Pseudomonas

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 bind effectively to complement receptor type 1 on human red blood cells . However, when this IgG was immune-complexed with 3H-labelled PM2 (bacteriophage directed against a marine Pseudomonas) dsDNA (Mr approximately 6 X 10(6), substantial binding of both the DNA antigen and IgG to the erythrocytes was demonstrable.

J Biol Chem, 1984 Dec 10, 259(23), 14840 - 2
Characterization of crystals of protocatechuate 3,4-dioxygenase from Pseudomonas cepacia; Ludwig ML et al.; Protocatechuate 3,4-dioxygenase (EC 1.13.11.3) from Pseudomonas cepacia (Mr = 198,000) has been crystallized from potassium phosphate at pH 7.5 . Of the two orthorhombic forms that have been characterized, one contains a single molecule of tetrameric protocatechuate dioxygenase, (alpha beta Fe3+)4, per asymmetric unit . The space group and cell dimensions for this form are: P2(1)2(1)2(1) with a = 125.9 A, b = 161.7 A, and c = 85.2 A.

J Biol Chem, 1984 Dec 10, 259(23), 14788 - 92
Immunocytochemical localization of carbon monoxide oxidase in Pseudomonas carboxydovorans . The enzyme is attached to the inner aspect of the cytoplasmic membrane; Rohde M et al.; The localization of carbon monoxide oxidase (CO oxidase), the key enzyme in CO metabolism of Pseudomonas carboxydovorans, was examined using modified immunoferritin and protein A-gold techniques . Cell extracts were incubated with specific immunoglobulin G antibodies raised against CO oxidase, followed by treatment with ferritin-conjugated goat-anti-rabbit immunoglobulin G antibodies (pre-embedding labeling) . Electron microscopic examination of ultrathin sections showed cytoplasmic membranes and inside-out vesicles labeled at the inner aspect, whereas the outer sides of protoplasts and membrane vesicles remained completely unlabeled . The highly sensitive protein A-gold method has been modified to allow labeling of CO oxidase with good ultrastructural preservation of the bacterial cell . Glutaraldehyde-fixed cells of P . carboxydovorans were osmificated and embedded in glycol methacrylate . Etched ultrathin sections were treated with sodium metaperiodate and incubated with the specific antibodies against CO oxidase . These antibodies were then allowed to react with protein A-gold complexes (postembedding labeling) . Exponentially grown cells showed 87% of CO oxidase associated with the cytoplasmic membrane and 13% of the enzyme in the cytoplasm . The results indicate that CO oxidase is attached in vivo to the inner aspect of the cytoplasmic membrane and suggest interaction of the enzyme with a membrane-bound electron acceptor . The ratio of enzyme associated with the cytoplasmic membrane decreased to 50% in the stationary growth phase.

Zh Mikrobiol Epidemiol Immunobiol, 1984 Dec, (12), 59 - 62
{Pleuropneumonia caused by Pseudomonas stutzeri}; Serkova GP et al.; Biological characteristics and antibiotic sensitivity of P . stutzeri strain, isolated from a child with pleuropneumonia, are presented . Formation of rugous colonies, growth at 41 degrees C and in the presence of 6.5% of NaCl, the positive results of the oxidase and nitrate reductase tests, the negative signs of arginine hydrolase and lysine decarboxylase activity permit the identification of this Pseudomonas species . The isolated culture has proved to be sensitive to amino glycoside antibiotics, carbonicillin and polymyxin.

Can J Microbiol, 1984 Dec, 30(12), 1429 - 36
Metabolism of alpha-terpineol by Pseudomonas incognita; Madyastha KM et al.; Details of the metabolism of alpha-terpineol by Pseudomonas incognita are presented . Degradation of alpha-terpineol by this organism resulted in the formation of a number of acidic and neutral metabolites . Among the acidic metabolites, beta-isopropyl pimelic acid, 1-hydroxy-4-isopropenyl-cyclohexane-1-carboxylic acid, 8-hydroxycumic acid, oleuropeic acid, cumic acid, and p-isopropenyl benzoic acid have been identified . Neutral metabolites identified were limonene, p-cymene-8-ol, 2-hydroxycineole, and uroterpenol . Cell-free extracts prepared from alpha-terpineol adapted cells were shown to convert alpha-terpineol, p-cymene-8-ol, and limonene to oleuropeic acid, 8-hydroxycumic acid, and perillic acid, respectively, in the presence of NADH . The same cell-free extract contained NAD+ -specific dehydrogenase(s) which converted oleuropyl alcohol, p-cymene-7,8-diol, and perillyl alcohol to their corresponding 7-carboxy acids . On the basis of various metabolites isolated from the culture medium, together with the supporting evidence obtained from enzymatic and growth studies, it appears that P . incognita degrades alpha-terpineol by at least three different routes . While one of the pathways seems to operate via oleuropeic acid, a second may be initiated through the aromatization of alpha-terpineol . The third pathway may involve the formation of limonene from alpha-terpineol and its further metabolism.

J Clin Microbiol, 1984 Dec, 20(6), 1225 - 6
Pseudomonas paucimobilis peritonitis in patients treated by peritoneal dialysis; Glupczynski Y et al.; Pseudomonas paucimobilis has rarely been reported as an opportunistic human pathogen . We report the isolation of this organism in two patients who developed peritonitis during the course of intermittent or continuous ambulatory peritoneal dialysis . The origin of the infection was related to contamination of the dialysate in the first patient but could not be determined in the second case.

Appl Environ Microbiol, 1984 Dec, 48(6), 1076 - 83
Validity of the tritiated thymidine method for estimating bacterial growth rates: measurement of isotope dilution during DNA synthesis; Pollard PC et al.; The rate of tritiated thymidine incorporation into DNA was used to estimate bacterial growth rates in aquatic environments . To be accurate, the calculation of growth rates has to include a factor for the dilution of isotope before incorporation . The validity of an isotope dilution analysis to determine this factor was verified in experiments reported here with cultures of a marine bacterium growing in a chemostat . Growth rates calculated from data on chemostat dilution rates and cell density agreed well with rates calculated by tritiated thymidine incorporation into DNA and isotope dilution analysis . With sufficiently high concentrations of exogenous thymidine, de novo synthesis of deoxythymidine monophosphate was inhibited, thereby preventing the endogenous dilution of isotope . The thymidine technique was also shown to be useful for measuring growth rates of mixed suspensions of bacteria growing anaerobically . Thymidine was incorporated into the DNA of a range of marine pseudomonads that were investigated . Three species did not take up thymidine . The common marine cyanobacterium Synechococcus species did not incorporate thymidine into DNA.

South Med J, 1984 Dec, 77(12), 1541 - 4
Malignant external otitis; Anon JB et al.; Malignant external otitis (MEO) is a potentially life-threatening Pseudomonas infection of the external auditory canal in which rapid diagnosis is the key to proper treatment . We present an anatomic description of the ear and skull base, with a review of the literature on MEO, in the hope that it will enable primary care physicians to work more closely with otolaryngologists in dealing with this illness.

Can Med Assoc J, 1984 Dec 1, 131(11), 1365 - 7
Pleuropulmonary melioidosis in a Cambodian refugee; Chan CK et al.; A 47-year-old Cambodian refugee presented with an acute respiratory illness that featured consolidation of the lower lobe of the left lung and progressive involvement of the adjacent pleura caused by Pseudomonas pseudomallei . Initial difficulty in identifying the organism resulted in an inadequate duration of therapy . Chronic pleural disease followed, and the organism became resistant to many antibiotics during therapy . A diagnosis of pleuropulmonary melioidosis should be entertained and the microbiology laboratory alerted when patients with pneumonia who are from endemic areas are encountered, so that a diagnosis can be made early and the appropriate treatment begun.

J Virol, 1984 Dec, 52(3), 1036 - 8
DNA synthesis in Pseudomonas acidovorans infected with mutants of bacteriophage phi W-14 defective in the synthesis of alpha-putrescinylthymine; Miller PB et al.; Normal levels of the hypermodified pyrimidine, alpha-putrescinylthymine, which is formed from hydhydroxymethyluracil at the polynucleotide level (Maltman et al., J . Virol . 34:354-359, 1984), are not required in bacteriophage luminal diameterW-14 DNA for the DNA to serve as a replicative template in luminal diameterW-14-infected cells.

J Gen Microbiol, 1984 Dec, 130 ( Pt 12), 3175 - 82
Discriminant analysis of volatile fatty acids produced in culture medium: a novel approach to the identification of Pseudomonas species; Peladan F et al.; The volatile fatty acids produced in culture medium by 357 Pseudomonas strains belonging to eight species were determined quantitatively by GLC . The resultant chromatograms were submitted to discriminant analysis . Stable discriminant functions were computed and included in a computerized identification system which also involved some distinctive volatile fatty acids regarded as two-state qualitative characters (presence or absence characters) . Using a test group of 249 strains belonging to the studied species, more than 89% of the identifications made by this system agreed with those made by conventional biochemical methods despite the relatively poor differentiation between P . putida and P . fluorescens . When the individual species within the matrices were weighted with prior probabilities reflecting results given by two simple biochemical tests, 96% of the 249 strains were correctly identified.

Arch Biochem Biophys, 1984 Nov 15, 235(1), 41 - 7
Detection of beta-carbanion formation during kynurenine hydrolysis catalyzed by Pseudomonas marginalis kynureninase; Bild GS et al.; 2-Amino-4-hydroxy-4-phenylbutyric acid has been shown to be formed during the Pseudomonas marginalis kynureninase-catalyzed hydrolysis of kynurenine in the presence of benzaldehyde and pyridoxal phosphate . The formation of 2-amino-4-hydroxy-4-phenylbutyric acid is the first demonstration, to our knowledge, of the controlled trapping of an amino acid beta-carbanion generated either chemically or enzymatically, and is perhaps the best empirical evidence to date that enzyme mechanisms can proceed through a beta-carbanionic intermediate . The lifetime of the beta-carbanionic alanyl intermediate generated by kynureninase is of sufficient duration to allow reaction with benzaldehyde . Other aromatic, but no aliphatic, aldehydes will undergo electrophilic addition with kynureninase-generated beta-carbanionic alanyl intermediates to form the corresponding amino acid.

J Bacteriol, 1984 Nov, 160(2), 618 - 21
Oxidation and dehalogenation of 4-chlorophenylacetate by a two-component enzyme system from Pseudomonas sp . strain CBS3; Markus A et al.; In cell-free extracts from Pseudomonas sp . strain CBS3 the conversion of 4-chlorophenylacetate to 3,4-dihydroxyphenylacetate was demonstrated . By Sephacryl S-200 chromatography two protein fractions, A and B, were obtained which both were essential for enzyme activity . Fe2+ and NADH were cofactors of the reaction . NADPH also activated the enzyme, but less effectively than NADH . FAD had no influence on enzyme activity . 4-Hydroxyphenylacetate, 4-chloro-3-hydroxyphenylacetate, and 3-chloro-4-hydroxyphenylacetate were poor substrates for the enzyme, suggesting that these substances are not intermediates of the reaction . We therefore suggest that the reaction proceeds via a dioxygenated intermediate.

Gene, 1984 Nov, 31(1-3), 31 - 8
The complete nucleotide sequence of the Pseudomonas gene coding for carboxypeptidase G2; Minton NP et al.; The complete nucleotide sequence of the Pseudomonas chromosomal gene coding for the enzyme carboxypeptidase G2 (CPG2) has been determined . The nucleotide sequence obtained has been confirmed by comparing the predicted amino acid sequence with that of randomly derived peptide fragments and by N-terminal sequencing of the purified protein . The gene has been shown to code for a 22 amino acid signal peptide at its N-terminus which closely resembles the signal peptides of other secreted proteins . An alternative 36 amino acid signal peptide which may function in Pseudomonas has also been identified . The codon utilisation of the gene is influenced by the high G + C (67.2%) content of the DNA and exhibits a 92.8% preference for codons ending in G or C . This unusual codon preference may contribute to the generally observed weak expression of Pseudomonas genes in Escherichia coli . A region of DNA upstream of the structural gene has also been sequenced and a ribosome binding site and two putative promoter sequences identified.

J Bacteriol, 1984 Nov, 160(2), 718 - 23
Molecular cloning of a malyl coenzyme A lyase gene from Pseudomonas sp . strain AM1, a facultative methylotroph; Fulton GL et al.; A genomic library containing HindIII partial digests of Pseudomonas sp . strain AM1 DNA was constructed in the broad-host-range cosmid pVK100 . PCT57, a Pseudomonas sp . strain AM1 methanol mutant deficient in malyl coenzyme A lyase activity, was complemented to a methanol-positive phenotype by mobilization of the pVK100 library into PCT57 recipients with the ColE1/RK2 mobilizing plasmid pRK2013 . Six different complemented isolates all contained a recombinant plasmid carrying the same 19.6-kilobase-pair Pseudomonas sp . strain AM1 DNA insert . Subcloning and complementation analysis demonstrated that the gene deficient in PCT57 (mcl-1) was located in a 1.6-kilobase-pair region within a 7.4-kilobase-pair EcoRI-HindIII fragment.

Biochemistry, 1984 Oct 23, 23(22), 5195 - 201
Inactivation of the Pseudomonas striata broad specificity amino acid racemase by D and L isomers of beta-substituted alanines: kinetics, stoichiometry, active site peptide, and mechanistic studies; Roise D et al.; Mechanism-based inactivators were used to probe the active site of the broad specificity amino acid racemase from Pseudomonas striata . Kinetic parameters for the inactivation of the racemase with both stereoisomers of beta-fluoroalanine, beta-chloroalanine, and O-acetylserine were determined . By use of 14C-labeled O-acetylserines, the stoichiometry of inactivator binding was found to be one inactivator bound per enzyme subunit . The PLP-dependent enzyme contains one coenzyme per subunit, and after NaB3H4 reduction of the PLP-imine bond, followed by trypsin digestion of the protein, the amino acid sequence of the PLP-binding peptide was determined . Trypsin digestion of the enzyme labeled with either L or D isomer of O-acetylserine and sequencing of the labeled peptide revealed that the inactivators bind to the same lysine residue which binds PLP in native enzyme . The characterization of a PLP adduct released from inactivated enzyme under some conditions is also described . Implications of the formation of this compound with respect to the overall reaction mechanism of inactivation are discussed.

Biochem Biophys Res Commun, 1984 Oct 15, 124(1), 178 - 82
Incorporation of {18O}water into 4-hydroxybenzoic acid in the reaction of 4-chlorobenzoate dehalogenase from pseudomonas spec . CBS 3; Muller R et al.; 4-Chlorobenzoate is dehalogenated by 4-chlorobenzoate dehalogenase from Pseudomonas spec . CBS 3 to form 4-hydroxybenzoate . In 18O enriched water the hydroxygroup of 4-hydroxybenzoate is quantitatively labelled with 18O . This result clearly shows, that 4-chlorobenzoate dehalogenase catalyzes the hydrolytic cleavage of the halogen-carbon bond, without the involvement of molecular oxygen, a reaction not yet described.

Biochem J, 1984 Oct 15, 223(2), 487 - 94
Purification and some properties of the D-lactate-2-sulphatase of Pseudomonas syringae GG; Crescenzi AM et al.; A soil bacterium grown on propan-2-yl sulphate as sole source of carbon and sulphur yielded extracts containing an enzyme capable of liberating sulphate from racemic lactate-2-sulphate . The enzyme was purified to homogeneity by a combination of streptomycin sulphate precipitation of nucleic acids, batch treatment with DEAE-cellulose, and chromatography on columns of DEAE-cellulose, Sephacryl S-300 and butyl-agarose . The protein was monomeric with an Mr of 55 000-60 000 . The enzyme activity was specific for D-lactate-2-sulphate (Km 6.6 nM; maximal specific activity 14.3 mumol/min per mg of protein) and showed no activity towards the L-isomer . The products of the enzyme's action were inorganic sulphate and D-lactate which were released in equimolar amounts and stoicheiometrically with the amount of ester hydrolysed . No L-lactate was formed . Retention of configuration implied cleavage of the O-S bond of the C-O-S ester link and this was confirmed by 18O-incorporation experiments in which 18O from 18O-enriched water in the incubation medium was incorporated exclusively and quantitatively into inorganic sulphate . Only two other esters (serine-O-sulphate and p-nitrophenyl sulphate) of a total of 29 compounds tested were substrates for the enzyme . D-Lactate, L-lactate-2-sulphate and the substrate analogues glycollate-2-sulphate and butyrate-2-sulphate were significantly inhibitory.

