recipients . All of the patients were receiving cyclosporine and azathioprine (Imuran) and had received prophylactic antithymocyte globulin or murine antihuman mature T cell (OKT3) . Eight of the patients were receiving steroids, and five of the patients had been weaned off steroids . A fever of greater than 38.5 degrees C was present in all 13 patients . Hypoxia, defined as PO2 less than 60 on room air, was present in 11 of the 13 patients . One patient had to be intubated before surgery for respiratory failure . The chest x-ray film most often revealed bilateral interstitial infiltrates . The operative approach was by a small anterior thoracotomy in 12 patients and a posterolateral thoracotomy in two patients . A definitive diagnosis was made in 11 of the 14 open lung biopsies: three patients had Legionella, one had cytomegalovirus (CMV), one had a Pseudomonas infection, three had Pneumocystis, one patient had CMV and Pneumocystis, one patient had a pulmonary infarct, and one patient had a pulmonary infarct and CMV . Specific therapy was instituted in all these patients . In the three patients in whom no specific diagnosis was made, therapy was also changed and the use of antibiotics discontinued . There was no mortality in this group . Morbidity was minimal . No patient had excessive bleeding, and none of the patients required the transfusion of blood or blood products . Three patients required mechanical ventilatory support for longer than 24 hours.(ABSTRACT TRUNCATED AT 250 WORDS)

Pediatr Infect Dis J, 1987 Mar, 6(3), 256 - 9
False positivity of Legionella serology in patients with cystic fibrosis; Wang EE et al.; Respiratory deterioration accounts for the morbidity and mortality observed in patients with cystic fibrosis . The role of Legionella in this deterioration was determined in a 2-year prospective study of 49 patients with cystic fibrosis and 19 sibling controls . Sera were obtained from participants on enrollment and at quarterly intervals . Legionella antibodies were measured in parallel using an indirect fluorescent assay . No seroconversions were observed . Eleven of 49 patients with cystic fibrosis (22%) were seropositive compared to none of 19 siblings (P less than 0.05) . Six of the 11 patients demonstrated high titers (greater than or equal to 1:512) that persisted throughout the study . Absorption with pools of various Pseudomonas species reduced the antibody titers such that only 3 remained positive after absorption . Legionella was not found to be an important cause of clinical deterioration during this study . The results of the absorption studies suggest that high titers to Legionella in this population are due to cross-reacting antibodies.

J Laryngol Otol, 1987 Mar, 101(3), 216 - 21
The diagnostic criteria of malignant external otitis; Cohen D et al.; The diagnostic criteria of malignant external otitis (MEO) have been reviewed . They were divided into two categories: obligatory and occasional . The obligatory criteria are: pain, edema, exudate, granulations, microabscess (when operated), positive bone scan or failure of local treatment often more than 1 week, and possibly pseudomonas in culture . The occasional criteria are diabetes, cranial nerve involvement, positive radiograph, debilitating condition and old age . All of the obligatory criteria must be present in order to establish the diagnosis . The presence of occasional criteria alone does not establish it . The importance of Tc99 scan in detecting osteomyelitis is stressed . When bone scan is not available, a trial of 1-3 weeks of local treatment is suggested . Failure to respond to such treatment may assist in making the diagnosis of MEO.

J Virol, 1987 Mar, 61(3), 883 - 8
Adenovirus-dependent changes in cell membrane permeability: role of Na+, K+-ATPase; Seth P et al.; Adenovirus-dependent release of choline phosphate from KB cells at pH 6.0 was partially blocked by ouabain . In K+-containing medium, maximum inhibition of release was obtained by 10(-5) M ouabain and half-maximal inhibition was achieved by about 0.5 X 10(-6)M ouabain . Ouabain did not block either the binding or the uptake of adenovirus by KB cells . Without K+, about 25% of cell-associated choline phosphate was released by adenovirus, whereas with 1 mM K+ about 50% was released . This activation by K+ was blocked by 0.1 mM ouabain . HeLa cells behaved like KB cells, but a mutant of HeLa cells resistant to ouabain (D98-OR) released much lower amounts of choline phosphate in response to human adenovirus type 2 (Ad2) . Wild-type D98-OR cells bound nearly the same amount of adenovirus as did normal HeLa cells . Ad2 also increased the activity of Na+,K+-ATPase in KB cells, with maximum activation at 50 micrograms of Ad2 per ml . In D98-OR cells, Ad2 failed to activate Na+,K+-ATPase activity . Ad2-dependent lysis of endocytic vesicles (receptosomes) was assayed by measuring Ad2-dependent enhancement of epidermal growth factor-Pseudomonas exotoxin toxicity . This action of adenovirus was increased when K+ was present in the medium . Under the conditions used, K+ had no effect on the amount of Ad2 or epidermal growth factor taken up by the cells . On the basis of these results, it is suggested that Ad2-dependent cellular efflux of choline phosphate and adenovirus-dependent lysis of receptosomes may require Na+,K+-ATPase activity.

Biochemistry, 1987 Feb 24, 26(4), 1099 - 104
Pseudomonas cepacia 3-hydroxybenzoate 6-hydroxylase: induction, purification, and characterization; Wang LH et al.; A single strain of Pseudomonas cepacia cells was differentially induced to synthesize salicylate hydroxylase, 3-hydroxybenzoate 6-hydroxylase, or 4-hydroxybenzoate 3-hydroxylase . A procedure was developed for the purification of 3-hydroxybenzoate 6-hydroxylase to apparent homogeneity . The purified hydroxylase appears to be a monomer with a molecular weight of about 44,000 and exhibits optimal activity near pH 8 . The hydroxylase contains one FAD per enzyme molecule and utilizes NADH and NADPH with similar efficiencies . The reaction stoichiometry for this enzyme has been determined . In comparison with other aromatic flavohydroxylases, this enzyme is unique in inserting a new hydroxyl group to the substrate at a position para to an existing one.

Biochemistry, 1987 Feb 24, 26(4), 1105 - 10
Pseudomonas cepacia 3-hydroxybenzoate 6-hydroxylase: stereochemistry, isotope effects, and kinetic mechanism; Yu YM et al.; A neutral flavin semiquinone species was formed upon photoreduction of Pseudomonas cepacia 3-hydroxybenzoate 6-hydroxylase whereas no flavin radical was detected by anaerobic reduction with NADH in the presence of m-hydroxybenzoate . In the latter case, the formation of flavin semiquinone is apparently thermodynamically unfavorable . A stereospecificity for the abstraction of the 4R-position hydrogen of NADH has been demonstrated for this hydroxylase . Deuterium and tritium isotope effects were observed with (4R)-{4-2H}NADH and (4R)-{4-3H}NADH as substrates . The DV effect indicates the existence of at least one slow step after the isotope-sensitive enzyme reduction by dihydropyridine nucleotide . A minimal kinetic mechanism has been deduced on the basis of initial velocity measurements and studies on deuterium and tritium isotope effects . Following this scheme, m-hydroxybenzoate and NADH bind to the hydroxylase in a random sequence . The flavohydroxylase is reduced by NADH, and NAD+ is released . Oxygen subsequently binds to and reacts with the reduced flavohydroxylase-m-hydroxybenzoate complex . Following the formation and release of water and gentisate, the oxidized holoenzyme is regenerated . The enzyme has a small (approximately 2-fold) preference for the release of NADH over m-hydroxybenzoate from the enzyme-substrates ternary complex.

J Biol Chem, 1987 Feb 15, 262(5), 2256 - 61
Effect of low pH on the conformation of Pseudomonas exotoxin A; Farahbakhsh ZT et al.; Previously we examined factors involved in the entry mechanism of Pseudomonas exotoxin A (PTx) at the level of lipid-protein interactions (Farahbakhsh, Z . T., Baldwin, R . L., and Wisnieski, B . J . (1986) J . Biol . Chem . 261, 11404-11408) . Exposure to a low pH environment appears to be an obligatory trigger of the entry pathway . In this report we describe the effect of pH upon the conformation of PTx . We have found that the intrinsic fluorescence of PTx is strongly dependent on pH, decreasing between pH 7.4 and 4.0 with a red shift in the emission lambda max . The changes are reversible and associated with the acquisition of a binding site for the fluorescent dye 1-anilino-8-naphthalenesulfonic acid (ANS) . The fluorescence intensity of ANS in the presence of PTx increases with decreasing pH and is accompanied by a blue shift in emission spectra, indicative of exposure of hydrophobic surfaces . These changes are also reversible . Both the intrinsic fluorescence and ANS binding profiles show a dramatic dependence on pH, with the transitions centered on pH 5.0 and 4.5, respectively . Circular dichroism studies reveal a 9% decrease in alpha-helicity between pH 7.7 and 4 . The susceptibility of toxin to trypsin cleavage is also a function of pH, increasing with decreasing pH . The pH 7.4 cleavage profile is regained when the acid-treated samples are brought back to pH 7.4 . The conformational changes observed in these pH shift experiments are likely to be physiologically significant because the conditions closely resemble those that the toxin would encounter if entry into the cytoplasm of a cell involves escape from an endosomal compartment.

Arch Biochem Biophys, 1987 Feb 15, 253(1), 100 - 7
P-450 binding to substrates camphor and linalool versus pressure; Marden MC et al.; The spin equilibrium of two bacterial cytochrome P-450 enzymes are compared by their visible spectra versus temperature and pressure . P-450 from Pseudomonas linalool shows a much weaker dependence on pressure than P-450 from P . putida which has camphor as substrate . The linalool system denatures at a higher pressure (3 kbar) than the camphor system (1 kbar) and shows a weaker dependence on external solvent conditions . The camphor system shows evidence of the binding of a second substrate molecule which reverses the effect of the first on the spin equilibrium . A model involving two substrate molecules is an alternative explanation of the apparent saturation with camphor of the spin equilibrium.

Biochem Biophys Res Commun, 1987 Feb 13, 142(3), 972 - 8
Pseudomonas diminuta LPS with a new endotoxic lipid A structure; Kasai N et al.; Lipid A that contains mainly 2,3-diamino-2,3-dideoxy-D-glucose, phosphate and fatty acids in the molar ratio 2:1:5-6 was found in Pseudomonas diminuta lipopolysaccharide . The lipid A was considered to have a diamino-sugar disaccharide structure that carries a nonglycosidic phosphomonoester group and amide-bound acyloxyacyl and 3-hydroxy fatty acyl groups . The lipopolysaccharide exhibited endotoxic activities including lethal toxicity, pyrogenicity, local Shwartzman activity, body weight-decreasing toxicity and Limulus activity . The free lipid A was also endotoxic.

J Biol Chem, 1987 Feb 5, 262(4), 1510 - 8
Purification and characterization of phthalate oxygenase and phthalate oxygenase reductase from Pseudomonas cepacia; Batie CJ et al.; An enzymatic system has been isolated that catalyzes dihydroxylation of phthalate to form 1,2-dihydroxy-4,5-dicarboxy-3,5-cyclohexadiene with consumption of NADH and O2 . This system is comprised of two proteins: a flavo-iron-sulfur protein with NADH-dependent oxidoreductase activity and a nonheme iron protein with oxygenase activity . Phthalate oxygenase is a large (approximately 217 kDa) protein composed of apparently identical 48-kDa monomers . The active enzyme has one Rieske-type {2Fe-2S} center and one mononuclear iron/monomer . Removal of the mononuclear iron by incubation with EDTA or with o-phenanthroline inhibits oxygenation; ferrous ion completely restores activity . No other metals are effective . Phthalate oxygenase is specific for phthalate or other closely related compounds . However, only phthalate is tightly coupled to NADH oxidation and O2 consumption with a stoichiometry of 1:1:1 . Phthalate oxygenase is chemically competent to oxygenate phthalate when artificially supplied with reducing equivalents and O2 . Phthalate oxygenase reductase is required, however, for efficient catalytic activity . The reductase is a monomeric 34-kDa flavo-iron-sulfur protein containing FMN and a plant-ferredoxin-type {2Fe-2S} center in a 1:1 ratio . Phthalate oxygenase reductase is specific for NADH but can pass electrons to a variety of acceptors, including: phthalate oxygenase, cytochrome c, ferricyanide, and dichlorophenolindophenol . This system is similar to other bacterial oxygenase systems involved in aromatic degradation including: benzoate dioxygenase, toluene dioxygenase, benzene dioxygenase, and 4-methoxybenzoate demethoxylase . However, phthalate oxygenase can be isolated in large quantities and is more stable than most other such systems.

J Biol Chem, 1987 Feb 5, 262(4), 1608 - 13
Inactivation of pea seed glutamine synthetase by the toxin, tabtoxinine-beta-lactam; Langston-Unkefer PJ et al.; Glutamine synthetase of plants is the physiological target of tabtoxinine-beta-lactam, a toxin produced by several disease-causing pathovars of Pseudomonas syringae . This toxin, a unique amino acid, is an active site-directed, irreversible inhibitor of glutamine synthetase from pea . ATP is required for inactivation . Neither ADP, AMP, nor adenosine 5'-(beta,gamma-methylene)triphosphate (AMP-PCP) supports inactivation . Adenyl-5'-yl imidophosphate (AMP-PNP) is slowly hydrolyzed by glutamine synthetase to produce adenyl-5'-yl phosphoramidate (AMP-PN) and inorganic phosphate as identified by 31P NMR spectroscopic analysis . AMP-PNP also supports a slow inactivation of glutamine synthetase by tabtoxinine-beta-lactam . These data are consistent with gamma-phosphate transfer being involved in the inactivation . Completely inactivated glutamine synthetase has 0.9 mumol of toxin bound/mumol of subunit . One mumol of ATP is bound per mumol of subunit of glutamine synthetase in the absence of either the toxin or another active site-directed inhibitor, methionine sulfoximine; whereas, a 2nd mumol of either {alpha- or gamma-32P}ATP is bound per mumol of subunit when glutamine synthetase is incubated in the presence of either toxin or methionine sulfoximine until all enzyme activity is lost . These data suggest that the gamma-phosphate hydrolyzed from ATP during inactivation remains with the enzyme-inhibitor complex, as well as the ADP . The open chain form, tabtoxinine, was neither a reversible nor an irreversible inhibitor of glutamine synthetase, suggesting that the beta-lactam ring is necessary for inhibition . The inactivation of glutamine synthetase with tabtoxinine-beta-lactam is pseudo-first-order when done in buffer containing 15% (v/v) ethylene glycol . The rate constant for this reaction is 3 X 10(-2) S-1, and the Ki for the toxin is 1 mM . Removal of the ethylene glycol from the buffer allows the reaction to proceed in a non-first-order manner with the apparent rate constant decreasing with time . As the enzyme is inactivated in these conditions, the binding affinity for the toxin appears to decrease, while the Km observed for glutamate does not change.

Eur J Biochem, 1987 Feb 2, 162(3), 691 - 8
Nitrile hydratase of Pseudomonas chlororaphis B23 . Purification and characterization; Nagasawa T et al.; Nitrile hydratase of Pseudomonas chlororaphis B23 was completely stabilized by the addition of 22 mM n-butyric acid . The enzyme was purified from extracts of methacrylamide-induced cells of P . chlororaphis B23 in eight steps . At the last step, the enzyme was crystallized by adding ammonium sulfate . The crystallized enzyme appeared to be homogeneous from analysis by polyacrylamide gel electrophoresis, analytical ultracentrifuge, and double diffusion in agarose . The enzyme has a molecular mass of about 100 kDa and consists of four subunits identical in molecular mass (approximately 25 kDa) . The enzyme contained approximately 4 mol iron/mol enzyme . The concentrated solution of highly purified nitrile hydratase had a pronounced greyish green color and exhibited a broad absorption in visible range with a absorption maxima at 720 nm . A loss of enzyme activity occurred in parallel with the disappearance of the absorption in the visible range under a variety of conditions . The enzyme catalyzed stoichiometrically the hydration of nitrile to amide, and no formation of acid and ammonia were detected . The enzyme was active toward various aliphatic nitriles, particularly, nitriles with 3-6 carbon atoms, e.g . propionitrile, n-butyronitrile, acrylonitrile and cyclopropyl cyanide, served as the most suitable substrates.

J Clin Microbiol, 1987 Feb, 25(2), 195 - 8
Characterization of hemolysin in extracellular products of Pseudomonas cepacia; Nakazawa T et al.; Pseudomonas cepacia is recognized as an opportunistic pathogen in immunocompromised patients . We screened 120 strains of P . cepacia isolated from clinical specimens for production of extracellular products . About 70% of these strains produced lipase, protease, and lecithinase, but only 4% produced hemolysin . A hemolysin produced by P . cepacia JN106 was characterized . The hemolysin was most active against human erythrocytes . Horse, sheep, chicken, and rabbit erythrocytes were also susceptible . The hemolysin was heat labile and was inhibited by sterols but was not activated by 2-mercaptoethanol and dithiothreitol . Four hemolysin-negative mutants obtained by N-methyl-N'-nitro-N-nitrosoguanidine treatment produced the other extracellular products . A 58-kilobase-pair plasmid found in the parent strain was also found in the mutant strains, suggesting that the hemolysin gene resides on the chromosome.

Ophthalmology, 1987 Feb, 94(2), 109 - 14
Corneal ulcers associated with cosmetic extended wear soft contact lenses; Cohen EJ et al.; In 52 patients with a history of using cosmetic extended wear soft contact lenses, corneal ulcers developed requiring admission to Wills Eye Hospital between January 1, 1982, and June 30, 1986 . A retrospective study was conducted to determine the contact lens history, clinical presentation, culture and sensitivity results, and final visual acuity of these patients . Pseudomonas organisms were isolated in 75% of culture-positive cases . The final visual acuity was 6/60 or less in 12% of all cases and 25% of the Pseudomonas ulcers . Patients and practitioners must be aware of this vision-threatening problem in patients wearing cosmetic extended wear lenses . The pathogenesis of corneal ulcers in contact lens patients has not been established . There may be an inherent increased risk in the use of extended wear, compared to daily wear, soft contact lenses for cosmetic purposes.

Mol Gen Genet, 1987 Feb, 206(2), 265 - 72
Identification and mapping of regions that confer plasmid functions and of sites for excisive recombination of plasmid pMMC7105; Poplawsky AR et al.; Strain PP808 of Pseudomonas syringae pv . phaseolicola contains pEXC8080 (34.6 kb), the smallest of several plasmids that originated by partial excision of the cryptic plasmid, pMMC7105 (150 kb), from the host chromosome . This excision plasmid is derived entirely of sequences from pMMC7105 and contains a 24 kb region referred to as common DNA, which is present in each of the other excision plasmids . A six enzyme restriction endonuclease map was constructed of pEXC8080 . The replication region was mapped by identifying small restriction fragments that conferred replication properties to pMB1 plasmids that otherwise fail to replicate in Pseudomonas . This region is located within the common DNA and is 0.8-3.8 kb in size . Sequences from pEXC8080 failed to stabilize pMB1 derivatives in Pseudomonas in the absence of antibiotic selection, but stability functions were mapped to a region of pMMC7105 that presumably remains integrated in the chromosome of strain PP808 . An incompatibility region was mapped to a 7.3 kb region on pEXC8080 that is closely linked to, but not included within, the replication region . The recombination site was mapped to a 1.2 kb region of the fusion fragment that was formed upon excision of pEXC8080 . RS-I, a repetitive sequence found on pMMC7105 was present in the fusion fragment at the site of recombination . RS-I was also mapped to BamHI fragments that recombined upon excision of pEXC8080 and suggest that it provides sites for homologous recombination.

J Biochem (Tokyo), 1987 Feb, 101(2), 441 - 5
The methylamine oxidizing system of Pseudomonas AM1 reconstituted with purified components; Fukumori Y et al.; The electron transport system coupled to the oxidation of methylamine in Pseudomonas AM1 was investigated by reconstituting it from the highly purified components . A mixture of methylamine dehydrogenase, cytochrome cH and cytochrome c oxidase (= cytochrome aa3) actively oxidized methylamine (161 mol of O2 consumed/mol of heme a of cytochrome c oxidase X min) . In this system, addition of amicyanin did not affect the oxygen consumption rate . The oxygen consumption rate of the cell-free extract prepared from the cells cultivated in a copper-deficient medium was directly proportional to the amount of amicyanin added, and extrapolation to zero copper concentration gave a value of 28 mol of O2 consumed/mol of heme a of cytochrome c oxidase X min . These results suggest that methylamine oxidation in the bacterium can occur at least to some extent without participation of amicyanin.

J Bacteriol, 1987 Feb, 169(2), 572 - 8
Molecular characterization and nucleic acid sequence of an avirulence gene from race 6 of Pseudomonas syringae pv . glycinea; Napoli C et al.; A gene was previously cloned from Pseudomonas syringae pv . glycinea race 6, designated avirulence gene A (avrA), that controls the expression of virulence by the pathogen on specific cultivars of soybean . A 3.2-kilobase (kb) AccI subclone from the cosmid clone pPg6L3 was shown to be active when cloned into the broad-host-range vector pRK404 . Transposon Tn5 mutagenesis and deletion analysis delineated a span of approximately 2.5 kb of DNA that was necessary for gene activity . The nucleotide sequence of a 3.409-kb segment of DNA which contained the avrA gene has been determined . An open reading frame of 2.721 kb of DNA, which correlates with the region of DNA defined by transposon mutagenesis and deletion analysis, was identified . The open reading frame would encode a protein of 100.866 kilodaltons, which is in good agreement with the 100-kilodalton protein expressed by Escherichia coli maxicells.

