Appl Environ Microbiol, 1985 Oct, 50(4), 1038 - 42 Ultrastructural localization of an extracellular protease in Pseudomonas fragi by using the peroxidase-antiperoxidase reaction; Thompson SS et al.; An extracellular protease, which previously has been found to correlate with the appearance of bleblike evaginations on the cell wall of Pseudomonas fragi ATCC 4973, was purified 38-fold by ammonium sulfate precipitation and Sephadex chromatography to yield a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Polyclonal rabbit antiserum raised against the purified enzyme had an enzyme-linked immunosorbent assay titer of 4 X 10(7) . The peroxidase antiperoxidase method was used to localize the neutral protease in P . fragi at the ultrastructural level . Electron microscopy of cell sections of this organism revealed that high concentrations of positive immunoperoxidase reaction product were located near the cell wall, whereas control sections stained with preimmune or heterologous serum did not show similar deposits to be present . These results are consistent with the hypothesis that blebs appearing on P . fragi contain high concentrations of neutral protease. J Biochem (Tokyo), 1985 Oct, 98(4), 1033 - 40 The reaction mechanisms and substrate-stereospecificities of L-alloisocitrate dehydrogenase and oxalosuccinate decarboxylase; Hoshiko S et al.; A sequential reaction was suggested for the conversion of L-alloisocitrate to alpha-oxoglutarate by an enzyme complex of L-alloisocitrate dehydrogenase and oxalosuccinate decarboxylase from Pseudomonas strain No . 2, during which oxalosuccinate was not released from the enzyme-substrate complex . The stereochemistry of oxalosuccinate formed by L-alloisocitrate dehydrogenase and decarboxylated by oxalosuccinate decarboxylase was opposite to that of the substrate for D-isocitrate dehydrogenase . Incubation of L-alloisocitrate with the dehydrogenase and decarboxylase in deuterium oxide provided {3-2H}-alpha-oxoglutarate, the configuration of which turned out to be the same as that produced by D-isocitrate dehydrogenase from D-isocitrate . The data suggested that enol form of alpha-oxoglutarate was involved as an intermediate in decarboxylation of oxalosuccinate by oxalosuccinate decarboxylase . L-Alloisocitrate dehydrogenase was shown to react with pro-S proton of NADH. J Bacteriol, 1985 Oct, 164(1), 14 - 8 Isolation and characterization of Tn5 insertion mutants of Pseudomonas syringae pv . syringae altered in the production of the peptide phytotoxin syringotoxin; Morgan MK et al.; A syringotoxin-producing strain of Pseudomonas syringae pv . syringae (B457) was subjected to Tn5 mutagenesis by the transposon vector pSUP1011 . Analyses of auxotrophs obtained suggested simple random insertions of Tn5 . Syringotoxin-negative mutants arose at a frequency of about 0.28% . In a Southern blot analysis, the loss of toxin production was associated with Tn5 insertions into chromosomal EcoRI fragments of about 10.5, 17.8, and 19.3 kilobases . Data from a Southern blot analysis of SstI-digested DNA from these mutants suggest that the 10.5- and 17.8-kilobase EcoRI fragments may be adjacent to or near each other . Mutants that produced only 3 to 4% wild-type toxin levels also were identified. Pediatr Pulmonol, 1985 Sep-Oct, 1(5), 238 - 43 Increased dosage requirements of tobramycin and gentamicin for treating Pseudomonas pneumonia in patients with cystic fibrosis; Mann HJ et al.; The pharmacokinetic behavior of tobramycin and gentamicin was evaluated in 27 patients who had cystic fibrosis (CF) . A previously studied, age-matched group of 334 patients who had been treated with gentamicin and who did not have CF served as controls . The CF patients, who ranged in age from 2 to 32 years and who had normal renal function, received 36 treatment courses with either tobramycin (19) or gentamicin (17) to treat Pseudomonas pneumonia . Serum concentrations were determined after a 1.5-mg/kg dose to compute half-life (t 1/2), elimination rate constant (k), and apparent volume of distribution (V) . From these values, doses were calculated to produce steady-state peak concentrations of 8.0 micrograms/ml with a dosing interval of every six hours . For tobramycin the mean (+/- SD) t1/2 was 1.0 (0.4) hours, V was 0.18 (0.06) l/kg, total body clearance (TBC) was 2.19 (0.71) ml/min/kg, and the calculated dose was 8.2 (2.1) mg/kg/day . For gentamicin t1/2 was 1.1 (0.5) hours, V was 0.20 (0.06) l/kg, TBC was 2.28 (0.89) ml/min/kg, and the calculated dose was 8.8 (2.4) mg/kg/day . The pharmacokinetic parameters were not statistically different between the two drugs, but the mean values of t1/2 and TBC of CF patients differed significantly from those of the control group . The calculated doses were larger than the manufacturer's maximum recommended dose of 7.5 mg/kg/day for 63% of tobramycin and 71% of gentamicin treatment courses . A dosing interval change to every four hours would have been appropriate in 28 of the 36 treatment courses (78%).(ABSTRACT TRUNCATED AT 250 WORDS) Arch Microbiol, 1985 Sep, 142(4), 365 - 9 Effects of myxothiazol and 5-undecyl-6-hydroxy-4,7-dioxobenzothiazole on the respiratory pathways of the phytopathogenic fluorescent bacteria Pseudomonas cichorii and Pseudomonas aptata; Cocchi S et al.; Myxothiazol inhibited the electron transport in the cytochrome b/c segment of membrane particles from Pseudomonas cichorii . A residual NADH-oxidation due to the presence of an alternative pathway via cytochrome o (Em, 7 = +250 mV) was sensitive to the quinone analog 5-undecyl-6-hydroxy-4,7-dioxobenzothiazole (UHDBT) . This latter inhibitor was equally effective in blocking the linear respiratory chain of Pseudomonas aptata, a strain deficient in cytochromes of c type and Rieske iron-sulphur centre . The analysis of the oxido-reduction kinetic patterns of cytochromes indicated that, among the b type haems present in P . aptata, only cyt . o could be reduced by ubiquinol-1 in a reaction insensitive to both antimycin A and myxothiazol but inhibited by UHDBT . This latter finding has been correlated to the fact that P . aptata exhibits a defective b/c complex . In membranes from P . cichorii, in which the absorption maximum of dithionite reduced cytochrome(s) b shifted by 2-3 nm in the presence of antimycin A and/or myxothiazol, the electron flow through the b/c oxidoreductase complex has tentatively been arranged in a proton motive "Q-cycle" like mechanism. J Steroid Biochem, 1985 Sep, 23(3), 327 - 32 The degradation of beta-sitosterol by Pseudomonas sp . NCIB 10590 under aerobic conditions; Owen RW et al.; The bacterial degradation of beta-sitosterol by Pseudomonas sp NCIB 10590 has been studied . Major biotransformation products included 24-ethylcholest-4-en-3-one, androsta-1,4-diene-3,17-dione, 3-oxochol-4-en-3-one-24-oic acid and 3-oxopregn-4-en-3-one-20-carboxylic acid . Minor products identified were 26-hydroxy-24-ethylcholest-4-en-3-one, androst-4-ene-3,17-dione, 3-oxo-24-ethylcholest-4-en-26-oic acid, 3-oxochola-1,4-dien-3-one-24-oic acid, 3-oxopregna-1,4-dien-3-one-20 carboxylic acid and 9 alpha-hydroxyandrosta-1,4-diene-3,17-dione . Studies with selected inhibitors have enabled the elucidation of a comprehensive pathway of beta-sitosterol degradation by bacteria. J Clin Microbiol, 1985 Sep, 22(3), 352 - 4 Pseudomonas pseudomallei infection from drowning: the first reported case in Taiwan; Lee N et al.; We report a case of Pseudomonas pseudomallei infection, in which the patient acquired the bacteria by aspiration of river water after a drowning incident near Manila, the Philippines . The pulmonary form of melioidosis was noted at the onset, but septicemia developed at a later stage . Positive blood cultures were obtained 17 days after the accident . The patient was treated successfully with a combination of amikacin and cephalothin . This is the first report of P . pseudomallei infection documented in Taiwan. Am Surg, 1985 Sep, 51(9), 534 - 6 Subclavian catheter changes every third day in high risk patients; Gregory JA et al.; The subclavian catheter is commonly used in burn and trauma patients but remains a significant source of bacterial invasion in this group of seriously compromised individuals . The authors arbitrarily decided to change the catheter routinely every 3 days in consecutive patients to evaluate whether early detection of colonization and lowering of the infection rate was possible with this technique . At the time of change, catheter tips and central venous blood samples were sent for culture . Twenty-four venous blood samples were sent for culture . Twenty-four patients were studied in whom a total of 143 catheter changes over a wire introducer were performed . There were 20 men and four women with an average age of 34.5 years . Twenty patients with burns (average 48% body surface area involved), two patients with abdominal gunshot wounds, and two patients with complicated blunt trauma were studied . Five patients (three burns, two trauma) died, four because of sepsis, one of respiratory failure . Forty of the 143 catheter tips yielded positive cultures; however, 26 of these were not associated with overt sepsis . This 28 per cent incidence of positive catheter cultures was significantly less than the 47 per cent incidence reported previously in an identical patient population in whom the catheter was changed and cultured on clinical suspicion of sepsis (P value less than 0.001) . There were 38 clinical episodes of sepsis, but the catheter could be implicated as the source in only five of these . Pseudomonas was the most common organism isolated from all sources . The authors conclude from this study that the incidence of positive catheters is significant in this high-risk group, but is decreased when the routine change protocol is implemented.(ABSTRACT TRUNCATED AT 250 WORDS) J Pediatr, 1985 Sep, 107(3), 382 - 7 Pseudomonas cepacia colonization in patients with cystic fibrosis: risk factors and clinical outcome; Tablan OC et al.; During the period of 1979 to 1983, 38 patients with cystic fibrosis (CF) at the CF center of St . Christopher's Hospital for Children in Pennsylvania developed respiratory tract colonization with Pseudomonas cepacia . Seventeen (45%) of the patients with colonization died . Yearly incidence rates of P . cepacia colonization fluctuated between 1.3% and 6.1%, suggesting an endemic phenomenon . Case-control studies showed that severe underlying CF, use of aminoglycosides, and having a sibling with CF and P . cepacia colonization were significant risk factors for P . cepacia colonization . Once colonized with P . cepacia, patients with CF were likely to be hospitalized longer (P = 0.008) and to die sooner (P = 0.0001) than control patients with CF . Environmental and microbiologic studies did not identify a common source or mode of transmission of P . cepacia among patients . The results of this investigation suggest that P . cepacia colonization of patients with CF was endemic in the hospital, occurred more frequently in those with severe disease, and was associated with adverse clinical outcome. Mikrobiologiia, 1985 Sep-Oct, 54(5), 854 - 6 {Plasmid participation in the degradation of alpha-methylstyrene}; Boronin AM et al.; Pseudomonas acidovorans 9 transforming alpha-methylstyrene into acetophenone contains four types of plasmid DNA with molecular masses of 130, 110, 36 and 54 MD . The loss of the "growth on alpha-methylstyrene" property by this strain correlates with the absence of plasmids with the molecular masses of 130 and 110 MD from the cells . All the types of plasmid DNA are found in transconjugants growing on alpha-methylstyrene and produced by crossing the parent P . acidovorans strain with the plasmidless variant of this strain incapable of alpha-methylstyrene transformation . Apparently, plasmids with the molecular masses of 130 and 110 MD participate in the genetic control of alpha-methylstyrene transformation into acetophenone by P . acidovorans 9. J Clin Invest, 1985 Sep, 76(3), 1261 - 7 Characterization of immunotoxins active against ovarian cancer cell lines; Pirker R et al.; The purpose of the present study was to develop immunotoxins directed against human ovarian carcinoma cells . Four monoclonal antibodies (260F9, 454C11, 280D11, and 245E7) were chosen because they were found to bind to various ovarian carcinoma cell lines . These antibodies were covalently linked to either Pseudomonas exotoxin (PE) or ricin A chain (RTA), and the conjugates were tested against five ovarian cancer cell lines (OVCAR-2, -3, -4, -5; A1847) . The ability of the immunotoxins to inhibit both protein synthesis and colony formation was evaluated . Qualitatively similar results were obtained for both types of assays . Usually, PE conjugates were more toxic than their corresponding RTA conjugates . 454C11-PE was very toxic for all ovarian carcinoma lines, whereas 454C11-RTA had low activity . Both 260F9-PE and 260F9-RTA were active in all OVCAR cell lines but not in A1847 cells . 