Carbohydr Res, 1984 Oct 15, 133(2), 289 - 96
Structural studies of the O-antigen of the lipopolysaccharide from an avirulent strain (M4S) of Pseudomonas solanacearum; Akiyama Y et al.; The structure of the O-antigen of the lipopolysaccharide from an avirulent strain (M4S) of Pseudomonas solanacearum has been investigated by methylation analysis, n.m.r . spectroscopy, and N-deacetylation-deamination, followed by analysis and controlled Smith-degradation of the product . These studies demonstrate that the O-antigen is composed of a tetrasaccharide repeating-unit having the following structure: ----3)-alpha-D-GlcpNAc-(1----2)-alpha-L-Rhap-(1----2)-alpha- L-Rhap-(1----3)- alpha-L-Rhap-(1----.

HNO, 1984 Oct, 32(10), 431 - 2
{Malignant external otitis in a young diabetic--successful azlocillin treatment}; Ganz H; The basic considerations for staging of malignant otitis are complimented by a case report . Azlocillin infusion was successful in the treatment of a 15 years old diabetic girl with early osteitis of the mastoid due to a pseudomonas infection of the external ear canal.

Eur J Clin Microbiol, 1984 Oct, 3(5), 411 - 8
Immunological aspects of humidifier fever; Forsgren A et al.; An epidemiological investigation was made of eight employees working in a defined section of an office building who complained of febrile reactions accompanied by respiratory tract symptoms and fatigue . Culture of the water from the humidifier for their section yielded strains of Pseudomonas acidovorans and an unidentified Pseudomonas species . In vitro studies showed that these strains activated the alternative pathway of the complement system in Mg-EGTA supplemented serum, as evidenced by properdin depletion and conversion of C3-proactivator (Factor B) and C3 . Whereas the employees with symptoms had significantly reduced properdin values during the symptomatic phase, a slight C3 conversion and antibodies to Pseudomonas spp., staff working in another part of the building with a different ventilation system did not . It was concluded that staff with humidifier fever symptoms had been exposed to Pseudomonas spp . through the faulty ventilation system, and their symptoms were most likely a result of complement activation.

Biochem J, 1984 Oct 1, 223(1), 119 - 27
Multiple forms of gamma-butyrobetaine hydroxylase (EC 1.14.11.1); Lindstedt S et al.; gamma-Butyrobetaine hydroxylase {4-trimethylaminobutyrate, 2-oxoglutarate:oxygen oxidoreductase (3-hydroxylating), EC 1.14.11.1} from human kidney was resolved into three forms by chromatofocusing . After further chromatography on an anion-exchanger, each form appeared as a single band on electrophoresis in polyacrylamide gel containing sodium dodecyl sulphate . The isoelectric points of isoenzymes 1, 2 and 3 were 5.6, 5.7 and 5.8 respectively, as estimated by isoelectric focusing . Their specific activities were 17-29 mu kat/g of protein . The concentrations of the three isoenzymes were about equal, possibly slightly lower for isoenzyme 1 . The requirement for Fe2+ and the Km values for gamma-butyrobetaine and 2-oxoglutarate were about the same for the different enzyme forms . L- and D-Carnitine caused decarboxylation of 2-oxoglutarate to the same extent (8 and 29%) with the three forms . The enzyme forms had the same mass, 64 kDa, as determined by gel filtration in nondenaturing media . The same subunit mass, 42 kDa, was obtained for the multiple forms by electrophoresis in polyacrylamide gels containing sodium dodecyl sulphate . Isoenzyme 2 was resolved into two protein bands by isoelectric focusing in polyacrylamide gels containing urea . Isoenzyme 1 contained only one of these bands and isoenzyme 3 the other . The three enzyme forms of gamma-butyrobetaine hydroxylase thus appear to be dimeric combinations of two subunits differing in charge but not in size . gamma-Butyrobetaine hydroxylase from crude extracts of human, rat and calf liver was also separated into multiple forms by a chromatofocusing technique . The isoenzyme pattern was the same in human liver and kidney . The technique used to resolve the mammalian enzymes gave no evidence for the presence of multiple forms of the bacterial enzyme from Pseudomonas sp . AK 1.

J Cell Biol, 1984 Oct, 99(4 Pt 1), 1296 - 308
A single mutation in Chinese hamster ovary cells impairs both Golgi and endosomal functions; Robbins AR et al.; A Chinese hamster ovary cell mutant DTG 1-5-4, was selected for pleiotropic defects in receptor-mediated endocytosis by methods previously described (Robbins, A . R., S . S . Peng, and J . L . Marshall, 1983, J . Cell Biol., 96:1064-1071) . DTG 1-5-4 exhibited increased resistance to modeccin, Pseudomonas toxin, diphtheria toxin, Sindbis virus, and vesicular stomatitis virus, as well as decreased uptake via the mannose 6-phosphate receptor . Fluorescein-dextran-labeled endosomes isolated from DTG 1-5-4 were deficient in ATP-dependent acidification in vitro . Endocytosis and endosome acidification were both restored in revertants of DTG 1-5-4 and in hybrids of DTG 1-5-4 with DTF 1-5-1, another endocytosis mutant exhibiting decreased ATP-dependent endosome acidification . Both DTG 1-5-4 and DTF 1-5-1 were blocked at two stages of infection with Sindbis virus: at low multiplicities of infecting virus, resistance reflected a block in viral penetration into the cytoplasm, but at higher multiplicities of infection the block was in virus release . Like endocytosis, release of Sindbis virus was increased in revertants of DTG 1-5-4 and in DTG 1-5-4 X DTF 1-5-1 hybrids . Decreased release of virus from DTG 1-5-4 correlated with defects in some of the Golgi apparatus-associated steps of Sindbis glycoprotein maturation: proteolytic processing of the precursor pE2, galactosylation, and transport to the cell surface all were inhibited . In contrast, mannosylation, fucosylation, and acylation of the Sindbis glycoproteins, and galactosylation of vesicular stomatitis virus and cellular glycoproteins occurred to similar respective extents in mutant and parent . Electron microscopic examination of Sindbis-infected DTG 1-5-4 showed a remarkable accumulation of nucleocapsids bound to cisternae adjacent to the Golgi apparatus; virions were observed in the lumina of some of these cisternae . That the alterations in both endocytosis and Golgi-associated steps of viral maturation result from a single genetic lesion indicates that these processes are dependent on a common biochemical mechanism . We suggest that endocytic and secretory pathways may share a common component involved in ion transport.

J Bacteriol, 1984 Oct, 160(1), 304 - 12
Specificity and mechanism of ferrioxamine-mediated iron transport in Streptomyces pilosus; Muller G et al.; Although the ferrioxamines are an important and well-characterized class of siderophores produced by several species of Nocardia, Streptomyces, Micromonospora, Arthrobacter, Chromobacterium, and Pseudomonas, no studies of the mechanism of ferrioxamine-mediated iron uptake have been performed for an organism which produces the siderophore . This is the first report of metal transport in Streptomyces pilosus mediated by the native ferrioxamines B, D1, D2, and E . 55Fe accumulation in these ferrioxamines was dependent on metabolic energy and was a saturable process with increasing complex concentration . The apparent Km for {55Fe}ferrioxamine B uptake was approximately 0.2 microM . Both chromic desferriferrioxamine B and {67Ga}desferriFerrioxamine B were transported at rates similar to those of the 55Fe complexes: this implies that no decomplexation or reduction of the metal ion is required for transport, since the chromic complexes are kinetically inert and the gallium complexes have no stable divalent state as a possible reduction product . In addition, isomers of inert chromic desferriferrioxamine B complexes were used to probe the stereospecificity of the ferrioxamine uptake system . The chromic complexes were separated into three fractions by cationic exchange chromatography and assigned as two cis and a (mixture of) trans geometrical isomer(s) by their visible spectra . {55Fe}ferrioxamine B uptake was equally inhibited by each isomer, suggesting that no differentiation between cis and trans geometrical isomers occurs . In the presence of chromic desferriferrioxamine B isomers, the uptake rates for 55Fe-labeled ferrioxamines E, D1, and D2 were even more strongly reduced than was that for {55Fe}ferrioxamine B itself . From these results we conclude that all the ferrioxamines tested are transported into the cells by the same uptake system.

J Bacteriol, 1984 Oct, 160(1), 294 - 8
Transport of mevalonate by Pseudomonas sp . strain M; Gill JF Jr et al.; Pseudomonas sp . M, isolated from soil by elective culture on R,S-mevalonate as the sole source of carbon, possessed an inducible transport system for mevalonate . This high-affinity system had a pH optimum of 7.0, a temperature optimum of 30 degrees C, a Km for R,S-mevalonate of 88 microM, and a V max of 26 nmol of mevalonate transported per min/mg of cells (dry weight) . Transport was energy dependent since azide, cyanide, or m-chlorophenylhydrazone caused complete cessation of transport activity . Transport of mevalonate was highly substrate specific . Of the 16 structural analogs of mevalonate tested, only acetoacetate, mevinolin, and mevaldehyde significantly inhibited transport . Growth of cells on mevalonate induced transport activity by 40- to 65-fold over that observed in cells grown on alternate carbon sources . A biphasic pattern for cell growth, as well as for induction of mevalonate transport activity, was observed when mevalonate was added to a culture actively growing on glucose . The induction of transport activity under these conditions began within 30 min after the addition of mevalonate and reached 60% of maximal activity during phase I . A further increase in mevalonate transport activity occurred during phase II of growth . Glucose was the preferred carbon source for growth during phase I, whereas mevalonate was preferred during phase II . Only one isomer of the R,S-mevalonate mixture appeared to be utilized, since growth ceased after 45 to 50% of the total mevalonate was depleted from the medium . However, nearly 30% of the preferred mevalonate isomer was depleted from the medium during phase I without significant metabolism to CO2 . These results suggest that mevalonate or a mevalonate catabolite may accumulate in cells of Pseudomonas sp . M during phase I and that glucose metabolism may inhibit or repress the expression of enzymes further along the mevalonate catabolic pathway.

Biochemistry, 1984 Sep 25, 23(20), 4761 - 7
Electron-transfer reactions of photoreduced flavin analogues with c-type cytochromes: quantitation of steric and electrostatic factors; Meyer TE et al.; We have found correlations between rate constants and the difference in redox potential of the reactants for electron-transfer reactions between oxidized cytochromes and either photoproduced riboflavin or flavin mononucleotide (FMN) semiquinones (the latter rate constants extrapolated to infinite ionic strength) . The riboflavin-cytochrome rate constants are about 70% of those for reduction by lumiflavin, probably because of steric interference by the ribityl side chain . Reduction of cytochromes by FMN semiquinone was ionic strength dependent in all cases, due to electrostatic interactions . Extrapolation of rate constants to infinite ionic strength shows that the phosphate exerts a significant steric effect as well (rate constants average about 27% of those for lumiflavin, although part of this decrease is due to a difference in the semiquinone pK value) . Differences in the magnitude of the FMN steric effect correlate well with surface topology differences for those cytochromes whose three-dimensional structures are known . Mitochondrial cytochromes c and the cytochromes c2 all showed attractive (plus-minus) interaction with FMN in spite of the fact that some of these proteins have large net negative charges . Four small c-type cytochromes (including Pseudomonas cytochrome c-551) show a weak repulsive interaction with FMN semiquinone . We conclude that flavosemiquinones interact at a site on the cytochromes that is near the exposed heme edge . There is a large positive electrostatic field at this site in mitochondrial cytochrome c and the cytochromes c2, but this region is primarily hydrophobic in Pseudomonas cytochrome c-551 and in the other small bacterial cytochromes.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1984 Sep 10, 259(17), 10669 - 74
Mechanism-based inactivation of bacterial kynureninase by beta-substituted amino acids; Kishore GM; Kynureninase from Pseudomonas marginalis has been shown to catalyze the elimination of beta-functionalities of beta-substituted amino acids such as beta-chloro-L-alanine, resulting in the formation of aminoacrylate-pyridoxal phosphate-enzyme complex . This intermediate can be processed further to produce either pyruvate, ammonia, and active enzyme or an inactive enzyme complex . Approximately 1 in 500 turnovers leads to inactivation of the enzyme . The mechanism of inactivation appears to involve nucleophilic addition of a carboxylate group at the active site to the beta-carbon of the aminoacrylate complex . Both subunits of kynureninase have been shown to be catalytically competent although the native enzyme contains only one pyridoxal phosphate per dimer . Since both aspartate beta-decarboxylase and kynureninase catalyze mechanistically similar reactions, these results further support the notion that the two active sites may have several common features.

J Biol Chem, 1984 Sep 10, 259(17), 10682 - 8
The kinetic mechanism of salicylate hydroxylase as studied by initial rate measurement, rapid reaction kinetics, and isotope effects; Wang LH et al.; The kinetic mechanism of Pseudomonas cepacia salicylate hydroxylase has been examined by steady state initial rate measurements, and stopped flow and equilibrium studies . Results indicate that salicylate and NADH bind to the hydroxylase randomly . The enzyme is reduced and NAD+ is released . Oxygen subsequently binds to the reduced enzyme . substrate complex, leading to the production of hydroxylated product, CO2, and water . Based on results of anaerobic rapid mixing experiments, the rate of enzyme reduction by NADH is enhanced 290- and 240-fold when the hydroxylase is complexed with salicylate and benzoate (a nonsubstrate effector), respectively . Salicylate enhances, whereas benzoate slightly weakens, the NADH binding to the enzyme . Primary isotope effects were observed with (4R)-{4-2H}- and (4R)-{4-3H}NADH but not with the (4S)-{4-2H}NADH . Using varying concentrations of benzoate, the pattern of tritium isotope effect on Vm/Km, T(V/K), also indicates that benzoate and NADH bind to the enzyme randomly . The intrinsic isotope effects, Dk, of (4R)-{4-2H}NADH on the reduction of enzyme . salicylate and enzyme . benzoate complexes were found to be 5.57 and 5.96, respectively . The former is much repressed but the latter is only slightly so in the expression of their corresponding deuterium isotope effects on Vm, DV . Furthermore, values of DV (1.69 to 5.07) show a rough correlation with the extents of uncoupling of substrate hydroxylation and H2O2 formation activities for a series of benzenoid effectors . These results indicate that relative to the step of enzyme reduction, the subsequent reaction(s) leading to H2O2 formation must be fast whereas that for substrate hydroxylation contains at least one slow step.

J Biochem (Tokyo), 1984 Sep, 96(3), 637 - 43
Isolation and characterization of N-long chain acyl aminoacylase from Pseudomonas diminuta; Shintani Y et al.; N-Long chain acyl aminoacylase II (Enzyme II) catalyzing the hydrolysis of N-long chain acyl amino acids was purified about 2,000-fold from the cell extracts of Pseudomonas diminuta with 1.8% of activity yield . The purified enzyme was homogeneous on polyacrylamide gel electrophoresis and the molecular weight was 220,000 . Enzyme II differed from N-long chain acyl aminoacylase I (Enzyme I) in molecular weight, in substrate specificity, and in behavior toward temperature and pH . Enzyme II showed broader substrate specificity than Enzyme I and catalyzed the hydrolysis of lipoamino acids containing various amino acid residues, although Enzyme I was almost specific to the lipoamino acids containing L-glutamate . The extent of hydrolysis by Enzyme II reaction varied depending on the kinds of lipoamino acids and were: 100% for palmitoyl-L-glutamate, 91% for myristoyl-L-glutamate, 85% for lauroyl-L-glutamate, 54% for lauroyl-L-aspartate, 28% for stearoyl-L-glutamate and 17.5% for lauroyl-glycine.

