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 i