280D11-PE was toxic for OVCAR-4; otherwise, 280D11-PE and RTA conjugates of both 280D11 and 245E7 had little activity . Specificity of immunotoxin action was shown by competition by excess antibody, nontoxicity in nontarget cells, and inactivity of an irrelevant immunotoxin . To investigate the basis of antibody-dependent differences in activity of the various immunotoxins, antibody uptake was studied in OVCAR-2 cells, and the results indicate that antibody internalization is one important factor in the activity of immunotoxins. Arch Intern Med, 1985 Sep, 145(9), 1621 - 9 Pseudomonas bacteremia . Retrospective analysis of 410 episodes; Bodey GP et al.; We reviewed 410 episodes of Pseudomonas bacteremia occurring in patients with cancer during a ten-year period . Pseudomonas bacteremia was most common among patients with acute leukemia . The majority of patients acquired their infections in the hospital, and 51% had received antibiotic therapy for other presumed or proved infection during the preceding week . Shock occurred in 33%, and 32% had concomitant pneumonia . The overall cure rate was 62%; it was 67% for patients receiving appropriate antibiotics but only 14% for those receiving inappropriate antibiotics . A one- to two-day delay in the administration of appropriate antibiotic therapy reduced the cure rate from 74% to 46% . Patients who received an antipseudomonal beta-lactam antibiotic with or without an aminoglycoside had a significantly higher cure rate than patients who received only an aminoglycoside (72% and 71% vs 29%) . Patients with shock, pneumonia, or persistent neutropenia had a substantially poorer prognosis. Somat Cell Mol Genet, 1985 Sep, 11(5), 421 - 31 Characterization of diphtheria-toxin-resistant mutants lacking receptor function or containing nonribosylatable elongation factor 2; Kohno K et al.; Stable mutants resistant to diphtheria toxin (DT) were isolated from Chinese hamster ovary cells (CHO-K1) by single-step mutations with various mutagens . All the mutants were classified into two major groups as reported by other workers (4-6): toxin-entry mutants (DTrI) and translational mutants (DTRII) at the level of elongation factor 2 (EF-2) . These mutants were further characterized by directly measuring the specific uptake of {125I}DT and the content of nonribosylatable EF-2 by two-dimensional gel analysis . DTrI mutants, which showed no cross-resistance to Pseudomonas exotoxin A (PA), had no ability to associate with {125I}DT and contained only ADP-ribosylatable EF-2, like wild-type cells . DTRIIb mutants maintained about 50% of the normal level of cellular protein synthesis in the presence of DT, and two-dimensional gel analysis directly showed that they contained equivalent amounts of ADP-ribosylatable and nonribosylatable EF-2 molecules . Fully toxin-resistant cells, named KEE1 (DTRIIa), were isolated from a DTRIIb mutant (KE1) by two-step mutation . KEE1 cells showed full resistance to DT and PA, the normal level of association with {125I}DT, and produced only nonribosylatable EF-2 . Biochemical analysis of somatic cell hybrids indicated that the DT-resistant character of class II behaved codominantly . These results strongly supported the hypothesis that two copies of the gene for EF-2 are functional in CHO-K1 cells. Mol Gen Mikrobiol Virusol, 1985 Sep, (9), 11 - 5 {Spontaneous genetic transformation in Pseudomonas pseudomallei}; Bulantsev AL et al.; Spontaneous genetic transformation has been registered in Pseudomonas pseudomallei cells . Transforming frequencies registered reach 10(-4)-10(-5) . Spontaneous transformation has been simulated for Pseudomonas pseudomallei in soil . Intrageneric spontaneous transformation has been demonstrated to occur between Pseudomonas pseudomallei and Pseudomonas mallei. J Bacteriol, 1985 Sep, 163(3), 1263 - 4 Suicide vector for transposon mutagenesis in Pseudomonas solanacearum; Morales VM et al.; A suicide vector was constructed by cloning the transfer genes of the wide-host-range (IncW group) plasmid R388 into the BamHI site of pBR325 . This plasmid can deliver Tn5 into Pseudomonas solanacearum at frequencies ranging from 10(-6) to 10(-9) per recipient. J Bacteriol, 1985 Sep, 163(3), 1222 - 8 Molecular cloning and structure of the gene for 7 beta-(4-carboxybutanamido)cephalosporanic acid acylase from a Pseudomonas strain; Matsuda A et al.; A Pseudomonas strain produced an enzyme capable of deacylating 7 beta-(4-carboxybutanamido)cephalosporanic acid to 7-aminocephalosporanic acid in response to glutaric acid . The gene for the enzyme was cloned within the PstI site of pBR325 as a 7.35-kilobase-pair DNA segment from a mutant of this strain whose enzyme is produced constitutively . The gene expression in the primary clone appeared to be low in Escherichia coli but was significantly enhanced by reducing the size of the initial segment coupled with E . coli promoters . Subsequent subcloning resulted in localization of the gene to a 2.45-kilobase-pair fragment . Three clone-specific polypeptides with molecular weights of ca . 16,000, 54,000, and 70,000 were shown by maxicell analysis . The former two corresponded to the small and large subunits of the purified enzyme from the Pseudomonas strain, and the third polypeptide was suggested to be their precursor . This was supported by DNA sequence study together with amino acid sequencing of the amino terminus of both subunits: the sequences for the small and large subunits were localized contiguously in this order on the structural gene without termination codons between them . The nucleotide sequence also disclosed the presence of a signallike sequence preceding that for the small subunit, consistent with the previous observation that the enzyme might be periplasmic in the Pseudomonas strain . Those results suggest a process for the formation of an active enzyme complex from a precursor through two steps of processing. Appl Environ Microbiol, 1985 Sep, 50(3), 605 - 10 Reassembly of a fimbrial hemagglutinin from Pseudomonas solanacearum after purification of the subunit by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Young DH et al.; Distilled water homogenates of Pseudomonas solanacearum B1, a highly fimbriated strain, strongly agglutinated human group A erythrocytes . The fimbriae and hemagglutinating activity were precipitated from the crude extract with 1% acetic acid, redissolved at pH 10, and precipitated again with 20 mM CaCl2 at pH 6.9 . Ca2+, Mg2+, and Zn2+ had similar ability to precipitate the fimbrial hemagglutinin, but Na+ and K+ were much less effective . The fimbrial protein in the precipitate was purified to homogeneity by preparative gel electrophoresis in sodium dodecyl sulfate . The major protein band was eluted, and sodium dodecyl sulfate was removed by chromatography on ion retardation resin (AG 11A8) in 6 M urea . After dialysis against 10 mM sodium acetate (pH 4.5) to remove the urea, the protein reassembled to yield long fibers . These fibers were identical to fimbriae in the crude extract in diameter (6 nm) and in their ability to cause hemagglutination . The purified fimbriae contained no carbohydrates and wee similar to other bacterial fimbriae in amino acid composition, with hydrophobic amino acids comprising 41.8% of the total. FEBS Lett, 1985 Aug 19, 188(1), 85 - 90 3 beta, 17 beta-hydroxysteroid dehydrogenase of Pseudomonas testosteroni . Kinetic evidence for the bifunctional activity at a common catalytic site; Minard P et al.; 3 beta, 17 beta-Hydroxysteroid dehydrogenase (3 beta 17 beta HSDH) is an NAD-dependent dehydrogenase which has a double specificity for the 3- and 17-positions on the steroid skeleton . When dehydroepiandrosterone (DHEA) is used as steroid substrate, and the assay coupled with ketosteroid-isomerase, the two reactions occur alternately and each reaction on the 3-position produces a chromophoric molecule . These two reactions can follow one another without dissociation of the coenzyme from the enzyme binding site . This is confirmed by competition experiments with another dehydrogenase. J Biol Chem, 1985 Aug 15, 260(17), 9818 - 9 Crystallization and preliminary x-ray crystallographic data of dienelactone hydrolase from Pseudomonas sp . B13; Ollis DL et al.; Dienelactone hydrolase (EC 3.1.1.45) from Pseudomonas sp . B13 has been crystallized in a form suitable for high resolution x-ray diffraction study . The crystals are orthorhombic, the space group being P212121, with unit cell dimensions a = 48.9 A, b = 71.2 A, and c = 77.5 A . There appears to be 1 molecule in the asymmetric unit. Nucleic Acids Res, 1985 Aug 12, 13(15), 5657 - 69 The nucleotide sequence of the tnpA gene completes the sequence of the Pseudomonas transposon Tn501; Brown NL et al.; The nucleotide sequence of the gene (tnpA) which codes for the transposase of transposon Tn501 has been determined . It contains an open reading frame for a polypeptide of Mr = 111,500, which terminates within the inverted repeat sequence of the transposon . The reading frame would be transcribed in the same direction as the mercury-resistance genes and the tnpR gene . The amino acid sequence predicted from this reading frame shows 32% identity with that of the transposase of the related transposon Tn3 . The C-terminal regions of these two polypeptides show slightly greater homology than the N-terminal regions when conservative amino acid substitutions are considered . With this sequence determination, the nucleotide sequence of Tn501 is fully defined . The main features of the sequence are briefly presented. J Biol Chem, 1985 Aug 5, 260(16), 9393 - 8 Mevalonate utilization in Pseudomonas sp . M . Purification and characterization of an inducible 3-hydroxy-3-methylglutaryl coenzyme A reductase; Gill JF Jr et al.; Pseudomonas sp . M grown on mevalonate as the sole source of carbon has 200- to 800-fold induced levels of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase . The enzyme, which was purified to a homogeneous state in 54% yield (final specific activity, 60.5 mumol of NAD+ reduced per min per mg of protein), converted R-mevalonate (Km = 0.15 mM) to S-HMG-CoA . Activity was sensitive to sulfhydryl modifying reagents . The apparent molecular weight of the holoenzyme was 178,000 and that of the subunit 43,000 . The enzyme thus appears to be a tetramer . Comparison of a 23-residue amino-terminal sequence with the cDNA-derived sequence of Chinese hamster ovary cell HMG-CoA reductase showed little homology and antibody raised against the Pseudomonas enzyme did not appear to cross-react with rat liver HMG-CoA reductase . Addition of mevalonate to cells growing on glucose was followed by a rapid and biphasic induction of HMG-CoA reductase activity . During phase I, mevalonate or its catabolites may accumulate in intact cells of Pseudomonas sp . M and acetoacetate, a competitive inhibitor of HMG-CoA reductase (Ki = 3.2 mM), may feedback inhibit the enzyme under these conditions. Am J Hosp Pharm, 1985 Aug, 42(8), 1745 - 9 Cost savings associated with use of gentamicin versus tobramycin; Schwinghammer TL et al.; A hospital's use and costs of tobramycin sulfate versus gentamicin sulfate before and after a tobramycin use review were compared . Retrospective audits of 100 charts of adult patients in a 515-bed hospital were performed for two six-month periods in 1983-84 . Tobramycin use was considered appropriate in patients with serum creatinine concentrations greater than 1.5 mg/dL or pre-existing renal disease, in any patient over 70 years of age, and in patients with neutropenia, documented pseudomonas infection, or infection with an organism shown to be resistant to gentamicin but sensitive to tobramycin . Tobramycin use was not justifiable in 37 (18.7%) of 198 patients whose charts were evaluable . Use of gentamicin in these 37 patients would have saved $14,300 . The infection control committee was notified of the audit results; the audit results and recommendations for tobramycin use were included in a letter to all physicians; and the infectious disease service held educational conferences on tobramycin use . In the first six months after the corrective measures, mean monthly tobramycin use decreased by 38% and gentamicin use increased by 48.9% . Total aminoglycoside costs decreased 30.2% and total aminoglycoside use decreased 12.5% . In the second six months after intervention, mean monthly tobramycin use was 11% less than before intervention, and mean monthly gentamicin use was 13% greater than before intervention . Total aminoglycoside costs were 3.6% less and total aminoglycoside use was 4% less than before the audit . The tobramycin use audit and subsequent interventions with prescribers were effective in reducing tobramycin use and costs for approximately six months; decreases in tobramycin use and costs were smaller during the second six months after intervention. J Biochem (Tokyo), 1985 Aug, 98(2), 493 - 9 Cytochrome c oxidase of Pseudomonas AM 1: purification, and molecular and enzymatic properties; Fukumori Y et al.; Cytochrome c oxidase (cytochrome aa3-type) {EC 1.9.3.1} was purified from Pseudomonas AM 1 to an electrophoretically homogeneous state and some of its properties were studied . The oxidase showed absorption peaks at 428 and 598 nm in the oxidized form, and at 442 and 604 nm in the reduced form . The CO compound of the reduced enzyme showed peaks at 432 and 602 nm . The enzyme molecule was composed of two kinds of subunits with molecular weights of 50,000 and 30,000 and it contained equimolar amounts of heme a and copper atom . The enzyme rapidly oxidized Candida krusei and horse ferrocytochromes c as well as Pseudomonas AM 1 ferrocytochrome c . The reactions catalyzed by the enzyme were strongly inhibited by KCN. J Cell Biol, 1985 Aug, 101(2), 350 - 7 Enhancement of ricin cytotoxicity in Chinese hamster ovary cells by depletion of intracellular K+: evidence for an Na+/H+ exchange system in Chinese hamster ovary cells; Ghosh PC et al.; Depletion of intracellular K+ has been reported to result in an arrest of the formation of coated pits in human fibroblasts (Larkin, J.M., M.S . Brown, J.L . Goldstein, and R.G.W . Anderson, 1983, Cell, 33:273-285) . We have studied the effects of K+ depletion on the cytotoxicities of ricin, Pseudomonas exotoxin A, and diphtheria toxin in Chinese hamster ovary (CHO) cells . The cytotoxicities of ricin and Pseudomonas toxin were enhanced in K+-depleted CHO cells whereas the cytotoxicity