Biochem J, 1984 Sep 1, 222(2), 315 - 20
Bacterial degradation of N-cyclopropylmelamine . The steps to ring cleavage; Cook AM et al.; The s-triazine cyclopropylmelamine (N-cyclopropyl-1,3,5-triazine-2,4,6-triamine) was degraded to about 6 mol of NH4+/mol of substrate by a mixture of two bacteria (strains A and D, both Pseudomonas spp.) Only strain A grew with cyclopropylmelamine as sole and limiting source of nitrogen . The organism obtained 2 mol of nitrogen/mol of substrate and excreted a product that was identified as cyclopropylammelide {6-cyclopropylamino-1,3,5-triazine-2,4(1 H,3 H)-dione} . Proteins in extracts from strain A were separated on a Sephadex G-200 column . Cyclopropylmelamine was found to be deaminated in two separable steps to cyclopropylammelide via cyclopropylammeline {4-amino-6-cyclopropylamino-1,3,5-triazine-2(1 H)-one}, which was identified . Strain D could not utilize cyclopropylmelamine or cyclopropylammeline, but could utilize cyclopropylammelide (or homologue) as sole and limiting source of nitrogen and obtain about 4 mol of nitrogen/mol of substrate . Proteins in cell extracts from strain D were separated on a DEAE-cellulose column . Alkylammelides were degraded quantitatively by one enzyme fraction to 1 mol of cyanuric acid plus 1 mol of alkylamine/mol of substrate . The specific activities of enzymes in extracts of the two strains were as high as the activities observed during growth . The three activities studied in the two strains were all active under aerobic and oxygen-free conditions . The reactions appear to be hydrolytic, yielding 2 mol of NH4+ plus 1 mol of cyclopropylamine and 1 mol of cyanuric acid/mol of substrate.

Infect Immun, 1984 Sep, 45(3), 596 - 603
Evidence for pseudomonas exotoxin A receptors on plasma membrane of toxin-sensitive lm fibroblasts; Manhart MD et al.; Pseudomonas exotoxin A enters mouse LM fibroblasts by receptor-mediated endocytosis and ultimately causes cell death . Here we present evidence for the existence of a specific receptor for the toxin . Toxin association with LM cells at 18 and 37 degrees C, but not at 4 degrees C, was highly specific . At 37 degrees C, the association increased with time, reaching a steady state by 5 h . Binding to paraformaldehyde-fixed cells at 37 degrees C was saturable (Kd = 5.4 nM), was reversible, and indicated ca . 100,000 binding sites per cell . It is believed that receptor-bound toxin is responsible for cell death . Once the kinetics of toxin entry were described, we examined the effect of reduced temperatures on the intracellular processing of toxin and thus its expression . Toxin-induced inhibition of protein synthesis was minimal at temperatures below 20 degrees C . This was seen even though at 20 degrees C sufficient toxin was internalized to kill cells, and toxin enzyme activity was maximal . Internalization of 125I-labeled toxin, but not of 125I-labeled horseradish peroxidase (marker of fluid-phase endocytosis), became rate limiting at 20 degrees C or below . These data suggest that reduced temperatures block a step in the receptor-mediated endocytic pathway essential for the expression of Pseudomonas toxin activity.

Clin Chem, 1984 Sep, 30(9), 1549 - 51
Quantification of salicylate in serum by use of salicylate hydroxylase; You K et al.; In this enzymatic method for salicylate in serum, Pseudomonas salicylate hydroxylase (EC 1.14.13.1) is used . This stable enzyme catalyzes the stoichiometric, unidirectional conversion of salicylate and NAD(P)H to catechol and NAD(P)+ in the presence of molecular oxygen . The concentration of salicylate in a clinical sample is determined by measuring the delta A at 340 nm as compared with a standard . This new method is rapid, highly specific, requires 40 microL or less of sample, and we saw no interference by any of 61 commonly used drugs . Lipemic, icteric, or hemolyzed samples can be used . Furthermore, this method does not involve extraction, deproteinization, or derivatization . Results are precise and agree well with those obtained by the Trinder test.

J Clin Invest, 1984 Sep, 74(3), 966 - 71
Pseudomonas exotoxin-anti-TAC . Cell-specific immunotoxin active against cells expressing the human T cell growth factor receptor; FitzGerald DJ et al.; An immunotoxin was constructed with an activity that discriminated between two T cell lines based on the expression of the T cell growth factor (TCGF) receptor on their cell surface . A toxic protein conjugate, designated PE-anti-TAC, was made by chemically coupling pseudomonas exotoxin (PE) to a monoclonal antibody (anti-TAC) that recognizes the human TCGF receptor . This conjugate was toxic to HUT-102 cells, a cell line that expresses the TCGF receptor, but was nontoxic for MOLT-4 cells, a receptor-negative line . The toxicity of PE-anti-TAC was enhanced 50-fold in the presence of human adenovirus type II and was reduced to control levels by adding excess anti-TAC antibody . The toxicity of PE-anti-TAC for HUT-102 cells was compared with PE-anti-transferrin receptor . To compare the route of entry for both anti-TAC and anti-TFR using electron microscopy, protein conjugates were made by coupling horseradish peroxidase (HRP) to each antibody . Anti-TFR-HRP entered HUT-102 cells by concentrative adsorptive endocytosis via coated pits, and the majority of the antibodies bound to the cell surface at 4 degrees C were seen in receptosomes by 10 min after warming to 37 degrees C . Anti-TAC-HRP was also found to enter HUT-102 cells via coated pits and receptosomes; but, in contrast to anti-TFR, anti-TAC did not selectively concentrate in coated pits, and therefore the majority of this surface-bound antibody were not internalized in HUT-102 cells by 10 min at 37 degrees C.

J Virol, 1984 Sep, 51(3), 650 - 5
Role of a low-pH environment in adenovirus enhancement of the toxicity of a Pseudomonas exotoxin-epidermal growth factor conjugate; Seth P et al.; A conjugate of Pseudomonas exotoxin and epidermal growth factor (PE-EGF) inhibits proteins synthesis in KB cells, and this inhibition is increased by adenovirus . Protein synthesis inhibition is dependent on the amount of adenovirus and PE-EGF used and the time of incubation of cells with these agents . With 1 microgram of adenovirus and 0.5 micrograms of PE-EGF per ml, protein synthesis is inhibited about 80% in a 60-min experiment . Under these conditions neither adenovirus nor PE-EGF alone has any effect . In the presence of several weak bases or monensin, the enhancement of toxicity was substantially inhibited; half-maximal inhibition was achieved with 40 microM chloroquine, 10 mM ammonium chloride, 5 mM methylamine, 0.1 mM N-hexylamine and 1 microM monensin . At the concentrations employed, none of the inhibitors affected the amount of virus taken up or bound to the cell surface, and chloroquine had no effect on the amount of EGF taken up in 60 min . Chloroquine did not prevent the toxicity of the PE-EGF (5 micrograms/ml) alone . Because these compounds are known to elevate the pH in receptosomes, it seems likely that the acidification of the receptosome either enhances the lysis of the membrane by adenovirus or enhances some other step in the release of PE-EGF.

Endocrinol Exp, 1984 Sep, 18(3), 141 - 50
Norethisterone and ethinylestradiol do not inhibit delta 5-3 beta-hydroxysteroid dehydrogenase in rat Leydig cells; Zeillinger R et al.; The effect of norethisterone, norethisterone acetate, ethinylestradiol and of a cyanoketone on the activity of HSD have been studied . Rat Leydig cells and extracts of Pseudomonas testosteroni were used as enzyme sources . Conversion of {3H}DHEA to A and {3H}pregnenolone to progesterone were used for the assay of enzyme activity . Km values for each substrate for Leydig cells and for bacterial enzyme were: 1.5 x 10(-5) mol l-1 and 6.0 x 10(-5) mol l-1 DHEA; 1.3 x 10(-5) mol and 7.7 x 10(-5) mol l-1 pregnenolone . For investigation of inhibition {3H}DHEA was used as substrate and apparent Ki values were calculated by Dixon plot analysis . In both systems norethisterone showed no inhibition of the enzyme . Norethisterone acetate proved also to be not inhibitory; only residual affinity to the enzyme was noted (Ki = 8.0 x 10(-5) mol l-1: Leydig cell preparation; Ki = 9.4 x 10(-5) mol l-1: bacterial enzyme) . Different result sin the two systems were obtained for ethinylestradiol and for a combination with norethisterone acetate . Ethinylestradiol or the combination with the progestin (Ki = 0.5 x 10(-5) mol; 1.4 x 10(-5) mol l-1) showed noteworthy inhibition of the bacterial enzyme but only weak competition for binding to the active site of the enzyme in rat Leydig cells (Ki = 6.0 x 10(-5) mol l-1; 8.3 x 10(-5) mol l-1) . It is concluded that there are greater differences between the two enzymes than considered previously . As expected, cyanoketone proved to be the most effective inhibitor in both systems . The most important result seems to be, that norethisterone does not inhibit HSD . This suggests that there is no causal relationship between hypospadias and progestins administered to females during pregnancy.

Clin Orthop, 1984 Sep, (188), 168 - 72
The potential value of the sedimentation rate in monitoring treatment outcome in puncture-wound-related Pseudomonas osteomyelitis; Crosby LA et al.; Recently, a recommendation was made to treat Pseudomonas osteomyelitis in children with only ten to 14 days of postsurgical antibiotics . However, there were no clinical or laboratory measurements to monitor the response . Fourteen new cases of puncture-wound-related Pseudomonas osteomyelitis of the foot in children were investigated to examine the response of erythrocyte sedimentation rate (ESR) to treatment . The ESR was increased in 12/12 patients with a mean peak of 41 mm/hr and a mean duration of 30 days . After appropriate treatment was begun, the ESR decreased at a mean rate of 1.1 mm/day . In four of five patients with a poor treatment response, the ESR remained elevated until antibiotics were changed (in 3) or surgery performed (in 1) . One patient with an abnormal ESR at completion of three weeks of postoperative antibiotics had a relapse after one year . These data suggest a need for careful assessment of treatment response, such as declining ESR, before antibiotics are discontinued at an arbitrary postoperative time period.

J Cell Physiol, 1984 Sep, 120(3), 271 - 9
Verapamil enhances the toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin and antitransferrin receptor with Pseudomonas exotoxin; Akiyama S et al.; Verapamil, a clinically important calcium channel blocker, has been found to cause a 40-fold enhancement of killing of the human KB cell line by a cytotoxic conjugate of epidermal growth factor with Pseudomonas exotoxin (EGF-PE) . Synergistic effects of verapamil and EGF-PE are also seen on HeLa D98 cells and a human epidermal carcinoma cell line, A431 . Verapamil also potentiates the effect of a toxic conjugate formed between Pseudomonas exotoxin and a monoclonal antibody to the human transferrin receptor (anti-TFR-PE) and enhances the effect of Pseudomonas exotoxin (PE) alone . Two other calcium antagonists were tested . Diltiazem enhances the cytotoxic effect of EGF-PE, but nifedipine does not . Verapamil does not affect the binding and uptake of 125I-EGF by KB cells, but it significantly delays the disappearance of internalized 125I-EGF from the cells . Density gradient fractionation studies using cell homogenates suggest that 125I-EGF accumulates in an undegraded form in lysosomes when cells are treated with verapamil . By immunofluorescence microscopy using an antibody to PE, EGF-PE was found to accumulate in lysosomes; by electron microscopy the lysosomes had an abnormal appearance . The effects of verapamil on toxicity of EGF-PE and lysosomal function appear to be related . However, it is not known whether the enhanced toxicity of EGF-PE in the presence of verapamil is due to its delayed degradation in lysosomes or some more general effect of verapamil on membrane permeability.

J Biochem (Tokyo), 1984 Aug, 96(2), 421 - 7
Oxidation and oxygenation of L-amino acids catalyzed by a L-phenylalanine oxidase (deaminating and decarboxylating) from Pseudomonas sp . P-501; Koyama H; A number of L-amino acids and derivatives were tested as substrates for the purified Pseudomonas L-phenylalanine oxidase . The reaction products of these amino acids were analyzed by high performance liquid chromatography and the kinetic properties of the reactions were partially characterized . In addition to L-phenylalanine, L-tyrosine, DL-o-tyrosine, DL-m-tyrosine, p-fluoro-DL-phenylalanine and beta-2-thienyl-DL-alanine served as substrates for both oxidation and oxygenation catalyzed by the enzyme . On the other hand, L-methionine and L-norleucine were enzymically converted to the corresponding alpha-keto acids with the consumption of oxygen and with the formation of ammonia and hydrogen peroxide in stoichiometric amounts . Kinetic studies showed that the Km values for oxidation and oxygenation of L-phenylalanine by the enzyme were 2.04 mM and 1.96 mM for oxygen, and 13.3 microM and 11.1 microM for L-phenylalanine, respectively . omega-Phenyl fatty acids such as phenylacetic acid, 3-phenylpropionic acid and 4-phenylbutyric acid were competitive inhibitors of the enzyme towards L-phenylalanine . Both oxidation and oxygenation of L-phenylalanine by the enzyme were also inhibited by phenylacetic acid competitively.

J Appl Bacteriol, 1984 Aug, 57(1), 75 - 81
Volatile compounds associated with the aerobic growth of some Pseudomonas species on beef; Dainty RH et al.; Five strains representing four clusters of meat spoilage pseudomonads were grown on sterile beef at 5 degrees C . After 7 days incubation sensory assessments were made and the chemical composition of the headspace gases determined by gas chromatography-mass spectrometry . There was good correlation between odour descriptions and chemical data for three of the strains . The most numerous types of product were esters and sulphur-containing compounds . Of 45 compounds identified only 1-undecene was common to all the tested strains.

J Appl Bacteriol, 1984 Aug, 57(1), 59 - 67
A study of the relative incidence of different Pseudomonas groups on meat using a computer-assisted identification technique employing only carbon source tests; Shaw BG et al.; A computer-assisted probabilistic identification technique employing 18 carbon source utilization tests has been developed and applied to 787 Pseudomonas strains isolated from beef, pork and lamb stored under aerobic conditions . Seven hundred and twelve (89.7%) were identified using these tests alone and a further six (0.8%) with extra tests . Taxa detected were Ps . fragi cluster 2, 390 strains (49.6% of all isolates); Ps . fragi cluster 1, 191 strains (24.9%); meat cluster 3, 87 strains (11.1%); Ps . fluorescens biotype I, 31 strains (3.9%); Ps . fluorescens biotype III, 7 strains (0.9%); and Ps . putida, 1 strain (0.1%) . The relative incidence of members of the various taxa was similar on beef, pork and lamb, and was unaffected by storage temperature in the range 0 degrees-10 degrees C . Each taxon was also detected at similar rates before and after spoilage . Meat origin (abattoir) affected the frequency of detection of meat cluster 3 and Ps . fluorescens biotype I strains but did not affect the incidence of detection of either cluster of Ps . fragi.

Arch Biochem Biophys, 1984 Aug 1, 232(2), 602 - 9
Inhibition of indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase by beta-carboline and indole derivatives; Eguchi N et al.; beta-Carboline derivatives inhibited both indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase activities from various sources . Among them, norharman is most potent for both enzymes from mammalian sources . Kinetic studies revealed that norharman is uncompetitive (Ki = 0.12 mM) with L-tryptophan for rabbit intestinal indoleamine 2,3-dioxygenase, and linearly competitive (Ki = 0.29 mM) with L-tryptophan for mouse liver tryptophan 2,3-dioxygenase . In addition, some beta-carbolines selectively inhibited one enzyme or the other . Pseudomonad tryptophan 2,3-dioxygenase was inhibited by a different spectrum of beta-carbolines . Such a selective inhibition by the structure of substrate analogs is more evident by the use of indole derivatives . Indole-3-acetamide, indole-3-acetonitrile and indole-3-acrylic acid exhibited a potent inhibition for mammalian tryptophan 2,3-dioxygenase, while they moderately inhibited the pseudomonad enzyme . However, they showed no inhibition for indoleamine 2,3-dioxygenase . These results suggest the difference of the structures of the active sites among these enzymes from various sources.

Mol Cell Biol, 1984 Aug, 4(8), 1528 - 33
Evidence that the penton base of adenovirus is involved in potentiation of toxicity of Pseudomonas exotoxin conjugated to epidermal growth factor; Seth P et al.; When KB cells are incubated for 1 h with human adenovirus type 2 or type 5 (1 microgram/ml) and a conjugate of epidermal growth factor and Pseudomonas exotoxin (EGF-PE), protein synthesis is inhibited by 80 to 90% . Under these conditions, neither adenovirus nor EGF-PE alone has any effect on host protein synthesis . Thus, adenovirus enhances the toxicity of EGF-PE . A number of antibodies to intact virus and capsid components were tested for their ability to block the enhancing activity and virus uptake . At appropriate dilutions, antibodies prepared against intact virus and penton base blocked the enhancing activity without affecting virus uptake . Antibodies against hexon and fiber blocked virus uptake and enhancing activity in parallel . These studies suggest that the penton base is important in lysis of the vesicles which contain adenovirus and EGF-PE, and this base allows virus and toxin to enter the cytoplasm.