J Bacteriol, 1987 Feb, 169(2), 470 - 4
Molecular cloning of copper resistance genes from Pseudomonas syringae pv . tomato; Bender CL et al.; A cosmid library of copper-resistant (Cur) Pseudomonas syringae pv . tomato PT23 plasmid DNA was constructed and mobilized into the copper-sensitive recipient P . syringae pv . syringae PS61 . One resultant cosmid clone, pCOP1 (46 kilobases), conferred copper resistance . The PT23 Cur gene(s) was located on pCOP1 by subcloning PstI restriction endonuclease fragments of pCOP1 in the broad-host-range vector pRK404 . A subclone containing a 4.4-kilobase PstI fragment conferred Cur on PS61 . The Cur gene(s) was further located by insertional inactivation with Tn5 . A subcloned fragment internal to the Cur determinant on pCOP2 was probed to plasmid and chromosomal DNA of four copper-resistant and three copper-sensitive strains of P . syringae pv . tomato . The probe hybridized to plasmids in resistant strains, but showed no detectable homology to copper-sensitive strains.

Cell, 1987 Jan 16, 48(1), 129 - 36
Functional domains of Pseudomonas exotoxin identified by deletion analysis of the gene expressed in E . coli; Hwang J et al.; Pseudomonas exotoxin A is a single chain toxin with three structural domains that inhibits protein synthesis in eukaryotic cells by catalyzing ADP ribosylation of elongation factor 2 . To study the function of these domains, we deleted different portions of the PE structural gene and expressed these constructs in E . coli using an inducible T7 promoter . These studies indicate that structural domain Ia is required for cell recognition, that structural domain II is required to translocate the toxin across a cellular membrane, and that structural domain III and a portion of domain Ib are required for ADP ribosylation activity . Toxin lacking domain Ia is about 100-fold less toxic to mice than intact PE and should be a useful molecule for the construction of immunotoxins.

Appl Environ Microbiol, 1987 Jan, 53(1), 156 - 62
Solid-state 13C nuclear magnetic resonance spectroscopy of simultaneously metabolized acetate and phenol in a soil Pseudomonas sp; Heiman AS et al.; We investigated concentration-dependent primary and secondary substrate relationships in the simultaneous metabolism of the ubiquitous pollutant phenol and the naturally occurring substrate acetate by a Pseudomonas sp . soil isolate capable of utilizing either substance as a sole source of carbon and energy . In addition to conventional analytical techniques, solid-state 13C nuclear magnetic resonance spectroscopy was used to follow the cellular distribution of {1-13C}acetate in the presence of unlabeled phenol . With 5 mM acetate as the primary substrate, Pseudomonas sp . 9S8D2 removed 1 mM phenol (secondary substrate) at a rate of 2 nmol/mg of total cell protein . Although extensive acetate metabolism was indicated by a significant redistribution of the carboxyl label, this redistribution was not affected by the presence of phenol as a secondary substrate . When the primary and secondary substrate roles were reversed, however, the presence of 1 mM phenol altered the metabolism of 0.1 mM acetate, as evidenced by both the two- to fourfold increases in carboxyl label that appeared in terminal methyl and acyl chain methylene carbon resonances and the decrease in label that occurred in the carbohydrate spectral region . These results suggest that, when phenol is present as the primary substrate, acetate is preferentially shuttled into fatty acyl chain synthesis, whereas phenol carbon is funnelled into the tricarboxylic acid cycle . Thus, simultaneous use of a xenobiotic compound and a natural substrate apparently does occur, and the relative concentrations of the two substrates do influence the rate and manner in which the compounds are utilized.(ABSTRACT TRUNCATED AT 250 WORDS)

Rev Infect Dis, 1987 Jan-Feb, 9(1), 124 - 9
Pseudomonas testosteroni infections: eighteen recent cases and a review of the literature; Barbaro DJ et al.; Pseudomonas testosteroni has been largely overlooked as a potential pathogen in humans . Ten cases of infection due to P . testosteroni were identified at a single metropolitan hospital in Texas during a three-year period . The organism was most often found in association with anatomic abnormalities of the gastrointestinal tract (six of 10 cases); perforation of the appendix was the commonest abnormality (five cases) . The infections were more often polymicrobial (seven cases) than monomicrobial (three cases) and usually involved other organisms that, like P . testosteroni, are of colonic origin . Eight additional cases of infection involving P . testosteroni were reported by other hospitals in Texas during the same period . The organism was isolated from the peritoneal cavity in five of these cases . The results of these surveys suggest that infections of humans with P . testosteroni, while not common, are not as rare as might be predicted on the basis of the number of cases reported in the literature.

J Basic Microbiol, 1987, 27(3), 173 - 6
{Demonstration of an NAD-dependent 6-phosphogluconate dehydrogenase in Pseudomonas syringae pv . phaseolicola}; Sauerstein J et al.; Crude extracts from cells of Pseudomonas syringae pv . phaseolicola, a fluorescent pseudomonad, when grown on glucose contain a NAD-linked 6-phosphogluconate dehydrogenase . The reaction of the enzyme, which produces 14CO2 from 1-14C-6-phosphogluconate, is not inhibited by NaF, a potent inhibitor of the Enter-Doudoroff (ED) pathway enzyme 6-phosphogluconate dehydratase . In the presence of phosphate or arsenate ions the NAD-linked glyceraldehyde-3-phosphate dehydrogenase reacts with glyceraldehyde-3-phosphate which, in the ED pathway, is produced from 6-phosphogluconate and overlaps the 6-phosphogluconate dehydrogenase reaction . Only a small proportion of glucose is metabolized via the 6-phosphogluconate dehydrogenase/oxidative pentose phosphate pathway.

Am J Nephrol, 1987, 7(1), 38 - 43
Successful treatment of Pseudomonas peritonitis during continuous ambulatory peritoneal dialysis; Nguyen V et al.; Successful eradication of Pseudomonas peritonitis is described in 12 (57%) of 21 cases from a large continuous ambulatory peritoneal dialysis (CAPD) program at a tertiary care center . In successful cases, cure was achieved within 17 days using therapy which included aminoglycoside started routinely at the onset of symptoms and an antipseudomonal penicillin or cephalosporin derivative added as soon as pseudomonas infection was identified on culture . Of the 9 treatment failures which required catheter removal, 2 had failure of peritoneal drainage, 4 had infection with multiple and/or drug-resistant Pseudomonas strains, and 3 had persistent catheter tunnel infection which resulted in recurrent Pseudomonas peritonitis . Factors such as diabetes mellitus and pediatric age group did not prevent successful medical therapy . Predisposing factors favoring development of Pseudomonas peritonitis included technical failures and in a few cases recent antibiotic therapy . We conclude that Pseudomonas peritonitis complicating CAPD can be successfully cured without catheter removal or discontinuation of CAPD in many cases, particularly when complicating factors are not present.

Toxicon, 1987, 25(2), 225 - 8
Production of tetrodotoxin and its derivatives by Pseudomonas sp . isolated from the skin of a pufferfish; Yotsu M et al.; Bacteria isolated from the skin of the pufferfish Fugu poecilonotus were screened for tetrodotoxin production . Tetrodotoxin and its derivatives were detected by HPLC analyses in broth cultures of a Pseudomonas sp . Upon alkali treatment the compounds yielded, as do tetrodotoxin and its derivatives, 2-amino-6-hydroxymethyl-8-hydroxyquinazoline, the identity of which was confirmed, after methylation or trimethylsilylation, by HPLC, mass spectrometry and gas chromatography - mass spectrometry.

Circ Shock, 1987, 21(3), 175 - 83
Ibuprofen and methylprednisolone in a pig Pseudomonas ARDS model; Harvey CF et al.; The effects of ibuprofen (I) and methylprednisolone (M) were studied in the Pseudomonas porcine model of adult respiratory distress syndrome (ARDS) . Four groups of animals were anesthetized and ventilated with 0.5 FIO2, 5 cm PEEP, and 20 cc/kg tidal volume: a control group given saline alone; Pseudomonas infusion alone (P); Pseudomonas with ibuprofen (I), 12.5 mg/kg given at 20 and 120 min; and P with methylprednisolone (M), 30 mg/kg given at 20 and 120 min . We compared the alteration in pulmonary hemodynamics with the alteration in the plasma concentration of thromboxane (TxB2) and 6-keto PGF1 alpha in the treated and untreated groups . Hemodynamic parameters measured included the pulmonary (PAP) and systemic (SAP) arterial pressures, cardiac index (C1), thermal-cardiogreen extravascular lung water (EVLW), and PaO2 . Albumin Flux was measured by a gamma scintigraphic method (slope index; SI) . P produced a dramatic increase in PAP (P less than 0.05) with a progressive increase in EVLW and SI (P less than 0.05) and fall (P less than 0.05) in PaO2, CI, and SAP . The acute pulmonary hypertension was associated with a significant rise in TxB2 and 6-keto PGF1 alpha in the Pseudomonas group . I effectively blocked the elevation of TxB2 and caused a significant but transient improvement in PAP and rise in PaO2 . Albumin flux and water leak as measured by SI and EVLW were not affected by ibuprofen . M reduced the elevation of TxB2 and 6-keto PGF1 alpha but failed to block these prostaglandins significantly.(ABSTRACT TRUNCATED AT 250 WORDS)

Pediatrics, 1987 Jan, 79(1), 138 - 46
Experience with heart transplantation in children; Fricker FJ et al.; Between March 1981 and March 1986, 200 orthotopic heart transplantations were performed at the University of Pittsburgh . Fourteen of those procedures were carried out in children 2 to 16 years of age . Two children received combined liver and heart transplants; one because of familial hypercholesterolemia with associated ischemic heart disease, and the other because of dilated cardiomyopathy associated with intrahepatic biliary atresia . Eight patients had dilated cardiomyopathy, and two had myocarditis . Two had heart transplantations for congenital heart disease: one had multiple muscular ventricular septal defects repaired in infancy and had an associated cardiomyopathy, and the other developed a cardiomyopathic ventricle from a congenital right coronary artery to right atrial fistula . Chronic immune suppression consisted 0.2 to 0.5 mg/kg/d of prednisone and 5 to 50 mg/kg/d cyclosporine, with the addition of antithymocyte globulin for unresolved moderate or severe acute rejection . There were three early postoperative deaths: one from intracranial bleeding, one from Pseudomonas mediastinitis, and one from ischemic injury to transplanted organs . Early postoperative complications included reversible renal failure, hypertension, and seizures . Late problems were related to allograft rejection and side effects of cyclosporine and corticosteroids . Significant rejection episodes occurred in all patients surviving longer than 2 weeks, with seven requiring antithymocyte globulin . Two patients died 8 months following transplantation of severe acute and chronic rejection; another patient required retransplantation for ischemic cardiomyopathy resulting from chronic rejection but subsequently died of recurring rejection 3 months after the second transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

J Cardiol Suppl, 1987, 14, 21 - 36
{Experimental production of mitral valve and papillary muscle lesions by cervical vagus stimulation in rabbits}; Imataka K et al.; Mitral valve and papillary muscle lesions were experimentally produced in rabbits by cervical vagal stimulation or compression of the abdominal aorta . The involvement induced the similar findings to mitral valve prolapse in human beings including mid-systolic clicks and a late systolic or holosystolic murmur . Extrasystoles were also common . Necropsies disclosed papillary muscle swelling, bleeding and edematous change in the mitral valve, swelling of the myocardial cells and interstitial fibrosis . Further, bacterial endocarditis was induced in such rabbits by injecting pseudomonas alkaligenes . Possible connection of experimentally-induced lesions of the mitral complex and mitral valve prolapse in human beings was discussed.

Trans R Soc Trop Med Hyg, 1987, 81(6), 1017 - 9
Melioidosis: a serological survey in a tuberculosis sanatorium in Hong Kong; So SY et al.; A serological survey of 275 Chinese patients with underlying pulmonary diseases in a tuberculosis sanatorium in Hong Kong showed that 39 (14%) had haemagglutinating antibody (HA) against Pseudomonas pseudomallei in a titre of 1: 80 or above . Only 9 of these 39 patients had travelled to endemic areas, suggesting that at least 30 patients (11%) had been exposed to Ps . pseudomallei locally . Females are affected as often as males, and the seropositive rate is the same whether patients are immunosuppressed or not . Because subclinical melioidosis is prevalent and HA may persist for a long time, even at a high titre, after infection, determination of HA alone cannot differentiate between active melioidosis and its masquerade--active tuberculosis.

Gene, 1987, 58(2-3), 257 - 64
Cloning of the gene for delta 5-3-ketosteroid isomerase from Pseudomonas testosteroni; Choi KY et al.; We have cloned an approx . 5-kb fragment of Pseudomonas testosteroni DNA containing the structural gene of delta 5-3-ketosteroid isomerase into the EcoRI site of the lambda gt11 genome . Escherichia coli infected with these recombinant phages produce a polypeptide which is recognized by antiserum raised against the purified isomerase . Four of the recombinant lambda gt11 clones contain significant levels of isomerase activity and produce an immunopositive polypeptide of the same apparent Mr as the native isomerase obtained from P . testosteroni . The approx . 5-kb fragment hybridizes to synthetic 21-mer and 17-mer oligodeoxynucleotide mixtures corresponding to the 5' and 3' regions, respectively, of the expected nucleotide sequence of the gene.

Methods Enzymol, 1987, 151, 139 - 45
Construction of immunotoxins using Pseudomonas exotoxin A; FitzGerald DJ; The above methodology has been used to prepare a variety of PE-immunotoxins . These reagents are potent cytotoxic agents for cells in culture that bind the appropriate antibody and nontoxic when cells do not bind the antibody . PE-immunotoxins can be used to select for mutant cultured cells which lack the target of the antibody (see chapter by Gottesman {9} on drug-resistant mutants) . Recently, we have also demonstrated in vivo activity when a PE-immunotoxin was used to inhibit the growth of human ovarian cancer cells in a nude mouse model of ovarian cancer . Also PE-immunotoxins are currently being evaluated in Phase 1 clinical trials for the treatment of adult T-cell leukemia.

Infection, 1987, 15 Suppl 2, S64 - 6
{Experiences with a Pseudomonas immunoglobulin in ventilated patients with Pseudomonas pneumonia in a surgical intensive care station}; Bohm D; In a clinical trial, the efficacy of a Pseudomonas immunoglobulin was studied in ten ventilated patients suffering from Pseudomonas pneumonia . Compared to ten patients of a previous study who had received a polyvalent immunoglobulin (control group), patients treated with Pseudomonas immunoglobulin fared better with respect to clinical success and duration of treatment, the period of antibiotic treatment being significantly shorter than in the control group.

Infection, 1987 Jan-Feb, 15(1), 73 - 5
{Experiences with a Pseudomonas immunoglobulin in artificially respirated patients with Pseudomonas pneumonia at a surgical intensive care unit}; Bohm D; In a clinical trial, the efficacy of a Pseudomonas immunoglobulin was studied in ten ventilated patients suffering from Pseudomonas pneumonia . Compared to ten patients of a previous study who had received a polyvalent immunoglobulin (control group), patients treated with Pseudomonas immunoglobulin fared better with respect to clinical success and duration of treatment, the period of antibiotic treatment being significantly shorter than in the control group.

J Bacteriol, 1987 Jan, 169(1), 8 - 13
Identification of transposable elements which activate gene expression in Pseudomonas cepacia; Scordilis GE et al.; This study demonstrated that transposable elements in Pseudomonas cepacia could be inserted upstream of a poorly expressed gene and increase its expression more than 30-fold . Five elements, TnPc1, IS402, IS403, IS404, and IS405, were isolated by their ability to increase expression of the beta-lactamase gene of the broad-host-range plasmid pRP1 . Increased expression resulted only from insertion of these elements, suggesting that insertional activation is an important means of elevating gene expression in this organism . Four of the elements inserted between a PstI site within the beta-lactamase gene and a BamHI site located 375 base pairs upstream of its promoter . The element IS403 inserted distal to the BamHI site within the coding region for the gene tnpR, suggesting that insertional activation can act over greater than expected distances . In addition, the element IS402 activated the beta-lactamase genes carried on plasmids pRP1 and pMR5 (temperature-sensitive pRP1) equally well in opposite orientations, demonstrating that insertional activation by this element occurs independent of its orientation.

J Bacteriol, 1987 Jan, 169(1), 394 - 402
Molecular cloning and expression of the 3-chlorobenzoate-degrading genes from Pseudomonas sp . strain B13; Weisshaar MP et al.; The genes specifying the utilization of 3-chlorobenzoate by Pseudomonas sp . strain B13 WR1 have been cloned by using a broad-host-range cosmid cloning system . Analysis of the catabolic products of the enzymatic reactions encoded by two hybrid cosmids, pMW65 and pMW90, by thin-layer and high-performance liquid chromatography demonstrated that both encoded the genes for the complete catabolism of 3-chlorobenzoate . Physical analysis of one of the cosmid derivatives, pMW65, by restriction endonuclease mapping and subcloning demonstrated that the pathway genes are encoded on a fragment no larger than 11 kilobases.

J Bacteriol, 1987 Jan, 169(1), 224 - 30
Insertion-sequence-dependent rearrangements of Pseudomonas cepacia plasmid pTGL1; Gaffney TD et al.; Pseudomonas cepacia 249 (ATCC 17616) harbors a 170-kilobase (kb) plasmid designated pTGL1 . We identified three insertion sequences, IS405, IS408, and IS411, on this plasmid . Various prototrophic and auxotrophic derivatives in our collection contained variants of pTGL1 formed by accretion and deletion of other elements . Plasmid pTGL6, the variant in one prototroph, evolved from pTGL1 by the addition of three copies of IS401 (1.3 kb) and one of IS402 (1 kb), to generate pTGL5, and recombination between two of the copies of IS401 on pTGL5 to form pTGL6 . The latter event entailed loss of one copy of IS401 and an additional 5.4 kb of plasmid DNA . Derivatives of the broad-host-range plasmid pRP1 carrying the above insertion sequences and recombinant plasmids carrying fragments of plasmids pTGL6 and pTGL5 were used as probes to ascertain the extent of reiteration of the various elements in the P . cepacia genome . The data indicate a high frequency of genomic rearrangements which presumably contributes to the extraordinary adaptability of this bacterium.

J Biol Chem, 1986 Dec 25, 261(36), 16785 - 7
Direct measurement of poly(beta-hydroxybutyrate) in a pseudomonad by solid-state 13C NMR; Jacob GS et al.; Four narrow lines are observed in the high-resolution cross-polarization magic-angle spinning 13C NMR spectra of intact, lyophilized samples of Pseudomonas sp . LBr, in addition to the broader lines normally associated with bacterial cellular material . These narrow lines arise from poly(beta-hydroxybutyrate) . The cellular carbon contained in this storage material can be measured quantitatively and nondestructively from the solid-state NMR spectra . We find that cells starved for phosphorus store up to 50% of their total carbon as poly(beta-hydroxybutyrate) . When such cells are used to inoculate medium containing a source of phosphorus, all of the poly(beta-hydroxybutyrate) is metabolized by the time the culture has reached midlogarithmic growth phase.

FEBS Lett, 1986 Dec 15, 209(2), 321 - 4
Detection of a new chloroperoxidase in Pseudomonas pyrrocinia; Wiesner W et al.; A new chloroperoxidase could be detected in Pseudomonas pyrrocinia ATCC 15,958, a bacterium that produces the antifungal antibiotic pyrrolnitrin . This enzyme was separated from a ferriprotoporphyrin IX containing bromoperoxidase which was also produced by this bacterium . The enzyme is capable of catalyzing the chorination of indole to 7-chloroindole . This procaryotic chloroperoxidase requires the presence of H2O2 and can also brominate monochlorodimedone, but cannot catalyze its chlorination . This enzyme is the first chloroperoxidase described from procaryotic sources.

JAMA, 1986 Dec 5, 256(21), 2991 - 5
The total artificial heart as a bridge to transplantation . A report of two cases; Copeland JG et al.; In 1985, at the University of Arizona, Tucson, two attempts were made to "bridge" patients from impending death to heart transplantation, using orthotopically positioned total artificial hearts . The first attempt, using an unapproved device on an emergency basis, failed after transplantation because of severe pulmonary edema and Pseudomonas pneumonia and the apparent transmission of a Pseudomonas infection from donor to recipient . The second experience, using a Jarvik-7 device, led to stable support for nine days with one major complication, a reversible neurologic deficit with no associated computed tomographic scan abnormality . This patient survived cardiac transplantation and, after being successfully treated for complications, has made a full recovery and returned to full-time work.