of diphtheria toxin was reduced by K+ depletion . The effects of NH4Cl on the cytotoxicities of ricin, Pseudomonas toxin, and diphtheria toxin were found to be similar to those of K+ depletion, and there were no additive or synergistic effects on ricin cytotoxicity by NH4Cl in K+-depleted medium . The enhancement of ricin cytotoxicity by K+ depletion could be completely reversed by the addition of K+, Rb+, and partially by the addition of Cs+, before the ricin treatment, whereas Li+ was ineffective . These protective effects of K+ or Rb+ requires a functional Na+/K+ ATPase . CHO cells grown in K+-depleted media were found to contain 6.3-fold increase in intracellular Na+ level, concomitant with a 10-fold reduction in intracellular K+ level . The enhanced cytotoxicity of ricin in K+-free medium and the increased uptake of Na+ could be abolished by amiloride or amiloride analogues, which are known to be potent inhibitors of the Na+/H+ antiport system . Our results suggest that a depletion of intracellular K+ results in an influx of Na+, which is accompanied by the extrusion of H+ . Consequently, there is an alkalinization of the cytosol and the ricin-containing endosomes . As a result, ricin is more efficiently released from the endosomes in-K+-depleted cells . Results from the studies of the binding, internalization, and degradation of 125I-ricin, and the kinetics of inhibition of protein synthesis by ricin in K+-depleted cells are consistent with this working hypothesis. J Cell Physiol, 1985 Jul, 124(1), 54 - 60 Effect of potassium depletion of cells on their sensitivity to diphtheria toxin and pseudomonas toxin; Sandvig K et al.; When Vero cells were depleted of potassium, the cells were protected against diphtheria toxin . Potassium depletion of Vero cells strongly reduced the binding of the toxin to cell surface receptors . Likewise, potassium depleted L-cells were protected against pseudomonas toxin . Diphtheria toxin binding was completely restored upon addition of potassium to the cells . This restoration was not prevented by inhibition of protein synthesis by cycloheximide . When cells were depleted of potassium in the presence of metabolic inhibitors, and then treated with diphtheria toxin, protein synthesis was reduced to the same extent as in cells with normal intracellular level of potassium . The results indicate that potassium depletion of Vero cells reduces the ability of the cells to bind diphtheria toxin by an ATP requiring process, and that binding, endocytosis and transfer of diphtheria fragment A across the membrane may occur at low intracellular levels of potassium. Am J Med, 1985 Jul, 79(1), 10 - 2 Superficial and systemic illness related to a hot tub; Kosatsky T et al.; In an outbreak of folliculitis in Alaska among bathers in a contaminated hot tub, one person was found in whom follicular lesions were preceded by deep, tender, peripheral nodules . Of nine affected bathers, five showed inflammation of Montgomery's follicles of the breast . Bathing longer in the tub and later in the day was associated with increased risk of disease . This investigation added serotype O-7,8 to the list of pseudomonads associated with hot tub infections. Appl Environ Microbiol, 1985 Jul, 50(1), 38 - 40 Influence of salts and temperature on the transfer of mercury resistance from a marine pseudomonad to Escherichia coli; Gauthier MJ et al.; Thirty-one strains of marine bacteria were examined for their ability to transfer mercury resistance to Escherichia coli in complex media; eight strains were able to transfer their resistance marker, with frequencies ranging from 10(-3) to 10(-8) . Frequencies generally increased with the increase of the mating period . Additional mating experiments were carried out with one strain, belonging to the pseudomonads, to estimate the influence of temperature, salinity, and time on the conjugal transfer frequency of mercury resistance markers . The higher frequencies occurred at 30 degrees C, in a salt medium (37%), after 24 h of mating. Appl Environ Microbiol, 1985 Jul, 50(1), 169 - 71 Construction of a cosmid clone library of Pseudomonas syringae pv . phaseolicola and isolation of genes by functional complementation; Ehrenshaft M et al.; A genomic library constructed from a wild-type strain of Pseudomonas syringae pv . phaseolicola in the broad-host-range cosmid vector pVK102 was used to isolate wild-type genes by complementation of Tn5-induced auxotrophic mutants . Selection pressure was required for maintenance of the vector and members of the library in strains of P . syringae. Biochemistry, 1985 Jun 18, 24(13), 3158 - 65 Alternate substrates and inhibitors of bacterial 4-hydroxyphenylpyruvate dioxygenase; Pascal RA Jr et al.; A variety of analogues of (4-hydroxyphenyl)pyruvic acid were synthesized, and the reactions of these compounds with the 4-hydroxyphenylpyruvate dioxygenase from Pseudomonas sp . P.J . 874 were examined . Several of the ring-substituted substrate analogues are reversible inhibitors of the enzyme, the most potent being the competitive inhibitor (2,6-difluoro-4-hydroxyphenyl) pyruvate (Ki = 1.3 microM) . Two substrate analogues (2-fluoro-4-hydroxyphenyl)pyruvate and {(4-hydroxyphenyl)thio}pyruvate proved to be alternate substrates for the enzyme . The former compound is converted to (3-fluoro-2,5-dihydroxyphenyl)acetate in an essentially normal catalytic sequence including oxidative decarboxylation, ring hydroxylation, and side-chain migration . The latter compound, however, undergoes oxidative decarboxylation and sulfoxidation to give {(4-hydroxyphenyl)sulfinyl}acetate; ring oxidation is not observed . The implications of these results with regard to the catalytic mechanism of 4-hydroxyphenylpyruvate dioxygenase are discussed. Am J Med, 1985 Jun 7, 78(6A), 85 - 91 Randomized trial of imipenem/cilastatin versus gentamicin and clindamycin in mixed flora infections; Solomkin JS et al.; Results of a randomized trial comparing imipenem/cilastatin versus the combination of gentamicin plus clindamycin for mixed flora surgical sepsis are reported herein . Seventy-four patients were evaluable, 50 of whom had intra-abdominal sepsis . No imipenem-resistant initially infecting isolates were encountered . When outcome was evaluated on the basis of severity scoring (APACHE II), no difference in mortality was noted . However, therapy in two patients with Pseudomonas emerging from a polymicrobial flora failed with gentamicin, whereas no Pseudomonas failures were noted with imipenem/cilastatin . The major difference noted was in toxicity . There was a 20 percent incidence of nephrotoxicity in gentamicin-treated patients despite serum level monitoring and multiple dose adjustments . The degree of efficacy and the relative tolerability of imipenem/cilastatin in seriously ill surgical patients is demonstrated. Biochem J, 1985 Jun 1, 228(2), 347 - 52 The inactivation of ornithine transcarbamoylase by N delta-(N'-sulpho-diaminophosphinyl)-L-ornithine; Templeton MD et al.; Phaseolotoxin, a tripeptide inhibitor of ornithine transcarbamoylase, is a phytotoxin produced by Pseudomonas syringae pv . phaseolicola, the causal agent of halo-blight in beans . In vivo the toxin is cleaved to release N delta-(N'-sulpho-diaminophosphinyl)-L-ornithine, the major toxic chemical species present in diseased leaf tissue . This paper reports on the interaction between N delta-(N'-sulpho-diaminophosphinyl)-L-ornithine and ornithine transcarbamoylase . N delta-(N'-Sulpho-diaminophosphinyl)-L-ornithine was found to be a potent inactivator of the enzyme, in contrast with phaseolotoxin, which previously has been reported to inhibit the enzyme reversibly . Inactivation by N delta-(N'-{35S}sulpho-diaminophosphinyl)-L-ornithine resulted in the incorporation of 35S into ethanol-precipitated protein . The stoicheiometry of 35S incorporation was approximately 1 mol/mol of active sites . Inactivation was second-order and a rate constant of 10(6) M-1 X s-1 at 0 degree C in 50 mM-Tris/HCl, pH 9.0, was obtained . Carbamoyl phosphate, a substrate of ornithine transcarbamoylase, protected the enzyme from inactivation . A dissociation constant of 3 microM for the enzyme-carbamoyl phosphate complex was calculated . L-Ornithine, the second substrate for ornithine transcarbamoylase, protected the enzyme only at high concentrations . The results are consistent with N delta-(N'-sulpho-diaminophosphinyl)-L-ornithine being a potent affinity label that binds via the carbamoyl phosphate-binding site of ornithine transcarbamoylase . Cleavage of phaseolotoxin to N delta-(N'-sulpho-diaminophosphinyl)-L-ornithine in vivo appears to be an important function in the physiology of the disease. Drug Intell Clin Pharm, 1985 Jun, 19(6), 427 - 9 Prolonged elimination of piperacillin in a patient with renal and liver failure; Green L et al.; A 22-year-old female with acute leukemia was admitted to the oncology center for evaluation and treatment . Piperacillin was used during her admission for treatment of Pseudomonal sepsis . The patient's renal and liver function deteriorated during the hospital course, requiring large dose adjustments of piperacillin . The elimination half-life of piperacillin was estimated to be 32 hours . Patients with both liver and renal disease require only small doses of piperacillin to achieve therapeutic plasma concentration. Appl Environ Microbiol, 1985 Jun, 49(6), 1442 - 7 Mixed carbon source utilization of meat-spoiling Pseudomonas fragi 72 in relation to oxygen limitation and carbon dioxide inhibition; Molin G; The growth of meat-spoiling Pseudomonas fragi 72 was studied on a defined salt medium supplemented with L-aspartate, citrate, creatine, creatinine, D-glucose, L-glutamate, and L-lactate . The utilization of the different carbon sources was followed in batch and continuous culture and under the influence of oxygen limitation and carbon dioxide inhibition (50% CO2 in air) . Under nonrestricted atmospheric conditions in batch culture, the organism showed a preference in the utilization of the carbon sources in the order glucose greater than lactate greater than citrate greater than aspartate-glutamate greater than creatine greater than creatinine . The first five sources were utilized simultaneously . The order of preference was changed in continuous culture to lactate-citrate-glutamate-aspartate greater than glucose greater than creatine greater than creatinine . All carbon sources were utilized at lower dilution rates, but as the rate was increased the concentration of the carbon sources started to increase in the effluent and the preference could be seen . Under conditions of oxygen limitation the preference for glucose was weakened, but for lactate it was slightly enhanced (batch and continuous culture) . Under conditions of CO2 inhibition, the preference for glucose was enhanced . However, lactate and amino acids were still preferred to glucose in the continuous culture . The utilization of creatine and creatinine was blocked by CO2 in batch culture, and only a slight utilization of creatine was noticed in a chemostat at lower dilution rates. J Pediatr, 1985 Jun, 106(6), 1030 - 4 Management of acute pulmonary exacerbations in cystic fibrosis: a critical appraisal; Nelson JD; Optimal management of acute exacerbations of pulmonary symptoms in patients with cystic fibrosis remains questionable . The underlying cause of such exacerbations has not been identified, and the microbiology of the sputum does not differ substantially during these exacerbations . Despite the absence of conclusive evidence that antibiotic therapy enhances treatment of cystic fibrosis, the consensus favors its use . Various combination and single-agent therapies with aminoglycosides, cephalosporins, and beta-lactam antibiotics are reviewed critically . The importance of high activity against Pseudomonas strains is addressed, as is the potential value of antibiotic prophylaxis . The drawbacks of aminoglycoside treatment are reported . No evidence proves the superiority of combination therapy over monotherapy . Recent results suggesting the effectiveness of monotherapy with piperacillin or ceftazidime are encouraging and deserve follow-up to test continued efficacy and the absence of development of resistant antibiotic strains . Further investigation into the prevention of acute pulmonary exacerbations in cystic fibrosis is essential. J Bacteriol, 1985 Jun, 162(3), 992 - 9 cDNA cloning of portions of the bacteriophage phi 6 genome; Mindich L et al.; Phage phi 6 has a genome consisting of three pieces of double-stranded RNA . Single-stranded RNA was prepared from phi 6 nucleocapsids by in vitro transcription with the phage RNA polymerase . These transcripts were polyadenylated and used as templates for the preparation of cDNA copies . The resulting DNA was cloned into the PstI restriction nuclease site of plasmid pBR322 . Insert-bearing plasmids were annealed to phi 6 RNA to assign the inserts to their proper segments . In this way we identified inserts corresponding to the large, medium, and small segments . Two large overlapping inserts of the small segment constitute the complete complement of the segment as determined by the sequence analysis of the DNA . In vitro coupled transcription and translation showed that the small segment inserts were able to direct the synthesis of the four known genes in the small segment . Two overlapping inserts in the medium segment constitute the entire segment and were shown to direct the in vitro synthesis of two of the three known proteins of the medium segment . Several inserts bearing about one-third the complement of the large segment were also isolated, and one of these directed the synthesis of a peptide that resembles protein P1 . Restriction endonuclease maps were prepared for the inserts, and by in vitro synthesis it was possible to refine the genetic map of phi 6 . A chimeric plasmid was constructed that combines plasmids pUC8 and RSF1010 . Inserts placed on this plasmid were transformed to Pseudomonas phaseolicola, the natural host of phage phi 6 . It was possible to refine further the genetic map by complementation of nonsense mutants of phi 6 with the cDNA. J Biol Chem, 1985 May 25, 260(10), 6281 - 7 Regulation of 3-indoleacetic acid production in Pseudomonas syringae pv . savastanoi . Purification and properties of tryptophan 2-monooxygenase; Hutcheson SW et al.; The oxidative decarboxylation of L-tryptophan to yield 3-indoleacetamide, catalyzed by tryptophan 2-monooxygenase, represents a controlling reaction in the synthesis of indoleacetic acid by Pseudomonas savastanoi (Pseudomonas syringae pv . savastanoi), a gall-forming pathogen of olive (Olea europea L.) and oleander (Nerium oleander L.) . Production of indoleacetic acid is essential for virulence of the bacterium in its hosts . Tryptophan 2-monooxygenase was characterized to determine its role in indoleacetic acid metabolism in the bacterium . The enzyme was purified to apparent homogeneity from Escherichia coli cells containing the genetic locus for this enzyme obtained from P . savastanoi . The preparation contained a single polypeptide with a mass of 62,000 that cross-reacted immunologically with a homologous protein in P . savastanoi . The holoenzyme contained one FAD moiety/subunit with properties consistent with a catalytic function . The enzyme preparation catalyzed an L-tryptophan-dependent O2 uptake and yielded 3-indoleacetamide as a product . Enzyme activity fit simple Michaelis Menten kinetics with a Km for L-tryptophan of 50 microM . 