J Biol Chem, 1984 Jul 25, 259(14), 9090 - 5
Enzymatic function in crystals of delta 5-3-ketosteroid isomerase . Catalytic activity and binding of competitive inhibitors; Westbrook EM et al.; Crystals of the steroid-metabolizing enzyme, delta 5-3-ketosteroid isomerase (EC 5.3.3.1) from Pseudomonas testosteroni, exhibit many enzymatic properties . Each enzyme subunit in the lattice binds a competitive inhibitor, progesterone, with the same stoichiometry (1:1) and affinity (KD = 6 X 10(-6) M) as the enzyme in solution . Another competitive inhibitor, 19-nortestosterone, competes with progesterone for the same binding sites in the crystal . The enzyme crystals catalyze the conversion of delta 5- to delta 4-ketosteroids, but because the enzyme is so efficient, and substrate diffusion into the crystal is so slow, substrate cannot penetrate deeply into the crystal before being converted to product . A general theoretical formulation is presented to account for the effects of substrate diffusion into enzyme crystals of different shapes and sizes . The dependence of apparent mean enzyme activity in steroid isomerase crystals as a function of crystal size is shown to be consistent with this theoretical formulation . These inhibitor binding and catalytic properties suggest that the enzyme is in an active conformation within these crystals.

J Biol Chem, 1984 Jul 25, 259(14), 9235 - 40
Human and murine class I MHC antigens share conserved serine 335, the site of HLA phosphorylation in vivo; Guild BC et al.; The site of phosphorylation of the HLA-B7 antigen in vivo is serine 335, which is located in the intracellular region . Pseudomonas fragi protease was used in limited proteolysis experiments of HLA antigens to identify the position of the phosphoserine residue . The intracellular region is composed of 30 amino acids at the carboxyl terminus of the heavy chain; up to nine serine residues are located in this region . Four of the serines are found at the distal end, and are encoded by exon 7 in both human and murine class I MHC antigens . Alignment of the protein and DNA sequences of the intracellular regions of human and murine class I antigens demonstrates conservation of the serine positions located within this exon . Realignment of exon 7, by introducing a gap in the murine sequences, increases homology across species, and reveals two conserved serines at 332 and 335 within the conserved sequence Ser-Asp/Glu-X-Ser(P)-Leu . The preservation of this sequence and the site of phosphorylation suggests that this modification of the intracellular region of histocompatibility antigens is functionally significant.

FEBS Lett, 1984 Jul 23, 173(1), 15 - 8
Proton magnetic resonance studies of 7Fe ferredoxins . Three redox states of the {4Fe-4S} cluster in a Pseudomonas ovalis ferredoxin; Nagayama K et al.; The oxidizability of a redox couple, {4Fe-4S}, in a 7Fe ferredoxin extracted from Pseudomonas ovalis was monitored by 1H-NMR . The iron-sulfur cluster in the ferredoxin was not only reducible (Nagayama et al., 1983) but also oxidizable in its native form . This result provided the first verification of 3 redox states for a redox center in ferredoxin, 4Fe, in the native form of the protein.

FEBS Lett, 1984 Jul 23, 173(1), 113 - 8
ADP-ribosyltransferase from beef liver which ADP-ribosylates elongation factor-2; Iglewski WJ et al.; Fragment A of diphtheria toxin and Pseudomonas toxin A intoxicate cells by ADP-ribosylating the diphthamide residue of elongation factor-2 (EF-2) resulting in an inhibition of protein synthesis {1-3} . A cellular enzyme from polyoma virus transformed baby hamster kidney (pyBHK) cells ADP-ribosylates EF-2 in an identical manner {4} . Here we describe a similar cellular enzyme from beef liver which transfers {adenosine-14C}ADP-ribose from NAD to EF-2 . The 14C-label can be removed from the EF-2 by snake venom phosphodiesterase as a soluble product which comigrates with AMP on TLC plates, indicating the 14C-label is present on EF-2 as monomeric units of ADP-ribose . Furthermore, the forward transferase reaction catalyzed by the beef liver ADP-ribosyltransferase is reversible by excess diphtheria toxin fragment A, with the formation of 14C-labeled NAD, indicating that both transferases ADP-ribosylate the same site on the diphthamide residue of EF-2 . Thus, beef liver and pyBHK mono(ADP-ribosyl)transferases both modify the diphthamide residue of EF-2, in a manner identical to diphtheria toxin fragment A and Pseudomonas toxin A . These results suggest the cellular enzyme is probably ubiquitous among eukaryotic cells.

Biochemistry, 1984 Jul 17, 23(15), 3534 - 40
Structure of pseudobactin 7SR1, a siderophore from a plant-deleterious Pseudomonas; Yang CC et al.; When grown in iron-limiting culture medium, sugar beet deleterious Pseudomonas 7SR1 produced extra-cellularly the yellow-green, fluorescent siderophore pseudobactin 7SR1 . Pseudobactin 7SR1 had a molecular formula of C46H63N13O23 and a molecular mass of 1166 g/mol . Pseudobactin 7SR1 contained a cyclic octapeptide with the amino acid sequence L-Ala-Gly-Ser-Ser-threo-beta-OH-Asp-Thr-Ser-N delta-OH-Orn . Since pseudobactin 7SR1 was not affected by nonspecific enzymes, it might contain D-amino acids . A yellow-green, fluorescent quinoline derivative is postulated to be attached via an ester bond to the serine residue following the glycine . A malamide group was attached to carbon 3 of the quinoline derivative . The three bidentate iron(III)-chelating groups consisted of an alpha-hydroxy acid group derived from beta-hydroxyaspartic acid, an omicron-dihydroxy aromatic group derived from the yellow-green, fluorescent chromophore, and a hydroxamate group derived from N delta-acetyl-N delta-hydroxyornithine . The chemical structure of pseudobactin 7SR1 is remarkably similar to that of pseudobactin, the siderophore of plant growth promoting Pseudomonas B10.

S Afr Med J, 1984 Jul 7, 66(1), 31 - 3
Piroxicam poisoning in a 2-year-old child . A case report; MacDougall LG et al.; This report describes the severe multisystem toxicity which followed ingestion of 5 piroxicam capsules (100 mg) by a 2-year-old child . Gastro-intestinal symptoms developed within 2 hours, resulting in severe fluid and electrolyte imbalance, mental confusion and a generalized seizure . Evidence of liver and renal dysfunction developed within 3 days . Haemopoietic toxicity was manifested by progressive peripheral pancytopenia, bone marrow aplasia and coagulopathy . Pseudomonas septicaemia developed during the period of neutropenia . Clinical, biochemical and haematological abnormalities slowly resolved over 3-4 weeks . In view of the increasing use of piroxicam as an anti-inflammatory agent it seemed important to draw attention to the potentially serious effects of accidental overdosage.

J Bacteriol, 1984 Jul, 159(1), 179 - 83
Purification and properties of the myo-inositol-binding protein from a Pseudomonas sp; Deshusses J et al.; A myo-inositol-binding protein was isolated from a Pseudomonas sp . soil isolate and was purified to homogeneity . Its molecular weight is 30,000, and it has a single binding site . The amino acid analysis showed that the protein contains three tryptophan residues and no cysteine . Tryptophan residues seem to be involved in the binding of the ligand, as shown by the modification of the fluorescence spectra and by the fact that oxidation of tryptophan residues with N-bromosuccinimide abolished the binding of myo-inositol . Sequence analysis of the N-terminal segment of 37 amino acids showed that 13 are conserved when compared with the galactose-binding protein of Escherichia coli.

J Cell Sci, 1984 Jul, 69, 87 - 105
Scintillation and autoradiographic studies on 63 nickel uptake in Pseudomonas tabaci; Al-Rabaee RH et al.; Scintillation studies on the uptake of 63Ni2+ by Pseudomonas tabaci demonstrate an incorporation of approximately 2.5 nmol per 10(10) bacterial cells, in medium containing 12 nmol (8.3 microCi) per ml . Over 80% of the incorporated Ni2+ is lost from the cells during washing, fixation and dehydration with ethanol . The remaining insoluble (bound) 63Ni2+ has the highest level in cells fixed in acetic acid/ethanol (0.4 nmol/10(10) cells), with smarter amounts in paraformaldehyde- and glutaraldehyde-fixed cells . The radioactive level in aldehyde-fixed cells represents a total Ni2+ uptake of about 10(-18) g or 10(4) atoms per cell . Light- and electron-microscope autoradiography corroborated the scintillation studies in demonstrating a higher retention of label by cells fixed in acetic acid/ethanol, possibly reflecting a higher retention of medium Mr proteins with this type of fixation . High-resolution electron-microscope autoradiography involving gold latensification with physical development demonstrated a clear localization of silver grains to the central nucleoid region (seen most clearly over the discrete nucleoid of aldehyde-fixed cells) and within this to the chromatin (seen most clearly over the condensed chromatin of acetic acid/ethanol-fixed cells) . It is suggested that the incorporated 63Ni2+ labels mainly central, genetically inactive DNA, while peripheral, actively transcribing DNA has little associated radioactivity . The pattern of cation association seen in this bacterium shows a number of close similarities to the situation seen in dinoflagellate cells.

Mikrobiologiia, 1984 Jul-Aug, 53(4), 547 - 50
{Exopolysaccharide of Mycobacterium flavum}; Kosenko LV et al.; Mycobacterium flavum 158a can produce exopolysaccharides whose quantity varies, depending on the culture age, from 88.2 to 186.8% of the cell biomass weight in a medium with sucrose and from 1.3 to 25.0% in a medium with a polysaccharide synthesized by the oligonitrophilic bacterium Pseudomonas sp . 158a . The absolute and relative content of exopolysaccharides in the cultural broth decreases during the intensive growth of M . flavum 158a . This appears to be caused by their assimilation as a carbon source in the processes of energy and constructive metabolism . The exopolysaccharides of M . flavum 158a contain galactose, glucose, mannose, fucose, xylose, ramnose and a lipophilic sugar X1 at the molar ratio 10:11:11:8:11:12:16 as well as a non-identified sugar X2 with the Rf below than that of ramnose . The exopolysaccharide of M . flavum 158a was shown to be heterogeneous . It is composed of at least 16 fractions differing in molecular mass (from 98,980 to 490 D), quantity (from 2 to 8), composition and percentage of the constituent monosaccharides.

Can J Microbiol, 1984 Jul, 30(7), 952 - 5
Dialysis culture for the production of Pseudomonas saccharophila alpha-amylase; Kadam SK et al.; Growth and alpha-amylase synthesis of Pseudomonas saccharophila was shown to be inhibited by the accumulation of a mixture of nonvolatile fatty acids during nondialysis cultivation . Using dialysis culture a 9-fold increase in the level of biomass and an 8.5-fold increase in alpha-amylase yield was achieved.

Lancet, 1984 Jun 16, 1(8390), 1338 - 9
Self-administered home intravenous antibiotic therapy in bronchiectasis and adult cystic fibrosis; Winter RJ et al.; 10 patients (mean age 23.1, range 17.1-40.5 years), 8 with cystic fibrosis (CF), and 2 with advanced bronchiectasis without CF, were taught, while being treated in hospital for exacerbations of pseudomonas infection, how to continue to give themselves intravenous antibiotics at home . They were discharged after satisfactory antibiotic levels had been achieved, and 22 courses were given at home over a total of 116 patient-days . In 14 of these, the greater part of the course was given at home (mean duration 6.6, range 5-10 days); 2 of these were given without admission to hospital . In the 8 patients with two or more infective exacerbations within a 12-month period there was no difference between home and hospital treatments in clinical improvement or in relapse time, defined as the interval between completion of treatment and subsequent antibiotic therapy . Self-administration of antibiotics intravenously at home for selected adults with cystic fibrosis and severe bronchiectasis reduces hospital stay and does not seem to be associated with an increased rate of recurrent infection.

Cancer, 1984 Jun 15, 53(12), 2592 - 600
NHL-3 protocol . Six-drug combination chemotherapy for non-Hodgkin's lymphoma; Koziner B et al.; Combination chemotherapy and radiotherapy (RT) were administered to 73 adults with non-Hodgkin's lymphoma (NHL) . Ten cycles of the following drugs were given: intravenous Adriamycin (doxorubicin) (25 mg/m2), cyclophosphamide (700 mg/m2) and vincristine (1.5 mg/m2) on day 1; arabinosylcytosine (100 mg/m2) and methotrexate (10 mg/m2) on days 3 to 5; and oral prednisone (60 mg/m2) on days 1 to 5 . Radiotherapy was given to resistant or initially bulky disease (2000 rad) . Patients were also randomized to receive pseudomonas vaccine or no immunotherapy . Of 61 evaluable patients, 33 (54%) achieved a complete response (CR) and 18 (30%) a partial response (PR) . Among 44 evaluable patients with diffuse histiocytic lymphoma (DHL), 22 (50%) had a CR, and 15 (34%) a PR . For 17 evaluable patients with nodular (4) and diffuse (11) mixed and poorly differentiated lymphocytic and diffuse "undifferentiated" (2) lymphomas, CR and PR rates were 65% and 18%, respectively . No statistically significant differences in response rate or duration and survival have been observed between the patients randomized to receive pseudomonas vaccine or no immunotherapy . Median follow-up time from start of treatment was 47.5 months . Median survival for all 73 patients (including inevaluables ) and for 52 DHL patients was 30.7 months . Poor prognostic features influencing survival included: female sex (P = 0.003), poor response to therapy (CR versus PR; P = 0.001), prior chemotherapy, (P = 0.01) and high levels of lactic dehydrogenase (P = 0.001) . It can be concluded that this combination of cycle and phase-active agents is of similar efficacy to other reported regimens in inducing major responses and that it has the potential to prolong disease-free survival . The analysis of prognostic factors has been used to dissect poor prognostic categories that might require different modalities of treatment.

Eur J Biochem, 1984 Jun 1, 141(2), 305 - 12
NMR and electron-paramagnetic-resonance studies of a dihaem cytochrome from Pseudomonas stutzeri (ATCC 11607) (cytochrome c peroxidase); Villalain J et al.; A dihaem cytochrome (Mr 37 400) with cytochrome c peroxidase activity was purified from Pseudomonas stutzeri (ATCC 11 607) . The haem redox potentials are far apart: one of the haems is completely ascorbate-reducible and the other is only reduced by dithionite . The coordination, spin states and redox properties of the covalently bound haems were probed by visible, NMR and electron paramagnetic resonance (EPR) spectroscopies in three oxidation states . In the oxidized state, the low-temperature EPR spectrum of the native enzyme is a complex superimposition of three components: (I) a low-spin haem indicating a histidinyl-methionyl coordination; (II) a low-spin haem indicating a histidinyl-histidinyl coordination; and (III) a minor high-spin haem component . At room temperature, NMR and optical studies indicate the presence of high-spin and low-spin haems, suggesting that for one of the haems a high-spin to low-spin transition is observed when temperature is decreased . In the half-reduced state, the component I (high redox potential) of the EPR spectrum disappears and induces a change in the g-values and linewidth of component II; the high-spin component II is no longer detected at low temperature . Visible and NMR studies reveal the presence of a high-spin ferric and a low-spin (methionyl-coordinated) ferrous state . The NMR data fully support the haem-haem interaction probed by EPR . In the reduced state, the NMR spectrum indicates that the low-potential haem is high-spin ferrous.

Eur J Biochem, 1984 Jun 1, 141(2), 297 - 303
NMR studies of a dihaem cytochrome from Pseudomonas perfectomarinus (ATCC 14405); Moura I et al.; Pseudomonas perfectomarinus (ATCC 14405) dihaem cytochrome c552 was studied by 300-MHz proton magnetic resonance . Some of the haem resonances were assigned in the fully reduced and fully oxidized states . No evidence was found for methionine haem axial coordination . The oxidation-reduction equilibrium was studied in detail . Due to the large difference in mid-point redox potential between the two haems (+174 mV, for haem II and -180 mV for haem I) an intermediate oxidation state could be obtained containing reduced haem I and oxidized haem II . In this way the total paramagnetic shift at different oxidation levels could be decomposed in the intrinsic and extrinsic contributions . It was found that the two haems interact . The rate of electron exchange is slow on the NMR time scale . The redox equilibria are discussed for four possible redox species in solution.