Asia Oceania J Obstet Gynaecol, 1986 Dec, 12(4), 489 - 92
Health implications of traditional female circumcision in pregnancy; Adetoro OO et al.; PIP: A case of traditional female circumcision during pregnancy, as practiced by her ethnic group, the Igbomina-Ekiti of Kawra State, with loss of the fetus as a result of infection, is presented . The woman was circumcised at age 20 at approximately 34 weeks' gestation . She had bled profusely during the procedure and was treated locally with herbs and snail juice . She had 5 days of pain and purulent bloody discharge . On hospital admission the patient was febrile and anemic, her vulva was hemorrhagic and edematous with partially excised clitoris and labia minora . Fetal heart sounds were present . She was given 2 units of blood, anti-tetanus toxoid, and prophylactic antibiotics . 2 days later the infecting organisms and their antibiotic sensitivity were identified, pseudomonas pyocyanea and Staph . Aureus, sensitive to erythromycin and gentamycin . Her fever abated, but she developed pre-eclampsia and she went into labor spontaneously . At 3 cm dilation, labor failed to progress despite artificial rupture of the membranes . A fresh stillborn female preterm infant was delivered by cesarean section . It was felt that the fetus died because of the infection . In Nigeria, female circumcision may be done in infancy by the Yorubas in the Western States, at puberty by the Igbos in Abakaliki, before marriage by the Isoko in Bendel States and the Hausas in the North, and during the 1st pregnancy by the Ogbaru in Anambra State and the Igbomina-Ekiti in Kawra State .

Eur J Clin Microbiol, 1986 Dec, 5(6), 655 - 6
Septic melioidosis after a visit to Southeast Asia; Bouvy JJ et al.; A case is reported of a fifty-seven year old man with fever, who was admitted to hospital after a recent visit to Southeast Asia . Among the clinical findings prostatitis and broncho-pneumonia were noted . Within twenty-four hours irreversible fulminant sepsis developed although he was treated with cefotaxime, tobramycin and erythromycin . Post mortem Pseudomonas pseudomallei was cultured from blood and aspirate collected by bronchoscopy . It is important to consider melioidosis as a cause of septic illness in patients who have been visiting Southeast Asia.

J Bioenerg Biomembr, 1986 Dec, 18(6), 461 - 70
Effect of oxygen limitation on the formation of the electron transport system of the phytopathogenic fluorescent bacterium Pseudomonas cichorii; Zannoni D; The composition of the membrane-bound electron transport system of the phytopathogenic bacterium Pseudomonas cichorii underwent modification in response to oxygen supply . Growth adaptation to low oxygen concentrations was characterized by repression of cytochromes involved in ubiquinol-cyt . c oxidoreductase and cyt . c oxidase activities . By contrast, cyto . o, i.e., the alternative cyanide-insensitive oxidase of P . cichorii, was unaffected by low oxygen tension . No a-type cytochromes could be detected at any stage of growth.

J Cell Biol, 1986 Dec, 103(6 Pt 1), 2283 - 97
Chinese hamster ovary cell mutants with temperature-sensitive defects in endocytosis . I . Loss of function on shifting to the nonpermissive temperature; Roff CF et al.; We have isolated three independent Chinese hamster ovary cell mutants (B3853, I223, and M311) with temperature-sensitive, pleiotropic defects in receptor-mediated endocytosis . Activities affected at 41 degrees C include uptake via the D-mannose 6-phosphate receptor, accumulation of Fe from diferric transferrin, uptake of alpha 2-macroglobulin, compartmentalization of newly synthesized acid hydrolases, resistance to ricin, and sensitivity to diphtheria and Pseudomonas toxins and modeccin . The three mutants also displayed decreased sialylation of some secreted glycoproteins at 41 degrees C, reminiscent of the nonconditional mutant DTG1-5-4 that showed both endocytic and Golgi-associated defects (Robbins, A.R., C . Oliver, J.L . Bateman, S.S . Krag, C.J . Galloway, and I . Mellman, 1984, J . Cell Biol., 99:1296-1308) . Phenotypic changes were detectable within 30 min after transfer of the mutants to 41 degrees C; maximal alteration of most susceptible functions was obtained 4 h after temperature shift . At 39 degrees C, the mutants exhibited many but not all of the changes manifested at 41 degrees C; resistance to diphtheria and Pseudomonas toxins required the higher temperature . Analysis of cell hybrids showed that B3853 and DTG1-5-4 are in one complementation group ("End1"); M311 and I223 are in another ("End2") . In the End1 mutants, loss of endocytosis correlated with complete loss of ATP-dependent endosomal acidification in vitro; in the End 2 mutants partial loss of acidification was observed . At the nonpermissive temperature, residual levels of endocytic activity in B3853 and M311 were nearly identical; thus, we conclude that the differences measured in endosomal acidification in vitro reflect the different genetic loci affected, rather than the relative severity of the genetic lesions . The mutations in M311 and I223 appear to have different effects on the same protein; in I223 (but not in M311) the full spectrum of phenotypic changes could be produced at the permissive temperature by inhibition of protein synthesis.

Mol Gen Mikrobiol Virusol, 1986 Dec, (12), 34 - 6
{Repression of the synthesis and allosteric inhibition of 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase in facultative methylotrophic bacterium Pseudomonas sp . M}; Olekhnovich IN et al.; The synthesis of 3-deoxy-D-arabinoheptulosonate-7-phosphate-synthase has been shown to be repressed by tyrosine and phenylalanine in the cells of facultative methylotrophic bacteria Pseudomonas sp . M . Activity of the enzyme is subjected to allosteric inhibition by tyrosine, tryptophane, anthranylate and phenylpyruvate.

Eur J Biochem, 1986 Nov 17, 161(1), 139 - 47
Mucopolysaccharidases from Pseudomonas sp . Isolation and partial characterization of constitutive enzymes involved in the degradation of keratan sulfate and chondroitin sulfate; Horton DS et al.; Four constitutive enzymes, capable of degrading keratan sulfate, were isolated from Pseudomonas sp.: a particulate endoglycosidase, a soluble endoglycosidase, a soluble exo-beta-D-galactosidase and a soluble exo-beta-D-N-acetylglucosaminidase . The endoglycosidases were shown to act only upon keratan sulfate forming beta-D-2-acetamido-2-deoxy-6-O-sulfoglucosyl-(1----3)-D-galactose, as the main product . This results indicates that the enzyme catalyses the hydrolysis of beta-D-galactose-(1----4)-N-acetylglucosamine linkages . It was also shown that this monosulfated disaccharide inhibits the particulate keratan sulfate endoglycosidase . The bovine nucleus pulposus keratan sulfate is depolymerized at a lower rate and extent when compared to the corneal keratan sulfate . The soluble endoglycosidase is very labile, in contrast to the particulate enzyme, which has been stored at -20 degrees C or at 4 degrees C for at least 12 months with no loss in activity . The particulate endoglycosidase and the soluble exo-beta-D-galactosidase and exo-beta-D-N-acetylglucosaminidase are induced when the bacteria is grown in adaptative media containing either 0.1% keratan sulfate or 0.1% chondroitin sulfate . Furthermore, particulate forms of the exoenzymes were detected . The soluble endoglycosidase specific activity, in contrast, is approximately the same in extracts of cells grown in glucose, keratan sulfate or chondroitin sulfate . A chondroitin sulfate lyase was also identified in the soluble extracts of Pseudomonas sp . cells . This enzyme depolymerizes chondroitin 4-sulfate, chondroitin 6-sulfate and hyaluronic acid forming unsaturated disaccharides as main products . It is also active upon the glucuronic-acid-containing regions of the dermatan sulfate molecules . The properties of the soluble enzymes, further purified by ion-exchange chromatography, and of the particulate keratan sulfate endoglycosidase are presented.

Carbohydr Res, 1986 Nov 15, 156, 165 - 72
Structural studies of a polysaccharide (S-88) elaborated by Pseudomonas ATCC 31554; Jansson PE et al.; The structure of the extracellular polysaccharide elaborated by Pseudomonas ATCC 31554 has been investigated, methylation analyses, specific degradations, and 1H-n.m.r . spectroscopy being the main methods used . It is concluded that the polysaccharide is composed of pentasaccharide repeating-units with the structure: ----3)-beta-D-Glcp-(1----4)-beta-D-GlcpA-(1----4)-beta-D-Glcp- (1----4)-alpha-L-{Rha or Man}-(1---- 3 increases 1 alpha-L-Rha . An unusual feature is that a sugar residue in the chain may be either L-rhamnose or L-mannose . The polysaccharide also contains O-acetyl groups (approximately 5%) which have not been located.

J Biol Chem, 1986 Nov 15, 261(32), 15112 - 4
Enzymes of vitamin B6 degradation . Purification and properties of 4- and 5-pyridoxolactonases; Jong YJ et al.; 4-Pyridoxolactone and 5-pyridoxolactone, formed by dehydrogenation of pyridoxal or isopyridoxal during the bacterial degradation of vitamin B6 by Pseudomonas MA-1 and Arthrobacter Cr-7, respectively, are hydrolyzed to the corresponding acids by distinct inducible lactonases which were purified to homogeneity . 4-Pyridoxolactonase from Pseudomonas MA-1 has an Mr of 54,000 and contains two probably identical subunits of Mr = 28,600 . It has a pH optimum of 7.0, a Km of 5.9 microM, and a Vmax at 25 degrees C of 35.2 mumol X min-1 X mg-1 . 5-Pyridoxolactonase from Arthrobacter Cr-7 has an Mr of 65,200 and also contains two probably identical subunits of Mr = 32,800 . It has a pH optimum of 7.1-7.7, a Km of 300 microM, and a Vmax at 25 degrees C of 21.5 mumol-1 X min-1 X mg-1 . The two lactonases require no added cofactors or metal ions; their activities are inhibited by sulfhydryl reagents but are not affected by metal-chelating reagents . Although the two lactonases are entirely specific for their respective substrates, 4-pyridoxolactone is a competitive inhibitor (KI = 52 microM) for 5-pyridoxolactonase, and 5-pyridoxolactone is a competitive inhibitor (KI = 48 microM) for 4-pyridoxolactonase.

J Biol Chem, 1986 Nov 15, 261(32), 15106 - 11
Enzymes of vitamin B6 degradation . Purification and properties of isopyridoxal dehydrogenase and 5-formyl-3-hydroxy-2-methylpyridine-4-carboxylic-acid dehydrogenase; Lee YC et al.; Two NAD+-dependent, highly specific pyridine-5-aldehyde dehydrogenases, 5-formyl-3-hydroxy-2-methylpyridine-4-carboxylic-acid (Compound 1) dehydrogenase and isopyridoxal dehydrogenase, were purified to homogeneity from Pseudomonas MA-1 and Arthrobacter Cr-7, respectively . Both enzymes are induced in response to growth of the organisms on pyridoxine and catalyze steps in the degradation of this compound by these organisms . Compound 1 dehydrogenase (Mr = 65,000) contains two subunits of equal size with methionine as the NH2-terminal amino acid and acts optimally at pH 7.8-8.5 . It catalyzes with equal facility (turnover number = 400-670 s-1 molecule-1) both the oxidation of Compound 1 (Km = 65 microM) by NAD+ (Km = 25 microM) to 3-hydroxy-2-methylpyridine-4,5-dicarboxylic acid and the reduction of Compound 1 by NADH (Km = 20 microM) to 4-pyridoxic acid and appears to act as a true dismutase . The possible advantage to the organism of its ability to act as a dismutase is discussed briefly . No oxidation of 4-pyridoxic acid by this enzyme was observed . Isopyridoxal dehydrogenase (Mr = 242,000) contains four subunits of equal size, again with methionine at the NH2 terminus . At its optimal pH of 8.0-8.6, it catalyzes the oxidation of isopyridoxal (Km = 40 microM, turnover number = 10 s-1 molecule-1) by NAD+ (Km = 40 microM) to a mixture of 5-pyridoxic acid and 5-pyridoxolactone, which are produced in constant ratio throughout the course of the reaction . Formation of the two products, although unusual, is readily understandable in terms of the structure of isopyridoxal in solution or the structure of a possible acyl-enzyme intermediate in the oxidative reaction.

Biochim Biophys Acta, 1986 Nov 7, 874(1), 23 - 9
Inactivation of Pseudomonas iron-superoxide dismutase by hydrogen peroxide; Yamakura F et al.; Pseudomonas Fe-superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) is inactivated by hydrogen peroxide by a mechanism which exhibits saturation kinetics . The pseudo-first-order rate constant of the inactivation increased with increasing pH, with an inflection point around pH 8.5 . Two parameters of the inactivation were measured in the pH range 7.8 to 9.0; the total H2O2 concentration at which the enzyme is half-saturated (K inact) was found to be independent of pH (30 mM) and the maximum rate constant for inactivation (k max) increased progressively with increasing pH, from 3.3 min-1 at pH 7.8 to 21 min-1 at pH 9.0 . This evidence suggests the presence of an ionization group (pKa approximately 8.5) which does not participate in the binding of H2O2 but which affects the maximum inactivation rate of the enzyme . The loss of dismutase activity of the Fe-superoxide dismutase is accompanied by a modification of 1.6, 1.1 and 0.9 residues of tryptophan, histidine and cysteine, respectively . Since the amino acid residues of the Cr-substituted enzyme, which has no enzymatic activity, were not modified by H2O2, the active iron of the enzyme is essential for the modification of the amino acid residues.

Br J Cancer, 1986 Nov, 54(5), 733 - 41
Influence of reduced concentration of L-glutamine on growth and viability of cells in monolayer, in spheroids, and in experimental tumours; Tannock IF et al.; L-Glutamine is a requirement for many cells in tissue culture, an intermediate in many metabolic pathways, and an alternative substrate to glucose for energy metabolism . These properties suggest that glutamine concentration might be a determinant of cell viability in tumours, especially in regions that are deficient in other metabolites . We have therefore studied the effects of glutamine depletion on single cells in culture, on spheroids and on experimental tumours . Absence of glutamine suppressed the growth rate of two cell lines, but cells cultured for up to 6 h in the absence of glutamine had no decrease in plating efficiency . There was little effect on growth of MGH-U1 (human bladder cancer) spheroids of varying the glutamine concentration in the range of 0.1 to 2 mM and spheroids exposed to these concentrations did not develop central necrosis . Lower concentration of glutamine suppressed the rate of spheroid growth, and spheroids did not grow in the absence of glutamine . Pseudomonas 7A glutaminase reduced the survival of cells in glutamine-free culture and prevented growth of spheroids . Glutaminase was injected into mice bearing experimental tumours to reduce blood levels of glutamine; some animals also received 15 Gy radiation to their tumours to assess the effects of glutamine levels on surviving nutrient-deprived (i.e . hypoxic) cells . Glutaminase had no effect on cell survival in the Lewis lung tumour or in MGH-U1 xenografts, with or without radiation; glutaminase caused dose-dependent growth delay of the KHT tumour, which was additive to that caused by radiation . The present results suggest that (i) short-term changes of glutamine concentration have small effects on cell viability; and (ii) depletion of glutamine levels in blood through the in vivo use of glutaminase is unlikely to produce major therapeutic effects against nutrient-deprived cells in solid tumours.

Appl Environ Microbiol, 1986 Nov, 52(5), 1195 - 202
Bacterial communities degrading amino- and hydroxynaphthalene-2-sulfonates; Nortemann B et al.; A 6-aminonaphthalene-2-sulfonic acid (6A2NS)-degrading mixed bacterial community was isolated from a sample of river Elbe water . The complete degradation of this xenobiotic compound may be described by a mutualistic interaction of two Pseudomonas strains isolated from this culture . One strain, BN6, could also grow on 6A2NS in monoculture, however, with accumulation of black polymers . This organism effected the initial conversion of 6A2NS into 5-aminosalicylate (5AS) through regioselective attack of the naphthalene skeleton in the 1,2-position . 5AS was totally degraded by another member of the community, strain BN9 . After prolonged adaptation of strain BN6 to growth on 6A2NS, this organism readily converted all naphthalene-2-sulfonates with OH- or NH2-substituents in the 5-, 6-, 7-, or 8-position . The corresponding hydroxy- or aminosalicylates were excreted in stoichiometric amounts, with the exception that the metabolite from 5A2NS oxidation was not identical with 6AS.

J Bacteriol, 1986 Nov, 168(2), 1019 - 22
Comparative study of refractile (R) bodies and their genetic determinants: relationship of type 51 R bodies to R bodies produced by Pseudomonas taeniospiralis; Kanabrocki JA et al.; The relationship of type 51 refractile (R) bodies to R bodies produced by Pseudomonas taeniospiralis was investigated . Proteins associated with type 51 R bodies were not serologically cross-reactive with proteins associated with R bodies from P . taeniospiralis . The genetic determinants for type 51 R bodies did not exhibit close homology with DNA sequences from P . taeniospiralis.

J Clin Microbiol, 1986 Nov, 24(5), 853 - 5
Community-acquired bloodstream infection caused by Pseudomonas paucimobilis: case report and review of the literature; Morrison AJ Jr et al.; Various sources of Pseudomonas paucimobilis bacterial infections have been documented . We report the third human case of bloodstream infection due to P . paucimobilis and review the literature in English regarding community-acquired and nosocomial infection due to this bacterium . Biochemical and genetic characteristics supporting the pathogenic potential of P . paucimobilis are presented, and the antibiotic susceptibility profile of the organism is summarized.

Pediatr Res, 1986 Nov, 20(11), 1174 - 7
Decreased baseline beta-lactamase production and inducibility associated with increased piperacillin susceptibility of Pseudomonas cepacia isolated from children with cystic fibrosis; Chiesa C et al.; The incidence of pulmonary infections in children with cystic fibrosis caused by Pseudomonas cepacia, an organism which may possess an inducible beta-lactamase, has increased since 1978 . Seven of 13 sputum isolates of P . cepacia from children with cystic fibrosis were classified as inducible by quantitative enzyme production following preincubation with 100, 200, or 400 micrograms/ml of cefoxitin . The recovery of inducible strains tended to be associated with recent ceftazidime therapy . Susceptibility to aztreonam, ceftazidime, and piperacillin alone or combined with the beta-lactamase inhibitors . YTR 830 or sulbactam, and isoelectric focusing for beta-lactamase were performed . Inducible isolates produced significantly more beta-lactamase than noninducible strains with or without the addition of cefoxitin . Noninducible isolates were more susceptible than inducible isolates to 8 micrograms/ml of piperacillin, a difference that was eliminated with the addition of either beta-lactamase inhibitor . Twelve of 13 strains produced a beta-lactamase band in the pH range of 7.9-8.1; no differences in satellite patterns were noted between the two groups of organisms . Increased production of beta-lactamase in the absence of an inducer may account for piperacillin resistance in P . cepacia in children with cystic fibrosis.

J Cardiovasc Surg (Torino), 1986 Nov-Dec, 27(6), 675 - 8
Surgical closure of persistent ductus arteriosus (PDA) in infants before 30 weeks gestation; Dasmahapatra HK et al.; Between October 1981 and December 1983 21 premature infants of mean gestational age 27.5 weeks (range 26-29 weeks) underwent surgical closure of persistent ductus arteriosus . Mean birth weight was 1080 g . There was no operative mortality . One death in an infant with pseudomonas septicaemia occurred two days after surgery . Twenty infants had features of idiopathic respiratory distress syndrome (IRDS) and required assisted ventilation prior to operation . Six infants had associated bronchopulmonary dysplasia (BPD) and 11 had signs of congestive cardiac failure . All infants presented with clinical features suggesting the diagnosis of PDA and in 18 the left atrial/aortic ratio was increased (mean 1.9:1) . In 18 infants a trial of Indomethacin therapy had failed . This experience supports the view that surgical closure of PDA in infants born before 30 weeks gestation can be accomplished safely . We believe that surgical treatment of PDA represents the optimal therapy in this high risk group of infants.

Diagn Microbiol Infect Dis, 1986 Nov, 5(4), 285 - 91
Bacterial adhesion in human upper gastrointestinal tract; Cerf M et al.; Small bowel biopsy specimens were taken from 21 patients undergoing gastrointestinal endoscopy to detect a possible adhesion of bacteria to the mucous layer of the upper gastrointestinal tract . In 30 control biopsy specimens taken from 10 patients free from gastrointestinal pathology, no associated bacteria were found, whereas in 23 biopsy specimens taken from eight gastrectomized patients an associated bacterial flora including E . coli or Pseudomonas was grown . Bacterial adhesion was confirmed by means of transmission electron microscopy of the eight patients yielding positive cultures . Bacterial adhesion induced local alterations of the brush border membrane . These results suggest that adherent bacteria may be present in hypochlorhydric patients . Pathophysiologic consequences require further studies.

Ann Ophthalmol, 1986 Nov, 18(11), 315 - 8
Conjunctival flaps in the treatment of refractory pseudomonas corneal abscess; Buxton JN et al.; Pseudomonas corneal abscess can result in a rapid downhill clinical course despite institution of appropriate medical measures . In this situation the clinician is faced with surgical intervention . A series of patients treated and stabilized through conjunctival flap therapy are presented . The role of conjunctival flap therapy in halting the progression of this entity is described.