3-Indoleacetamide and 3-indoleacetic acid were identified as regulatory effectors . The apparent Ki for 3-indoleacetamide was 7 microM; that for indoleacetic acid was 225 microM . At Km concentrations of tryptophan, enzyme activity was inhibited 50% by 25 microM 3-indoleacetamide . In contrast, 230 microM indoleacetic acid was required to effect a similar inhibition . Phenylalanine and tyrosine were ineffective as regulatory metabolites . These results indicate that IAA synthesis in P . savastanoi is regulated by limiting tryptophan and by feedback inhibition from indoleacetamide and indoleacetic acid. Biochemistry, 1985 May 21, 24(11), 2606 - 9 Mechanism of inactivation of 3-oxosteroid delta 5-isomerase by 17 beta-oxiranes; Bantia S et al.; The affinity label (17S)-spiro{estra-1,3,5(10),6,8-pentaene-17,2'-oxiran}-3-ol (5 beta) inactivates 3-oxosteroid delta 5-isomerase from Pseudomonas testosteroni by formation of a covalent bond between Asp-38 of the enzyme and the steroid . High-performance liquid chromatography (HPLC) analysis of tryptic digests of inactivated enzyme shows that two isomeric steroid-containing peptides are formed in a ratio of 9:1 at pH 7 (TPS1 and TPS2) . Hydrolysis of each of these peptides produces a different steroid: TPS1 releases 17 alpha-(hydroxymethyl)estra-1,3,5(10),6,8-pentaene-3,17 beta-diol (S1) whereas TPS2 yields 17 beta-(hydroxymethyl)estra-1,3,5(10),6,8-pentaene-3,17 alpha-diol (S2) . Inactivation of the enzyme by (17S)-spiro{estra-1,3,5(10),6,8-pentaene-17,2'-oxiran-18O}-3-ol, followed by mass spectral analysis of the diacetate of the steroid released upon hydrolysis of the enzyme-inhibitor bond, reveals that TPS1 is formed by attack of Asp-38 at the methylene carbon of the oxirane . In contrast, TPS2 is produced by Asp-38 attack at the tertiary carbon . These results imply that inactivation occurs through concurrent SN1 and SN2 reactions of Asp-38 with the protonated inhibitor and that Asp-38 is located on the alpha face of the steroid when it is bound to the active site in the correct manner to react for both the SN1 and SN2 processes. Eur J Biochem, 1985 May 2, 148(3), 447 - 53 Purification and properties of carboxypeptidase G2 from Pseudomonas sp . strain RS-16 . Use of a novel triazine dye affinity method; Sherwood RF et al.; A folate-degrading enzyme, carboxypeptidase G2, has been purified on a large scale from Pseudomonas sp . strain RS-16 . Homogeneous enzyme was obtained by a three-step procedure involving ion-exchange chromatography and a novel triazine dye (affinity) chromatography step which utilizes Zn2+ to promote adsorption of the enzyme . Enzyme was selectively eluted by the use of a chelating agent (EDTA) and a step change in pH . The enzyme is a dimeric protein (Mr 83000) with two identical subunits of 41800 and contains four atoms of zinc per enzyme molecule, which are required for full activity . The enzyme follows Michaelis-Menten kinetics with Km values of 4.0 microM for folate, 8.0 microM for methotrexate and 34.0 microM for 5-methyltetrahydrofolate, the predominant form of reduced folate found in plasma. Vopr Med Khim, 1985 May-Jun, 31(3), 31 - 7 {Thermostabilization of glutamin(asparagin)ase from Pseudomonas aurantica BKMB-548}; Kabanova EA et al.; In studies on kinetics of thermoinactivation of glutaminase (asparaginase) from Ps . arantiaca BKMB-548 at 50 degrees and pH 7.0 in presence or in absence of L-glutamate the enzyme inactivation was found to obey the first order equation . Both the glutaminase and asparaginase activities decreased at a similar rate . L-Glutamate stabilized the enzyme due to direct interaction with its molecule . Stability of the complex formed was evaluated quantitatively . L-Glutamate reacted apparently with a specific site on the surface of the enzyme molecule; Kdiss was 0.42 +/- 0.03 mM at pH 7.0 and 50 degrees . No cooperative effect was found . L-Aspartate protected the enzyme completely; stabilizing effects of L-cysteine, L-serine and glycine were similar to the effect of L-glutamate (94%, 84%, 83% and 82%, respectively) . At the same time, glutarate, succinate, alpha-ketobutyrate, alpha-ketoglutarate, gamma-aminobutyrate and N-benzoyl glutamate did not exhibit the stabilization effect . The data obtained suggest that the high stabilizing effect might exhibit only the substances containing simultaneously free alpha-NH2 and alpha-COOH groups in a molecule, whereas presence of COOH groups at beta--or gamma-carbon atoms was not essential for the stabilizing effect. Biull Eksp Biol Med, 1985 May, 99(5), 557 - 60 {Isolation, purification and physicochemical properties of glutamin-asparaginase from Pseudomonas boreopolis 526}; Pekhov AA et al.; A new homogeneous enzyme which is capable of catalyzing the hydrolysis of both glutamine and asparaginase has been purified from extracts of Pseudomonas boreopolis 526 by the improved method . Purification involves few stages . The ratio of glutaminase to asparaginase activity is approximately 1.5:1.0 . The enzyme is stable on storage and has a wide pH optimum of action (6-8.5) . The molecular weight is about 134 000-145 000 D and the subunit molecular weight is about 34 000 D . No free SH-groups have been detected in the enzyme molecule. Am Rev Respir Dis, 1985 May, 131(5), 791 - 6 Pseudomonas cepacia colonization among patients with cystic fibrosis . A new opportunist; Thomassen MJ et al.; Pseudomonas cepacia colonization among patients with cystic fibrosis (CF) at our center has increased from 7% (of 419 patients) to 15% (of 450 patients) over the past 5 yr (July 1978 through June 1983) . The proportion of patients dying with P . cepacia colonization has increased over this 5-yr period (Year 1, 9% (1/11) of the deaths were associated with P . cepacia; Year 5, 55% (16/29) were associated with P . cepacia) . These observations have led to a heightened concern regarding the presence of P . cepacia in the CF population . Characteristics of the patient population that might relate to P . cepacia colonization were reviewed . Increasing numbers of patients in good clinical condition became colonized with P . cepacia . Females in good clinical condition who acquire P . cepacia appear to be at special risk of developing severe and unexpected pulmonary complications that often end in death . In contrast, males, regardless of clinical condition, appear less likely to experience an immediate decline in clinical status . Hospitalization is potentially implicated in contributing to the increase in P . cepacia colonization because many patients' initial positive cultures were concurrent with or followed a hospital stay . Sixteen patients with CF and P . cepacia had siblings with CF, 6 of whom subsequently acquired P . cepacia . This frequency is more than double that in our overall CF population. Pathol Biol (Paris), 1985 May, 33(5), 309 - 12 {Pharmacokinetics of apalcillin in pediatrics}; Akbaraly JP et al.; Apalcillin is a semi-synthetic broad-spectrum penicillin which is particularly active against Pseudomonas . Pharmacokinetic studies were carried out after slow (3 mn) intravenous administration of a single dose of 20 or 30 mg/kg in five categories of infants: premature, dysmature, full term newborn up to 8 days of age, infant aged 8 days to 1 month and infant beyond 1 month . Elimination half-lives in these five groups are respectively: 6.83, 5.44, 5.01, 2.28 and 1.28 hours . A decrease in elimination half-life and increase in total clearance are observed as the infant matures . Renal clearance represents a quarter of total clearance suggesting than there is considerable extrarenal clearance . Pharmacokinetic findings beyond one month of age are comparable to those demonstrated in adults . The dysmature infant is characterized by a significantly lower steady-state distribution volume (p less than or equal to 0.025) as compared to the full term neonate . From these results the recommended dosage is: 20 mg/kg twice daily for premature, dysmature and full term neonates up to 8 days, 30 mg/kg twice daily for infants from 8 days to one month of age, and 30 mg/kg three or four times a day for infants above one month of age. J Bacteriol, 1985 May, 162(2), 773 - 6 Some mercurial resistance plasmids from different incompatibility groups specify merR regulatory functions that both repress and induce the mer operon of plasmid R100; Foster TJ et al.; Transcription of the mer genes of plasmid R100 is regulated by the product of the merR gene . The merR gene negatively regulates its own expression and also controls the transcription of the merTCA operon both negatively (in the absence of inducer) and positively (in the presence of inducer) . We used transcriptional mer-lac fusions of R100-1 in complementation tests to measure the ability of the merR products of different mercury-resistant transposons and plasmids to functionally interact with R100-1 . Plasmids from incompatibility groups C, B, S, L, and P, as well as the Pseudomonas transposons Tn501 and Tn3401, regulated the expression of the R100 mer genes in a similar fashion to the R100-1 merR product itself, suggesting that these elements are closely related . Only plasmid R391 (IncJ) did not complement. Pathol Biol (Paris), 1985 May, 33(5), 412 - 5 {Comparison of the in vitro activity of 6 quinolones on Pseudomonas sp.}; Lesage D et al.; Activity of six quinolones (nalidixic acid, pipemidic acid, oxolinic acid, pefloxacin, ofloxacin and norfloxacin) against one-hundred and ten Pseudomonas strains was studied in vitro . Five species of Pseudomonas were represented, i.e . aeruginosa, maltophilia, cepacia, stutzeri and paucimobilis . Isolates came from two Paris hospitals . Minimal inhibitory concentrations (MICs) were determined using gelose dilution according to WHO recommendations (Mueller Hinton medium, multiple inoculator, controlled inoculum) . Modal CMIs classify activities of the six tested quinolones against P . aeruginosa in the following order: nalidixic acid: 64 mg/l; pipemidic acid: 16-32 mg/l; oxolinic acid: 16 mg/l; pefloxacin: 2 mg/l; ofloxacin: 2 mg/l; norfloxacin: 1 mg/l . The other Pseudomonas species exhibit a variety of resistance phenotypes which are described in detail . High CMIs are found for certain P . aeruginosa strains . Two of these, i.e . DL 55 and DL 59, are highly resistant to all the tested quinolones . Their pattern of resistance is comparable to that of a mutant, PAO 38-02, obtained in vitro in the presence of pefloxacin . This fact suggests that quinolones may induce in vivo selection of resistant P . aeruginosa mutants. Plasmid, 1985 May, 13(3), 200 - 4 A vehicle for the introduction of transposons into plant-associated pseudomonads; Lam ST et al.; A recombinant plasmid with wide-host-range transfer functions, narrow-host-range replication functions, and carrying a kanamycin-resistant transposon transferred kanamycin resistance to a number of plant-associated pseudomonads . Southern hybridization studies suggest that only a small portion of the plasmid, coinciding with the location of the transposon, is present in the kanamycin-resistant Pseudomonas derivatives . The plasmid sequences appear to be inserted at a number of different sites in the recipient genome . This plasmid can thus be used as a vehicle for the introduction of transposons into some plant-associated pseudomonads and should be useful in both genetic and ecological studies of these bacteria. Thorax, 1985 May, 40(5), 358 - 63 Ceftazidime compared with gentamicin and carbenicillin in patients with cystic fibrosis, pulmonary pseudomonas infection, and an exacerbation of respiratory symptoms . British Thoracic Society Research Committee; Sedimentation analysis of ribonucleotide reductase activity in extracts of Pseudomonas stutzeri; Analysis of ribonucleotide reductase and DNA polymerase activities in extracts of Pseudomonas stutzeri by centrifugation in discontinuous sucrose gradients indicated that