J Biol Chem, 1984 May 10, 259(9), 5612 - 7
Selenite binding to carbon monoxide oxidase from Pseudomonas carboxydovorans . Selenium binds covalently to the protein and activates specifically the CO----methylene blue reaction; Meyer O et al.; The CO----methylene blue and CO----dichlorophenol indophenol activities of carbon monoxide oxidase were specifically activated upon aerobic incubation with selenite, whereas the NADH----methylene blue activity was not altered . Fully active enzyme contained selenium, molybdenum, and flavin adenine dinucleotide in a 1:1:1 ratio . Selenium was covalently bound to the protein, probably between the sulfurs of half-cystine residues, and not a constituent of the molybdenum cofactor . The action of selenite was directed to the cytoplasmic species of carbon monoxide oxidase exclusively, whereas the CO----methylene blue activity of the membrane-bound enzyme remained unaffected.

Arch Microbiol, 1984 May, 138(1), 37 - 43
Comparison of two bacterial azoreductases acquired during adaptation to growth on azo dyes; Zimmermann T et al.; Selection for utilization of carboxy-Orange I {1-(4'-carboxyphenylazo)-4-naphthol} in the chemostat yielded Pseudomonas strain K24 which was unable to grow on carboxy-Orange II {1-(4'-carboxyphenylazo)-2-naphthol} while selection for growth on carboxy-Orange II had previously led to strain KF46 which did not utilize carboxy-Orange I . Orange I azoreductase of strain K24, the key enzyme of dye degradation, was purified 80-fold with 17% yield to electrophoretic homogeneity and compared to the previously purified Orange II azoreductase of strain KF46 . Common properties of the two enzymes were their monomeric structure, their specificity for NADPH and NADH as cosubstrates, the range of their Km values for substrates and cosubstrates as well as their reactivity towards a series of substrate analogs . They differed from each other with respect to molecular weight (21,000 and 30,000) and in the absolute requirement of Orange I azoreductase for a hydroxy group in the 4'position of the naphthol ring of the substrate molecule as compared to the requirement for substrates with a 2-naphthol moiety by Orange II azoreductase . The pure enzymes did not exhibit immunological cross-reaction with each other . Crude extracts of strains K24 and KF46 and of azoreductase-negative strains isolated at different stages of the adaptation experiments, however, contained material which cross-reacted (CRM) with both anti Orange I azoreductase serum and anti Orange II azoreductase serum . The CRM may represent a common precursor protein of the azoreductases in strains K24 and KF46.

Arch Dis Child, 1984 May, 59(5), 435 - 8
Serial study of C reactive protein in neonatal septicaemia; Hindocha P et al.; Serial C reactive protein concentrations were assayed by electroimmunoassay in 41 infants . Values in most of the non-infected infants were below 0.3 mg/dl, the lower limit of detection of C reactive protein by electroimmunoassay . Eleven of 12 infants with proved sepsis (positive blood cultures) had significantly raised concentrations and one infant with recurrent pseudomonas chest infection had a raised C reactive protein concentration . High C reactive protein concentrations were also found in infants with suspected infection . Successful treatment was followed by a decrease in the C reactive protein concentration . Total white blood cell count was not as appropriate as C reactive protein determination in the early identification of bacterial infection in the newborn.

Am J Trop Med Hyg, 1984 May, 33(3), 474 - 8
The prevalence of human melioidosis in Northern Queensland; Ashdown LR et al.; Sera from 9,047 individuals from Northern Queensland were examined for the presence of hemagglutinating antibodies to Pseudomonas pseudomallei, the causative agent of melioidosis, and 512 (5.7%) were found to have titers of 1:40 or greater . The distribution of positive reactors in various groups was uneven, and significantly higher prevalences of positive antibody titers were found in the sera from Aborigines (7.9-10.6%), Torres Strait Islanders (7.8%), Vietnamese refugees (29%) and from persons with certain medical conditions including chronic alcoholism (15%), chronic infections (14.8%), diabetes mellitus (8.6%) and liver disease (12.9%) . There were significantly fewer positive reactors (1.4%) amongst the armed forces stationed in Northern Queensland . At present, the boundaries of the major endemic region of Australia extend north from Rockhampton along the coast to Darwin and inland, west from Rockhampton to Tennant Creek in central Australia . Townsville was found to have the highest prevalence (5.2%) of positive reactors of all urban populations of Northern Queensland . The extent of the disease is such that it can no longer be considered a rare infection in Northern Queensland.

Am J Trop Med Hyg, 1984 May, 33(3), 467 - 73
Melioidosis in Far North Queensland . A clinical and epidemiological review of twenty cases; Guard RW et al.; During the last 4 years, 20 cases of clinical melioidosis were diagnosed in the geographical area between Tully and Thursday Island . Sixteen were diagnosed by culture of Pseudomonas pseudomallei, and four by positive serology with appropriate clinical features . Most cases occurred during or after a heavy wet season . All patients were adult, and males predominated . Farmers and stockmen represented predisposed populations due to their prolonged soil contact . Ten patients were white Australians, six were Aborigines and four were Torres Strait Islanders . Twelve cases were first diagnosed by positive blood culture and four by sputum culture . The primary site of infection was pulmonary in 14 cases, genitourinary tract in one case, subcutaneous tissues in one case, and joints in two cases . In cases of fulminating infection metastatic abscesses were commonly found in many organs; typically lungs, liver, kidneys and spleen . Six patients had acute fulminating disease and died . Fourteen patients successfully responded to appropriate therapy, but relapse occurred in three, all of whom had an alcohol problem and showed poor drug compliance . The presence of diabetes mellitus in six patients confirmed the important known association of these two diseases . In three fulminating and four subacute infections the serology was negative at the time of diagnosis by culture . Antibiotic therapy for the different forms of this disease is reviewed, and a laboratory protocol for the rapid reporting of positive culture results is included.

Arch Intern Med, 1984 May, 144(5), 1005 - 10
Empiric therapy for infections in patients with granulocytopenia . Continuous v interrupted infusion of tobramycin plus cefamandole; Feld R et al.; A combination of tobramycin sulfate and cefamandole nafate was used as initial empiric therapy for the treatment of 71 evaluable febrile (temperature greater than 38.5 degrees C) episodes in 64 (neutrophils, less than 1,000/microL) adult patients with cancer and granulocytopenia . Carbenicillin sodium or ticarcillin disodium was substituted for cefamandole in patients with Pseudomonas infections and in patients in whom the initial regimen was unsuccessful . Twenty-nine episodes were randomized to receive tobramycin by continuous infusion, while 42 were randomized to receive tobramycin by interrupted infusion . Twenty-seven (79%) of the 34 documented infections responded to the initial empiric antibiotic combination, ten (83%) of 12 being given continuous infusion and 17 (77%) of 22 being given interrupted infusion of tobramycin . Nephrotoxic reaction occurred in 7% of patients treated with continuous infusion and 15% treated with interrupted infusion, mostly patients older than 60 years . Tobramycin, by either continuous or interrupted infusion, plus cefamandole is safe and efficacious empiric therapy for infections in patients with cancer and granulocytopenia.

J Virol, 1984 May, 50(2), 529 - 32
Polypeptide synthesis directed by bacteriophage phi W-14 and by mutants defective in DNA synthesis; Miller PB et al.; The latent period of bacteriophage phi W-14 is approximately 65 min when the doubling time of its host, Pseudomonas acidovorans, is 85 min . Host protein synthesis is shut off relatively slowly, stopping approximately 25 min after infection . There are several phases of phage-specific polypeptide synthesis during the latent period: early polypeptides appear within 10 min after infection; middle polypeptides start to appear between 10 nd 30 min; late polypeptides appear after 30 min . The lengths of time for which individual polypeptides are synthesized vary widely . Several late polypeptides do not appear in the virion . DNA replication is not required for late gene expression . The hypermodified pyrimidine, alpha-putrescinylthymine, appears not to be required in both strands of the DNA duplex for transcription.

Infect Control, 1984 May, 5(5), 219 - 22
Persistent isolation of an unusual Pseudomonas species from a phenolic disinfectant system; Newman KA et al.; A well-characterized unusual Pseudomonas species contaminated the piped disinfectant system of a newly-opened laminar air flow intensive care facility . This organism was frequently isolated (10(4)-10(6) cfu/ml) from phenolic diluted 1:256 in the system, and could also be recovered (0.01-0.2 cfu/ml) from undiluted phenolic . During the 20-month period when this unusual Pseudomonas was present, none of the severely compromised, granulocytopenic oncology patients treated in the intensive care facility were either colonized or infected with this Pseudomonas sp . Eradication of the organism from the system proved difficult and was accomplished by removing a contaminated reservoir of diluted phenolic disinfectant followed by transient cleansing of the system with very high concentrations (84,000 ppm) of chlorine . This experience demonstrates that phenolics should be added to the list of disinfectants which can harbor Pseudomonas spp . in the clinical setting.

Ann Microbiol (Paris), 1984 May-Jun, 135A(3), 411 - 25
{Taxonomic value of the characterization of Pseudomonas by the analysis of volatile fatty acids produced in culture}; Peladan F et al.; Gas liquid-chromatography was used in order to characterize the volatile fatty acids produced in culture supernatants by 632 Pseudomonas strains . Statistical analysis of these results allowed testing of the discriminatory power of this analytical methodology (factor analysis) to point out the presence of subgroups (clustering according to the variance) and to show the relationship between species and subspecies (three-dimensional plot) . Fourteen Pseudomonas species could be accurately characterized by this methodology; the classification of Pseudomonas build up, on the basis of qualitative and quantitative aspects of their volatile fatty acids production, agrees with the current classification based on rRNA/DNA homology complexes.

Ann Microbiol (Paris), 1984 May-Jun, 135A(3), 399 - 410
{Gas chromatographic study of volatile fatty acids produced by 14 species of Pseudomonas}; Peladan F et al.; A quantitative gas-liquid chromatographic method for the determination of the volatile fatty acids produced in standardized liquid culture media by Pseudomonas reference strains belonging to 14 species, some of medical interest, is proposed here . This method, using 14 reference strains, permitted a precise identification of Pseudomonas species according to characteristic chromatograms . The key used for this identification gave us a classification corresponding to the DNA homology groups.

Mikrobiologiia, 1984 May-Jun, 53(3), 483 - 8
{Conditions for initiating heat stress in Pseudomonas geniculata}; Elisashvili VI et al.; The heating of Pseudomonas geniculata 338 at an elevated temperature causes a heat stress in the culture . The extent of the stress depends on the temperature and duration of heating . The incubation of the bacterium at 40 and 45 degrees C did not inhibit its growth after 30 min of heating, and no essential quantities of intracellular compounds absorbing at 260 nm were lost (E260 increased by 12-19%) . When the bacterium was heated at 50 degrees C for the same period of time, a three-hour lag-phase appeared during the subsequent cultivation of the bacterium whereas . E260 rose by a factor of 1.7 . The resistance of the bacterium to heating depended on the physiological state of the culture: cells at the logarithmic growth phase were most susceptible to heating while the bacterium became more resistant to heating in the course of aging . The addition of NaCl at a concentration of 1.5% or of 10(-3)-10(-4) M EDTA to the reparation medium makes it possible to estimate the population of bacterial cells in the state of stress.

Biosci Rep, 1984 May, 4(5), 433 - 40
Uncoupling and isotope effects in gamma-butyrobetaine hydroxylation; Holme E et al.; Replacement of unlabeled gamma-butyrobetaine with gamma-{2,3,4-2H6}butyrobetaine has a profound effect on the stoichiometry between decarboxylation of 2-oxoglutarate and hydroxylation in the reaction catalyzed by human gamma-butyrobetaine hydroxylase . The ratios between decarboxylation and hydroxylation are 1.16 with unlabeled and 7.48 with deuterated gamma-butyrobetaine as substrate . From these ratios an internal isotope effect of 41 has been calculated . DV in the overall reaction measured as 2- oxoglutarate decarboxylation is 2.5 and DV/K is 1.0 . For gamma-butyrobetaine hydroxylase from Pseudomonas sp . AK 1, 2-oxoglutarate decarboxylation exceeds hydroxylation with 10% when deuterated gamma-butyrobetaine is used . No excess was found with unlabeled substrate and no internal isotope effect could be calculated . DV for the bacterial enzyme is 6.

Proc Natl Acad Sci U S A, 1984 May, 81(10), 3158 - 62
Chromosomal assignment of the gene for human elongation factor 2; Kaneda Y et al.; Elongation factor 2 (EF-2), polypeptidyl -tRNA translocase, is an essential factor for protein synthesis in eukaryotic cells and Archebacteria . We isolated diphtheria toxin-resistant human primary embryo cells that contain EF-2 that cannot be ADP-ribosylated by diphtheria toxin and Pseudomonas exotoxin A (PA toxin) . Somatic cell hybrids were constructed from mouse L cells and toxin-resistant human embryo cell mutants . Forty-one hybrid clones were isolated, of which 15 clones were resistant to PA toxin . Karyotypic analysis and isozyme studies revealed that there was an absolute correlation between human chromosome 19 and resistance to PA toxin in the hybrids . On subcloning of PA toxin-resistant hybrid cells, we obtained one PA toxin-resistant hybrid subclone containing human chromosome 19 as the only human chromosome . Furthermore, the resistance to PA toxin of hybrid cell strains was lost after infection with poliovirus, for which sensitivity is conferred by human chromosome 19 . It was confirmed by using two-dimensional gel electrophoresis that PA toxin resistance in hybrid cells was caused by the presence of EF-2 resistant to ADP-ribosylation by fragment A of diphtheria toxin . These facts suggest that the gene encoding EF-2 is located on human chromosome 19.

J Leukoc Biol, 1984 May, 35(5), 475 - 87
Functional resistance of inflammatory macrophages to methotrexate in vitro; Zeller JM et al.; The purpose of this study was to evaluate the effects of high and low therapeutic doses of methotrexate (MTX) on macrophage metabolism and function in vitro . Monolayers of elicited rat peritoneal macrophages (PM) were exposed to a wide range of MTX concentrations (10(-8) M-10(-3) M) for 24 or 48 hr and macrophage RNA and protein metabolism were evaluated by the incorporation of {3H}5-uridine and {14C}1-leucine, respectively, into trichloroacetic acid (TCA)-precipitable material . Macrophage functional activity was examined by measuring the uptake of {14C}Pseudomonas aeruginosa to assess phagocytosis and the release of 51Cr from antibody-coated {51Cr}sheep red blood cells (SRBC) to assess antibody-dependent cell-mediated cytotoxicity . Following a 24-hr incubation with 10(-3) M MTX, incorporation of {3H}5-uridine into PM monolayers was enhanced 79% when compared to control monolayers (p less than 0.005) . Washout studies revealed that the stimulation of uridine incorporation was no longer observed by 24 hr following the removal of MTX from the culture medium . Incubation with 10(-3) M MTX for 48 hr returned uridine incorporation to control levels, although leucine incorporation into protein was reduced by 22% (p less than 0.01) . The depression in leucine incorporation in the presence of 10(-3) M MTX was not reversed after the removal of MTX from the culture medium . Uptake of {14C}P . aeruginosa was not altered following a 24- or 48-hr incubation with either 10(-7) M or 10(-3) M MTX . Similarly, {51Cr}SRBC cytolysis was not affected by a 2-hr preincubation with and continuous exposure to between 10(-8) M and 10(-3) M MTX . These results demonstrate that incubation of inflammatory macrophages with clinically high doses of MTX can alter macrophage RNA and protein metabolism without producing demonstrable changes in macrophage functional activity.

Proc Natl Acad Sci U S A, 1984 May, 81(9), 2703 - 7
Cellular ADP-ribosyltransferase with the same mechanism of action as diphtheria toxin and Pseudomonas toxin A; Lee H et al.; An ADP-ribosyltransferase was found in elongation factor 2 (EF-2) preparations from polyoma virus-transformed baby hamster kidney (pyBHK) cells . Like fragment A of diphtheria toxin and Pseudomonas toxin A, this eukaryotic cellular enzyme transfers {14C}adenosine from NAD+ to EF-2 . However, the cellular transferase is immunologically distinct from fragment A . The transferase also can be distinguished from fragment A and Pseudomonas toxin A by the inhibition of the activity of the former by cytoplasmic extracts and by histamine . Snake venom phosphodiesterase digestion of the {14C}adenosine-labeled EF-2 product of the cellular transferase reaction yielded {14C}AMP, indicating that the cellular enzyme is a mono(ADP-ribosyl)transferase . The forward ADP-ribosylation reaction catalyzed by the cellular enzyme is reversed by fragment A, yielding {14C}NAD+ . The results strongly suggest that the cellular transferase is a mono(ADP-ribosyl)transferase, which ADP-ribosylates the same diphthamide residue of EF-2 as does fragment A and Pseudomonas toxin A.