Pediatr Pulmonol, 1986 Nov-Dec, 2(6), 368 - 72
Differences in drug susceptibility between isolates of Pseudomonas cepacia recovered from patients with cystic fibrosis and other sources and its relationship to beta-lactamase focusing pattern; Aronoff SC et al.; Pseudomonas cepacia, a significant pulmonary pathogen among children with cystic fibrosis (CF), often possesses an inducible beta-lactamase . The beta-lactamase isoelectric focusing pattern and beta-lactam susceptibility of CF and non-CF isolates of P . cepacia were compared . Against all of the test strains, ceftazidime and piperacillin were more effective than aztreonam . More CF isolates were resistant to 8 micrograms/ml of ceftazidime than non-CF isolates . Isoelectric focusing of cefoxitin-induced, cell-free preparations of the CF isolates produced significantly more bands than comparable preparations of non-CF isolates . Organisms producing a beta-lactamase band that focused in the pH range of 8.5 to 8.7 were significantly more resistant to 8 micrograms/ml of ceftazidime than other isolates . The increased resistance of CF isolates of P . cepacia to ceftazidime may be the result of the production of a specific bacterial beta-lactamase.

J Bacteriol, 1986 Nov, 168(2), 878 - 85
Naturally occurring TOL plasmids in Pseudomonas strains carry either two homologous or two nonhomologous catechol 2,3-oxygenase genes; Chatfield LK et al.; Structural genes for catechol 2,3-oxygenase (C23O) were cloned from the TOL plasmids pWW5, pWW14, pWW74, pWW84, and pWW88 isolated from Pseudomonas strains of diverse geographical origins . Each pKT230-based C23O+ recombinant plasmid carried a 2.05-kilobase XhoI insert which showed strong homology in Southern hybridizations with the xylE gene from the archetype TOL plasmid pWW0 . Fragments were mapped for restriction endonuclease sites and were classified into two closely related groups on the basis of restriction maps . C23O structural genes were located on cloned fragments by a combination of subcloning and site-specific mutagenesis . All five TOL plasmids examined yielded clones whose maps differed from that of xylE of pWW0 by only a single XbaI site, but in addition plasmids pWW5, pWW74, and pWW88 carried a second, homologous C23O gene with seven further restriction site differences . The remaining plasmids, pWW14 and pWW84, carried a second nonhomologous C23O gene related to the second C23O gene (C23OII) of TOL plasmid pWW15 described previously (H . Keil, M . R . Lebens, and P . A . Williams, J . Bacteriol . 163:248-255, 1985) . Thus, each naturally occurring TOL plasmid in this study appears to carry genes for two meta cleavage dioxygenases.

J Bacteriol, 1986 Nov, 168(2), 512 - 22
Gene cluster of Pseudomonas syringae pv . "phaseolicola" controls pathogenicity of bean plants and hypersensitivity of nonhost plants; Lindgren PB et al.; Loss of the ability of Pseudomonas syringae pv . "phaseolicola" NPS3121 to elicit a hypersensitive response on tobacco and other nonhost plants was associated with loss of pathogenicity on the susceptible host bean . Eight independent, prototrophic transposon Tn5 insertion mutants which had lost the ability to elicit a hypersensitive response on tobacco plants were identified . Six of these mutants no longer produced disease lesions on primary leaves of the susceptible bean cultivar Red Kidney and failed to elicit a hypersensitive response on the resistant bean cultivar Red Mexican and on the nonhost plants tomato, cowpea, and soybean . The two remaining mutants had reduced pathogenicity on Red Kidney bean and elicited variable hypersensitive responses on the other plants tested . Southern blot analysis indicated that each mutant carried a single independent Tn5 insertion in one of three EcoRI fragments of about 17, 7, and 5 kilobases . Marker exchange mutagenesis further supported the conclusion that the pleiotropic mutant phenotype was not associated with multiple Tn5 insertions . A genomic library of the wild-type strain was constructed in the cosmid vector pLAFR3 . A recombinant plasmid, designated pPL6, that carried P . syringae pv . "phaseolicola" genomic sequences was identified by colony hybridization . This plasmid restored the wild-type phenotype to all but one mutant, suggesting that genes affected by the insertions were clustered . Structural analysis of pPL6 and the wild-type genome indicated that the 17- and 5-kilobase EcoRI fragments were contiguous in the strain NPS3121 genome.

Proc Natl Acad Sci U S A, 1986 Nov, 83(22), 8467 - 71
Altered effector specificities in regulators of gene expression: TOL plasmid xylS mutants and their use to engineer expansion of the range of aromatics degraded by bacteria; Ramos JL et al.; Stimulation of transcription from positively regulated promoters involves regulatory proteins that have been activated, generally as a consequence of binding low molecular weight effector molecules . To define essential structural features of effectors for one positively acting gene regulator, the xylS-encoded protein, which activates the TOL plasmid meta-cleavage pathway operon promoters, effector activities of a wide range of benzoate derivatives have been systematically analyzed, and mutant xylS-encoded proteins exhibiting altered effector specificities have been generated and characterized . Cloned mutant xylS genes were trans dominant in partial diploids containing the wild-type xylS allele and could therefore be used to effect expansion of the range of aromatic compounds completely or partially degraded by Pseudomonas bacteria . The method developed to isolate mutant xylS-encoded proteins has general applicability and could in principle be used to isolate gene regulator specificity mutants of any inducible regulatory system.

J Bacteriol, 1986 Nov, 168(2), 702 - 7
Glyphosate catabolism by Pseudomonas sp . strain PG2982; Shinabarger DL et al.; The pathway for the degradation of glyphosate (N-phosphonomethylglycine) by Pseudomonas sp . PG2982 has been determined by using metabolic radiolabeling experiments . Radiorespirometry experiments utilizing {3-14C}glyphosate revealed that approximately 50 to 59% of the C-3 carbon was oxidized to CO2 . Fractionation of stationary-phase cells labeled with {3-14C}glyphosate revealed that from 45 to 47% of the assimilated label is distributed to proteins and that the amino acids methionine and serine are highly labeled . Adenine and guanine received 90% of the C-3 label found in the nucleic acid fraction, and the only pyrimidine base labeled was thymine . These results indicated that C-3 of glyphosate was at some point metabolized to a C-1 compound whose ultimate fate could be both oxidation to CO2 and distribution to amino acids and nucleic acid bases that receive a C-1 group from the C-1-donating coenzyme tetrahydrofolate . Pulse-labeling of PG2982 cells with {3-14C}glyphosate resulted in the isolation of {3-14C}sarcosine as an intermediate in glyphosate degradation . Examination of crude extracts prepared from PG2982 cells revealed the presence of a sarcosine-oxidizing enzyme that oxidizes sarcosine to glycine and formaldehyde . These results indicate that the first step in glyphosate degradation by PG2982 is cleavage of the carbon-phosphorus bond, resulting in the release of sarcosine and a phosphate group . The phosphate group is utilized as a source of phosphorus, and the sarcosine is degraded to glycine and formaldehyde . This pathway is supported by the results of {1,2-14C}glyphosate metabolism studies, which show that radioactivity in the proteins of labeled cells is found only in the glycine and serine residues.

Biochemistry, 1986 Oct 21, 25(21), 6711 - 5
Immunochemical detection of guanine nucleotide binding proteins mono-ADP-ribosylated by bacterial toxins; Eide B et al.; Rabbits immunized with ADP-ribose chemically conjugated to carrier proteins developed antibodies reactive against guanine nucleotide binding proteins (G proteins) that had been mono-ADP-ribosylated by bacterial toxins . Antibody reactivity on immunoblots was strictly dependent on incubation of substrate proteins with both toxin and NAD and was quantitatively related to the extent of ADP-ribosylation . Gi, Go, and transducin (ADP-ribosylated by pertussis toxin) and elongation factor II (EF-II) (ADP-ribosylated by pseudomonas exotoxin) all reacted with ADP-ribose antibodies . ADP-ribose antibodies detected the ADP-ribosylation of an approximately 40-kilodalton (kDa) membrane protein related to Gi in intact human neutrophils incubated with pertussis toxin and the ADP-ribosylation of an approximately 90-kDa cytosolic protein, presumably EF-II, in intact HUT-102 cells incubated with pseudomonas exotoxin . ADP-ribose antibodies represent a novel tool for the identification and study of G proteins and other substrates for bacterial toxin ADP-ribosylation.

Carbohydr Res, 1986 Oct 15, 154, 81 - 92
Synthesis of branched cyclomalto-oligosaccharides using Pseudomonas isoamylase; Abe J et al.; Branched cyclomalto-oligosaccharides (cyclodextrins) were synthesised from cyclomalto-oligosaccharides and maltose or maltotriose through the reverse action of Pseudomonas isoamylase . The reaction rate was greater with maltotriose than with maltose, and with increasing size of the cyclomalto-oligosaccharide (cG6 less than cG7 less than cG8) . Maltotriose is effective as both a side-chain donor and acceptor, and three isomers of 6-O-alpha-maltotriosylmaltotriose (branched G6) were formed through mutual condensation, but maltose was effective only as a side-chain donor . Each branched cyclomalto-oligosaccharide and G6 was purified by liquid chromatography, and their structures were determined by chemical, enzymic, and 13C-n.m.r . spectroscopic analyses.

Carbohydr Res, 1986 Oct 15, 154, 239 - 50
Monomer sequence and acetylation pattern in some bacterial alginates; Skjak-Braek G et al.; The sequential structures and acetylation patterns of alginates from several strains of Azotobacter vinelandii and Pseudomonas species, including P . aeruginosa, P . putida, P . fluorescens, and P . mendocina, have been studied by 1H-n.m.r . spectroscopy . O-Acetyl groups were exclusively associated with the D-mannuronic acid residues and the degree of acetylation varied in the range 4-57%, depending upon the proportion of this acid in the polymer . 1H-N.m.r . spectroscopy of a naturally occurring and an artificially acetylated D-mannuronan made it possible to determine the degrees of acetylation at O-2, O-3, and O-2,3 . The most conspicuous difference between alginates from A . vinelandii and the four Pseudomonas species was the complete absence of consecutive L-guluronic acid residues in the latter.

J Biochem (Tokyo), 1986 Oct, 100(4), 859 - 66
Spectral and kinetic studies on Pseudomonas L-phenylalanine oxidase (deaminating and decarboxylating); Koyama H et al.; Pseudomonas L-phenylalanine oxidase (deaminating and decarboxylating) contains two FAD molecules in one molecule of the enzyme (Koyama, H . (1983) J . Biochem . 93, 1313-1319) . When the enzyme was mixed anaerobically with L-phenylalanine, beta-2-thienylalanine, L-tyrosine, or L-methionine, a spectral species (purple intermediate) with a broad absorption band around 540 nm was observed with each substrate, and decayed slowly . From the data on the overall reaction kinetics, the rate of the L-phenylalanine oxidase reaction was expressed as follows . e/v = e/Vm + A/{S} + B/{O2} where e represents the concentration of enzyme unit, v the rate of the overall reaction, Vm the maximum velocity, and A and B are constants . Furthermore, the reactions of the enzyme with beta-2-thienylalanine (mostly an oxygenase substrate) and L-methionine (an oxidase substrate) were analyzed by the "stopped flow" method . The following scheme for the mechanism of L-phenylalanine oxidase reaction with both substrates is proposed, based on the data obtained . (formula; see text) Where Eox represents the oxidized form of the enzyme unit, EoxS the enzyme unit (oxidized form)-substrate compound, X the purple intermediate with a characteristic broad absorption band around 540 nm, S the substrate and P the product.

Cell Immunol, 1986 Oct 1, 102(1), 187 - 97
Active target cell processes, possibly involving receptor-mediated endocytosis, are critical for expression of cytotoxicity by natural killer cell-derived cytolytic factor; Deem RL et al.; Inhibitors of energy metabolism, 2-deoxyglucose and cyanide were shown to inhibit NKCF-mediated lysis of L929 target cells at the same molar concentrations that effectively inhibited cellular ATP levels and the toxic effects of pseudomonas toxin A . In addition, inhibitors of receptor-mediated endocytosis, cytochalasin B, a microtubule disrupter, and trifluoperazine, an inhibitor of clathrin-coat formation, inhibited NKCF-mediated lysis and expression of pseudomonas toxin activity, but had little effect upon cellular ATP . Lysomotropic agents chloroquine, ammonium chloride, and dansylcadavarine also inhibited both NKCF-mediated lysis and pseudomonas toxin activity . These results are similar to those involving diphtheria toxin and the plant toxins abrin, modeccin, and ricin, whose mode of action involves inhibition of protein synthesis following receptor-mediated endocytosis . However, it was determined that NKCF did not cause a decrease in the rate of protein synthesis up to the time of cell death . These results suggest that active target cell processes (possibly involving receptor-mediated endocytosis of NKCF) must occur for target cell lysis to be completed.

Am Rev Respir Dis, 1986 Oct, 134(4), 669 - 71
Pseudomonas cepacia: decrease in colonization in patients with cystic fibrosis; Thomassen MJ et al.; The incidence and prevalence of Pseudomonas cepacia pulmonary colonization were noted to be increasing in patients with cystic fibrosis (CF) . Previous work had indicated a greater prevalence of P . cepacia among siblings (with CF) of patients colonized by P . cepacia as well as an association of initial positive P . cepacia cultures with a hospitalization . Because of uncertainty regarding the source and mode of transmission, limited precautionary measures were instituted in 1983, including physical separation of hospitalized patients colonized with P . cepacia from non-colonized patients, reeducation of staff concerning basic infection control procedures, explanation to families regarding these precautionary efforts, and institution of separate summer camp sessions . Repeated environmental cultures throughout the hospital were negative for P . cepacia . Coincident with the institution of control measures, a sharp decline in incidence occurred (8.2% in 1983 versus 1.7% in 1984) . These results are suggestive of patient-to-patient transmission . Because P . cepacia infections have been associated with shorter survival in some patients with CF, we will continue our current segregation measures.

J Inorg Biochem, 1986 Oct-Nov, 28(2-3), 253 - 61
Gold-resistant bacteria: excretion of a cystine-rich protein by Pseudomonas cepacia induced by an antiarthritic drug; Higham DP et al.; P . cepacia bacteria adapted to growth in a chemically defined medium containing millimolar concentrations of Au(I) thiolates including the antiarthritic drug Au(I) thiomalate . The bacteria became very large, accumulated polyhydroxybutyrate and gold, and excreted a yellow protein ("thiorin"), which caused foaming of the culture medium . Thiorin was shown by 1H-NMR, amino acid analysis, and gel filtration chromatography to be of low molecular weight (ca . 9500) and to contain predominantly Cys (oxidized), Glx, and Gly.

Proc Natl Acad Sci U S A, 1986 Oct, 83(19), 7256 - 60
Identification and purification of a bacterial ice-nucleation protein; Wolber PK et al.; The protein product of a gene (inaZ) responsible for ice nucleation by Pseudomonas syringae S203 has been identified and purified after overexpression in Escherichia coli . The amino acid composition and the N-terminal sequence of the purified, denatured protein corresponded well with that predicted from the sequence of the inaZ gene . The product of inaZ was also found to be the major component in preparations of ice-nucleating, proteinaceous particles, obtained after extraction with and gel filtration in a mixture of urea and the nondenaturing detergent octyl beta-D-thioglucopyranoside . The activity of these preparations in the absence of added lipid implies that the protein participates directly in the nucleation process.

J Biol Chem, 1986 Sep 25, 261(27), 12883 - 8
Purification and some properties of component A of the 4-chlorophenylacetate 3,4-dioxygenase from Pseudomonas species strain CBS; Markus A et al.; Pseudomonas sp . strain CBS 3 possesses a two-component enzyme system which converts 4-chlorophenylacetate to 3,4-dihydroxyphenylacetate by the incorporation of 2 atoms of molecular oxygen . Component A of this enzyme system was purified to homogeneity by a 5-step procedure . After the last purification step the enzyme was homogeneous in analytical and sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The molecular weight of the native protein was determined to be 140,000 by Sephadex G-200 and 144,000 by analytical ultracentrifugation . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that component A consists of three identical subunits with a molecular weight determined to range between 46,000 and 52,000 . The isoelectric point was estimated to be 5.0 . Component A shows an intensive red-brown color, and in the oxidized state it exhibits a visible absorption spectrum with a maximum at 458 nm and a shoulder at 560 nm . By reduction with sodium dithionite a new peak with a maximum at 518-520 nm is observed . The enzyme contains iron (1.6-1.8 mol/subunit) and acid-labile sulfide (1.6-1.9 mol/subunit) which suggests that component A is an iron-sulfur protein.

Biochim Biophys Acta, 1986 Sep 25, 861(1), 201 - 4
Mechanism of action of Pseudomonas syringae phytotoxin, syringomycin . Interaction with the plasma membrane of wild-type and respiratory-deficient strains of Saccharomyces cerevisiae; Zhang L et al.; The effects of the phytotoxin, syringomycin, produced by Pseudomonas syringae pv . syringae, were examined on cells of a wild-type and a respiratory-deficient (rho0) mutant of Saccharomyces cerevisiae . The growth of both strains in liquid culture was inhibited by 0.5 micrograms syringomycin per ml and higher . Uptake rates of tetraphenylphosphonium and dimethyloxazolidine ions in cell suspensions of both strains increased when 1.5 micrograms per ml syringomycin was added . These responses were kinetically and quantitatively similar in the two strains and indicated increases in electrical potential (cell interior negative) and pH differences (cell interior alkaline) across the plasma membrane . Glucose (0.1 M) enhanced the effect on the electrical potential, was required for the pH changes, and increased the cellular ATP levels . These results show that the effects of syringomycin are energy-dependent and are due to alterations of plasma membrane and not to mitochondrial function.

J Biol Chem, 1986 Sep 5, 261(25), 11693 - 6
4-Hydroxyphenylpyruvate dioxygenase is an iron-tyrosinate protein; Bradley FC et al.; A resonance Raman investigation into the blue chromophore of 4-hydroxyphenylpyruvate dioxygenase, a non-heme iron enzyme from Pseudomonas P . J . 874, reveals the presence of enhanced vibrations characteristic of tyrosinate coordination to the iron center . The excitation profiles for these features show that they are associated with the 595 nm absorption feature . EPR studies of this enzyme indicate the presence of a high-spin ferric center in a rhombic environment, as evidenced by a signal at g = 4.3 with the correct intensity for the measured iron content . This enzyme thus belongs to the emerging class of iron-tyrosinate proteins.

J Gen Microbiol, 1986 Sep, 132 ( Pt 9), 2583 - 6
A generalized transducing phage of Pseudomonas cepacia; Matsumoto H et al.; A generalized transducing phage, named CP75, was derived from a lysogenic strain of Pseudomonas cepacia . The frequency of transduction per phage particle ranged from 1.0 X 10(-6) to 2.0 X 10(-6) for a given marker . About half of the 105 P . cepacia strains tested were sensitive to the phage . The molecular size of the CP75 genome was approximately 52 kb.

J Infect, 1986 Sep, 13(2), 157 - 8
Pseudomonas cepacia--fatal pulmonary infection in a patient with cystic fibrosis; Glass S et al.; A 9-year-old girl with cystic fibrosis (CF) was admitted with an exacerbation of respiratory infection and subsequently died . At death Pseudomonas cepacia was cultured from her sputum, in large numbers, 10(7) colony forming units/ml.

Proc Natl Acad Sci U S A, 1986 Sep, 83(17), 6627 - 30
Antitumor effects of an immunotoxin made with Pseudomonas exotoxin in a nude mouse model of human ovarian cancer; FitzGerald DJ et al.; An immunotoxin composed of Pseudomonas toxin coupled to an antibody to the human transferrin receptor was evaluated for its effect on ovarian cancer . In the tumor model employed, 60 million human ovarian cancer cells were injected into the peritoneal cavity of an immunodeficient nude mouse . By day 5, cancer cells were implanted and growing in small clusters throughout the peritoneal cavity . On days 5-8, 0.3-2 micrograms of immunotoxin was injected into the peritoneal cavity . Control mice died with malignant ascites at 34-58 days after the implantation of tumor cells, whereas immunotoxin-treated mice lived to 100 days or longer . Irrelevant immunotoxins or antibody alone had no antitumor activity . These findings suggest that intraperitoneal injection of immunotoxins may have a role in the treatment of ovarian cancer.

Arch Microbiol, 1986 Aug, 145(3), 220 - 7
The production and release of an extracellular polysaccharide during starvation of a marine Pseudomonas sp . and the effect thereof on adhesion; Wrangstadh M et al.; A marine Pseudomonas sp . S9 produced and released an extracellular polysaccharide during complete energy and nutrient starvation in static conditions . The presence of the polysaccharide on the cell surface, demonstrable by immune transmission electron microscopy, correlated with changes in the degree of adhesion to hydrophobic surfaces . Polysaccharide coated cells showed a lower degree of adhesion than did cells devoid of the polymer . After 10 h of starvation, no ruthenium red stained antibody stabilized polysaccharides could be observed on the cell surface . The polysaccharide was not produced during growth since lysates of mid-log phase cells did not precipitate the antiserum . The relative proportions of sugars in the polysaccharide were 28% glucose, 35% N-acetyl-glucosamine and 37% N-acetylgalactosamine . The released polysaccharide did not significantly alter the physical parameters of surface tension and viscosity of the starvation regime . Cells starved in agitated conditions did not produce any extracellular polysaccharides and exhibited a different adhesion pattern to hydrophobic surfaces.