these two enzymes are associated with two different high molecular weight cellular components . In addition, 95% of the ribonucleotide reductase activity was pelleted by centrifugation of extracts for 4 hr at 200,000 X G . The reductase activity remained particulate (sedimentable) following sonication whereas some 90% of the DNA polymerase activity was rendered soluble (non-sedimentable) by this technique . This data indicate that the P . stutzeri ribonucleotide reductase is not a cytosolic enzyme, but is associated with a macromolecular component in the cell. Biochem Biophys Res Commun, 1985 Apr 16, 128(1), 271 - 7 Purification and properties of gamma-oxalomesaconate hydratase from Pseudomonas ochraceae grown with phthalate; Maruyama K; Pseudomonas ochraceae produced inducibly a hydro-lyase which catalyzes the reversible conversion of gamma-oxalomesaconate into (-)-gamma-oxalocitramalate . The enzyme has been purified to homogeneity from the bacteria grown with phthalate . The enzyme was a dimeric protein (pI=4.9) with a Mr of 68,000 and showed a high specificity for gamma-oxalomesaconate (Km=14 microM) and (-)-gamma-oxalocitramalate (Km=6.4 microM) . Equilibrium constant for the hydration of gamma-oxalomesaconate at pH 8.0 and 24 degrees C was 2.5 . Various thiols activated the enzyme. J Bacteriol, 1985 Apr, 162(1), 324 - 7 Cloning of genes involved in myo-inositol transport in a Pseudomonas sp; Gauchat-Feiss D et al.; A soil isolate of a Pseudomonas sp . can utilize myo-inositol (MI) as the sole carbon source . In this strain, MI is transported through the membrane by a high-affinity transport system in which a periplasmic binding protein is involved . Mutants impaired in the transport system were obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine and subsequently identified by their slow growth rate at low MI concentrations . Strains with a low linear initial rate of MI uptake were analyzed . Using a broad-host-range cosmid cloning system, we have constructed a gene bank of the wild-type Pseudomonas sp . in an Escherichia coli recA-host . A rapid mating technique enabled us to screen the gene library for clones which are able to restore the active transport of MI in the mutant . An 11.5-kilobase segment containing genes involved in the MI transport has been isolated, and its restriction enzyme cleavage map has been determined. J Gen Microbiol, 1985 Apr, 131 ( Pt 4), 897 - 903 Expression of the Pseudomonas gene coding for carboxypeptidase G2 in Saccharomyces cerevisiae; Clarke LE et al.; The Pseudomonas gene coding for carboxypeptidase G2 was introduced into Saccharomyces cerevisiae on an Escherichia coli/yeast shuttle vector pROG5 . The level of enzyme activity obtained was independent of the orientation of the gene within the pBR322-derived tetracycline resistance gene of the vector, indicating that expression can occur from a Pseudomonas promoter in yeast. Appl Environ Microbiol, 1985 Apr, 49(4), 761 - 4 Plasmid-borne Tn5 insertion mutation resulting in accumulation of gentisate from salicylate; Monticello DJ et al.; Plasmid-borne Tn5 insertion mutants of a Pseudomonas species which accumulated 2,5-dihydroxybenzoate (gentisate) following growth on 2-hydroxybenzoate (salicylate) were obtained from a pool of mutants that were unable to grow on naphthalene . One such mutant was characterized further . The ability of this mutant to oxidize gentisate was 100-fold less than the ability of a Nah+ Sal+ strain harboring the unmutagenized plasmid, although both strains oxidized and grew on salicylate . These bacteria were presumably able to metabolize salicylate via catechol, since they possessed an inducible, plasmid-encoded catechol 2,3-dioxygenase . Our results suggest that there is an alternate, plasmid-encoded route of salicylate degradation via gentisate and that some plasmid-associated relationship between this pathway and naphthalene oxidation exists. Bioorg Khim, 1985 Apr, 11(4), 536 - 8 {Antigenic bacterial polysaccharides . 13 . The structure of the O-specific polysaccharide chain of the lipopolysaccharide from Pseudomonas cepacia strain IMV 4137}; Knirel' IuA et al.; On mild acid degradation of a lipopolysaccharide from Pseudomonas cepacia strain IMV 4137, a serologically active O-specific polysaccharide was obtained and shown to contain L-rhamnose and D-galactose . According to 1H- and 13C-NMR data as well as methylation analysis, the polysaccharide is made up of disaccharide repeating units of the following structure:----2)-alpha-L-Rhap-(1----4)-alpha-D-Galp-(1----. Southeast Asian J Trop Med Public Health, 1985 Mar, 16(1), 83 - 7 Pseudomonas pseudomallei in southern Thailand; Nachiangmai N et al.; The distribution of Pseudomonas pseudomallei was studied in soil and water from various sources of Songkla province, Southern Thailand . P . pseudomallei was isolated from the surface soil of rubber plantations (60.9%) and from the bottom sediments of rice fields (78.1%) . Farmers and plantation workers will have a greater risk of contracting melioidosis. Mikrobiologiia, 1985 Mar-Apr, 54(2), 186 - 90 {Chemical nature of the autoregulating factor d1 in Pseudomonas carboxydoflava}; Osipov GA et al.; A chromatographically individual fraction with the biological activity of factor d1 was isolated from Pseudomonas carboxydoflava by the techniques of column and thin-layer chromatography . Alkyl hydroxybenzoles were shown to be the active principle of factor d1 inducing the transition of P . carboxydoflava vegetative cells into the hypometabolic or anabiotic state . As was found from the data of PMR, UV and IR spectroscopy and gas chromatography in combination with mass spectrometry, the active principle of factor d1 included alkylresorcins: 5-n-nonadecylresorcin and 5-n-heneicosylresorcin at a ratio of 1:3. Arch Dis Child, 1985 Mar, 60(3), 219 - 24 Humidification of incubators; Harpin VA et al.; The effect of increasing the humidity in incubators was examined in 62 infants of less than 30 weeks' gestation . Thirty three infants nursed in high humidity for two weeks were compared retrospectively with 29 infants from an earlier study who were nursed under plastic bubble blankets or with topical paraffin but without raised humidity . Humidification reduced skin water loss and improved maintenance of body temperature from birth, but did not delay the normal postnatal maturation of the skin . Infants nursed without humidity frequently became hypothermic in spite of a high incubator air temperature . These advantages must be weighed against the finding that overheating was more common and Pseudomonas was more commonly isolated from the infants . It is recommended that incubator humidity is raised for babies under 30 weeks' gestation in the first days of life but meticulous attention should be paid to fluid balance, avoiding overheating, and cleansing of the humidifier reservoir. J Clin Microbiol, 1985 Mar, 21(3), 445 - 6 Cervical osteomyelitis caused by Pseudomonas cepacia in an intravenous-drug abuser; Smith MA et al.; We report a case of vertebral osteomyelitis caused by Pseudomonas cepacia in a patient with a history of intravenous-drug abuse . P . cepacia infections are usually nosocomial, although community-acquired infections occur more commonly in intravenous-drug abusers than in the general population . Vertebral osteomyelitis caused by P . cepacia has not previously been reported. J Clin Microbiol, 1985 Mar, 21(3), 314 - 7 Pseudomonas mesophilica infections in humans; Smith SM et al.; Reported here is the case of a patient with metastatic adenocarcinoma of the lung who had bacteremia involving Pseudomonas mesophilica . Of the common laboratory media tested at 35 degrees C, buffered charcoal yeast extract agar and nutrient agar provided the best growth; however, other media supported growth at lower temperatures . Since blood cultures are routinely subcultured onto chocolate agar and then incubated at 35 degrees C, awareness of the characteristics of P . mesophilica and of the isolation techniques as outlined may enhance the recovery of this and related bacterial species. J Bacteriol, 1985 Mar, 161(3), 1171 - 5 Purification of bromoperoxidase from Pseudomonas aureofaciens; van Pee KH et al.; A Bromoperoxidase has been isolated and purified from Pseudomonas aureofaciens ATCC 15926 mutant strain ACN . The purified enzyme was homogeneous as determined by polyacrylamide gel electrophoresis and ultracentrifugation . This bromoperoxidase can utilize bromide ions in the presence of hydrogen peroxide and a halogen acceptor for the catalytic formation of carbon-halogen bonds . The homogeneous enzyme also has peroxidase and catalase activity . Based on the results from gel filtration and ultracentrifugation, the molecular weight of this procaryotic bromoperoxidase is 155,000 to 158,000 . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows a single band having the mobility of a 77,000-molecular-weight species . We thus conclude that this bromoperoxidase exists in solution as a dimeric species . The heme prosthetic group of bromoperoxidase is ferriprotoporphyrin IX . The spectral properties of the native and reduced enzyme are reported . This bromoperoxidase is the first halogenating enzyme purified from procaryotic sources. Prikl Biokhim Mikrobiol, 1985 Mar-Apr, 21(2), 177 - 83 {Production and properties of an immobilized enzyme preparation with peptidase activity}; Deniakina EK et al.; A heterogeneous multienzyme preparation with the peptidase activity, isolated from the cells of Pseudomonadacea bacteria, was immobilized on alumina . The specific activity of the immobilized enzyme complex is not a simple function of the bound protein quantity, but depends on immobilization conditions . An additional glutaraldehyde treatment results in higher thermostability of the immobilized enzyme preparation . The substrate specificity of the preparation retains after immobilization, and it becomes less sensitive to pH changes. Gan To Kagaku Ryoho, 1985 Mar, 12(3 Pt 2), 660 - 8 {The effect of calcium antagonists on intracellular transport of epidermal growth factor and the conjugate of epidermal growth factor with pseudomonas exotoxin}; Akiyama S; Calcium antagonists, verapamil and D600, enhance the toxicity of EGF-PE and anti-TFR-PE . EGF and EGF-PE are accumulated in their intact form in the lysosomes of cells treated with D600 or verapamil . Verapamil and D600 decrease the activity of lysosomal enzymes, especially of cathepsin B . Verapamil may enhance the cytotoxicity of EGF-PE by blocking lysosomal degradation of EGF-PE. Br J Ophthalmol, 1985 Mar, 69(3), 189 - 91 Gentamicin-resistant Pseudomonas endophthalmitis after penetrating keratoplasty; Insler MS et al.; A case of pseudomonas endophthalmitis after penetrating keratoplasty is described . Bacterial sensitivities in this case and in five of six previously reported cases of endophthalmitis after corneal transplant showed resistance to gentamicin, an antibiotic routinely used in the transport medium . The recovery of antibiotic resistant bacteria in antibiotic supplemented media is discussed with emphasis on prevention. Biochem J, 1985 Mar 1, 226(2), 469 - 76 Irreversible inhibition of delta 5-3-oxosteroid isomerase by 2-substituted progesterones; Penning TM; 2 alpha-Cyanoprogesterone (I) and 2-hydroxymethyleneprogesterone (II) were synthesized and screened as irreversible active-site-directed inhibitors of the delta 5-3-oxosteroid isomerase (EC 5.3.3.1) from Pseudomonas testosteroni . Both compounds were found to inhibit the purified bacterial enzyme in a time-dependent manner . In either case the inactivated enzyme could be dialysed without return of activity, indicating that a stable covalent bond had formed between the inhibitor and the enzyme . Inactivation mediated by compounds (I) and (II) followed pseudo-first-order kinetics, and at higher inhibitor concentrations saturation was observed . The competitive inhibitor 17 beta-oestradiol offered protection against the inactivation mediated by both compounds, and initial-rate studies indicated that compounds (I) and (II) can also act as competitive inhibitors yielding Ki values identical with those generated during inactivation experiments . 2 alpha-Cyanoprogesterone (I) and 2-hydroxymethyleneprogesterone (II) thus appear to be active-site-directed . To compare the reactivity of these 2-substituted progesterones with other irreversible inhibitors of the isomerase, 3 beta-spiro-oxiranyl-5 alpha-pregnan-20 beta-ol (III) was synthesized as the C21 analogue of 3 beta-spiro-oxiranyl-5 alpha-androstan-17 beta-ol, which is a potent inactivator of the isomerase {Pollack, Kayser & Bevins (1979) Biochem . Biophys . Res . Commun . 91, 783-790} . Comparison of the bimolecular rate constants for inactivation (k+3/Ki) mediated by compounds (I)-(III) indicated the following order of reactivity: (III) greater than (II) greater than (I) . 