J Biol Chem, 1984 Apr 10, 259(7), 4452 - 7
Enzymatic dehydrogenation of tryptophan residues of human globins by tryptophan side chain oxidase II; Takai K et al.; Enzymatic dehydrogenation of tryptophan residues in human alpha- and beta-globins has been demonstrated with tryptophan side chain oxidase II, a hemoprotein from Pseudomonas . In 1 M ammonium acetate, pH 4, the enzyme catalyzed the conversion of Trp-14 of alpha-globin to alpha, beta-dehydro-Trp . Under identical conditions, Trp-15 of beta-globin was converted to alpha, beta-dehydro-Trp, whereas Trp-37 was unmodified . No modification was obtained in 0.4% sodium dodecyl sulfate or 8 M urea . The dehydrogenase component of tryptophan side chain oxidase II was also active toward globins in the presence of ferricyanide, whereas tryptophan side chain oxidase I was inert . Upon analyses of the tryptic fragments of the modified beta-globin, two forms of alpha, beta-dehydro-Trp-15 distinguishable by ultraviolet absorption and fluorescence were identified . Each of them rapidly equilibrated with another form on irradiation at 300-380 nm . We tentatively assigned them to two isomers of alpha, beta-dehydro-Trp distinguishable by the configuration around its beta-carbon atom . alpha, beta-Dehydro-Trp in globins and certain polypeptides used herein showed a blue fluorescence at 420 nm . In {alpha, beta-dehydro-Trp 15}beta-globin, the fluorescence was exceptionally intense, the fluorescence yield being comparable to that of NADH . From several lines of evidence, the result was ascribed to the excitation energy transfer from Trp-37 to alpha, beta-dehydro-Trp-15 in the modified beta-globin.

Appl Environ Microbiol, 1984 Apr, 47(4), 825 - 30
Secondary substrate utilization of methylene chloride by an isolated strain of Pseudomonas sp; LaPat-Polasko LT et al.; Secondary substrate utilization of methylene chloride was analyzed by using Pseudomonas sp . strain LP . Both batch and continuously fed reactors demonstrated that this strain was capable of simultaneously consuming two substrates at different concentrations: the primary substrate at the higher concentration (milligrams per liter) and the secondary substrate at the lower concentration (micrograms per liter) . The rate of methylene chloride utilization at trace concentrations was greater in the presence of the primary substrate, acetate, than without it . However, when the substrate roles were changed, the acetate secondary substrate utilization rate was less when methylene chloride was present . Thus, substrate interactions are important in the kinetics of secondary substrate utilization . Pseudomonas sp . strain LP showed a preference toward degrading methylene chloride over acetate, whether it was the primary or secondary substrate, providing it was below an inhibitory concentration of ca . 10 mg/liter.

Can J Microbiol, 1984 Mar, 30(3), 306 - 15
Use of DNA hybridization values to construct three-dimensional models of fluorescent pseudomonad relationships; Hildebrand DC et al.; Three-dimensional models of the relationships among fluorescent pseudomonads were prepared from appropriately transformed percent DNA homology values . The transformation selected was f(x) = ( (1 - HOM/HOM)200, where HOM = fractional DNA homology and 200 is a scaling factor . Model accuracy was quite good as a plot of transformed DNA homology values versus model distances was essentially linear for homology values greater than 30% . The model suggested that bacterial strains within the fluorescent pseudomonads appear to be related in a three-dimensional continuum with no clear and easy "natural" demarcation into groups (i.e., species).

Antimicrob Agents Chemother, 1984 Mar, 25(3), 362 - 5
Purification and properties of an inducible cephalosporinase from Pseudomonas maltophilia GN12873; Saino Y et al.; An inducible cephalosporinase was purified from Pseudomonas maltophilia GN12873 . The pI was 8.4, and the molecular weight was ca . 56,000 by gel filtration or 27,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that this enzyme had two subunits . The optimal pH and optimal temperature were 7.5 and 45 degrees C, respectively . Enzyme activity was inhibited by clavulanic acid, sulbactam, cephamycin derivatives, carbapenem antibiotics, iodine, HgCl2, and p-chloromercuribenzoate . The enzyme showed a broad substrate profile, hydrolyzing cephaloridine, cefazolin, cefsulodin, penicillin G, ceftizoxime, and ampicillin at a high rate.

Hum Pathol, 1984 Mar, 15(3), 244 - 7
Circulating immune complexes and the nephropathy of cystic fibrosis; Davis CA et al.; To explore the putative nephropathic role of Pseudomonas-associated immune complexes, the authors measured the quantity of immune complexes in sera obtained, before death, from 20 patients with cystic fibrosis, and compared these findings with the histologic features of the lesions and with immunofluorescence patterns of kidney tissue obtained at autopsy . The immune complexes were measured by solid-phase C1q (C1q immune complex) and conglutinin to detect complexes containing IgM, IgA, and IgG . Elevated levels of C1q immune complex (13 patients) suggested the possibility of renal deposition of C3 (P less than 0.005) and IgM (P less than 0.05) . The only three patients with IgA tissue deposits had elevated levels of C1q immune complex with normal IgA immune complexes . No other assay findings correlated with the immunofluorescence findings . Despite the prominent C3 in tissue deposits, the histologic features were not significantly associated with the results of the immune complex assays . This study indicates that complement-activating IgM-containing complexes can be deposited in renal tissues of patients with cystic fibrosis, but their nephropathogenicity is doubtful . These observations of kidney lesions, which diminish the injurious role of immune complexes in cystic fibrosis, may be relevant to an understanding of the pathogenesis of the lung lesions, which recent studies have linked to the presence of immune complexes.

Mikrobiologiia, 1984 Mar-Apr, 53(2), 218 - 22
{Breakdown of alkyl sulfonate by Pseudomonas rathonis}; Stavskaia SS et al.; Pseudomonas rathonis T/1 utilizing alkyl sulfonate, an anionic surfactant, as a sole source of carbon and energy was isolated from the soil taken near a plant producing synthetic detergents . Pseudomonas rathonis T, a more active variant of the above strain, was obtained by selection under the conditions of continuous cultivation . As was shown by identification of intermediate products in the destruction of alkyl sulfonate, its molecule is first cleaved at the C--S bond yielding higher aliphatic alcohols which are then oxidized to the corresponding carboxylic acids . The strain can be used for purification of industrial sewage containing anionic surfactants.

J Pediatr, 1984 Mar, 104(3), 460 - 6
Allergy to semisynthetic penicillins in cystic fibrosis; Moss RB et al.; Allergic reactions to anti-Pseudomonal penicillin derivatives are an increasing problem in therapy of cystic fibrosis lung disease . We evaluated 15 patients, ages 12 to 37 years, with documented allergic reactions to carbenicillin, ticarcillin, or piperacillin . Intradermal skin test reactions were positive for benzylpenicillin in seven patients, penicilloyl-polylysine in one, and ticarcillin or piperacillin in eight, for a total of 11 of 11 tested . Results of radioallergosorbent testing to penicilloyl conjugates were positive in eight of 14 patients and equivocal in four others . Overall, skin tests or RAST results were positive in 13 of 15 patients . All patients were desensitized with a semisynthetic penicillin by continuous serial intravenous infusion of 10-fold dose increments, beginning with 10(-6) of the therapeutic dose . Desensitization was successful in 25 of 26 instances . After intravenously administered therapy, maintenance of desensitization with dicloxacillin orally was unsuccessful in four of six patients . We conclude that (1) allergy to semisynthetic penicillins in cystic fibrosis usually is IgE mediated; (2) such allergy can be evaluated by skin testing; (3) it can be safely and in most cases successfully treated by intravenous desensitization; and (4) allergic patients should be desensitized on each subsequent admission for intravenously administered therapy.

J Bacteriol, 1984 Mar, 157(3), 821 - 7
Integration and excision of pMC7105 in Pseudomonas syringae pv . phaseolicola: involvement of repetitive sequences; Szabo LJ et al.; The site for integration of pMC7105 into the chromosome of Pseudomonas syringae pv . phaseolicola has been mapped to a 2.6-kilobase-pair (kb) Bg/II-EcoRI fragment on this 150-kb indigenous plasmid . Selected excision plasmids resulting from imprecise excision of pMC7105 were used to identify one of the plasmid-chromosome juncture fragments and to characterize the mechanism of recombination from the chromosome . A 14.2-kb BamHI plasmid-chromosome juncture fragment has been identified in pEX8060 (234 kb), an excision plasmid which carries approximately 90 kb of chromosomal sequences to the left of the site of integration . This fragment contains a portion of the 2.6-kb Bg/II-EcoRI fragment as well as chromosomal sequences . Blot hybridization with a probe made from selected fragments of pMC7105 revealed three distinct repetitive sequences, RS-I, RS-II, and RS-III, on this plasmid . The 2.6-kb fragment, to which the site of integration maps, also contains RS-II . Five copies of RS-II are present in pMC7105, and more than 20 copies are present in the chromosome . Eight small excision plasmids were shown to result from recombination among fragments of pMC7105 that contain common repetitive sequences . The results indicate that integration and excision of pMC7105 occur through general recombination at homologous repetitive sequences.

J Clin Microbiol, 1984 Feb, 19(2), 291 - 3
Characterization of Pseudomonas cepacia isolates from patients with cystic fibrosis; McKevitt AI et al.; Pseudomonas cepacia infections which follow a fulminant course and which include septicemia are being reported with increasing frequency from cystic fibrosis patients . Forty-eight P . cepacia isolates from cystic fibrosis patients were screened for production of potential virulence factors . A majority of strains tested produced protease and lipase . Eleven strains harbored plasmids of approximate molecular weights in the range 50 X 10(6) to 100 X 10(6) . Twenty-two strains produced a smooth lipopolysaccharide . Studies are presently under way to determine the role of these potential virulence factors in the pathogenesis of P . cepacia disease.

Laryngoscope, 1984 Feb, 94(2 Pt 1), 192 - 6
Pseudomonas rhinosinusitis; Fried MP et al.; Although rare in the otherwise healthy patient, pseudomonas rhinosinusitis is encountered most frequently in the immunocompromised host or severely traumatized patient . Intravenous antibiotic therapy in conjunction with aggressive surgical drainage is required . Two cases are documented that are typical of this philosophy . A third patient with isolated nasal involvement, because of multiple medical disabilities, was treated with local debridement and topical therapy alone and responded.

J Bacteriol, 1984 Feb, 157(2), 435 - 9
NADP-dependent glutamate dehydrogenase from a facultative methylotroph, Pseudomonas sp . strain AM1; Bellion E et al.; The NADP-dependent glutamate dehydrogenase (EC 1.4.1.4.) elaborated by the methylotrophic bacterium Pseudomonas sp . strain AM1 when growing on succinate and ammonium chloride was studied . The enzyme, which has a pH optimum of 9.0, was purified 140-fold and shown to have Km values of 20.2 mM, 0.76 mM, 0.033 mM, and 31.6 mM for ammonia, alpha-ketoglutarate, NADPH, and glutamate, respectively . The native molecular weight was determined by polyacrylamide gel electrophoresis to be 190,000, and electrophoresis under denaturing conditions in the presence of sodium dodecyl sulfate revealed a minimum molecular weight of 50,000 . The enzyme was highly specific; NADH was unable to replace NADPH in the reaction, various alpha-keto acids could not replace alpha-ketoglutarate, and neither methylamine nor hydroxylamine could substitute for ammonia . Glutamate dehydrogenase was synthesized by the bacteria only when ammonia was its nitrogen source and was repressed if methylamine or nitrate were provided as sources of nitrogen instead of ammonia.

J Bacteriol, 1984 Feb, 157(2), 643 - 8
Molybdopterin in carbon monoxide oxidase from carboxydotrophic bacteria; Meyer O et al.; The carbon monoxide oxidases (COXs) purified from the carboxydotrophic bacteria Pseudomonas carboxydohydrogena and Pseudomonas carboxydoflava were found to be molybdenum hydroxylases, identical in cofactor composition and spectral properties to the recently characterized enzyme from Pseudomonas carboxydovorans (O . Meyer, J . Biol . Chem . 257:1333-1341, 1982) . All three enzymes exhibited a cofactor composition of two flavin adenine dinucleotides, two molybdenums, eight irons and eight labile sulfides per dimeric molecule, typical for molybdenum-containing iron-sulfur flavoproteins . The millimolar extinction coefficient of the COXs at 450 nm was 72 (per two flavin adenine dinucleotides), a value similar to that of milk xanthine oxidase and chicken liver xanthine dehydrogenase at 450 nm . That molybdopterin, the novel prosthetic group of the molybdenum cofactor of a variety of molybdoenzymes (J . Johnson and K . V . Rajagopalan, Proc . Natl . Acad . Sci . U.S.A . 79:6856-6860, 1982) is also a constituent of COXs from carboxydotrophic bacteria is indicated by the formation of identical fluorescent cofactor derivatives, by complementation of the nitrate reductase activity in extracts of Neurospora crassa nit-l, and by the presence of organic phosphate additional to flavin adenine dinucleotides . Molybdopterin is tightly but noncovalently bound to the protein . COX, sulfite oxidase, xanthine oxidase, and xanthine dehydrogenase each contains 2 mol of molybdopterin per mol of enzyme . The presence of a trichloroacetic acid-releasable, so-far-unidentified, phosphorous-containing moiety in COX is suggested by the results of phosphate analysis.

J Infect Dis, 1984 Feb, 149(2), 257 - 63
Renal and auditory toxicity of high-dose, prolonged therapy with gentamicin and tobramycin in pseudomonas endocarditis; Tablan OC et al.; The nephrotoxicity and auditory toxicity of high-dose (mean, 28.6 g) and prolonged (mean, 61.6 days) courses of gentamicin and tobramycin were monitored in 15 patients receiving 17 courses of treatment for pseudomonas endocarditis . Doses were adjusted in a manner that maintained peak levels of aminoglycoside in serum at 12-15 micrograms/ml and trough levels at less than 2 micrograms/ml . Drug-related renal dysfunction and auditory toxicity occurred in 63% and 44%, respectively, of gentamicin-treated patients and in 43% and 25%, respectively, of tobramycin-treated patients . Mean maximal rises (+/- SEM) in serum creatinine levels were 0.8 (+/- 0.4) mg/dl in the group given gentamicin and 1.6 (+/- 0.7) mg/dl in the group given tobramycin . Mean maximal decreases in pure-tone hearing threshold levels were greater in gentamicin-treated patients (58.3 dB) than in those given tobramycin (22.5 dB) . Both forms of toxicity appeared earlier and at a smaller dose with gentamicin than with tobramycin.

J Pediatr, 1984 Feb, 104(2), 206 - 10
Pseudomonas cepacia infection in cystic fibrosis: an emerging problem; Isles A et al.; The prevalence of Pseudomonas cepacia infection increased from 10% in 1971 to 18% by 1981 in a population of approximately 500 patients with cystic fibrosis . Carriage of P . aeruginosa has remained unchanged at 70% to 80% over the same period . Patients infected with P . cepacia have greater impairment of pulmonary function than those with P . aeruginosa . A syndrome characterized by high fever, severe progressive respiratory failure, leukocytosis, and elevated erythrocyte sedimentation rate has occurred in eight patients over the past 3 years, with a 62% fatality rate . Because P . cepacia strains are uniformly resistant to ticarcillin, piperacillin, and aminoglycosides, and because ceftazidime is ineffective despite in vitro activity, treatment of these infections is very difficult . Prevention of acquisition and effective treatment of P . cepacia in patients with cystic fibrosis are now major clinical problems in our clinic.