Carbohydr Res, 1986 Aug 1, 150, 187 - 97
Synthesis of the tetrasaccharide repeating-unit of the lipopolysaccharide isolated from Pseudomonas maltophilia; Liptak A et al.; The title tetrasaccharide having the structure 3-O-Me-beta-L-Xylp-(1----4)- alpha-L-Rhap-(1----4)-alpha-L-Rhap-(1----2)-L-Rhap was obtained by reaction of the alpha-acetobromo derivative of 4-O-(3-O-methyl-beta-L-xylopyranosyl)-L-rhamnopyranose and benzyl 3,4-di-O-benzyl-2-O-(2,3-O-isopropylidene-alpha-L-rhamnopyranosyl)- alpha-L-rhamnopyranoside, followed by removal of the protecting groups . The synthesised compounds were characterised on the basis of n.m.r . data.

Eur J Biochem, 1986 Aug 1, 158(3), 469 - 75
Purification and characterization of N,N-dimethylformamidase from Pseudomonas DMF 3/3; Schar HP et al.; The N,N-dimethylformamide-hydrolyzing enzyme (DMFase) from Pseudomonas DMF 3/3 has been purified to apparent electrophoretic homogeneity with an overall 49-fold purification, a 24% yield and a final specific activity of 1.98 mumol N,N-dimethylformamide (DMF) hydrolyzed min-1 (mg protein)-1 . The native DMFase has a relative molecular mass of 250 000 and is composed of two light-chain (Mr = 15 000) and two heavy-chain (Mr = 105 000) subunits . The stability of DMFase is optimal at pH values above 7.5 and at temperatures below 20 degrees C . The activity of the enzyme is inhibited by metal-chelating agents such as EDTA and 2,2'-dipyridyl . Emission and atomic absorption spectroscopy measurements showed that iron is present in significant amounts in DMFase, indicating that it is an iron-containing amidohydrolase . In the ultraviolet/visible spectrum prominent bands were observed at 224 nm, 280 nm and 396 nm and shoulders are present at 418 nm and 467 nm . DMFase from Ps . DMF 3/3 has an isoelectric point of 7.7 . The enzyme exhibits optimal activity between pH 5 and 6 and at 40 degrees C . The substrate spectrum is rather narrow . The enzyme hydrolyzes preferentially substituted short-chain aliphatic amides such as DMF, N-ethylformamide and N-methylformamide . N,N-dimethylformamide, N,N-dimethylacetamide and unsubstituted amides, e.g . formamide, prolinamide, acetamide, acrylamide and butyramide are substrates as well, but are hydrolysed at significantly lower rates . DMFase obeys Michaelis-Menten kinetics and its Km and Vmax values for DMF are 13.8 mM and 1.89 U/mg, respectively, as determined from a Lineweaver-Burk plot.

Cancer Res, 1986 Aug, 46(8), 4047 - 52
Alteration of chemotherapy toxicity using a chemically defined liquid diet in rats; Kehoe JE et al.; Controversy exists as to whether administration of a chemically defined diet alters toxicity to chemotherapy . The purpose of this study was to evaluate toxicity to methotrexate in rats fed a chemically defined liquid diet or a regular chow diet . In the first study, 48 adult rats were randomized to be fed a chemically defined liquid diet or a regular diet for 14 days when methotrexate (25 or 50 mg/kg) was given . All liquid diet rats became anorexic and died within 96 h, while no deaths were observed in rats fed regular diet . When 20 liquid diet and regular diet rats were pair-fed to equalize caloric intake before and after methotrexate administration, similar mortality results occurred . In a second study, methotrexate (50 mg/kg) or saline was given and 60 h later all animals were sacrificed to obtain small bowel luminal cultures and tissue sections for histological evaluation . Administration of the liquid diet altered small bowel flora to predominantly Escherichia coli and Pseudomonas sp . and histology showed severe small bowel mucosal enteritis in comparison with regular diet rats . To evaluate whether the changes in intestinal flora or alterations in drug pharmacokinetics were responsible for the increased mortality, two additional studies were done . Gentamicin (4.8 mg/kg/day) was given p.o . or i.m . to the rats on the chemically defined liquid diet . A significant reduction of intraluminal bacteria occurred, but survival time was not improved in animals receiving antibiotics . When mean serum methotrexate levels were analyzed in non-antibiotic-treated rats, drug concentrations were significantly increased at 24, 36, and 48 h after methotrexate injection in the elemental liquid diet rats compared with chow diet rats . Administration of a chemically defined liquid diet to rats receiving methotrexate increased the occurrence and severity of intestinal enteritis, altered intraluminal bowel flora, and decreased clearance of methotrexate from the serum.

Acta Chem Scand B, 1986 Aug, 40(7), 515 - 21
High affinity protein-binding and enzyme-inducing activity of methyltrienolone in Pseudomonas testosteroni; Pousette A et al.; The synthetic androgen methyltrienolone (R 1881) was shown to increase steroid delta 1 dehydrogenase activity when added to cultures of Pseudomonas testosteroni at concentrations of 10(-10)-10(-8)M . Incubation with a soluble extract of P . testosteroni showed that (3H)-R 1881 was bound to a macromolecule with high affinity (Kd 0.6 X 10(-9)M) and low capacity (number of binding sites 120 X 10(-15) mol/mg of protein) . The (3H)-R 1881-macromolecule complex was partially destroyed following treatment with protease, was precipitated by addition of ammonium sulfate at 20% of saturation, sedimented at 6.3 S both in 0.01 and 0.4 M KCl solutions, and had an isoelectric point of pH 6.3 . The complex was partially bound to DNA-cellulose . Analysis by sucrose gradient centrifugation indicated that neither (3H)-testosterone and (3H)-estradiol-17 beta nor (3H)-corticosterone were bound with high affinity to the (3H)-R 1881-binding macromolecule . It is suggested that the partially characterized R 1881-binding macromolecule, which at least in certain respects resembles androgen receptors described in mammalian cells, is involved in the inductive effect of R 1881 on the delta 1 dehydrogenase activity in P . testosteroni.

J Urol, 1986 Aug, 136(2), 454 - 5
Hemorrhagic complications of piperacillin therapy; Lee M et al.; Clinically significant hemorrhage associated with extended spectrum anti-Pseudomonas penicillins is uncommon . We describe 2 patients with bleeding complications owing to piperacillin . In addition, we review the incidence and mechanism of this adverse reaction, and make suggestions for the management of these patients.

Arch Ophthalmol, 1986 Aug, 104(8), 1230 - 2
Treatment of experimental Pseudomonas corneal ulcers with enoxacin, a quinolone antibiotic; Sugar A et al.; Enoxacin is a broad-spectrum quinolone-derivative antibiotic . In a rabbit model of keratitis caused by a Pseudomonas species, enoxacin (3 mg/mL) was as effective as gentamicin sulfate (3 mg/mL) and enoxacin (10 mg/mL) in reducing viable bacterial counts in corneas after 24 hours of hourly therapy with eye drops . Bacterial counts were reduced by about 5000-fold by enoxacin treatment when compared with placebo-treated controls . Penetration studies of topical enoxacin (3 mg/mL) showed that concentrations in cornea and aqueous humor reached levels above reported minimal inhibitory concentrations when an epithelial defect was present . Further investigation of enoxacin for treatment of ocular disease is warranted.

J Antibiot (Tokyo), 1986 Aug, 39(8), 1160 - 6
Studies on lipoxygenase inhibitors . II . KF8940 (2-n-heptyl-4-hydroxyquinoline-N-oxide), a potent and selective inhibitor of 5-lipoxygenase, produced by Pseudomonas methanica; Kitamura S et al.; Pseudomonas methanica KY4634 was found to produce 5-lipoxygenase inhibitor designated KF8940, MY12-62a and MY12-62c . The inhibitors were purified by solvent extraction, silica gel column chromatography, reversed-phase low pressure liquid chromatography and crystallization . The chemical structures of KF8940, MY12-62a and MY12-62c were determined to be 2-n-heptyl-4-hydroxyquinoline-N-oxide, 2-n-heptyl-4-hydroxyquinoline and 3-n-heptyl-3-hydroxy-1,2,3,4-tetrahydroquinoline-2,4-dione, respectively, on the basis of their physico-chemical properties . Among them, KF8940 was the most potent inhibitor . The compound inhibited 5-lipoxygenase of rat basophilic leukemia cells in a dose-dependent manner and the half maximal inhibitory concentration (IC50) was 1.5 X 10(-7) M . At this concentration, KF8940 did not inhibit bovine platelet 12-lipoxygenase and cyclooxygenase, and the IC50 values for these enzyme were 3.5 X 10(-5) M and 1.7 X 10(-4) M, respectively . The results indicated that KF8940 is a potent and selective inhibitor of 5-lipoxygenase . The IC50 value of MY12-62c for 5-lipoxygenase was 1.9 X 10(-5) M and that of MY12-62a was 1.9 X 10(-5) M.

Mol Gen Genet, 1986 Aug, 204(2), 273 - 80
Replicon fusions promoted by insertion sequences on Pseudomonas cepacia plasmid pTGL6; Barsomian G et al.; Plasmid pMR5 (pRP1ts) failed to replicate in Pseudomonas cepacia at 47 degrees C . Selection at this temperature for maintenance of tetracycline resistance associated with this plasmid allowed isolation of cointegrate plasmids formed by fusion of pMR5 with pTGL6, a 170 kb plasmid harbored by P . cepacia 249 . In the cointegrate plasmids pTGL100, pTGL101, and pTGL102, different regions of pTGL6 were involved in fusion with the same tra-2-containing region of pMR5 . Formation of all three plasmids was promoted by insertion sequences on pTGL6, which were also represented in the chromosome . Two different copies of a 1.3 kb element, IS401, were involved in formation of pTGL100 and pTGL101 . Another insertion sequence, IS402 (1 kb), promoted the fusion which formed pTGL102 . Southern hybridization experiments indicated that each of the cointegrate plasmids contained an additional copy of the fusion mediating element . Plasmid pTGL100 was observed to resolve into two independent replicons: pTGL6 and pTGL105 (pMR5::IS401), a novel derivative of pMR5 containing a copy of IS401 . The third cointegrate plasmid, pTGL102, evolved in two steps: fusion of pTGL6 and pMR5 mediated by IS402, and transposition of IS411 (1.9 kb) to a region of pMR5 distinct from that involved in the fusion . Plasmid pTGL6 contained one copy of IS402 and IS411 while pTGL102 contained two copies of each of these elements.

Appl Environ Microbiol, 1986 Aug, 52(2), 281 - 9
Inhibitor studies of dissimilative Fe(III) reduction by Pseudomonas sp . strain 200 ("Pseudomonas ferrireductans")
Arnold RG, DiChristina TJ, Hoffmann MR.
Aerobic respiration and dissimilative iron reduction were studied in pure, batch cultures of Pseudomonas sp . strain 200 ("Pseudomonas ferrireductans") . Specific respiratory inhibitors were used to identify elements of electron transport chains involved in the reduction of molecular oxygen and Fe(III) . When cells were grown at a high oxygen concentration, dissimilative iron reduction occurred via an abbreviated electron transport chain . The induction of alternative respiratory pathways resulted from growth at low oxygen tension (less than 0.01 atm {1 atm = 101.29 kPa}) . Induced cells were capable of O2 utilization at moderately increased rates; dissimilative iron reduction was accelerated by a factor of 6 to 8 . In cells grown at low oxygen tension, dissimilative iron reduction appeared to be uncoupled from oxidative phosphorylation . Models of induced and uninduced electron transport chains, including a mathematical treatment of chemical inhibition within the uninduced, aerobic electron transport system, are presented . In uninduced cells respiring anaerobically, electron transport was limited by ferrireductase activity . This limitation may disappear among induced cells.

Klin Wochenschr, 1986 Jul 15, 64(14), 633 - 5
{IgG antibodies against toxic shock syndrome toxin 1 in human immunoglobulins}; Dickgiesser N et al.; IgG antibodies against toxic shock syndrome toxin-1 in human immunoglobulins were determined using the ELISA technique . Of the drugs for intramuscular application, hemogamma and beriglobin contained the highest amount of antibodies . The highest concentration of antibodies in drugs for intravenous application was found in Pseudomonas polyglobin and in Venimmun.

Biull Eksp Biol Med, 1986 Jul, 102(7), 71 - 4
{Biological properties of glutamin-(asparagin-)ase from Pseudomonas boreopolis 526}; Pekhov AA et al.; Glutamine(asparagine)ase from Ps . boreopolis 526 has an antineoplastic effect on lymphoid leukemia P-388 . The enzyme half-life in the mouse serum is 8.5 hours . Glutamine(asparagine)ase has no cross-antigenicity with L-asparaginase from E . coli (Bayer, FRG) . Specific antibodies against L-asparaginase (Bayer, FRG) do not influence the activity of glutamine(asparagine)ase.

J Pediatr, 1986 Jul, 109(1), 51 - 4
Pseudomonas cepacia in the hospital setting: lack of transmission between cystic fibrosis patients; Hardy KA et al.; Possible mechanisms of transmission of Pseudomonas cepacia in hospitalized patients with cystic fibrosis were examined . Twelve patients were colonized with P . cepacia prior to admission (group 1), and 15 patients were not (group 2) . Daily contact occurred between both groups . Sputum cultures were obtained from all patients at admission and discharge, and 3 and 6 months after discharge in group 2 patients . Environmental cultures included cough plates, settle plates, spirometry tubing, stethoscopes, sinks, wall outlets, and hands of patients, physicians, nurses, and respiratory therapists . Specimens were plated on a P . cepacia-selective medium . All group 1 patients remained colonized with P . cepacia at discharge . None of the group 2 patients acquired P . cepacia during hospitalization or follow-up . P . cepacia was recovered from three of 151 environmental cultures; two of these were presumably patient related . P . cepacia contamination of pulmonary function equipment, wall outlets, hands, and stethoscopes was not documented . We conclude that transient aerosol environmental contamination is uncommon and restricted to the immediate patient environment.

J Clin Microbiol, 1986 Jul, 24(1), 152 - 4
Serological classification of Pseudomonas cepacia by somatic antigen; Nakamura Y et al.; Ten samples of antiserum against Pseudomonas cepacia were prepared by the intravenous immunization of rabbits with heat-killed organisms . Ten P . cepacia strains used for immunization were proven unique antigenic strains . Using these antisera, we serogrouped 127 strains of P . cepacia, and 114 strains (89.8%) fell under one of the ten serogroup . The most prevalent serogroup was C (26.8), the second most prevalent being D (18.1%) . When we compared our serogroups with the serogroups of Monteil et al . (H . Monteil, C . Richard, and A . Heidt, Med . Mal . Infect . 11:544-547, 1981) and Heidt et al . (A . Heidt, H . Monteil, and C . Richard, J . Clin . Microbiol . 18:738-740, 1983), five out of seven of their serogroups were represented by our antisera.

Cancer Res, 1986 Jul, 46(7), 3262 - 7
Antibody-Pseudomonas exotoxin A conjugates cytotoxic to human breast cancer cells in vitro; Bjorn MJ et al.; Breast tumor selective antibodies (MAB) conjugated to Pseudomonas exotoxin A (PE) formed immunotoxins with potent cytotoxicities for human breast tumor cell lines . The most effective of the MAB-PE conjugates were about 500-fold more toxic to the breast tumor target cell lines than to the nontarget human fibroblast cell line . Specificity of cytotoxicity by MAB-PE conjugates was demonstrated by protection of target cells in the presence of excess unconjugated homologous antibody . A MAB-PE conjugate inhibited protein synthesis more rapidly than the corresponding MAB-ricin toxin A chain (RTA) conjugate . Generally, there was a direct correlation between the cytotoxicity of RTA and PE when conjugated to the same MAB: MABs that made highly cytotoxic RTA conjugates made effective PE conjugates and MABs that made poorly cytotoxic RTA conjugates made PE conjugates of low cytotoxicity; however, one MAB-PE immunotoxin was cytotoxic to the human breast tumor cell lines, whereas the corresponding MAB-RTA immunotoxin was noncytotoxic at the highest dose tested . In contrast to MAB-RTA conjugates, which require a cleavable (disulfide) linkage for maximal in vitro cytotoxicity, MAB-PE conjugates were about equally cytotoxic when linked by either a cleavable or noncleavable (thioether) bond.

Biochim Biophys Acta, 1986 Jun 5, 871(2), 142 - 8
Perturbation of Pseudomonas cytochrome oxidase by guanidine hydrochloride to detect differential stabilization of the heme d1 and heme c moieties; Horowitz P et al.; The optical properties of Pseudomonas cytochrome oxidase (ferrocytochrome-c:oxygen oxidoreductase, EC 1.9.3.2) were monitored as a function of guanidine hydrochloride (Gdn X HCl) concentration to probe for differential stabilization of its prosthetic groups, heme d1 and heme c . The protein fluorescence intensity increased with the Gdn X HCl concentration, revealing two transitions, a sharp one between 1.3 and 1.5 M Gdn X HCl, and a second less well defined extending from 2.5 to 4.5 M . Only the transition at the lower Gdn X HCl concentrations was present in titrations followed using the emission maxima . The spectral maximum for native Pseudomonas cytochrome oxidase was at approx . 335 nm and shifted to approx . 350 nm above 2 M Gdn X HCl . The heme d1 absorbance at 638 nm decreased with increasing {Gdn X HCl}, giving a transition at 1.3-1.5 M, and no transition up to 4 M Gdn X HCl when the heme c was monitored at 525 nm . Along with the decrease at 638 nm, an absorption band appeared at 681 nm, suggesting heme d1 release into solution . Fluorescence titration of heme d1-depleted enzyme, prepared by gel filtration, showed a single transition similar to the transition occurring in the intact enzyme at high Gdn X HCl concentrations . Circular dichroism spectra revealed clearly distinguishable transitions for the heme d1 and heme c near 1.5 and 3.0 M Gdn X HCl, respectively . These results suggest that the two hemes are in regions of the protein with different stabilities which may represent distinct structural domains.

Pathol Biol (Paris), 1986 Jun, 34(5 Pt 2), 653 - 6
{Diffusion of piperacillin into bronchial secretions}; Bergogne-Berezin E et al.; Piperacillin is a ureido-penicillin characterized by the presence of a piperazine group at the position 6 of the beta-lactam ring . This group confers a broader spectrum that includes Pseudomonas . In vitro studies have shown that piperacillin is active against clinical strains recovered from intensive care unit patients with severe lower respiratory tract infections . The purpose of our study was to investigate the usefulness of piperacillin in such patients by evaluating the drug's diffusion into bronchial secretions . A single 4 g dose of piperacillin was given intravenously over three minutes to each of 6 intensive care unit patients . Serum and bronchial secretion samples (obtained through a tracheal intubation or tracheostomy tube) were taken 1/2 hour, 2 h, 4 h and 6 h after the injection to evaluate piperacillin kinetics . Serum piperacillin concentrations were maximal 30 mn after the IV (mean value: 90.6 +/- 23.8 micrograms/ml) and thereafter fell gradually (mean value after 4 hours: 40.4 +/- 27.5 micrograms/ml) . Peak concentrations in bronchial secretions were recorded at 2 hours (12.2 +/- 8.5 micrograms/ml); the mean residual value at 6 hours was 4.9 +/- 8.7 micrograms/ml . The diffusion ratio (ratio of bronchial secretion concentration to simultaneous serum concentration) was 15.5% to 24.5% . Our results show that the diffusion of piperacillin into bronchial secretions is outstanding and support the use of this drug in severe respiratory tract infections.

Bioorg Khim, 1986 Jun, 12(6), 780 - 8
{Experimental and theoretical conformation analysis of the O-antigenic polysaccharide from Pseudomonas cepacia strain 3181 . II . The polysaccharide}; Lipkind GM et al.; The spatial structure of Pseudomonas cepacia 3181 polysaccharide in aqueous solution is discussed basing on the data of nuclear Overhauser effect, observed with preirradiation of anomeric protons of all 3 D-rhamnose residues in the repeat unit, and theoretical conformational analysis . It is shown that conformational states of the free disaccharides and corresponding disaccharide units of the polysaccharide are similar . All conformations of the polysaccharide may be described by one shape representing an extended structure with characteristic turns in the D-Rha alpha 1-2-D-Rha beta 1-3-D-Rha units.

Bioorg Khim, 1986 Jun, 12(6), 771 - 9
{Experimental and theoretical conformation analysis of the O-antigenic polysaccharide from Pseudomonas cepacia strain 3181 . I . Disaccharide links}; Lipkind GM et al.; Nuclear Overhauser effects, with preirradiation of glycoside bond anomeric protons, coupling constants 3J C3, H1' and 3J C1', H4 and linkage optical rotations A were measured for L-Rha beta 1-3-L-Rha alpha 1-OMe and L-Rha alpha 1-3-L-Rha alpha 1-OMe which are the models of the disaccharide units of the Pseudomonas cepacia polysaccharide . Theoretical conformational analysis was carried out in terms of a mechanical molecular model approximation . The spatial structures of these disaccharides as well as of D-Rha alpha 1-2-D-Rha beta 1-OMe in aqueous solutions were discussed basing on the obtained results.