2-Mercaptoethanol offers complete protection against the inactivation of the isomerase mediated by 2 alpha-cyanoprogesterone (I) . Under the conditions of inactivation compound (I) appears to be completely stable, and no evidence could be obtained for enolate ion formation in the presence or absence of enzyme . It is suggested that cyanoprogesterone inactivates the isomerase after direct nucleophilic attack at the electropositive 2-position, and that tautomerization plays no role in the inactivation event . By contrast, 2-mercaptoethanol offers no protection against the inactivation mediated by 2-hydroxymethyleneprogesterone, and under the conditions of inactivation this compound appears to exist in the semi-enolized form. J Bacteriol, 1985 Mar, 161(3), 1103 - 11 Comparison of physical and genetic properties of palindromic DNA sequences; Warren GJ et al.; Some viable palindromic DNA sequences were found to cause an increase in the recovery of genetic recombinants . Although these palindromes contained no Chi sites, their presence in cis caused apparent recA+-dependent recombination to increase severalfold . This biological property did not correlate with the physical properties of the palindromes' extrusion of cruciform structures in vitro . Thus, two unrelated palindromes with similar effects on recombination in both Escherichia coli and Pseudomonas syringae displayed quite different kinetics of cruciform formation . In plasmids of native superhelical density, one palindrome underwent rapid cruciform formation at 55 degrees C, whereas the other did not form detectable cruciforms at any temperature . A shorter palindrome with similarly rapid kinetics of cruciform formation did not affect recombination detectably . The lack of a clear relationship between physical and genetic properties was also demonstrated in the case of longer, inviable palindromes . Here we found that the degree of asymmetry required in vivo to rescue a long palindrome from inviability far exceeded that required to kinetically prohibit cruciform extrusion in vitro. Cancer Res, 1985 Mar, 45(3), 1005 - 7 Potentiation of cytotoxic activity of immunotoxins on cultured human cells; Akiyama S et al.; The cytotoxic activity against human tumor cells of toxic conjugates of Pseudomonas exotoxin with anti-transferrin receptor antibody or epidermal growth factor was potentiated up to 10 to 20-fold by the calcium antagonists verapamil, D-600, and diltiazem and by the lysosomotropic agent beta-glycylphenyl-naphthylamide . The potentiating activity of these agents could be predicted by measuring the inhibition of protein synthesis by the immunotoxins on various cell lines . The use of potentiating agents such as these in combination with immunotoxins may prove useful in the treatment of some human cancers. Mol Gen Mikrobiol Virusol, 1985 Mar, (3), 27 - 30 {Transfer of prophage Mu into methylotrophic bacteria in the plasmid RP4}; Naumov GN et al.; Bacteriophage Mu genome has been transferred into the cells of Pseudomonas methanolica and Methylobacterium sp . SKF240, that are naturally resistant to the bacteriophage, as a fragment of a hybrid plasmid RP4::Mu cts62 . Temperature induction of the bacteriophage results in host cell lysis . Plasmid RP::Mu cis62 is maintained in methylotrophic cells presenting a cointegrative structure.The genetic and electrophoretic, analyses of the DNA isolated from transconjugant cells have confirmed the conclusion . Bacteriophage Mu propagation has been shown to be restricted in methylotrophic cells. Biochemistry, 1985 Feb 26, 24(5), 1110 - 6 Gamma-butyrobetaine hydroxylase: stereochemical course of the hydroxylation reaction; Englard S et al.; The stereochemical course of the aliphatic hydroxylation of gamma-butyrobetaine by calf liver and by Pseudomonas sp AK1 gamma-butyrobetaine hydroxylases has been determined . With {3(RS)-3-3H}-gamma-butyrobetaine or {3(R)-3-3H}-gamma-butyrobetaine as substrate, a rapid and significant loss of tritium to the medium occurred . On the other hand, with {3(S)-3-3H}-gamma-butyrobetaine, only a negligible release of tritium to the aqueous medium was observed . Indeed, on hydroxylation of {3(S)-3-2H}-gamma-butyrobetaine by either the calf liver or bacterial hydroxylase, the isolated product L-carnitine was found to have retained all of the deuterium initially present in the 3(S) position . Since the absolute configuration of the product L-carnitine has been determined to be R, such results are only compatible with a hydroxylation reaction that proceeded with retention of configuration . With {methyl-14C,3(R)-3-3H}-gamma-butyrobetaine as substrate for the calf liver hydroxylase, the percentage of tritium retained in the {methyl-14C}-L-carnitine product was determined as a function of percent reaction . The results of these studies indicated that pro-R hydrogen atom abstraction exceeded 99.9% . Experiments using racemic {methyl-14C,3(RS)-3-3H}-gamma-butyrobetaine as substrate yielded similar results and additionally allowed us to estimate alpha-secondary tritium kinetic isotope effects of 1.10 and 1.31 for the bacterial and calf liver enzymes, respectively . These results are discussed within the context of the radical mechanism for gamma-butyrobetaine hydroxylase previously proposed {Blanchard, J . S., & Englard, S . (1983) Biochemistry 22, 5922}, and the required topographical arrangement of enzymic oxidant and substrate is illustrated. J Biol Chem, 1985 Feb 25, 260(4), 2379 - 83 Enzymes of vitamin B6 degradation . Purification and properties of two N-acetylamidohydrolases; Huynh MS et al.; alpha-(N-Acetylaminomethylene)succinic acid hydrolase (Compound A hydrolase, EC 3.5.1-) and alpha-hydroxymethyl-alpha'-(N-acetylaminomethylene)succinic acid hydrolase (Compound B hydrolase, EC 3.5.1-) were purified to homogeneity from Pseudomonas MA-1 and Arthrobacter Cr-7, respectively . The two inducible enzymes catalyze Reactions 1 and 2, respectively, which release the first generally useful anabolic intermediates during growth of these organisms with (formula; see text) pyridoxine as a sole source of carbon and nitrogen . Compound A hydrolase is a monomeric protein of Mr 32,500 with aspartic acid as its NH2-terminal residue . Compound B hydrolase (Mr congruent to 205,000) is a multimer containing probably six identical subunits with glycine as the NH2 terminus . The two enzymes have quite different amino acid analyses, although both are high in Asx and Glx, lack tryptophan, and show similar stabilities to pH and temperature . Compound A hydrolase has a pI of 4.4, a Km of 3.3 microM, and a Vmax of 3.1 mumol X min-1 X mg-1 at pH 6.5 and 25 degrees C; no analogue substrates were found . Compound B hydrolase has a pI of 4.2, a Km of 25 microM, and a Vmax of 3.8 mumol X min-1 X mg-1 at 25 degrees C and pH 7.0; it also hydrolyzes Compound A slowly . Both enzymes are inhibited competitively by di- and tricarboxylic acids, itaconic acid being among the most effective . Sulfite inhibits both enzymes by a time-dependent mechanism not yet understood . The two amidases appear to differ greatly in architecture despite the similarity in properties and in the overall reactions they catalyze. Biochem J, 1985 Feb 15, 226(1), 147 - 53 Purification and properties of S-adenosylmethionine: aldoxime O-methyltransferase from Pseudomonas sp . N.C.I.B . 11652; Harper DB et al.; An enzyme catalysing the O-methylation of isobutyraldoxime by S-adenosyl-L-methionine was isolated from Pseudomonas sp . N.C.I.B . 11652 . The enzyme was purified 220-fold by DEAE-cellulose chromatography, (NH4)2SO4 fractionation, gel filtration on Sephadex G-100 and chromatography on calcium phosphate gel . Homogeneity of the enzyme preparation was confirmed by isoelectric focusing on polyacrylamide gel and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis . The enzyme showed a narrow pH optimum at 10.25, required thiol-protecting agents for activity and was rapidly denatured at temperatures above 35 degrees C . The Km values for isobutyraldoxime and S-adenosyl-L-methionine were respectively 0.24 mM and 0.15 mM . Studies on substrate specificity indicated that attack was mainly restricted to oximes of C4-C6 aldehydes, with preference being shown for those with branching in the 2- or 3-position . Ketoximes were not substrates for the enzyme . Gel filtration on Sephadex G-100 gave an Mr of 84 000 for the intact enzyme, and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated an Mr of 37 500, suggesting the presence of two subunits in the intact enzyme . S-Adenosylhomocysteine was a powerful competitive inhibitor of S-adenosylmethionine, with a Ki of 0.027 mM . The enzyme was also susceptible to inhibition by thiol-blocking reagents and heavy-metal ions . Mg2+ was not required for maximum activity. J Biol Chem, 1985 Feb 10, 260(3), 1400 - 6 Infrared spectra of carbon monoxide complexes of indoleamine 2,3-dioxygenase and L-tryptophan 2,3-dioxygenases . Effects of substrates on the CO-stretching frequencies; Uchida K et al.; Carbonmonoxy indoleamine 2,3-dioxygenase from rabbit small intestine exhibited two CO stretch bands at 1953 and 1933 cm-1 with half-band widths (delta v 1/2) of both approximately 15 cm-1 . Upon addition of an excess amount of L-tryptophan, the substrate, the spectrum changed into that with an intense single band at 1902 cm-1 with the delta v 1/2 of 15 cm-1 . Carbonmonoxy L-tryptophan 2,3-dioxygenase of Pseudomonas acidovorans in the absence of L-tryptophan showed a fused CO stretch band which consists of two components at 1965 and 1958 cm-1 (delta v 1/2 for the fused band; 25 cm-1), which was converted into a sharp single band at 1968 cm-1 (delta v 1/2; 10 cm-1) upon addition of excess L-tryptophan . On the other hand, CO complex of rat liver L-tryptophan 2,3-dioxygenase in the absence of L-tryptophan gave a spectrum with a poorly defined peak around 1961 cm-1 . By the addition of L-tryptophan, the spectrum changed into that with two distinct bands at 1972 and 1920 cm-1 (delta v 1/2; 6 and 13 cm-1, respectively) . These spectra were insensitive to pH in a range where the enzymes were not denatured (neutral to near pH 9) . The infrared spectra of the carbonmonoxy enzymes were also affected by the addition of certain effectors such as skatole and alpha-methyl-DL-tryptophan, which facilitate the binding of L-tryptophan to the catalytic site of intestinal and Pseudomonas enzymes, respectively . However, the changes were of different types from those by the saturating amount of L-tryptophan . Possible mechanisms for these phenomena are discussed in relation to the structure of the heme-CO complex in these heme-containing dioxygenases. J Appl Bacteriol, 1985 Feb, 58(2), 167 - 74 Regulation of syringomycin synthesis in Pseudomonas syringae pv . syringae and defined conditions for its production; Gross DC; Production of the phytotoxin, syringomycin (SR), by Pseudomonas syringae pv . syringae strain B301D was regulated by both iron and inorganic phosphate similar to that of many bacterial secondary metabolites . Iron concentrations of 2 mumol/l or more in deferrated potato-dextrose broth (PDB) resulted in the production of 1024 SR units/ml, a yield comparable to that produced in non-deferrated PDB . Moreover, production of one SR unit required approximately 0.4 ng of available FeCl3 . No SR was produced by strain B301D in deferrated PDB despite growth nearly identical with that of B301D in deferrated PDB supplemented with 10 mumol/l FeCl3 . Furthermore, a phosphate concentration of 1 mmol/l or more was suppressive to SR production . Of the amino acids tested, L-histidine at a concentration of ca 20 mmol/l was the most effective nitrogen source for SR synthesis under defined conditions . Based on these observations, a synthetic medium, SR minimal, was formulated for SR or syringotoxin production by representative strains of Ps . syringae pv . syringae . The regulation of phytotoxin production is discussed in relation to pathogen survival and virulence. Arch Ophthalmol, 1985 Feb, 103(2), 216 - 8 Gentamicin, tobramycin, amikacin, and netilmicin levels in tears following intravenous administration; Woo FL et al.; Peak and trough tear and serum concentrations were determined in 27 human volunteers undergoing intravenous (IV) gentamicin sulfate, tobramycin sulfate, amikacin sulfate, and netilmicin sulfate therapy . Although effective serum concentrations were achieved, tear levels were subtherapeutic . The mean peak tear concentrations were 0.4 microgram/mL, 0.5 microgram/mL, 1.7 micrograms/mL, and 0.3 microgram/mL for gentamicin, tobramycin, amikacin, and netilmicin, respectively . These levels did not approach the minimum inhibitory concentrations for Pseudomonas and raise some concern regarding the risk-benefit ratio of IV antibiotics for bacterial keratitis. J Clin Microbiol, 1985 Feb, 21(2), 278 - 9 Pseudomonas pickettii as a cause of pseudobacteremia; Verschraegen G et al.; An outbreak of pseudobacteremia caused by Pseudomonas pickettii biovariant 1 is reported . The common source was the aqueous chlorhexidine solution prepared by the hospital pharmacy . The contamination problem caused by the antiseptic solution was eventually solved by a series of preventive measures. J Appl Bacteriol, 1985 Feb, 58(2), 175 - 85 The effect of pH on the selection of carbaryl-degrading bacteria from garden soil; Larkin MJ et al.; At an alkaline pH and in an aqueous solution carbaryl hydrolyses to form 1-naphthol, methylamine and carbon dioxide, but it is much more stable at an acid pH . Soil perfusion column experiments indicated that the rate of carbaryl degradation at pH 6.0 to 7.0 was limited by the rate of chemical hydrolysis . Bacterial communities of at least 12 and 14 members were selected in continuous cultures using carbaryl as the sole carbon and nitrogen source at pH 6.0 . These communities were supported by the slow formation of hydrolysis products and a carbaryl-degrading bacterium was not selected after greater than 2000 h . A bacterial community of at least eight members was selected using equimolar 1-naphthol and methylamine as its sole carbon and nitrogen source . In contrast, after a lag of between 10 and 50 days, soil perfusion column and continuous culture enrichments at pH 5.2 and 5.0, respectively, led to the selection of a Pseudomonas sp . which could utilize carbaryl as its sole carbon and nitrogen source. J Dairy Res, 1985 Feb, 52(1), 77 - 89 Isolation and characterization of heat stable proteinases from Pseudomonas isolate AFT 21; Stepaniak L et al.; Pseudomonas strain AFT 21 produced three