Am J Gastroenterol, 1984 Feb, 79(2), 104 - 8
Cell-wall defective bacteria and Crohn's disease: studies using athymic nude mice; Zuckerman MJ et al.; Indirect immunofluorescence studies were performed to determine whether the antigenic recognition by Crohn's disease (CD) sera of lymphoid tissue from nude (nu/nu) mice injected with CD filtrate is related to cell wall defective pseudomonas-like bacterial revertant forms (CWD-RF) . Antisera raised in rabbits against two CWD-RF isolates from CD tissues did not stain lymphomas or hyperplastic lymph nodes produced by CD filtrates, although these tissues demonstrated positive immunofluorescence with CD sera . Pre-absorption of reactive CD sera with CWD-RF did not block this immunofluorescence . Formalinized suspensions of CWD-RF were injected into 36 nu/nu, 12 nu/+, and 31 conventional mice . Thirty other mice were fed suspensions of bacteria . Several nu/nu injected with CWD-RF developed lymphoma (n = 2) and plasma cell hyperplasia (n = 5), none of which reacted with CD sera . Mice fed bacteria did not show intestinal pathology . These studies demonstrate that CWD-RF are not directly related to lymphomagenesis in nu/nu induced by CD filtrates and that different CD serum antibodies are involved in recognition of CWD-RF and lymphoid tissues from nu/nu previously injected with CD filtrates.

Radiology, 1984 Feb, 150(2), 541 - 5
Malignant external otitis: early scintigraphic detection; Strashun AM et al.; Pseudomonas otitis externa in elderly diabetics may extend aggressively to adjacent bone, cranial nerves, meninges, and vessels, leading to a clinical diagnosis of "malignant" external otitis . Early diagnosis is necessary for successful treatment . This study compares the findings of initial radiographs, thin-section tomography of temporal bone, CT scans of head and neck, technetium-99m methylene diphosphonate (MDP) and gallium-67 citrate scintigraphy, and single-photon emission computed tomography (SPECT) for detection of temporal bone osteomyelitis in ten patients fulfilling the clinical diagnostic criteria of malignant external otitis . Skull radiographs were negative in all of the eight patients studied . Thin-section tomography was positive in one of the seven patients studied using this modality . CT scanning suggested osteomyelitis in three of nine patients . Both Tc-99m and Ga-67 citrate scintigraphy were positive in 10 of 10 patients . Three patients who were considered to be in clinical remission had positive Tc-99m scans and normal Ga-67 scans . These results suggest that technetium and gallium scintigraphy are more sensitive than radiographs and CT scans for early detection of malignant external otitis . Gallium scintigraphy appears to be more specific for follow-up evaluation of these patients.

Clin Chim Acta, 1984 Jan 31, 136(2-3), 131 - 6
A simple and rapid enzymatic determination of L-phenylalanine with a novel L-phenylalanine oxidase (deaminating and decarboxylating) from Pseudomonas sp . P-501; Koyama H; A simple and rapid method for the determination of L-phenylalanine using Pseudomonas L-phenylalanine oxidase (deaminating and decarboxylating) has been developed . This method involves the oxidative deamination of L-phenylalanine using L-phenylalanine oxidase from Pseudomonas sp . P-501, and the colorimetric determination of hydrogen peroxide formed from L-phenylalanine with 4-aminoantipyrine and N,N'-dimethylaniline in the presence of peroxidase . This method requires a smaller quantity of sample and is less time-consuming than those previously reported . The method can be used for the direct assay of serum L-phenylalanine levels without pretreatment.

J Biol Chem, 1984 Jan 25, 259(2), 1136 - 42
Apoenzyme of Pseudomonas cepacia salicylate hydroxylase . Preparation, fluorescence property, and nature of flavin binding; Wang LH et al.; The apoenzyme of Pseudomonas cepacia salicylate hydroxylase was prepared by a dialysis method . The apoprotein retains a dimeric structure and binds one FAD per monomer . Flavin binding results in both 81 and 60% of quenching and 15- and 5-nm blue shifts of FAD and protein fluorescence, respectively . A hydrophobic environment for the flavin site and a conformational difference between apoprotein and holoenzyme are thus indicated . Prior binding of NADH markedly retards the holoenzyme activity development upon a subsequent FAD addition . Flavin 1,N6-ethenoadenine dinucleotide binds to the apoenzyme much more weakly than FAD but this reconstituted holoenzyme and the FAD X enzyme both exhibit similar activities . The adenine moiety appears to be important to binding . The formation of holoenzyme from apoprotein and FAD involves minimally a two-step reversible process, an initial flavin-binding step followed by a conformational transition . At both 6 and 23 degrees C, the rates of hydroxylase activity recovery can be correlated with the rates of FAD binding, indicating that the initial FAD X apoenzyme complex is fully active and the subsequent slow conformational change has no significant effect on the catalytic efficiency . Overall dissociation constants calculated based on kinetic data are essentially identical with those determined by equilibrium measurements.

Biochim Biophys Acta, 1984 Jan 18, 784(1), 62 - 7
Anion binding to resting and half-reduced Pseudomonas cytochrome c peroxidase; Ellfolk N et al.; The anion-binding characteristics of resting and half-reduced Pseudomonas cytochrome c peroxidase (ferrocytochrome c-551: hydrogen peroxide oxidoreductase, EC 1.11.1.5) have been examined by EPR and optical spectroscopy with cyanide, azide and fluoride as ligands . The resting enzyme was found to be essentially inaccessible for ligation, which indicates that it has a closed conformation . In contrast, the half-reduced enzyme has a conformation in which the low-potential heme is easily accessible for ligands, a behavior parallel to that towards the substrate hydrogen peroxide (Ronnberg, M., Araiso, T., Ellfolk, N . and Dunford, H.B . (1981) Arch . Biochem . Biophys . 207, 197-204) . Cyanide and azide caused distinct changes in the low-potential heme c moiety, and the gz values of the two low-spin derivatives were 3.14 and 3.22, respectively . Fluoride binds to the same heme, giving rise to a high-spin signal at g = 6 . The dissociation constants of the anions differ widely from each other, the values for the cyanide, azide and fluoride being 23 microM, 2.5 mM and 0.13 M, respectively . In addition, a partial shift of the low-spin peak at g = 2.84 of the half-reduced species to 3.24 was observed even at low concentrations of fluoride.

Clin Allergy, 1984 Jan, 14(1), 109 - 12
Airborne endotoxins and humidifier disease; Rylander R et al.; The presence of humidifier disease in a printing factory was investigated with particular emphasis on airborne endotoxins . The water in the humidifier was contaminated with Pseudomonas . The amount of airborne endotoxin when the humidifier was operating was 0.13-0.39 microgram/m3 . Twenty of the fifty workers investigated reported typical symptoms of fever, chills, and chest tightness when the humidifiers were operating . Symptoms were more frequent among non-smokers . The estimated inhaled dose of endotoxin was found to be sufficient to cause the observed symptoms . The determination of airborne endotoxins in future episodes of humidifier disease is recommended.

Surg Gastroenterol, 1984, 3(2), 3 - 7
Endoscopic papillotomy (EPT) in acute obstructive suppurative cholangitis; Solhaug JH et al.; Eight high risk patients, median age 79 years, with a distal obstruction of the common bile duct and serious clinical symptoms of acute obstructive cholangitis were treated by EPT . In seven patients, impaction of a stone in the common bile duct was found and in one patient, an obstructing cancer . EPT was performed without immediate complications and followed by obvious drainage of purulent bile in all patients . Repapillotomy with stone extraction was necessary in three patients 5, 6, and 10 days after the first EPT . The papillotomy was followed by immediate symptom relief, normalization of body temperature, and a decrease in leukocytes and bilirubin and alkaline phosphatase values within the first several postoperative days . Average hospitalization time was 8 days, ranging from 4-17 days . The patient with pancreatic cancer died 3 months after the EPT . One other patient died in pseudomonas sepsis 17 days after an uncomplicated EPT . ERCP controls in the other six patients have been normal and they all remain symptom free . Since early decompression is mandatory in these patients and laparotomy with internal decompression is associated with a high morbidity and mortality, endoscopic decompression should probably be the recommended treatment in patients with obstructive, septic cholangitis prior to employing this therapeutic option.

Neurology, 1984 Jan, 34(1), 105 - 7
Melioidosis and bilateral third-nerve palsies; Beck RW et al.; Central nervous system involvement in melioidosis is unusual . We describe a 34-year-old man who developed a Pseudomonas pseudomallei meningitis, manifested as bilateral third-nerve palsies, 13 years after having been in southeast Asia . Diagnosis was established by a fourfold rise in the serum antibody titer for the bacterium . Recovery occurred after treatment with rifampin, isoniazid hydrochloride, ethambutol, and trimethoprim-sulfamethoxazole . Since a long latent period from exposure to overt infection is possible, additional cases of melioidosis in the United States can still be expected in veterans of the Vietnam War.

Arch Immunol Ther Exp (Warsz), 1984, 32(5), 523 - 8
Anti-Pseudomonas immunoglobulin . IV . Combined anti-Pseudomonas immunoglobulin and Pseudomonas vaccine immunotherapy of burned patients--clinical investigations; Sakiel S et al.; Sheep anti-Pseudomonas immunoglobulin and Pseudomonas vaccine were used for combined immunotherapy in 55 severely burned patients . The results of clinical investigations show that active and passive immunization in the early state of thermal injury increases the anti-Pseudomonas agglutinin titer, diminish the threat of Pseudomonas sepsis and death and significantly accelerate the recovery.

J Pediatr Gastroenterol Nutr, 1984, 3 Suppl 1, S99 - 105
Improved prognosis in CF patients with normal fat absorption; Corey M et al.; CF patients with normal fat absorption, as a group, have lower mean sweat chloride levels, maintain better pulmonary function and weight for their age, and appear to survive longer than CF patients with steatorrhea . The prognostic advantage for CF males in general is not seen in the pulmonary function data for patients with normal fat absorption, but may be reflected in the smaller number of females in this group . Males in both groups are clearly better at maintaining good weight than are females . Whether this means that nutritional intervention can improve pulmonary course or that other factors (genetic, endocrinological, environmental) dictate nutritional and pulmonary state, as well as sex differences, remains to be shown . Non-steatorrheic patients are far less likely than steatorrheic patients to have Pseudomonas infecting their lungs . This is a significant prognostic advantage since the progressive lung disease and eventual mortality of most CF patients can be charted by their acquisition of P . aeruginosa and the increasing frequency of exacerbation and attempts to eradicate this organism . CF has been called a lethal genetic disease because affected homozygotes did not generally survive to procreate . However, increasing numbers of young women with CF are surviving to an age where pregnancy and child-rearing are options for them . The majority of patients who reach this stage with sufficiently good health to embark on a pregnancy are patients with normal fat absorption . They also appear to be more likely to remain well throughout pregnancy and as young mothers . There is no doubt that CF patients with normal fat absorption have a better prognosis than those with typical CF malabsorption.(ABSTRACT TRUNCATED AT 250 WORDS)

Nauchnye Doki Vyss Shkoly Biol Nauki, 1984, (8), 23 - 6
{Spectrophotometric method of assessing molecular DNA-DNA hybridization applicable to bacteria of the genus Pseudomonas}; Levanova GF et al.; A variant of unisotopic optical DNA-DNA molecular hybridization method basing on the study of reassociation rate of DNA renaturation fragments is described . The choice of working part of the renaturation curve has been standardized . Using the species of Pseudomonas genus it has been shown that this method permits to obtain reproductive results.

Z Allg Mikrobiol, 1984, 24(7), 427 - 35
Dynamic model of discontinuous and continuous phaseolotoxin production of Pseudomonas syringae pv . phaseolicola; Guthke R et al.; From experimental data of kinetics of growth, glucose consumption and product formation of Pseudomonas syringae pv . phaseolicola the development and parameter estimation of a mathematical model is presented . The model describes the behaviour of both, batch and chemostat culture, as well as for different temperatures . The model is favoured for dynamic optimization studies . Maximal productivity is reached in the chemostat for a dilution rate which is only a little bit smaller than the wash out point.

Prep Biochem, 1984, 14(4), 363 - 72
A method for the synthesis of {14C}-kynurenine; Bild G et al.; A new method is described for the synthesis of {14C}-labelled L-kynurenine from {14C}-L-tryptophan, using extracts of tryptophan-adapted cells of Pseudomonas marginalis . It is based on the selective, rapid inactivation of kynureninase by a newly discovered inhibitor of this enzyme, 3-chloro-L-alanine . The yield of {14C}-kynurenine produced in this manner is 76% theoretical.

Zentralbl Mikrobiol, 1984, 139(2), 109 - 18
{Studies on the variability of the phaseolotoxin production by Pseudomonas syringae pv . phaseolicola}; Volksch B et al.; Isolation of bacteria from a field of bean plants (Phaseolus vulgaris L.) with conspicuous symptoms of halo blight disease resulted in 123 bacterial strains from which 57 were identified as Pseudomonas syringae pv . phaseolicola . At 18 degrees C the phaseolotoxin production of the isolated strains differs widely in submerse culture . Only few strains produce high amounts of phaseolotoxin being comparable with those of the reference strain 1321, while most of the strains show a low capability of phaseolotoxin production . Furthermore, we proved the stability of phaseolotoxin production of 29 strains . After one year about 50% of the strains were extremely reduced in their capability of phaseolotoxin production . We found that the reason for this reduction of toxin production is the appearance of Tox- segregants within a Tox+ clone . At 24 degrees C all strains (with one exception) show considerable lower amounts of toxin production or none at all: maximally 30% of the toxin amounts synthesized at 18 degrees C were produced . At 28 degrees C none of the isolated strains produce phaseolotoxin . The present data allow the conclusion to be drawn that in natural environments there exists a wide spread regarding the amount and stability of the phaseolotoxin production.

Mol Gen Genet, 1984, 197(3), 373 - 83
Physical structure, genetic content and expression of the alkBAC operon; Owen DJ et al.; We cloned sequences of the alk (alkane utilization) operon of Pseudomonas and characterized them physically and genetically . These sequences were used to construct a DNA restriction map of the alkBAC region . We physically mapped alk::Tn7 insertions and delta alkBA deletions, and we were able to show complementation or marker rescue of alk point mutations by cloned DNA sequences . Our results confirmed the existence of an operon containing structural loci encoding activities for membrane alkane hydroxylase component (alkB), soluble alkane hydroxylase component (alkA) and membrane alcohol dehydrogenase (alkC) . Physical mapping of alkC::Tn7 insertions and complementation of alkC point mutations by cloned sequences from the alkBA region showed that we were previously mistaken in inferring the existence of a separate unlinked alkC cluster . Studies with an alkB-lacZ transcription fusion construct established that the operon is transcribed in the order alkBAC and is under positive regulation by alkR regulatory functions.

Res Vet Sci, 1984 Jan, 36(1), 16 - 20
Preliminary study on the use of ceftazidime, a broad spectrum cephalosporin antibiotic, in snakes; Lawrence K et al.; Ceftazidime, a broad spectrum cephalosporin antibiotic with an enhanced anti-pseudomonal activity, was tested in vitro against a variety of reptilian bacterial isolates . Blood concentrations of this antibiotic were determined in clinically ill snakes following an intramuscular injection at a dose rate of 20 mg kg-1 . Peak plasma levels of up to 70.5 micrograms ml-1 were reached one to eight hours after the injection and therapeutic plasma levels were maintained for at least 96 hours . A series of snakes treated with ceftazidime at a dose rate of 20 mg kg-1 every 72 hours showed a rapid and obvious clinical response to treatment . The snakes were maintained at 30 degrees C during treatment and the effect of environmental temperature on antibiotic half-life is discussed . Ceftazidime proved to be a highly active antibiotic against the bacteria known to cause disease in reptiles, with no obvious adverse effects having been so far described.

Acta Chem Scand B, 1984, 38(1), 79 - 83
The formation of the primary hydrogen peroxide compound (compound I) of Pseudomonas cytochrome c peroxidase as a function of pH; Ronnberg M et al.; The effect of pH on the stability and overall catalytic activity of half-reduced Pseudomonas cytochrome c peroxidase was studied over the pH range 3.5-8 . The stability of the enzyme as deduced from 40 s incubation experiments is virtually unaffected by pH . However, there is a bell-shaped pH dependence for the overall catalytic reaction using H2O2 as oxidizing substrate and cytochrome c-551 as reducing substrate with maximum turnover rate of pH 6 . The effects of pH on (1) rate of reduction of the totally ferric enzyme by reduced azurin over the pH range 3.5-8 and (2) the rate of compound I formation from the half-reduced enzyme and hydrogen peroxide over the pH range 4-8 were also investigated . The reduction reaction rate also appears bell-shaped with optimum rate at pH 5.6 . The rate of compound I formation is virtually pH independent above pH 5 but drops dramatically as the pH is lowered from 5 to 4 . The influence of an ionization with apparent pKa value of 4.4 is implicated in compound I formation . This enzyme acid group must be deprotonated for compound I formation to occur suggesting the importance of base catalysis.