Am J Med, 1986 May 30, 80(5C), 75 - 8
In vitro considerations in monotherapy and combination therapy for severe infections; Gaya H; The introduction of the many new penicillins and cephalosporins has not radically altered the results of therapy in infected patients with impaired host defenses . Pseudomonas-active beta-lactam combinations with aminoglycosides remain the mainstay of treatment . Among the new cephalosporins, only ceftazidime shows promise in clinical trial, and the results of the fourth European Organization for Research on Treatment of Cancer (EORTC) trial, which includes the combination of amikacin plus ceftazidime, are awaited with interest . Whichever regimen is chosen, care must be taken to detect the emergence of resistant strains . This may entail changing the empiric regimen used on a cyclic basis in order to minimize the problem and maintain an acceptable level of clinical response.

Eur J Biochem, 1986 May 15, 157(1), 121 - 8
The pterin (bactopterin) of carbon monoxide dehydrogenase from Pseudomonas carboxydoflava; Kruger B et al.; Radioactively labeled carbon monoxide (CO) dehydrogenase has been obtained in good yield and purity from Pseudomonas carboxydoflava grown in the presence of {32P}phosphate . One enzyme molecule contained an average of 8.32 molecules of phosphate . The entire phosphate content was confined to 2 molecules of FAD and 2 molecules of a pterin . These were noncovalently bound . Molybdoenzyme cofactors could be extracted into N-methyl formamide; pterins were isolated by thin-layer chromatography . CO dehydrogenase contained a novel pterin, different from molybdopterin, which was also resolved in other bacterial molybdoenzymes . Therefore, it was tentatively named bactopterin . The characteristic features of bactopterin were as follows . A relative molecular mass, Mr, of 730 which was much greater than that of molybdopterin (330) (Mr values refer to molybdenum-free forms of the cofactors; presumably, the latter were also devoid of the sulfhydryl groups contained in the native compounds) . A content of 2 molecules of phosphate/molecule compared to only 1 phosphate in molybdopterin . Bactopterin was three times less susceptible to air oxidation than molybdopterin . Native bactopterin was cleaved by perchloric acid into two phosphorous-containing fragments with Mr of 330 and 420 . The smaller one is believed to be very similar to molybdopterin, the larger one was not a pterin but probably contained an aromatic structure.

Aust Vet J, 1986 May, 63(5), 146 - 9
Melioidosis in intensive piggeries in south eastern Queensland; Ketterer PJ et al.; The epidemiology of melioidosis was investigated in 8 intensive piggery units which used water from the same river in south eastern Queensland . In 3 consecutive years cases of disease followed heavy rainfall and flooding . Although Pseudomonas pseudomallei was not isolated from water or soil samples the water supply was suspected as the source of infection . Affected pigs were detected at slaughter by the presence of abscesses most commonly in the bronchial lymph nodes (40%) and spleen (34%) . One hundred and fifty nine cases were observed at slaughter from a total of 17,397 animals at risk . Infection by inhalation of water aerosols derived from nipple drinkers, hose sprays and a water misting cooler was considered to be responsible for the bronchial lymph node lesions . These outbreaks occurred outside the area in which melioidosis is generally regarded as being endemic.

J Clin Microbiol . 1986 May;23(5):962.
Pseudomonas cepacia isolated from crystal violet solution in a hospital laboratory; Walsh DM et al.; Pseudomonas cepacia was isolated from crystal violet solution in a hospital laboratory . A similar strain was isolated from the cerebrospinal fluid of a patient . It was difficult to determine whether these strains were identical, but they possessed similar biochemical patterns.

Infect Immun, 1986 May, 52(2), 445 - 53
Reduced temperature alters Pseudomonas exotoxin A entry into the mouse LM cell; Morris RE et al.; The movement of Pseudomonas exotoxin A (PE) into the cytoplasm of mouse LM fibroblasts was followed by using inhibition of protein synthesis as a biochemical index of toxin activity; biotinyl-PE and avidin-gold colloids were used for electron microscopy . At 37 degrees C both specific antitoxin and pronase-trypsin protected cells against PE toxicity when added within seconds of warming cells, whereas methylamine was protective when added during the first 7 min of endocytosis . Lowering the temperature to 19 degrees C afforded protection when the temperature transition was accomplished within 15 min of the original endocytic event . These data suggest that PE enters an acidic compartment before reaching a step blocked by shifting cells from 37 to 19 degrees C . PE expressed toxicity for LM cells at 19 degrees C, but at a concentration 1 order of magnitude higher than that required at 37 degrees C . At 19 degrees C, antitoxin or trypsin-pronase protection was rapidly ablated . In contrast cells were fully protected by methylamine for 90 min . Using electron microscopy we demonstrated that toxin moved normally (30 s) to coated areas at 19 degrees C, but remained at this site for up to 20 min before being internalized . The majority of the toxin internalized at 19 degrees C remained in endosomes or in Golgi-associated vesicles and was not delivered to lysosomes . The results suggest that, under physiological conditions (37 degrees C), PE rapidly enters cells through coated areas, moves to an acidic compartment (i.e., the endosome), and then probably to the Golgi region en route to lysosomes . The evidence suggests that movement of toxin from endosomes or Golgi vesicles to lysosomes is blocked at 19 degrees C . We hypothesize that the active form of PE enters the cytosol, where it expresses its toxicity during fusion of Golgi-derived, toxin-laden vesicles with lysosomes.

J Appl Bacteriol, 1986 May, 60(5), 455 - 62
Factors affecting the measurement of antibiotic resistance in bacteria isolated from lake water; Jones JG et al.; It is more difficult to obtain a reliable assessment of antibiotic resistance in populations of aquatic bacteria than in those populations which are well characterized (e.g . bacteria of medical and veterinary significance) . Factors which influence the results include the bacterial taxa involved, their site of origin and the methods and media used to isolate and subculture the bacteria, and to perform the sensitivity tests . Examples of these effects are provided . The resistance profiles obtained with populations of aquatic pseudomonads depend on the species composition of the population . Resistance patterns in aquatic bacteria varied with the site from which they were isolated; a higher incidence of resistance was recorded along shorelines and in sheltered bays than in the open water . The inclusion of antibiotics in the media employed for primary isolation increased the number of individual and multiple resistances recorded . A similar effect was observed with increased inoculum size in the sensitivity disc method but this could be reversed by raising the incubation temperature . The medium used to conduct the test also affected the results and many aquatic bacteria failed to grow on media such as Iso-Sensitest Agar . It is recommended that the sensitivity disc method is adopted for aquatic bacteria because it permits interpretation of a wider range of response . Comparison of the incidence of antibiotic resistance in different habitats will remain meaningless, however, until comprehensive methods for the identification of bacteria are developed and the techniques used for sensitivity testing are standardized.

Pathol Biol (Paris), 1986 May, 34(5), 424 - 9
{Evaluation of the antibiotic sensitivity of Pseudomonas sp . using the API-ATB-PSE system . Comparison with diffusion and the reference agar dilution method}; Delisle F et al.; In vitro activity of twelve antibiotics (ticarcillin, mezlocillin, azlocillin, piperacillin, cefoperazone, cefsulodin, ceftazidime, gentamicin, netilmicin, pefloxacin and ciprofloxacin) was determined by measuring minimum inhibitory concentrations (MICs) using agar dilution according to WHO recommendations, agar diffusion and the API-ATB-PSE system . One-hundred and forty-two Pseudomonas strains were studied . Five species of Pseudomonas were represented, i.e . aeruginosa, maltophilia, cepacia, stutzeri and paucimobilis . Isolates came from two Paris hospitals . Percentages of total agreement, minor discrepancies and major discrepancies between the results obtained with the API-ATB-PSE method and those recorded with the reference methods were determined . After checking the discordant strains, the results are discussed in order to evaluate the reliability of the ATB method for antibiotic susceptibility testing with Pseudomonas sp.

Infect Control, 1986 May, 7(5), 281 - 4
Pseudomonas cepacia; Gregory WJ et al.; P . cepacia is reported to be an increasing cause of infection and colonization of patients in hospitals . Historically it is an important contaminant in the pharmaceutical industry . Its nutritional versatility, ability to survive and multiply in water, high intrinsic resistance to antibiotics, and ability to multiply in the majority of traditional disinfectants make it a superb agent for causing nosocomial infection . Recognition of its differences from P . aeruginosa and its ability to contaminate agents used in hospitals is important in proper treatment and infection control.

J Heart Transplant, 1986 May-Jun, 5(3), 215 - 28
Three recent cases of the total artificial heart before transplantation; Levinson MM et al.; Three recent cases from one institution using the total artificial heart (TAH) before transplantation are reviewed . The first patient was implanted for 12 hours with the pneumatic Phoenix total artificial heart after failure of a donor heart 1 day after transplant . Following retransplantation the patient died from severe pulmonary edema, pulmonary hypertension, right ventricular failure, and Pseudomonas septicemia . The second patient was implanted with the Jarvik-7 total artificial heart for rapidly deteriorating idiopathic cardiomyopathy . Major complications during the 9 1/2-day implant consisted of severe pulmonary edema for the first 4 days and a multifocal cerebral embolic event on the seventh day after implantation from which he fully recovered . Major problems after transplant included disseminated toxoplasmosis and two mild episodes of rejection . The patient was discharged 68 days after surgery and remains well . The third patient was a 40-year-old woman with rapidly progressing acute influenza A viral myocarditis . Despite immunosuppressive and antiviral therapy, cardiogenic shock with multiple organ failure developed . The 70 ml Jarvik-7 was implanted for 4 1/2 days . Acute humoral rejection from autoantibodies and alloantibodies led to a cardiac arrest on the second day after transplantation . A second 70 ml Jarvik-7 implant was followed by severe multisystem and infectious complications . After prolonged intensive care support, the patient recovered and is now awaiting transplantation . Nearly 100% cytotoxic antibody reactivity caused by multiple antigenic stimuli is preventing ready access to donor hearts for this patient . We view the current role of the total artificial heart as a tool to preserve life until a suitable donor heart can be found, reverse the end-organ effects of progressive heart failure and low output, and restore transplant candidacy in selected patients with temporary reversible contraindications to transplantation.

J Surg Res, 1986 May, 40(5), 438 - 44
Effects of ibuprofen on a pig Pseudomonas ARDS model; Lee CC et al.; The effects of ibuprofen (I) were studied in the Pseudomonas (P) porcine ARDS model . Pigs, 14-26 kg (5 in each group), were anesthetized and ventilated with 0.5 FiO2 and 5 cm H2O PEEP . A control (C) group received saline only, a second group was given P, 1 X 10(8) org/ml at 0.3 cc/20 kg/min, and a third group was given P followed by 12.5 mg I at 20 and 120 min . Pulmonary arterial (PAP), wedge (PWP) and systemic arterial pressures, cardiac output (CO), and thermal-cardiogreen extravascular lung water (EVLW), thromboxane (TxB2), 6-keto-PGF1 alpha, PaO2, PaCO2 were determined every 30 min . Albumin flux was measured with scintigraphic determination of lung:heart radioactivity ratios versus time, called slope index (SI) . At 3 hr, P produced marked (P less than 0.05) increases in PAP (18 +/- 7 to 37 +/- 2 mm Hg), TxB2 (471 +/- 513 to 9216 +/- 3615 pg/ml), 6-keto-PGF1 alpha, EVLW (6.4 +/- 1.4 to 14.6 +/- 5.7 mg/kg), and SI (0.4 +/- 0.2 to 1.7 +/- 0.5 X 10(-3) U/min) with decreases in PaO2 (214 +/- 47 to 101 +/- 41 torr), CO and SAP . Ibuprofen caused a rapid clearing of TxB2 and 6-keto-PGF1 alpha associated with a transient decrease in PAP; PaO2 was considerably improved compared to P; however, CO, SAP, EVLW, and SI were unaffected . Prostaglandin blockage temporarily ameliorated the pulmonary hypertension and markedly improved oxygenation in this porcine septic ARDS model, but failed to alter increased permeability, confirming other studies that the increased pulmonary shunt in ARDS is not only dependent upon capillary leak.

J Bacteriol, 1986 May, 166(2), 598 - 603
Cloning of the gene for indoleacetic acid-lysine synthetase from 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 on bacterial production of the phytohormone indoleacetic acid (IAA) . The genetic determinants for IAA synthesis are located on a plasmid (pIAA) and are organized in an operon in oleander strains of the bacterium . P . syringae subsp . savastanoi further converts IAA to an amino acid conjugate, 3-indole-acetyl-epsilon-L-lysine (IAA-lysine) . The gene for IAA-lysine synthetase (iaaL) was found on the IAA plasmid by screening pIAA deletion mutants for the ability to convert IAA to IAA-lysine . The iaaL locus was then cloned in the vector pUC8 from a bank of P . syringae subsp . savastanoi EW2009 plasmid DNA to construct recombinant plasmid pLG87 . The specific activity of IAA-lysine synthetase in Escherichia coli transformed with pLG87 was 47 times higher than that of the enzyme extract from P . syringae subsp . savastanoi . The direction of transcription of the iaaL gene was determined to be opposite to that of the IAA operon . The location of the iaaL gene on pIAA1 was mapped by Tn5 insertion mutagenesis to a 2.5-kilobase-pair fragment 2 kilobase pairs from the IAA operon.

J Bacteriol, 1986 May, 166(2), 581 - 90
Isolation and complementation analysis of 10 methanol oxidation mutant classes and identification of the methanol dehydrogenase structural gene of Methylobacterium sp . strain AM1; Nunn DN et al.; A method has been developed for the direct selection of methanol oxidation mutants of the facultative methylotroph Methylobacterium sp . strain AM1 (formerly Pseudomonas sp . strain AM1) . Using this direct selection technique, we have isolated mutants of Methylobacterium sp . strain AM1 that are no longer capable of growth on methanol but retain the ability to grow on methylamine . These methanol oxidation (Mox) mutants were complemented with a genomic clone bank of this organism constructed in the broad-host-range cosmid pVK100, and subcloning and Tn5 mutagenesis experiments have assigned the Mox mutants to 10 distinct complementation groups . Using an open reading frame beta-galactosidase fusion vector and antibodies specific for Methylobacterium sp . strain AM1 methanol dehydrogenase, we have identified the methanol dehydrogenase structural gene and determined the direction of transcription . The results suggest that the synthesis and utilization of an active methanol dehydrogenase in this organism requires at least 10 different gene functions.

Genetika, 1986 Apr, 22(4), 705 - 8
{Isolation of Tyr- mutants of facultative methylotrophic Pseudomonas sp . M.}; Olekhnovich IN et al.; A method for isolation of Tyr- mutants of facultative methylotrophic bacteria Pseudomonas sp . M which possess two tyrosine synthesis pathways is presented . The method is based on the two-step blocking of the tyrosine synthesis: the first step of the supplementary pathway of synthesis from phenylalanine, the second being the main pathway from 4-hydroxyphenylpyruvate.

J Wildl Dis, 1986 Apr, 22(2), 238 - 44
Lead exposure in an "urban" peregrine falcon and its avian prey; DeMent SH et al.; Necropsy of a 7-yr-resident peregrine falcon (Falco peregrinis) from Baltimore showed a Pseudomonas infection involving the pharynx as the immediate cause of death . Concentrations of lead in liver and kidney measured 0.74 and 1.40 ppm, respectively . A survey of lead exposure was performed on 40 urban rock doves (Columbia livia) . Thirteen additional rock doves were collected from sites removed from lead contamination and served as controls . The mean concentration of lead in the blood of the urban rock doves was 0.96 ppm (range 0.29-17.0 ppm) compared to 0.05 ppm (0.01-0.07 ppm) for control birds . Ninety-eight percent (39/40) of the urban rock doves had elevated concentrations of lead in their blood, while 27% (11/40) had sublethal concentrations . None of the control birds had increased concentrations of lead in their blood . Concentrations of lead in liver and kidney of 13 urban rock doves were 3.48 ppm and 9.53 ppm, respectively, compared to concentrations of 0.43 ppm and 0.50 ppm for four control rock doves . From these data a mean total concentration of lead per rock dove was calculated at 4.60 ppm for urban birds and 0.33 ppm for control birds.

Mol Gen Genet, 1986 Apr, 203(1), 64 - 72
Molecular cloning and characterization of sequences from the regulatory cluster of the Pseudomonas plasmid alk system; Owen DJ; Alkane oxidation functions encoded by the Pseudomonas plasmid CAM-OCT are positively regulated by one or more products of a locus designated alkR . To characterize this locus in greater detail, molecular cloning and restriction mapping of sequences covering the alkR region have been carried out in Escherichia coli, followed by mobilization to Pseudomonas recipients for analysis of genetic content . Inserts from Pseudomonas (CAM-OCT) strains were cloned into vectors pLAFR1, the pLAFR1::Tn7S derivative pXJS5403, and the transposon vector Tn3 delta 596 . This has made it possible to: (1) construct a detailed restriction map of cloned fragments and the alkR region of CAM-OCT; (2) map insertion sites of the transposon Tn7S into alkR cistrons; and (3) analyze the ability of cloned sequences to complement or effect marker rescue of alkR nitrosoguanidine- and Tn7S-induced mutations . In addition, transcription of an alkB'-lacZ transcription fusion in the presence of a cloned 18.5 kb EcoRI alkR fragment and an inducer of the alk system confirmed that our cloned sequences contain functional alkR cistrons . The complementation/marker rescue results indicate that alkR is a complex locus and that the products of at least three cistrons are required for the complete AlkR+ phenotype . One of these cistrons is identified by mutations which alter a component of the inducer recognition system.

Microb Pathog, 1986 Apr, 1(2), 139 - 48
The causal agent of halo blight in bean, Pseudomonas syringae pv . phaseolicola, attaches to stomata via its pili; Romantschuk M et al.; The phytopathogenic pseudomonad Pseudomonas syringae pv . phaseolicola causes halo blight of bean (Phaseolus vulgaris L.) . Initiation of infection depends on the ability of the cells to adhere to the target cell surface . P . syringae pv . phaseolicola expresses pili, which are the receptors of the lipid-containing dsRNA bacteriophage phi 6 . phi 6-resistant bacterial strains can be divided into different piliation types . It was possible to show that the adhesion of the bacteria onto plant cell surface was dependent on the pili . Non-piliated bacterial stains showed a much lower adherence to the leaf surface than strains expressing phi 6 specific pili . Scanning electron microscopy showed that the piliated bacteria attached to the leaf surface at the site of stomata . Non-piliated bacteria were evenly distributed on the leaf surface . All bacterial strains used in this study were capable of causing halo blight if injected into the plant . If the bacteria were sprayed on the plants, followed by spraying of sterile buffer, only piliated bacteria caused symptoms.

J Bacteriol, 1986 Apr, 166(1), 224 - 9
Role of glutamine synthetase adenylylation in the self-protection of Pseudomonas syringae subsp . "tabaci" from its toxin, tabtoxinine-beta-lactam; Knight TJ et al.; Selected pathovars of Pseudomonas syringae produce an extracellular phytotoxin, tabtoxinine-beta-lactam, that irreversibly inhibits its known physiological target, glutamine synthetase (GS) . Pseudomonas syringae subsp . "tabaci" retains significant amounts of glutamine synthetase activity during toxin production in culture . As part of our investigation of the self-protection mechanism(s) used by these pathovars, we have determined that GS becomes adenylylated after toxin production is initiated and that the serine released from the zinc-activated hydrolysis of tabtoxin is a factor in the initiation of this adenylylation . The adenylylation state of this GS was estimated to range from E5.0-7.5 . The irreversible inactivation by tabtoxinine-beta-lactam of unadenylylated and adenylylated glutamine synthetase purified from P . syringae subsp . "tabaci" was investigated . Adenylylated GS was inactivated by tabtoxinine-beta-lactam at a slower rate than was unadenylylated enzyme . Adenylylated GS (E7.5-10.5) was significantly protected from this inactivation in the presence of the enzyme effectors, AMP, Ala, Gly, His, and Ser . Thus, the combination of the adenylylation of GS after toxin production is initiated and the presence of the enzyme effectors in vivo could provide part of the self-protection mechanism used by subsp . "tabaci".

Br J Cancer, 1986 Mar, 53(3), 377 - 84
The potential of carboxypeptidase G2-antibody conjugates as anti-tumour agents . I . Preparation of antihuman chorionic gonadotrophin-carboxypeptidase G2 and cytotoxicity of the conjugate against JAR choriocarcinoma cells in vitro; Searle F et al.; Carboxypeptidase G2, a zinc metalloenzyme isolated from Pseudomonas sp . strain RS-16, which catalyses the hydrolytic cleavage of reduced and non-reduced folates to pteroates and L-glutamate, has been linked to a monoclonal antibody (W14A) raised to human chorionic gonadotrophin . The coupling efficiency and retention of antibody and enzymatic activities are compared for three separate methods of preparing 1:1 conjugates . Preliminary in vitro studies on the cytotoxicity of the free enzyme and the conjugated enzyme towards JAR choriocarcinoma cells are reported . Despite the limitations of the in vitro model, it could be demonstrated that a significant proportion of 10(6) choriocarcinoma cells lost viability when exposed to either free or conjugated enzyme for 72 hours at concentrations of carboxypeptidase G2 of 1-3 units ml-1 of medium.