heat stable extracellular proteinases in milk and nutrient broth at 7 or 21 degrees C, but the proportions depended on medium and cultivation temperature . The three proteinases were EDTA- and o-phenanthroline-sensitive metalloenzymes and were not inhibited by N-ethylmaleimide or phosphoramidon . Proteinases I and II showed maximum activity at pH 7-7.5 and proteinase III at pH 8.5 . All three enzymes showed maximum activity at 45-47.5 degrees C, but had relatively high (19-27% of maximum) activity at 4 degrees C . They were unstable at 55 degrees C in phosphate buffer, pH 6.6, or synthetic milk ultrafiltrate (SMUF) containing 12 mmol Ca2+, but were stabilized by short preheating at 100 degrees C . They were extremely heat stable in both phosphate buffer and SMUF, pH 6.6, at 70-150 degrees C . Their D-values at 140 degrees C were 69, 54 and 80 s respectively . The Z-values for Pseudomonas AFT 21 proteinase III in phosphate buffer and SMUF were 29.7 and 30.3 degrees C respectively; the corresponding activation energies for inactivation were 8.7 x 10(4) J mol-1 and 9.2 X 10(4) J mol-1. Biochimie, 1985 Feb, 67(2), 191 - 7 Catabolism of arylglycerol-beta-aryl ethers lignin model compounds by Pseudomonas cepacia 122; Odier E et al.; Pseudomonas cepacia 122 can grow on several lignin model compounds including the arylglycerol-beta-aryl ethers guaiacylglycerol-beta-coniferyl ether and guaiacylglycerol-beta-guaiacyl ether . Non-phenolic lignin model compounds are not degraded by this bacterium . The enzyme system catalyzing guaiacylglycerol-beta-guaiacyl ether dissimilation in Pseudomonas cepacia 122 is inducible and repressed by glucose . Guaiacylglycerol and guaiacylglycerol-beta-guaiacyl ether were identified as intermediates in guaiacylglycerol-beta-coniferyl ether catabolism . Guaiacol, guaiacoxyethanol, vanillin and vanillic acid were identified as intermediates of guaiacylglycerol-beta-guaiacyl ether breakdown indicating that a C alpha-C beta splitting mechanism is involved in the degradation of aryl-alkyl ethers by this bacterium. Cancer Res, 1985 Feb, 45(2), 751 - 7 Anti-transferrin receptor antibody linked to Pseudomonas exotoxin as a model immunotoxin in human ovarian carcinoma cell lines; Pirker R et al.; The present in vitro study was performed to evaluate the potential usefulness of immunotoxins in treating human ovarian carcinomas . A monoclonal antibody against the human transferrin receptor was covalently linked to Pseudomonas exotoxin . The activity of this immunotoxin (anti-TFR-PE) was studied in five ovarian carcinoma cell lines, a breast carcinoma cell line (MCF-7), and in KB cells . The ovarian carcinoma cell lines included one previously established cell line (A1847) and four recent isolates obtained from the malignant ascites of patients with metastatic ovarian carcinoma (OVCAR cell lines) . While all cell lines showed inhibition of protein synthesis by anti-TFR-PE, there were quantitative differences when the level of protein synthesis was assayed after a 12-hr incubation with the immunotoxin . These differences resulted from different kinetics of anti-TFR-PE activity in the various cell lines . Higher levels of cellular binding and internalization of anti-TFR were shown to contribute to increased toxicity of anti-TFR-PE . Verapamil increased the rate of protein synthesis inhibition and thus enhanced the toxicity of anti-TFR-PE in the OVCAR cell lines. Biochim Biophys Acta, 1985 Jan 21, 827(1), 63 - 72 Purification and properties of extracellular poly(3-hydroxybutyrate) depolymerases from Pseudomonas lemoignei; Nakayama K et al.; Extracellular poly(3-hydroxybutyrate) depolymerase was purified from the culture medium of Peudomonas lemoignei and separated into four isozymes (A1, A2, B1 and B2) by CM-Sepharose CL-6B chromatography . The molecular weights of A1 and A2 and those of B1 and B2 were estimated to be 54 000 and 58 000, respectively, by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . The isoelectric points of A1, A2, B1 and B2 were found to be approximately pH 9.7, 10.0, 10.0 and 10.6, respectively, by isoelectric focusing . All four enzymes hydrolyzed poly(3-hydroxybutyrate) and oligomeric esters of D-(-)-3-hydroxybutyrate, but showed no activity toward the dimeric ester . Analysis of hydrolytic products of the oligomeric esters with A1 and B2 suggested that the enzymes hydrolyzed mainly the second and third ester bonds from the free hydroxy terminus at different frequencies, depending upon the chain length of the substrates. Biochem J, 1985 Jan 15, 225(2), 435 - 9 Inhibition of class C beta-lactamases by (1'R,6R)-6-(1'-hydroxy)benzylpenicillanic acid SS-dioxide; Knight GC et al.; beta-Lactamases, enzymes that catalyse the hydrolysis of the beta-lactam ring in beta-lactam antibiotics, are divided into three classes, A, B and C, on the basis of the structures so far determined . There are relatively few effective inhibitors of class C beta-lactamases . A beta-lactam sulphone with a hydroxybenzyl side chain, namely (1'R,6R)-6-(1'-hydroxy)benzylpenicillanic acid SS-dioxide (I), has now been studied . The sulphone is a good mechanism-based inhibitor of class C beta-lactamases . At pH8, the inhibition of a Pseudomonas beta-lactamase is irreversible, and proceeds at a rate that is about one-tenth the rate of concurrent hydrolysis . The labelled enzyme has enhanced u.v . absorption and is probably an enamine . At a lower pH, however, inhibition is transitory. J Basic Microbiol, 1985, 25(8), 543 - 6 Detection of two ornithine carbamoyltransferases in a phaseolotoxin-producing strain Pseudomonas syringae pv . phaseolicola; Jahn O et al.; The phaseolotoxin-producing Pseudomonas syringae pv . phaseolicola strain 1321 contains two ornithine carbamoyltransferases which differ in resistance to phaseolotoxin . Whereas ornithine carbamoyltransferase 1 (OCT 1) is inhibited at low concentrations of phaseolotoxin, ornithine carbamoyltransferase 2 (OCT 2) is insensitive to phaseolotoxin . The activity of the insensitive enzyme is correlated with the amount of toxin formed. Circ Shock, 1985, 17(2), 121 - 36 Lung fluid and protein flux during postoperative sepsis; Holman JM Jr et al.; Pulmonary microvascular injury during sepsis after injury appears to be amplified with plasma fibronectin deficiency, but the degree of injury relative to the extent of sepsis has not been defined . We evaluated pulmonary vascular permeability in sheep as influenced by various levels of postoperative Pseudomonas sepsis during a period of plasma fibronectin deficiency . The hemodynamic response to Pseudomonas was very similar regardless of the intensity of septic challenge and characterized by systemic arterial hypotension, decreased cardiac output, and pulmonary arterial hypertension . In contrast, increased pulmonary microvascular permeability was observed with increments in the bacterial challenge . Thus, lung protein clearance (LPC) or so called pulmonary transvascular protein clearance (TPC) used as an index of lung vascular permeability was 9.1 +/- 1.9 ml/hr, 15.1 +/- 1.7 ml/hr, and 19.3 +/- 3.0 ml/hr 2 hr after low (3 X 10(9) i.v.; 1 X 10(10) i.p.), medium (3 X 10(9) i.v.; 3 X 10(10) i.p.), and high (5 X 10(9) i.v.; 5 X 10(10) i.p.) dose Pseudomonas challenges, respectively . Thus, the extent of the altered pulmonary microvascular integrity in sheep during sepsis after surgery in the presence of fibronectin deficiency is dependent on the degree of bacterial sepsis . In addition, infusion of cryoprecipitate was an effective means of reversing the plasma fibronectin deficiency . Accordingly, this may be used as a model to investigate the mechanism of altered lung fluid balance during postoperative septic shock and the effect of fibronectin on this response. Z Naturforsch {C}, 1985 Jan-Feb, 40(1-2), 51 - 60 Chemical and physical characterization of four interfacial-active rhamnolipids from Pseudomonas spec . DSM 2874 grown on n-alkanes; Syldatk C et al.; Four extracellular glycolipids produced under growth-limiting conditions were isolated from the culture broth of Pseudomonas spec . DSM 2874 . After purification by column and thick-layer chromatography they were identified as anionic rhamnolipids . 1H and 13C-NMR studies showed that two of these, beta(beta(2-O-alpha-L-rhamnopyranosyloxy)decanoyl)decanoic acid and beta(beta(2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosylox y)decanoyloxy) decanoic acid, were identical with compounds described previously, while the other more hydrophilic compounds, beta(2-O-alpha-L-rhamnopyranosyloxy)decanoic acid and beta(2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosyloxy) decanoic acid, were new compounds . Surface and interfacial activity of the organic crude extract and of the purified components were determined in different aqueous solutions . The pH-dependence of surface and interfacial properties of the two previously described rhamnolipids (4, 20, 23) were examined in Teorell-Stenhagen-buffer (supplemented with 10% NaCl) at pH 3.0 and pH 9.0 . All rhamnolipids reduced the surface-tension from 72 to about 30 mN/m and the interfacial-tension from 42 to about 1 mN/m . The critical micelle concentrations were of the order of 5 to 200 mg/l depending on the structure of the molecule. Scand J Infect Dis, 1985, 17(1), 63 - 6 Pseudomonas cepacia septicemia in patients with burns: report of two cases; Brauner A et al.; Pseudomonas cepacia has been ascribed to low pathogenicity in man . Within a 10-day period this organism caused 2 cases of septicemia in the Karolinska Hospital burn unit, one with fatal outcome . Both cases were severely burned patients . A serological response to Ps . cepacia was observed in the surviving patient . The blood isolates from the patients showed a very high degree of similarity in biochemical tests, indicating a common origin although the source was not found . The characteristic antibiogram with resistance to aminoglycosides as well as ampicillin and most cephalosporins causes therapeutic problems, since many septicemias of unknown origin are treated with a combination of ampicillin and an aminoglycoside. Acta Paediatr Scand, 1985 Jan, 74(1), 152 - 7 Chronic granulomatous disease associated with chronic glomerulonephritis; Frifelt JJ et al.; A boy with chronic granulomatous disease (CGD) developed glomerulonephritis at the age of 12 years . The glomerulonephritis progressed to terminal uraemia at age 15 when maintenance haemodialysis was started . The clinical course was complicated by pulmonary aspergillosis and Pseudomonas septicaemia from which he eventually died . The glomerulonephritis was of unknown origin, and a possible relationship between CGD and glomerulonephritis is discussed. Mikrobiologiia, 1985 Jan-Feb, 54(1), 136 - 40 {Oxidation of alpha-methylstyrene by Pseudomonas cultures}; Dzhusupova DB et al.; The work was aimed at studying alpha-methylstyrene oxidation by three active Pseudomonas cultures . P . acidovorans 9 oxidized alpha-methylstyrene only to acetophenone whereas two P . aeruginosa cultures assimilated the compound entirely . Analysis of the intermediate products and enzymes catalyzing the aromatic ring cleavage suggests that these strains oxidize alpha-methylstyrene via a new unknown pathway. J Basic Microbiol, 1985, 25(3), 187 - 95 {Characterization and differentiation of fluorescent pseudemonads by substrate utilization studies}; Schopp W et al.; A rapid procedure is described for detecting and differentiating pseudomonads by their ability to degrade substrates with diverse physico-chemical properties, including volatile and non-volatile water-insoluble compounds, strong organic acids and bases, surfactants, disinfectants, and other toxic substances . The bacteria are embedded in an agar medium containing mineral salts and exposed to about 5 mg amounts of six to eight different substrates placed on the surface of an agar plate . After incubation at 25 degrees C for 24 hrs, growth zones around utilizable substrates may be used to differentiate various Pseudomonas strains and species. J Basic Microbiol, 1985, 25(1), 43 - 9 {Enhanced formation of plaque in RNA phages/bacteria systems under the effect of surface-active preparations}; Menzel G; The effect of 21 surfactants on the plaque formation was tested in three RNA-phages/Escherichia coli systems and in one RNA-phage/Pseudomonas aeruginosa system . Especially anionic detergents proved to be able to influence the plaque formation substantially . In high concentrations of Metaupon, Fekunil 602, Fekunil S-BA, Emulgator W 270, and Emulgator O-BA the plaque formation by the phages M 12 and f2 was inhibited and in low concentrations it was promoted . In the system Q beta/E . coli AB 301 the effect of the detergents mentioned was restricted to the prevention of the appearance of plaques . The detergent E 30 brought about only plaque inhibitions, too, in all used RNA-phages/E . coli systems . The treatment with effective detergents in the system PP7/P . aeruginosa, however, increased only the number of plaques . This phenomenon was evident in the formation of radiate plaque patterns in the lawn of bacteria . In the case of RNA-phages of E . coli the formation of such trains of lysis depends on the choice of the nutritive medium . The addition of the detergent Fekunil 602 at different times after the contact between phages and host bacteria affects the length of the beams of lysis . The ionogenity of surfactants seems to be of importance for the formation of radiate plaque patterns, since the tested cationic compound and the nonionic surfactants, contrary to anionic surfactants, did not cause any beams of lysis. Zh Mikrobiol Epidemiol Immunobiol, 1985 Jan, (1), 55 - 9 {Immunoenzyme analysis method for detecting