Laryngoscope, 1986 Mar, 96(3), 264 - 6
Necrotizing otitis externa occurring concurrently with epidermoid carcinoma; Mattucci KF et al.; Malignant external otitis (necrotizing otitis externa) is an infectious process which occurs most often in middle-aged and elderly diabetic patients and is characterized by cultures positive for Pseudomonas and granulation tissue at the junction of the bony and cartilaginous portion of the external auditory canal . Epidermoid carcinoma of the external auditory canal is also seen most often in the middle-aged and elderly population . To the best of our knowledge, no case of acute necrotizing otitis externa occurring concomitantly with epidermoid carcinoma of the external auditory canal has been documented . A case is presented and discussed here and the importance of biopsy of the external auditory canal is stressed . Theoretical considerations of the possible relationship between these two disorders are discussed . This case illustrates the need to discontinue the use of the term "malignant external otitis" and replace it with the term "necrotizing otitis externa."

Mikrobiologiia, 1986 Mar-Apr, 55(2), 231 - 6
{Characteristics of plasmid pBS271 controlling epsilon-caprolactam degradation by bacteria in the genus Pseudomonas}; Boronin AM et al.; Conjugative plasmids control the ability of five Pseudomonas strains isolated from the rectifiers of chemical plants to grow on epsilon-caprolactam as a sole carbon and nitrogen source . All the plasmids have a high molecular mass of their DNA (ca . 300 MDa) and control epsilon-caprolactam degradation at least to succinate . One of the plasmids (pBS271) belongs to the incompatibility group P-2 and suppresses the growth of a broad spectrum of temperate and virulent P . aeruginosa bacteriophages as well as that of some P . putida bacteriophages.

J Biochem (Tokyo), 1986 Mar, 99(3), 607 - 13
Oxidation of 1,5-anhydro-D-glucitol to 1,5-anhydro-D-fructose catalyzed by an enzyme from bacterial membranes; Nakamura T et al.; Bacteria which grow on 1,5-anhydro-D-glucitol (AG) were isolated from soil . One such strain showing the highest AG-assimilating activity was further characterized and identified as a new strain of the Pseudomonas family (named Pseudomonas sp . NK-85001) . A subcellular membranous fraction obtained from this strain catalyzed the oxidation of AG to 1,5-anhydro-D-fructose . This oxidation reaction consumed molecular oxygen as the terminal electron acceptor . The AG-oxidizing activity was further purified after solubilization . The AG oxidation catalyzed by this solubilized enzyme utilized molecular oxygen only in the presence of an electron mediator such as 2,6-dichlorophenolindophenol or phenazine methosulfate . Thus, the enzyme was suggested to be a dehydrogenase rather than an oxidase . The solubilized enzyme preparation also showed a strict substrate specificity . The observed specificity indicated that application of the enzyme for AG assay in clinical samples might be possible.

Biochem Int, 1986 Mar, 12(3), 413 - 20
6-diazo-5-oxo-L-norleucine and azaserine as affinity inhibitors of glutamin(asparagin)ase; Lebedeva ZI et al.; Incubation of homogeneous glutamin(asparagin)ase from Pseudomonas aurantiaca with 6-diazo-5-oxo-L-norleucine (DON) and azaserine leads to an almost complete inactivation of the enzyme . The inactivation process in both cases involves the step of reversible binding of the enzyme with the inhibitor into a complex and subsequent modification of the enzyme within this complex . The data on saturation of the enzyme by low concentrations of inhibitors, the protective effect of substrate and its analogs as well as of the competitive inhibitor and product of the enzymatic reaction, L-aspartate, suggest that the modification of functional groups takes place in the enzyme active site . The presence of essential threonine hydroxyl groups in/or near the enzyme active site is surmised.

Biochem Cell Biol, 1986 Mar, 64(3), 215 - 22
Gastric proteases of the Greenland cod Gadus ogac . II . Structural properties; Squires EJ et al.; Three gastric proteases were isolated from the stomach mucosa of the Greenland cod (Gadus ogac) . The cod proteases were all less stable to heating and protease 1 retained less activity at 5 degrees C when the pH was greater than 5 in comparison with porcine pepsin . The activities of cod proteases 1 and 2, with hemoglobin as the substrate, were doubled in the presence of 25 mM NaCl, while cod protease 3 and porcine pepsin were not stimulated by the salt . The cod proteases did not cross-react with antibodies raised against porcine pepsin . However, some cross-reactivity was noted with antibodies raised against proteases from psychotrophic pseudomonads . The molecular weights of all the cod proteases were in the range of 36,000-38,000 . The amino acid compositions of the cod proteases as compared by the Metzger difference index differed from the mammalian gastric proteases by about the same extent that pepsin, gastricsin, and chymosin differ from each other . Of the cod enzymes, protease 1 differed from mammalian gastric proteases, while cod proteases 3 was more like chymosin with respect to amino acid composition . Cod protease 1 had the lowest hydrophobicity index and chymosin had the highest . The hydrophobicity indices of cod proteases 2 and 3 were intermediate between that of porcine pepsin and bovine chymosin . It is suggested that the Greenland cod proteases represent less differentiated forms of gastric proteases than the mammalian pepsins, gastricsins, and chymosins.

J Gen Microbiol, 1986 Mar, 132 ( Pt 3), 737 - 42
Molecular size diversity of citrate synthases from Pseudomonas species; Mitchell CG et al.; Two forms of citrate synthase (EC 4.1.3.7) have been found in several species of Pseudomonas, a 'large' form (Mr congruent to 250,000) which is generally inhibited by NADH and reactivated by AMP, and a 'small' form (Mr congruent to 100,000) which is insensitive to these nucleotide effectors . Other species of Pseudomonas were found to contain either the 'large' or the 'small' form . Gel filtration and ion-exchange with the technique of fast protein liquid chromatography were used to resolve the enzymes . Where both citrate synthases were present, there did not appear to be an equilibrium between the two forms . The results reveal a new and complex diversity of citrate synthase within the genus Pseudomonas.

J Clin Microbiol, 1986 Mar, 23(3), 560 - 2
Production and utilization of pyochelin by clinical isolates of Pseudomonas cepacia; Sokol PA; Forty-three Pseudomonas cepacia isolates were screened for the production of pyochelin . Twenty-one (49%) produced pyochelin, and 22 (51%) were pyochelin negative . Of the 21 strains producing pyochelin, 18 were from patients with severe infections, 11 of which resulted in death . Of the 22 strains which did not produce pyochelin, 13 were from patients with mild or moderate infections . Pyochelin production by P . cepacia isolates infecting cystic fibrosis patients correlates with morbidity and mortality in these patients . Pyochelin was shown to stimulate the in vitro growth of P . cepacia in the presence of transferrin . P . cepacia isolates were able to accumulate 59Fe-pyochelin regardless of whether they produced this siderophore.

J Hosp Infect, 1986 Mar, 7(2), 189 - 92
Pseudomonas paucimobilis contamination of cool mist tents on a paediatric ward; Dale BA et al.; Investigations resulting from the isolation of Pseudomonas paucimobilis from the atomizer units of paediatric cool mist tents implicated the hospital water supply as the potential source of contamination . The pathogenic potential of the organism and its role in nosocomial infection is discussed.

J Biol Chem, 1986 Feb 15, 261(5), 2170 - 8
Binding of 17O-labeled substrate and inhibitors to protocatechuate 4,5-dioxygenase-nitrosyl complex . Evidence for direct substrate binding to the active site Fe2+ of extradiol dioxygenases; Arciero DM et al.; Pseudomonas testosteroni protocatechuate 4,5-dioxygenase catalyzes extradiol-type oxygenolytic cleavage of the aromatic ring of its substrate . The essential active site Fe2+ binds nitric oxide (NO) to produce an EPR active complex with an electronic spin of S = 3/2 . Hyperfine broadening of the EPR resonances of the nitrosyl complex of the enzyme by protocatechuate (3,4-(OH)2-benzoate, PCA) enriched specifically with 17O (I = 5/2) in either the 3 or the 4 hydroxyl group shows that both groups can bind directly to the Fe2+ in the ternary complex . Analogous results are obtained for PCA binding to catechol 2,3-dioxygenase-NO complex suggesting that substrate binding by the Fe2+ may be a general property of extradiol dioxygenases . The protocatechuate 4,5-dioxygenase inhibitor, 4-17OH-benzoate binds directly to the Fe of the nitrosyl adduct of the enzyme through the OH group . Since previous studies have shown that water also is bound to the Fe in this ternary complex, but not in the ternary complex with PCA, the data strongly imply that there are 3 sites in the Fe coordination which can be occupied by exogenous ligands . 3-17OH-benzoate is an inhibitor of the enzyme but does not elicit detectable hyperfine broadening in the EPR spectrum of the nitrosyl adduct suggesting that it binds to the enzyme, but not to the Fe . The EPR spectra of ternary enzyme-NO complexes with PCA or 4-OH-benzoate labeled with 17O exclusively in the carboxylate substituent are not broadened, suggesting that this moiety does not bind to the Fe.

Nucleic Acids Res, 1986 Feb 11, 14(3), 1293 - 301
Identification and characterisation of PmaCI an endonuclease of novel specificity from Pseudomonas maltophila; Walker JN et al.; We report the use of MonoQ FPLC (Fast Protein Liquid Chromatography) for the rapid purification of a novel Type II restriction endonuclease PmaCI, from Pseudomonas maltophila, which recognises the sequence 5'-CAC decreases GTG-3' . The resulting enzyme is free of other nucleases to a level suitable for its characterisation by multiple-substrate digestion and DNA sequencing techniques . This method appears to be widely applicable and we have used it for the isolation of restriction endonucleases of comparable purity from a range of other organisms . Also described is a rapid method for screening a library of small inserted regions in recombinant M13 molecules for the presence and subsequent screening of restriction sites of interest.

Appl Environ Microbiol, 1986 Feb, 51(2), 446 - 8
Susceptibility of psychrotrophic pseudomonads of milk origin to psychrotrophic bacteriophages; Patel TR et al.; A total of 47 psychrotrophic pseudomonads isolated from raw milk came from Newfoundland (19 isolates), British Columbia (6 isolates), Ontario (19 isolates), and Cork, Ireland (3 isolates) . The susceptibility of these was tested against 30 bacteriophages isolated from cold-storage beef . Distinct lysotypes were observed with isolates representing different geographic regions . Phages as agents in the control of bacterial populations in milk is discussed.

Appl Environ Microbiol, 1986 Feb, 51(2), 268 - 75
Existence of a novel enzyme, pyrroloquinoline quinone-dependent polyvinyl alcohol dehydrogenase, in a bacterial symbiont, Pseudomonas sp . strain VM15C; Shimao M et al.; A novel enzyme, pyrroloquinoline quinone (PQQ)-dependent polyvinyl alcohol (PVA) dehydrogenase, was found in and partially purified from the membrane fraction of a PVA-degrading symbiont, Pseudomonas sp . strain VM15C . The enzyme required PQQ for PVA dehydrogenation with phenazine methosulfate, phenazine ethosulfate, and 2,6-dichlorophenolindophenol as electron acceptors and did not show PVA oxidase activity leading to H2O2 formation . The enzyme was active toward low-molecular-weight secondary alcohols rather than primary alcohols . A membrane-bound PVA oxidase was also present in cells of VM15C . Although the purified oxidase showed a substrate specificity similar to that of PQQ-dependent PVA dehydrogenase and about threefold-higher PVA-dehydrogenating activity with phenazine methosulfate or phenazine ethosulfate than PVA oxidase activity with H2O2 formation, it was shown that the enzyme does not contain PQQ as the coenzyme, and PQQ did not affect its activity . Incubation of the membrane fraction of cells with PVA caused a reduction in the cytochrome(s) of the fraction.

Exp Cell Res, 1986 Feb, 162(2), 436 - 48
Thioridazine enhances lysosomal accumulation of epidermal growth factor and toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin; Kuratomi Y et al.; Thioridazine, a phenothiazine calmodulin inhibitor, aggravated the cytotoxic effect of a conjugate (EGF-PE) of epidermal growth factor (EGF) coupled with Pseudomonas exotoxin against cultured HeLa cells . Other phenothiazine calmodulin inhibitors, trifluoperazine and chlorpromazine, also intensified the cytotoxic effect of EGF-PE, whereas N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W7) had no such effect . By using iodinated epidermal growth factor ( {125I}EGF), the effect of thioridazine on intracellular transport of EGF was examined . The release of radioactivity associated with {125I}EGF into medium was slow in the presence of thioridazine . The Percoll gradient centrifugation pattern showed that thioridazine delayed both the appearance of {125I}EGF in lysosomes and the disappearance of {125I}EGF from the lysosomes . The pH value in lysosomes was 5.28 in thioridazine-treated HeLa cells, while that in untreated cells was 5.15 . Thioridazine was found to inhibit lysosomal enzyme activities of cathepsin B and acid phosphatase, but not beta-hexosaminidase when cell extracts were treated with the drug . Electron microscopy showed an increased number of electron-dense bodies, possibly autophagosomes/lysosomes in HeLa cells grown for 48 h with 3 micrograms/ml thioridazine . The potentiating action of EGF-PE by thioridazine is discussed in relation to the altered lysosomal function in treated cells.

Aust N Z J Med, 1986 Feb, 16(1), 75 - 7
Brain abscess due to Pseudomonas pseudomallei; Lee MK et al.; A brain abscess due to Pseudomonas pseudomallei is presented . Central nervous system involvement in melioidosis is rare and generally occurs in association with disseminated infection . The importance of early diagnosis and prompt treatment with prolonged antibiotic therapy is emphasised.

J Bacteriol, 1986 Feb, 165(2), 585 - 94
Siderophores and outer membrane proteins of antagonistic, plant-growth-stimulating, root-colonizing Pseudomonas spp; de Weger LA et al.; As an approach to understanding the molecular basis of the reduction in plant yield depression by root-colonizing Pseudomonas spp . and especially of the role of the bacterial cell surfaces in this process, we characterized 30 plant-root-colonizing Pseudomonas spp . with respect to siderophore production, antagonistic activity, plasmid content, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis patterns of their cell envelope proteins . The results showed that all strains produce hydroxamate-type siderophores which, because of the correlation with Fe3+ limitation, are thought to be the major factor responsible for antagonistic activity . Siderophore-negative mutants of two strains had a strongly decreased antagonistic activity . Five strains maintained their antagonistic activity under conditions of iron excess . Analysis of cell envelope protein patterns of cells grown in excess Fe3+ showed that most strains differed from each other, although two classes of similar or identical strains were found . In one case such a class was subdivided on the basis of the patterns of proteins derepressed by iron limitation . Small plasmids were not detected in any of the strains, and only one of the four tested strains contained a large plasmid . Therefore, it is unlikely that the Fe3+ uptake system of the antagonistic strains is usually plasmid encoded.

J Bacteriol, 1986 Feb, 165(2), 534 - 41
Indigenous plasmids in Pseudomonas syringae pv . tomato: conjugative transfer and role in copper resistance; Bender CL et al.; Twenty strains of Pseudomonas syringae pv . tomato were examined for the presence of plasmid DNA . P . syringae pv . tomato plasmids were grouped into five size classes: class A ranged from 95 to 103 kilobases (kb); class B ranged from 71 to 83 kb; class C ranged from 59 to 67 kb; class D ranged from 37 to 39 kb; and class E was 29 kb . All strains contained at least two plasmids in classes A and B . The conjugative ability of P . syringae pv . tomato plasmids in three strains was demonstrated by mobilization of the nonconjugative plasmid RSF1010 into Pseudomonas syringae pv . syringae recipients . Plasmids from the three conjugative strains were labeled with Tn5 . Four conjugative plasmids were identified by their repeated transfer to P . syringae pv . syringae recipients . P . syringae pv . tomato strains varied in sensitivity to copper sulfate (CuSO4): MICs were 0.4 to 0.6 mM for sensitive strains, 1.2 mM for moderately resistant strains, and 1.6 to 2.0 mM for very resistant strains . One very resistant strain, PT23, functioned as a donor of copper resistance . Recipient P . syringae pv . syringae strains PS51 and PS61 were inhibited by 0.1 mM CuSO4, whereas the CuSO4 MICs for transconjugant strains PS51(pPT23A) and PS61(pPT23C) were 1.8 and 2.6 mM, respectively . P . syringae pv . tomato strains PT12.2 and PT17.2 were inhibited by 0.6 mM copper sulfate, but their copper sulfate MICs were 2.6 and 1.8 mM, respectively, when they acquired pPT23C . Therefore, copper resistance in PT23 was controlled by two conjugative plasmids, designated pPT23A (101 kb) and pPT23C (67 kb).

Am J Ophthalmol, 1986 Jan 15, 101(1), 41 - 3
Pseudomonas keratitis and folliculitis from whirlpool exposure; Insler MS et al.; A local outbreak of pseudomonal folliculitis from whirlpool exposure occurred in 12 persons . A corneal ulcer developed in one patient within 48 hours of using the whirlpool . Treatment with fortified gentamicin resolved the corneal infiltrate and vision returned to normal . A second patient, who had not showered immediately after leaving the whirlpool, was left with areas of skin hyperpigmentation and scarring despite treatment with ultraviolet rays and tetracycline . Although the skin rash may be self-limited, the potential for visual loss from pseudomonal keratitis emphasizes the need for proper disinfection of swimming and whirlpool water.

Biochem J, 1986 Jan 15, 233(2), 493 - 7
Inactivation of 3 alpha-hydroxysteroid dehydrogenase by superoxide radicals . Modification of histidine and cysteine residues causes the conformational change; Kim HS et al.; 3 alpha-Hydroxysteroid dehydrogenase (EC 1.1.1.50) from Pseudomonas testosterone was inactivated by superoxide radicals generated by the aerobic xanthine oxidase reaction . Superoxide dismutase, NAD+, bovine serum albumin and histidine and cysteine as free amino acids partially protected the enzyme from inactivation . NADH-binding properties were determined by fluorescence spectroscopy, and no variation was found between native enzyme and the unmodified fraction of the partly inactivated one . The fluorescence emission maximum for the completely inactivated enzyme was shifted 10 nm to a longer wavelength when compared with the native one, and it seems possible that the modification of histidine and cysteine residues by superoxide radicals causes the conformational change of the enzyme and the consequent loss of catalytic activity.

J Adolesc Health Care, 1986 Jan, 7(1), 57 - 9
Malignant external otitis in a diabetic adolescent; Benuck I et al.; Malignant external otitis (MEO) is an unusual medical problem . The case reported is that of a diabetic adolescent who presented with severe ear pain unresponsive to oral antibiotics and analgesics . The diagnosis of MEO was made, and he was successfully treated with a combination of intravenous anti-Pseudomonas agents . A review of the pediatric cases, guidelines for diagnosis, length of treatment, and prognosis are presented.

J Trauma, 1986 Jan, 26(1), 34 - 9
Early protein alteration in blister fluid and serum associated with burn injury; Deitch EA et al.; Since the most common site of infection in burned patients is the burn wound, we have previously studied the biologic effect of burn wound blister fluid (BF) on control lymphocyte and neutrophil activity . BF will not support the phagocytosis of Pseudomonas by normal neutrophils, and a subset of the BF samples suppressed the maximal mitogen response of control lymphocytes by more than 50% . The current work was carried out to analyze in depth the composition of BF using crossed immunoelectrophoresis . Twenty BF and six serum specimens collected from 20 patients between 6 and 18 hours postburn had 19 separate proteins measured . These proteins included immunologic proteins, antiproteases, acute-phase reactants, carrier proteins, and lipoproteins . A total of 546 protein measurements were made . The concentrations of all subgroups of proteins were significantly lower in the BF and burn serum specimens than in control serum . When the blister fluids were stratified according to their effect on normal lymphocyte activity, the suppressive blister fluid samples had higher levels of C3 and lower levels of I alpha I than the nonsuppressive samples . The elevation in C3 was secondary to the local activation of C3 and the generation of multiple C3 breakdown products . These changes in C3 are of potential biologic importance, since evidence has accumulated indicating that the various fragments of C3 can modulate both neutrophil and lymphocyte function . Thus, the results of this study suggest that local changes in the blister fluid may adversely affect local immunity and predispose the patient to burn wound sepsis.

Pediatr Pathol, 1986, 6(2-3), 131 - 7
Perinatal mortality associated with intrauterine infection due to pseudomonads; Turkel SB et al.; Pseudomonads are common causes of nosocomial infections but are rarely implicated in perinatal disease . In a retrospective autopsy study we found that 9% of all acute congenital bacterial infections were due to Pseudomonas species . Premature rupture of membranes occurred in half the cases and clinical maternal amnionitis in two-thirds . One case was apparently nosocomial in origin . No known risk factors were implicated in any other case . Seven infants were stillborn and two died within a few hours . Congenital pneumonia, funisitis, and chorioamnionitis were found at autopsy . Intrauterine infection due to the pseudomonads poses a serious problem that has not been previously recognized.

Mikrobiologiia, 1986 Jan-Feb, 55(1), 96 - 9
{Numerical taxonomy of pseudomonads of the diminuta group}; Pavlenko GV et al.; The taxonomy of the "diminuta" group is discussed . The method of numerical taxonomy was used to characterize 135 strains of 10 species belonging to Pseudomonas, Gluconobacter and Acetobacter in terms of sixty phenotypical features . The similarity coefficients of the strains were calculated with computers . According to the data of numerical analysis, the species P . diminuta and P . vesiculare represent a single phenon different from Pseudomonas, Gluconobacter and Acetobacter species.

Cold Spring Harb Symp Quant Biol, 1986, 51 Pt 2, 879 - 90
P450 genes: evolution, regulation, and relationship to human cancer and pharmacogenetics; Gonzalez FJ et al.; Three pharmacogenetic differences (the acetylation, debrisoquine 4-hydroxylase, and Ah locus polymorphisms) appear to be associated with increased risk of environmentally caused human cancer . The latter two polymorphisms represent differences in P450 gene expression . It is predicted that molecular biological studies will soon provide a means (RFLP patterns or expression vector assays) of predicting individual cancer risk related to these polymorphisms . At the present time, only the Ah locus polymorphism has been explored at the molecular biological level . More than 30 P450 genes, including eight from the human, have been isolated and sequenced to date . The P450 gene superfamily comprises at least eight families, including one gene family that has diverged more recently into at least five subfamilies . The P450 gene superfamily can be regarded as ancient, with Pseudomonas and human amino acid sequences displaying a small but significant resemblance in the region of the enzyme active-site . P450 enzymic activity can be depressed 30-60% by immunosuppressive agents . The mechanism of this effect is unknown . This phenomenon can become clinically important if a cancer patient who is taking any chemotherapeutic drug that is detoxified by P450 then receives one of these agents . The Ah receptor-controlled gene expression, positive and negative control elements, and negative autoregulatory loop--best characterized in the mouse P(1)450 gene and upstream sequences, plus the receptor-defective and the P(1)450 enzymic activity-negative mutants in mouse cell cultures--provide the molecular geneticist with an exciting model system for studying the regulation of a clinically relevant gene family . The high degree of homology in a 220-bp segment between mouse and human P(1)450 sequences (about 1140 to 920 bases upstream from the mRNA cap site) suggests that this positive regulatory element (dioxin-inducible enhancer which also controls constitutive gene expression) is conserved between Mus domesticus and Homo sapiens.

Acta Otorhinolaryngol Belg, 1986, 40(4), 635 - 43
{Malignant otitis externa}; Ninove B et al.; Malignant external otitis is a progressive Pseudomonas infection starting in the external auditory canal . It tends to occur in the elderly diabetic patient and to invade cartilage, bone, nerves and adjacent soft tissues . Aggressive medical management and surgical debridement is curative in most cases as it appears in the case reported in the following communication.

J Pediatr Gastroenterol Nutr, 1986 Jan, 5(1), 97 - 102
Prognostic factors associated with patient survival during nutritional rehabilitation in malnourished children and adolescents with cystic fibrosis; Levy L et al.; Nineteen children and adolescents with cystic fibrosis and malnutrition were given intensive nutritional support in an effort to reverse malnutrition . Standard techniques of enteral and parenteral feeding were used . As 10 of 19 patients died shortly after nutritional intervention began, we retrospectively analyzed patient data in order to discover whether or not any patient characteristics were associated with survival . Using a multivariate analysis, a linear discriminant function was derived employing average heart rate, the presence or absence of Pseudomonas cepacia in the sputum culture, PaCO2, and the patient's age at the time of intervention . This function correctly classified these 19 patients . A further 10 nonselected CF patients who also received nutritional support were similarly assessed using the function, and again, survival was accurately predicted . Therefore, this function can be used to predict the likelihood of patient survival during the provision of advanced nutritional support . It may be helpful in deciding whether or not advanced nutritional support is warranted in certain CF patients.

Trans Ophthalmol Soc U K, 1986, 105 ( Pt 1), 26 - 31
Pathogenic mechanisms of organisms virulent to the eye; Watt PJ; Gonococci possess long range adhesins in the form of pili permitting initial contact with conjunctival cells . Subsequently sticky surface protein (Protein II) bonds the gonococcus close to the host cell surface . Damage is mediated both by the intracellular uptake and the introduction of pores in the host cell membrane . Unlike gonococci, pseudomonas can only attach to damaged cells but once the eye is invaded a wide range of enzymes and toxins leads to rapid tissue destruction . The mechanisms by which Chlamydia trachomatis induce trachoma are ill-understood but involve intense antigenic stimulation . Methods are now available to investigate the antigenic structure of chlamydia at sub-molecular levels; such an approach is required for the development of a vaccine against trachoma.

Biochemistry, 1985 Dec 17, 24(26), 7511 - 6
Use of a solid-phase photoaffinity reagent to label a steroid binding site: application to the delta 5-3-ketosteroid isomerase of Pseudomonas testosteroni; Hearne M et al.; In order to extend our analysis of the reactions that occur during the active site directed photoinactivation of delta 5-3-ketosteroid isomerase sensitized by unsaturated steroid ketone photoaffinity reagents, the site of covalent attachment has been identified . A solid-phase photoaffinity reagent, delta 6-testosterone-agarose, has been employed for this purpose; this type of reagent, in contrast to solution-phase reagents, facilitated the recovery of a peptide fragment of the isomerase bearing the residue at which covalent attachment had occurred . Amino acid analysis and sequence determination of the peptide provided evidence that the site of attachment was aspartate-38 . This result, in combination with the low-resolution crystallographic structure of the enzyme {Westbrook, E . M., Piro, O . E., & Sigler, P . B . (1984) J . Biol . Chem . 259, 9096-9103}, suggests that aspartate-38 is located in the vicinity of the bottom of the steroid-binding pit . The potential usefulness of solid-phase photoaffinity reagents in the identification of sites of covalent attachment on target proteins such as hormone receptors is discussed.

Biol Chem Hoppe Seyler, 1985 Dec, 366(12), 1085 - 91
Purification and properties of bromoperoxidase from Pseudomonas pyrrocinia; Wiesner W et al.; A bromoperoxidase was purified and partially characterized from Pseudomonas pyrrocinia ATCC 15958, a bacterium that produces the antifungal antibiotic pyrrolnitrin . The purified enzyme preparation was homogeneous as determined by polyacrylamide gel electrophoresis and ultracentrifugation . The molecular mass of the enzyme was estimated to be 154 kDa +/- 3 kDa as determined by gel filtration and ultracentrifugation . Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed a single band with the mobility of a 76-kDa species . Therefore, in solution at neutral pH, bromoperoxidase exists as a dimeric species . The isoelectric point was 5.0 . The prosthetic group of this procaryotic bromoperoxidase was ferriprotoporphyrin IX . The spectral properties of the native and reduced enzyme are reported . The purified enzyme showed brominating as well as peroxidase and catalase activity.

Asian Pac J Allergy Immunol, 1985 Dec, 3(2), 200 - 4
A case comparison of acquired immune deficiency syndrome (AIDS) in homosexual males with spindle-endothelial cell abnormalities and with recrudescent melioidosis; Tanphaichitra D et al.; The AIDS syndrome includes cases of biopsy-proven Kaposi's sarcoma in persons under 60 years of age, or biopsy- or culture-proven Pneumocystis carinii pneumonia, or either of the life-threatening opportunistic infections in young previously healthy persons with no underlying cause of immunodeficiency (Center for Disease Control criteria) . Here we described the first case of AIDS with early Kaposi's sarcoma-like lesions in homosexual male drug addict and have compared the clinical and laboratory findings with those of another homosexual male having recrudescent melioidosis due to Pseudomonas pseudomallei.

Ann Ophthalmol, 1985 Dec, 17(12), 753 - 6
Pseudomonas cepacia endophthalmitis; Del Piero E et al.; A 72-year-old white man who had undergone surgical trabeculectomy and extracapsular cataract extraction with a posterior-chamber lens implantation in the left eye suffered from chronic iridocyclitis for eight months . He subsequently presented with acute hypopyon and vitritis . Anterior-chamber and vitreous cultures were positive for Pseudomonas cepacia . The infection was successfully treated with subconjunctival piperacillin, intravitreal cefotaxime, and intravenous piperacillin and gentamicin . To our knowledge, this is the first documented case of Pseudomonas cepacia endophthalmitis.

J Biochem (Tokyo), 1985 Dec, 98(6), 1719 - 22
One of two copper atoms is not necessary for the cytochrome c oxidase activity of Pseudomonas AM 1 cytochrome aa3; Fukumori Y et al.; From Pseudomonas AM 1 grown in a medium deficient in Cu, aa3-type cytochrome c oxidase was purified which contained 2 molecules of haem a and one atom of Cu per molecule . The enzyme showed absorption peaks at 428 and 595 nm in the oxidized form and at 442 and 604 nm in the reduced form, and its CO complex showed peaks at 432 and 602 nm . The enzyme in the oxidized state showed an obscure absorption peak around 800 nm instead of a peak at 820 nm . One mol of the enzyme oxidized maximally 76, 75, and 98 mol of the ferrocytochromes c of Candida krusei, horse and Pseudomonas AM 1 per sec, respectively . These reactions were 50% inhibited by 7 microM KCN . The product of reduction of O2 catalyzed by the enzyme was concluded to be H2O on the basis of the ratio of ferrocytochrome c oxidized to O2 consumed.

J Biol Chem, 1985 Nov 25, 260(27), 14633 - 5
Crystallographic characterization of phthalate oxygenase reductase, an iron-sulfur flavoprotein from Pseudomonas cepacia; Correll CC et al.; Pseudomonads grown on phthalate synthesize a series of enzymes that metabolize this aromatic substrate . Among the inducible enzymes is a reductase which transfers electrons from NADH to the terminal dioxygenase that converts phthalate to the corresponding cis-1,2-dihydrodiol (Keyser, P . (1976) Ph . D . thesis, University of Miami; Ribbons, D . W., and Evans, W . C . (1960) Biochem . J . 76, 310-318) . The phthalate oxygenase reductase induced in Pseudomonas cepacia is a single polypeptide chain (Mr approximately equal to 33,000) with two prosthetic groups, FMN and {2Fe-2S} . This oxidoreductase has been crystallized at pH 6.7 from polyethylene glycol 6000 in space group R3 with a = b = 113.4 A and c = 77.7 A (hexagonal indexing).

Eur J Biochem, 1985 Nov 15, 153(1), 75 - 80
Isolation and characterization of a blue copper protein from Thiobacillus versutus; van Houwelingen T et al.; The blue copper protein induced during growth of Thiobacillus versutus on methylamine was purified and characterized . It is an acidic protein (isoelectric point 4.7), contains one Cu2+ ion/enzyme molecule, is a monomeric protein (molecular mass about 14 kDa), has a maximum in its absorption spectrum at 596 nm (molar absorption coefficient 3.9 X 10(3) M-1 cm-1), shows an axial type-I electron paramagnetic resonance spectrum (g parallel = 2.239, g perpendicular = 2.046 and A parallel = 5.6 mT) and has a redox potential (Eo) of + 260 mV . In view of these properties and in view of the fact that the protein is active as an electron carrier between methylamine dehydrogenase and cytochrome c, it is concluded that it is similar to the amicyanins isolated from Methylomonas sp . strain J and Pseudomonas sp . strain AM 1.

Infect Control, 1985 Nov, 6(11), 451 - 5
Biological sterilization monitors: a four-year in-use evaluation of two systems; Kotilainen HR et al.; Biological monitors (BI) are considered to be the best monitor of the sterilization process yet false positives may result in recalls and quality assurance difficulties . To assess the frequency, type and reasons for questionable results, we undertook a 4-year in-use study of two commonly used BI types--spore strips (Spordi) and a self-contained crushable ampule (Attest)--for both steam and ethylene oxide (EO) . After laboratory verification of time/kill ratios for a portion of each involved lot, 2 BI of each type were placed in test pack within a randomly selected load run at standard time and temperature . All resulting positive BI were subcultured . Steam cycle positives were uncommon (32/1,1710 positive Spordi, 1.9%; 20/1,710 positive Attest, 1.2%) and could be related to chamber temperature or steam quality . All of the 4 BI per load were positive in only three loads; physical monitors indicated gross malfunction . Five positive Spordi were due to either contaminants or a malfunctioning incubator . EO-related positives were more common (53/1,109 positive Spordi, 4.8%; 25/1,109 positive Attest, 2.3%) . One-half of the Spordi tests became positive after 48 hours of incubation . Organisms other than B . subtilis were recovered from 49.1% of the positive tests (26/53) . The Attest was remarkable for its lack of contamination; 1/25 was positive for Pseudomonas stutzeri only . More positives were observed during the winter months when relative humidity was below 20% . This finding was more commonly observed with the EO Attest . In summary, we found no significant difference in the performance of either BI.(ABSTRACT TRUNCATED AT 250 WORDS)

Ophthalmology, 1985 Nov, 92(11), 1542 - 9
Necrotizing scleritis . A clinico-pathologic study of 41 cases; Rao NA et al.; Scleritis is not a single clinical or pathologic entity . It has several distinct forms . A clinico-pathologic study of 41 cases of necrotizing scleritis suggests that these different histopathologic forms of disease may reflect different mechanisms of immunopathogenesis . These cases were divided into three main groups: (1) scleral inflammations associated with various systemic autoimmune diseases, including 11 cases of rheumatoid arthritis, three cases of Wegener's granulomatosis, one case of polychondritis, and one case of Goodpasture's syndrome; (2) infectious scleritis, consisting of four cases of herpes zoster ophthalmicus and two cases of pseudomonas scleritis; and (3) idiopathic scleritis, without evidence of systematic disorder, consisting of 19 cases . The first group exhibited predominantly necrosis of the sclera surrounded by granulomatous inflammation and vasculitis . None of these cases showed lymphoid follicles, or healing attempts manifested by proliferation of fibroblasts and blood vessels at the site of inflammation . The idiopathic group revealed few small foci of scleral necrosis and mainly non-granulomatous inflammation . In addition, there was evidence of proliferation of granulation tissue and lymphoid follicles in this group of eyes.

J Bacteriol, 1985 Nov, 164(2), 823 - 30
Nitrous oxide reduction by members of the family Rhodospirillaceae and the nitrous oxide reductase of Rhodopseudomonas capsulata; McEwan AG et al.; After growth in the absence of nitrogenous oxides under anaerobic phototrophic conditions, several strains of Rhodopseudomonas capsulata were shown to possess a nitrous oxide reductase activity . The enzyme responsible for this activity had a periplasmic location and resembled a nitrous oxide reductase purified from Pseudomonas perfectomarinus . Electron flow to nitrous oxide reductase was coupled to generation of a membrane potential and inhibited by rotenone but not antimycin . It is suggested that electron flow to nitrous oxide reductase branches at the level of ubiquinone from the previously characterized electron transfer components of R . capsulata . This pathway of electron transport could include cytochrome c', a component hitherto without a recognized function . R . capsulata grew under dark anaerobic conditions in the presence of malate as carbon source and nitrous oxide as electron acceptor . This confirms that nitrous oxide respiration is linked to ATP synthesis . Phototrophically and anaerobically grown cultures of nondenitrifying strains of Rhodopseudomonas sphaeroides, Rhodopseudomonas palustris, and Rhodospirillum rubrum also possessed nitrous oxide reductase activity.

J Gen Virol, 1985 Nov, 66 ( Pt 11), 2461 - 9
Function of pili in bacteriophage phi 6 penetration; Romantschuk M et al.; The genome of bacteriophage phi 6, which has a lipid protein envelope, consists of three pieces of dsRNA . Virus infection is initiated by attachment to a phi 6-specific host pilus followed by fusion of the phage membrane and the bacterial outer membrane . In this study we analysed several different phi 6 hosts as well as more than 200 independently isolated phi 6-resistant variants derived from Pseudomonas syringae pv . phaseolicola . It is shown that phi 6-specific pili are coded by genes located in the host chromosome . It appears that pilus reaction is needed to pull the pilus-associated virus through the extracellular polysaccharide of the host and thus to bring it into contact with the outer membrane where membrane fusion can take place.

J Bacteriol, 1985 Nov, 164(2), 882 - 6
Occurrence of succinyl derivatives in the catabolism of arginine in Pseudomonas cepacia; Vander Wauven C et al.; Pseudomonas cepacia NCTC 10743 utilizes arginine as the sole source of carbon and nitrogen for growth . Arginine is degraded to glutamate via succinyl derivatives . The catabolic sequence in this pathway is L-arginine----N2-succinylarginine----N2-succinylornithine--- -N2-succinylglutamate semialdehyde----N2-succinylglutamate----glutamate + succinate . The formation of the enzymes responsible for arginine degradation is regulated not only by induction but also by both carbon and nitrogen catabolite repression.

Zh Mikrobiol Epidemiol Immunobiol, 1985 Nov, (11), 23 - 6
{The polysaccharide component of Pseudomonas pseudomallei slime}; Denisov II; The polysaccharide component contained in the slime of the causative agent of melioidosis was obtained . This component was found to be a biopolymer, mainly of the carbohydrate nature, consisting of galactose, glucose, mannose, rhamnose and two unidentified carbohydrates . The slime polysaccharide component contained two thermostable and acid-resistant antigens . The action of alkali led to the loss of one of these antigens . Rabbit antisera to the preparations of the slime polysaccharide component with titers of 1:64 to 1:256, determined in the immunodiffusion test, were obtained . In the precipitation test the slime polysaccharide component reacted with antisera from sick experimental animals, thus confirming the suggestion of its secretion in the process of the development of melioidosis infection.

J Clin Microbiol, 1985 Oct, 22(4), 490 - 4
Typing of Pseudomonas cepacia by bacteriocin susceptibility and production; Govan JR et al.; The significance of Pseudomonas cepacia as an opportunistic pathogen in immunocompromised patients has become increasingly recognized . Particularly disturbing is its increased incidence, reported by several North American centers, in respiratory tract cultures from patients with cystic fibrosis . Epidemiological studies of P . cepacia have been hampered by a lack of typing methods . In this paper we report the development of a typing scheme based on bacteriocin production and susceptibility . For bacteriocin production, test isolates of P . cepacia were rapidly applied to the surfaces of agar plates with a multiple inoculator . After incubation of these test isolates for 5.5 h and their exposure to chloroform, indicator strains were applied in agar overlays without prior removal of the test strain growth . After 18 h of incubation, inhibition zones caused by bacteriocin activity were recognized . A similar procedure was used to examine the bacteriocin susceptibility of the test strain . The bacteriocin type of the test strain was defined based on its bacteriocin production as judged by zones of inhibition against a set of eight indicator strains and by susceptibility or resistance of the test strain to bacteriocin produced by six producer strains . Of 373 strains of P . cepacia, 95.2% were typed into a total of 44 type combinations . Bacteriocin typing provided a suitable procedure for epidemiological studies of colonization or infection by P . cepacia . The technique described in this paper was simple to perform, gave a result within 24 h, provided good strain discrimination, and was suitable for clinical, environmental, and phytopathogenic strains.

J Steroid Biochem, 1985 Oct, 23(4), 523 - 8
Extraction of a steroid transport system from Pseudomonas testosteroni membranes and incorporation into synthetic liposomes; Francis MM et al.; A steroid binding protein has been extracted from Pseudomonas testosteroni membranes with an organic solvent system . This protection binds some C19 and C21 steroids but not C18 steroids . When this protein is incorporated into synthetic lipid vesicles constructed from P . testosteroni phospholipids, the vesicles perform concentrative uptake of testosterone in the presence of the ionophore valinomycin . This steroid binding protein is thus believed to be the steroid permease of this organism.

Genetika, 1985 Oct, 21(10), 1627 - 33
{Tryptophan of facultative methylotrophic bacteria Pseudomonas sp . M . II . Regulation of tryptophan biosynthesis}; Olekhnovich IN et al.; Regulation of tryptophan biosynthesis of facultative methylotrophic Pseudomonas sp . M was studied . Repression of the trpE, trpD and trpC genes by tryptophan was demonstrated . It was also shown that the trpE and trpDC genes are derepressed noncoordinately . No regulation of the trpF gene product could be demonstrated, indicating that its synthesis is constitutive . The trpA and trpB genes are inducible by indol-3-glycerophosphate . Anthranilate synthase and tryptophan synthase were sensitive to the feedback inhibition . The tryptophan concentrations giving 50% inhibition were estimated to be 9 microM and 1 microM, respectively . Experimental evidence for activation of the N-5-phosphoribosyl anthranilate isomerase and for inhibition of the indol-3-glycerophosphate synthase by some tryptophan intermediates was obtained.

J Biol Stand, 1985 Oct, 13(4), 321 - 6
Serological failure of Pseudomonas vaccination in patients receiving multiple chemotherapy; Bannister P et al.; Publication Types:

	Clinical Trial
	Controlled Clinical Trial

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