anti-Pseudomonas aeruginosa antibodies in persons inoculated with pyoimmunogen}; Severtsova MK et al.; The possibility of using the indirect ELISA techniques for evaluating the level of the post-vaccinal production of humoral antibodies in donors immunized with Pyoimmunogen . P . aeruginosa vaccine, has been studied . The specificity and high resolution of this test system, based on the immobilization of the antigens of the vaccinal preparation on a solid-phase carrier, have been demonstrated . A rational method for the evaluation of specific antibody titers with due regard to the spectrophotometric data indicating the results of the reaction and the degree of the dilution of the serum under test has been proposed. J Antibiot (Tokyo), 1985 Jan, 38(1), 17 - 23 Isolation and characterization of bulgecins, new bacterial metabolites with bulge-inducing activity; Shinagawa S et al.; Bulgecins A, B and C, new bacterial metabolites which induce the formation of bulges by cooperation with beta-lactam antibiotics, were isolated from the culture broth of Pseudomonas mesoacidophila . The three components, separated by column chromatography on QAE-Sephadex A-25, are water-soluble acidic compounds containing a sulfate group in the molecule . Acid hydrolysis showed that D-glucosamine and a new proline derivative are common constituents of the three components . In addition, taurine and beta-alanine are constituents of bulgecins A and B, respectively. Biochem Soc Symp, 1985, 50, 171 - 91 Entry mechanisms of protein toxins and picornaviruses; Olsnes S et al.; The mode of entry into cells of a number of protein toxins with intracellular sites of action and of three picornaviruses is discussed . Of the different toxins in this group, diphtheria toxin has been most thoroughly studied with respect to its uptake mechanism . This toxin binds to cell surface receptors which are possibly part of the major anion-transport system in the cells . The bound toxin is then endocytosed and, when the pH drops below pH 5, a normally hidden hydrophobic domain is exposed and inserted into the membrane . By a process which, in addition to low pH, requires chloride transport and a proton gradient across the membrane, the toxin A fragment is translocated to the cytosol . When diphtheria toxin is bound at the cell surface, rapid entry through the surface membrane can be induced by treatment with low pH . Modeccin and Pseudomonas exotoxin A also require low pH for entry, but low pH is not able to induce rapid entry of these toxins from the cell surface . Another group of toxins, abrin, ricin and viscumin, is characterized by the fact that low pH in the medium prevents the toxins from entering the cytosol, but not from entering endocytic vesicles . However, when the pH is subsequently returned to neutrality the endocytosed toxins are able to enter the cytosol . In the picornaviruses the entry of a single hydrophilic macromolecule per cell is also sufficient to induce maximal biological effect . Poliovirus, like diphtheria toxin, appears to enter the cytosol from an acidic intracellular compartment which may be the endosome . Also human rhinovirus 2 requires low pH for entry, whereas encephalomyocarditis virus does not enter at low pH . The similarities and differences between the uptake mechanisms of toxins and viruses are discussed. Scand J Urol Nephrol Suppl, 1985, 92, 49 - 51 Infections after transplantation of a contaminated kidney; Bijnen AB et al.; In a retrospective study of 350 consecutive cadaver renal transplants, at least 83 (24%) of the donor kidneys were found to be contaminated . This was associated with three perinephric abscesses, one mycotic aneurysm, one pseudomonas sepsis, and four superficial wound infections . As a result, three patients lost their grafts, and one patient died from septic complications . The one year graft survival of the remainder of the recipients of contaminated grafts did not differ from that of the whole series . It is concluded that contamination of donor kidneys is a frequent event, but serious complications are relatively rare. J Int Med Res, 1985, 13(5), 270 - 5 Pseudomonas cepacia: the sensitivity of nosocomial strains to new antibiotics; Santos Ferreira MO et al.; Pseudomonas cepacia, considered a phytopathogenic organism for many years, has been shown recently to be widely distributed geographically . The hospital environment has become an important source of this organism but the resistance of Ps . cepacia to most antibiotics has made the treatment of infections a problem . One hundred per cent of the strains tested have proved to be sensitive to the sulphonamides and to novobiocin, 93.0% to the combination of trimethoprim and sulfamethoxazole (co-trimoxazole); 85.2% to minocycline; 77.8% to chloramphenicol and dibekacin and 44.4% to nalidixic acid . One hundred per cent of the strains exhibit resistance to ampicillin, cephalothin, cefamandole, cefoxitin, colistin, cefuroxime, tetracycline and cefazolin; 88.9% to amikacin, tobramycin and sisomycin; 85.2% to carbenicillin . The new beta-lactams, apalcillin, ceftazidime, N-formimidoyl-thienamycin, piperacillin, cefotaxime and azlocillin proved to be the most potent of the molecules tested, inhibiting 90% of the strains, at concentrations of 4, 8, 8, 8, 32 and 16 mg/l and 100% of the strains at 8, 16, 16, 32, 32 and 64 mg/l, respectively . In contrast to the usual sensitivity patterns of Pseudomonas spp, Ps . cepacia has been shown to be resistant to colistin, cefsulodin and the aminoglycosides . However, unlike Ps . aeruginosa, Ps . cepacia has been shown, by the dilution method, to be sensitive to co-trimoxazole, 92.3% of the strains being inhibited by 16 mg/l. Scand J Infect Dis Suppl, 1985, 44, 63 - 7 Penetration of ceftazidime into the normal rabbit and human eye; Walstad RA et al.; The penetration of ceftazidime into the aqueous humour and the vitreous body of the rabbit eye, after intravenous (i.v.) bolus or subconjunctival injection, was investigated . A dose of 50 mg/kg body weight was administered . After i.v . administration the mean penetration into the aqueous humour was 13% of the plasma values . After subconjunctival injection into the left eye, mean levels of 14% and 25% of the plasma concentrations were found in the right and left eye, respectively . The concentrations in the vitreous body were in all cases below the ceftazidime detection limit (1 mg/l), i.e . less than 1% of the plasma levels . The mean penetration of ceftazidime into human aqueous humour (measured during cataract extraction) was 19% after 2 g i.v . bolus injection . Ceftazidime levels sufficient to inhibit the growth of most pathogens commonly responsible for intraocular infections, including Pseudomonas spp., were consistently found in the aqueous humour . However, inadequate concentrations were achieved in the vitreous body. Acta Paediatr Scand, 1985 Jan, 74(1), 107 - 13 Intensive treatment of pseudomonas chest infection in cystic fibrosis: a comparison of tobramycin and ticarcillin, and netilmicin and ticarcillin; Conway SP et al.; Seventeen cystic fibrosis patients aged 3.1 years to 19.8 years had 30 courses of intensive treatment for relapse of their pseudomonas chest infection . The combination of netilmicin and ticarcillin was compared with tobramycin and ticarcillin in an open study . A significant subjective and objective improvement occurred in all patients . Pseudomonas was cleared temporarily from the sputum in 11 out of the 30 courses of treatment (37%) . There was no significant difference between the netilmicin and tobramycin groups, nor evidence of sustained renal or ototoxicity . Intensive therapy of pseudomonas chest infection in cystic fibrosis patients is described in detail. Laryngoscope, 1985 Jan, 95(1), 29 - 33 Sinusitis in immunodeficient and immunosuppressed patients; Berlinger NT; Sinusitis tends to occur in immunodeficient and immunosuppressed patients during periods of severe leukopenia . This group of patients includes those with primary immunodeficiency diseases, patients with leukemia receiving chemotherapy, and those undergoing bone marrow transplantation or kidney transplantation . The clinical and radiographic signs may be minimal or initially unimpressive . Sinusitis due to Aspergillus, Phycomycetes, or Pseudomonas may be fulminant and even fatal, requiring extensive surgical procedures for control. Crit Care Med, 1985 Jan, 13(1), 46 - 8 Postextubation hypoxemia treated with a continuous positive airway pressure mask; Dehaven CB Jr et al.; Twenty-seven surgical patients who developed post-extubation hypoxemia unresponsive to routine respiration therapy (incentive spirometry and chest physical therapy) received continuous positive airway pressure (CPAP) delivered through a mask at an inspired oxygen fraction (FIO2) of 0.45 . All patients responded with an increased PaO2 and achieved a PaO2/FIO2 ratio of at least 300 with a mean CPAP of 8.3 +/- 2.8 cm H2O . Mean duration of treatment was 23 +/- 14 h . Two (7%) patients required reintubation, one for control of excessive secretions and the other for persistent Pseudomonas pneumonia . Mask CPAP was an effective treatment for postextubation hypoxemia in this group of surgical patients. Mol Gen Mikrobiol Virusol, 1985 Jan, (1), 22 - 5 {Conjugative R-plasmid of facultative methylotrophic bacteria Pseudomonas sp . M}; Zheldakova RA et al.; 50 Md conjugative plasmid, designated pM3, has been found in the cells from natural isolates of Pseudomonas sp M . The plasmid determines the resistance to tetracycline and streptomycin and is capable of conjugative transfer between the cells of Pseudomonas and Escherichia coli . The conjugative derivatives of pM3 deleted for 14 Md of molecular mass were isolated after acridine dyes treatment of cells harbouring plasmid pM3 . The discovered plasmid was not shown to belong to IncP1 incompatibility group. Gene, 1985, 36(1-2), 27 - 36 Cloning vectors derived from the Pseudomonas plasmid pVS1; Itoh Y et al.; The Pseudomonas plasmid pVS1, which has about seven copies, was reduced to a minimal replicon and used to construct stable gene-cloning vehicles . The host for all cloning experiments was P . aeruginosa strain PAO . Two nonmobilizable plasmids, pME260 and pME290, and one RP1-mobilizable plasmid, pME285, were constructed . The vectors pME260 (6.3 kb) and pME290 (6.8 kb) carry the Tn801 bla gene specifying carbenicillin (Cb) resistance, a good selective marker in Pseudomonas, and the Tn903 aph gene encoding kanamycin (Km) resistance, with useful restriction sites for insertional inactivation . The Mob+ vector pME285 (10.6 kb) carries the aph gene and the Tn501-derived merRTCA genes coding for mercuric ion resistance, another good selective marker in Pseudomonas . The hypothetical merD gene, which may follow the merA gene in Tn501 but is absent from pME285, appeared to be dispensable for mercuric ion resistance in P . aeruginosa . The Mob- vector pME290 could be introduced by transformation and maintained in strains of P . aeruginosa, P . fluorescens, P . putida, P . acidovorans, P . stutzeri, P . mendocina, P . cepacia, and P . syringae . The plasmid was compatible with IncP-1 and IncP-4 replicons. Neurochirurgia (Stuttg), 1985 Jan, 28(1), 12 - 6 {Current aspects of pseudomonas meningitis}; Bruckner O et al.; The article reports on the incidence, the conditions of occurrence, possibilities and successes of treatment with certain (combinations of) antibiotics, in dealing with cases of pseudomonas meningitis . The various possible substances used for treatment are discussed . Rates of penetration and CSF concentrations of azlocillin and cefsulodin are stated . Alternative possibilities for treatment are pointed out. Antibiot Chemother, 1985, 36, 49 - 57 Pseudomonas pili . Studies on antigenic determinants and mammalian cell receptors; Paranchych W et al.; P . aeruginosa PAK pili are thin 5.2 nm diameter filaments containing a single 15-kd polypeptide subunit which is 144 amino acid residues in length . Studies on pili binding to a variety of synthetic sugars representing many di- tri- and tetra-saccharide structures found in mammalian glycoproteins and glycolipids failed to reveal any significant binding activity . On the other hand, a wide spectrum of binding activities was observed when a variety of structural proteins and enzymes were used as binding substrates . Of 30 proteins tested, phosphorylase b, pyruvate kinase and aldolase showed highest pilus binding activity . It was concluded that the PAK pilus receptor is probably a polypeptide rather than an oligosaccharide . Using arginine-specific cleavage to produce four large peptides, several proteases to produce subfragments of the large peptides, and antipilus rabbit antiserum, PAK pilin was found to contain four antigenic determinants . Epitopes near the NH2- and COOH-termini were only weakly immunogenic, whereas two epitopes near the center of the pilus protein titrated about 85% of the antipilus antibodies . Cleavage of the pilus protein into smaller peptides resulted in marked decreases in the affinity of antigenic peptides for their specific antibodies, suggesting that the immunodominant epitopes of PAK pilin are conformation-specific. Biochem J, 1984 Dec 15, 224(3), 755 - 9 Complement-component-C3-opsonized immunoglobulin G anti-DNA antibodies do not bind effectively to red blood cells unless aggregated on a high-Mr DNA matrix; Horgan C et al.; Large, soluble ds (double-stranded) DNA-IgG (immunoglobulin G) anti-dsDNA immune complexes (greater than or equal to 200 S) that were previously opsonized with complement were digested with DNAase . The small complement-component-C3-fragment-labelled IgG (11-14 S) that was then isolated did not bi