Acta Microbiol Pol, 1984, 33(2), 103 - 10
Interaction between bacterial metabolites and some pesticides . I . Interaction between the phenolic compounds produced by Pseudomonas acidovorans and the herbicide Venzar; Kosinkiewicz B; The phenolic compounds produced by Pseudomonas acidovorans interacted with the herbicide Venzar changing its phytotoxicity . Bioassay with Chlorella vulgaris and lettuce showed that the interaction of the bacterial phenols and Venzar was synergistic or antagonistic depending on the concentration of the phenolic compounds its chemical structure and pH of the environment . The humic-like polymers isolated from the bacterial culture reduced to a small extent the phytotoxicity of Venzar . It was found that {C14}-labelled Venzer was incorporated into the bacterial humic-like compounds due to the action of the enzymatic system of the bacteria.

Mol Gen Genet, 1984, 196(2), 254 - 7
Use of phenotypic suppression for direct selection of loss of aminoglycoside antibiotic resistance determinants; Hector ML; A strain of Escherichia coli containing a conditional drug dependent arginine auxotrophy was used to select for the loss of plasmid and/or transposon encoded kanamycin (Km) or streptomycin (Sm) resistance determinants . Because these determinants inactivate the corresponding drug thus eliminating drug suppression, loss of the drug-resistance determinants was selected directly by growth on minimal media plates containing sub-lethal dosages of the drug . This method was used to select loss of Km or Sm resistance determinants due to loss of plasmids, transposition from plasmid to chromosome, and education of transposons from the chromosome . Drug suppression was compared to phage PRD1 resistance in selecting for loss of plasmid vehicles during transposition and was found to be 10-1,000 times more efficient . Eighty percent of the eductant clones had undergone imprecise eductions suggesting that this method may be useful in selecting stable deletion mutants . An antibiotic suppressible strain of Pseudomonas stutzeri was obtained implying a broad utility of this selection procedure.

Mol Gen Genet, 1984, 195(1-2), 90 - 5
Characterization of eight excision plasmids of Pseudomonas syringae pv . phaseolicola; Szabo LJ et al.; Pseudomonas syringae pv . phaseolicola strain LR719 contains a 150 kilobase pair (kb) plasmid pMC7105, stably integrated into its chromosome . Occasionally, single colony isolates of this strain contain an excision plasmid . Eight unique excision plasmids were selected and characterized by BamHI restriction endonuclease and blot hybridization analyses . These plasmids ranged in size from 35 to 270 kb; the largest contained approximately 130 kb of chromosomal DNA sequences . Restriction maps of pMC7105 were developed to deduce the site of integration and to identify the fragments in which recombination occurred to produce each excision plasmid . The eight excision plasmids were arranged into five classes based on the sites where excision occurs . A 20 kb region of pMC7105, which includes BamHI fragment 9 and portions of adjacent fragments, is present in all excision plasmids and thought to contain the origin of replication . The site of integration on pMC7105 maps within BamHI fragment 8 . This fragment shows homology with seven other BamHI fragments of pMC7105 and with five chromosomal fragments identified among the excision plasmids . The data strongly suggest that the integration of pMC7105 may have occurred at a repetitive sequence present on the chromosome and on the plasmid.

J Biol Chem, 1983 Dec 25, 258(24), 14981 - 91
EPR and Mössbauer studies of protocatechuate 4,5-dioxygenase . Characterization of a new Fe2+ environment; Arciero DM et al.; Protocatechuate 4,5-dioxygenase from Pseudomonas testosteroni has been purified to homogeneity and crystallized . The iron containing, extradiol dioxygenase is shown to be composed of two subunit types (alpha, Mr = 17,700 and beta, Mr = 33,800) in a 1:1 ratio; such a composition has not been observed for other extradiol dioxygenases . The 4.2 K Mossbauer spectrum of native protocatechuate 4,5-dioxygenase prepared from cells grown in 57Fe-enriched media consists of a doublet with quadrupole splitting, delta EQ = 2.22 mm/s, and isomer shift delta Fe = 1.28 mm/s, demonstrating a high spin Fe2+ site . These parameters, and the temperature dependence of delta EQ, are unique among enzymes but are strikingly similar to those reported for the reaction center of the photosynthetic bacterium Rhodopseudomonas sphaeroides R-26, suggesting very similar ligand environments . The Fe2+ of protocatechuate 4,5-dioxygenase can be oxidized, for instance by H2O2, to yield high spin Fe3+ with EPR g values around g = 6 (and g = 4.3) . In the oxidized state, protocatechuate 4,5-dioxygenase is inactive; the iron, however, can be rereduced by ascorbate to yield active enzyme . Our data suggest that protocatechuate binds to Fe2+; the spectra indicate that the ligand binding is heterogenous . The Mossbauer spectra observed here are fundamentally different from those reported earlier (Zabinski, R., Munck, E., Champion, P., and Wood, J . M . (1972) Biochemistry 11, 3212-3219) . The spectra of the earlier (reconstituted) preparations, which had substantially lower specific activities, probably reflect adventitiously bound Fe3+ . We discuss here how adventitiously bound iron can be identified and removed . The Fe2+ which is present in native protocatechuate 4,5-dioxygenase and its complexes with substrates and inhibitors reacts quantitatively with nitric oxide to produce a species with electronic spin S = 3/2 . The EPR and Mossbauer spectra of these complexes compare favorably with EDTA . Fe(II) . NO . We have studied the latter complex extensively and have analyzed the Mossbauer spectra with an S = 3/2 spin Hamiltonian . EPR spectra show that protocatechuate 4,5-dioxygenase-NO complexes with substrates or inhibitors are heterogeneous and consist of several well defined subspecies . The data show that NO, and presumably also O2, has access to the active site Fe2+ in the enzyme-substrate complex . The use of EPR-detectable NO complexes as a rapid and sensitive tool for the study of the EPR silent active site iron of extradiol dioxygenases is discussed.

Biochem J, 1983 Dec 15, 216(3), 641 - 54
The degradation of cholic acid by Pseudomonas sp . N.C.I.B . 10590 under anaerobic conditions; Owen RW et al.; The bacterial degradation of cholic acid under anaerobic conditions by Pseudomonas sp . N.C.I.B . 10590 was studied . The major unsaturated neutral compound was identified as 12 beta-hydroxyandrosta-4,6-diene-3,17-dione, and the major unsaturated acidic metabolite was identified as 12 alpha-hydroxy-3-oxochola-4,6-dien-24-oic acid . Eight minor unsaturated metabolites were isolated and evidence is given for the following structures: 12 alpha-hydroxyandrosta-4,6-diene-3,17-dione, 12 beta,17 beta-dihydroxyandrosta-4,6-dien-3-one, 12 beta-hydroxyandrosta-1,4,6-triene-3,17-dione, 12 beta,17 beta-dihydroxyandrosta-1,4,6-trien-3-one, 12 beta-hydroxyandrosta-1,4,6-triene-3,17-dione, 12 beta,17 beta-dihydroxyandrosta-1,4,6-trien-3-one, 12 alpha-hydroxyandrosta-1,4-diene-3,17-dione, 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione, 3,12-dioxochola-4,6-dien-24-oic acid and 12 alpha-hydroxy-3-oxopregna-4,6-diene-20-carboxylic acid . In addition, a major saturated neutral compound was isolated and identified as 3 beta,12 beta-dihydroxy-5 beta-androstan-17-one, and the only saturated acidic metabolite was 7 alpha,12 alpha-dihydroxy-3-oxo-5 beta-cholan-24-oic acid . Nine minor saturated neutral compounds were also isolated, and evidence is presented for the following structures: 12 beta-hydroxy-5 beta-androstane-3,17-dione, 12 alpha-hydroxy-5 beta-androstane-3,17-dione, 3 beta,12 alpha-dihydroxy-5 beta-androstan-17-one, 3 alpha,12 beta-androstan-17-one, 3 alpha,12 alpha-dihydroxy-5 beta-androstan-17-one, 5 beta-androstane-3 beta,12 beta,17 beta-triol, 5 beta-androstane-3 beta,12 alpha,17 beta-triol, 5 beta-androstane-3 alpha,12 beta,17 beta-triol and 5 beta-androstane-3 alpha,12 alpha,17 beta-triol . The induction of 7 alpha-dehydroxylase and 12 alpha-dehydroxylase enzymes is discussed, together with the significance of dehydrogenation and ring fission under anaerobic conditions.

J Biol Chem, 1983 Dec 10, 258(23), 14422 - 7
Rapid reaction studies on the oxygenation reactions of catechol dioxygenase; Walsh TA et al.; The reaction of oxygen with catechol 1,2-dioxygenase from Pseudomonas arvilla ATCC 23974 in complex with catechol, 4-methylcatechol, and 4-fluorocatechol has been studied using single turnover stopped flow spectrophotometry . Two sequential enzyme intermediates have been resolved and their visible spectra characterized by computer-assisted methods . These intermediates are spectrally similar to those observed in a similar study with protocatechuate dioxygenase (Bull, C., Ballou, D . P., and Otsuka, S . J . Biol . Chem . 256, 12681-12686 (1981), although the first intermediate seen with the latter enzyme was not observed in this study . The rate of formation of intermediate I is oxygen-dependent and also accelerated by electron-donating substituents on the C-4 of the substrate . This is consistent with the proposed substrate reduction of dioxygen to form a hydroperoxide . Intermediate I is thus suggested to be a 6-hydroperoxycyclohexa-3,5-diene-1-one . The decay of intermediate I is also accelerated by electron donors and is consistent with the rearrangement of intermediate hydroperoxide via an acyl migration mechanism . It is inconsistent with mechanisms involving nucleophilic attack at the carbonyl carbon . Intermediate II is proposed to be an enzyme-product complex based on the resemblance of its visible spectra to those of the benzoate complex of catechol 1,2-dioxygenase and enzyme-product complexes of protocatechuate dioxygenase . Careful 18O2-labeling experiments have shown that no label is lost to the solvent, implying that no free hydroxide forms during catalysis.

J Biol Chem, 1983 Dec 10, 258(23), 14413 - 21
Halogenated protocatechuates as substrates for protocatechuate dioxygenase from Pseudomonas cepacia; Walsh TA et al.; Substrates containing electron-withdrawing groups were reacted with protocatechuate 3,4-dioxygenase and oxygen . Haloprotocatechuates (5-fluoro-, 5-chloro-, 5-bromo-, 2-chloro-, and 6-chloroprotocatechuates) are oxygenated by the enzyme at rates 28- to 3000-fold lower than that with the native substrate . These lower rates are due to both deactivation of substrate to O2 attack, and to the formation of abortive enzyme-substrate (ES) complexes . Such ES complexes with haloprotocatechuates are spectrally distinct from the normal ES complex . 6-Chloroprotocatechuate produces changes more like those due to protocatechuate . The abortive ES complexes, when rapidly mixed with oxygen, decay to free enzyme and product monophasically, and the dependence of the rates on O2 concentration shows that a rate-limiting step precedes reaction with O2 . Thus these complexes are rather unreactive toward O2, and the rate-limiting step in oxygenation is their conversion to active complexes . In contrast, the reaction of O2 with the enzyme and 6-chloroprotocatechuate is biphasic, the first phase being dependent on O2 concentration (2 X 10(4) M-1 S-1) and the second not (7 S-1) . The intermediate formed after the first phase strongly resembles the second intermediate seen in the reaction of enzyme with protocatechuate and O2 (Bull, C., Ballou, D . P., and Otsuka, S., (1981) J . Biol . Chem . 256, 12681-12686), implying that the electron-withdrawing effect of the chlorine slows the O2 addition step considerably while the conversion to the second intermediate is hardly affected . When the enzyme cycles through several turnovers with 6-chloroprotocatechuate, an enzyme species is formed that resembles the unreactive ES complexes seen with the other haloprotocatechuates, indicating that a small amount of the unreactive complex is in equilibrium with the reactive complex and that during successive turnovers the enzyme is slowly converted into the unreactive form . The formation of this form correlates with the observation that in assays the rate of product formation gradually decreases with time.

Biochemistry, 1983 Dec 6, 22(25), 5922 - 9
gamma-butyrobetaine hydroxylase: primary and secondary tritium kinetic isotope effects; Blanchard JS et al.; Primary and secondary tritium kinetic isotope effects have been determined for the reactions catalyzed by purified preparations of gamma-butyrobetaine hydroxylase obtained from Pseudomonas sp AK 1 and from calf liver . With {methyl-14C,(3R)-3-3H}-gamma-butyrobetaine as substrate, the bacterial hydroxylase was found to exhibit a primary T(V/K) of 1.3-1.5 . This value was determined from measurements of either the specific activity of the medium 3H2O or of the ratio of 3H/14C in the residual gamma-butyrobetaine . Under identical conditions of analysis, the calf liver enzyme exhibited a primary T(V/K) of approximately 15 . With {methyl-14C,(4R)-4-3H} gamma-butyrobetaine as substrate, a beta-secondary T(V/K) of 1.10 has been determined for the calf liver hydroxylase; this supports the existence in the reaction mechanism of an sp2-hybridized transition state . A large normal value of 1.31 for the alpha-secondary T(V/K), as derived from measurements with {methyl-14C,2,3-3H}-gamma-butyrobetaine, suggests that the motions of the primary and alpha-secondary hydrogens are coupled in the C-H cleavage step and resulting synchronous rehybridization . A chemical mechanism involving homolytic cleavage of the C-H bond at the position undergoing hydroxylation is proposed and discussed.

J Appl Biochem, 1983 Dec, 5(6), 375 - 81
Taurocyamine-utilizing mutants from a wild-type strain of Pseudomonas; Yorifuji T et al.; Pseudomonas sp . ATCC 14676 produces glycocyaminase (EC 3.5.3.2) and guanidinobutyrase (EC 3.5.3.7) . Taurocyamine (2-guanidinoethane sulfonate) is a gratuitous inducer of both of these amidinohydrolases . Mutants of this organism capable of utilizing taurocyamine as a nitrogen source were isolated directly from the wild-type cells after uv irradiation or treatment with N-methyl-N'-nitro-N-nitrosoguanidine; frequencies of mutations observed under appropriate conditions were above 10(-7) . Strain U2-3-3, which was selected from the 11 isolated taurocyamine-utilizing strains, was proved to be derived from the wild-type strain . Both taurocyamine and 4-guanidinobutyrate were able to induce an enzyme of strain U2-3-3 that liberated urea from taurocyamine, whereas glycocyamine failed to induce the system . The activity of the enzyme toward taurocyamine was found to be about one-third of that toward guanidinobutyrate when both taurocyamine and guanidinobutyrate were used as inducer . These observations suggest that the enzyme of the mutant capable of hydrolyzing taurocyamine has emerged from guanidinobutyrase of the wild-type strain which hydrolyzes taurocyamine at a very low rate, probably as a result of a point mutation in the structural gene.

J Antimicrob Chemother, 1983 Dec, 12(6), 555 - 63
Synergy with double and triple antibiotic combinations compared; Berenbaum MC et al.; Combinations of two sets of antibiotics (gentamicin-carbenicillin-rifampicin and trimethoprim/sulphamethoxazole-carbenicillin-rifampicin) were tested against isolates of Pseudomonas maltophilia . An abbreviated checker-board method permitted tests with combinations of all three antibiotics and of each of the possible three pairs in each set . Triple combinations were usually synergistic, whereas 25% of combinations of pairs were antagonistic or additive . The modal degree of synergy was greater with combinations of three antibiotics than with combinations of any two . For any isolate, greater synergy was generally obtained with a triple than with a double combination, and the mean degree of synergy was always greater with triple than with double combinations . It was advantageous to add to double combinations even an antibiotic to which the organism was resistant . Organisms multiply resistant to three antibiotics were also resistant to 25% of combinations of pairs, but never resistant to combinations of all three . Synergy was more likely to be present at all antibiotic ratios with triple than with double combinations . Since ratios at the site of action may differ greatly from that of the administered combination, there would be substantial clinical benefit in identifying sets of antibiotics of which combinations at all ratios showed synergy . The search for these may entail exploring multicomponent combinations and, with conventional methods, this raises a considerable logistic problem . A strategy is therefore proposed, designed to find antibiotic sets showing synergy at all ratios.

J Antibiot (Tokyo), 1983 Dec, 36(12), 1735 - 42
The biosynthesis of brominated pyrrolnitrin derivatives by Pseudomonas aureofaciens; van Pee KH et al.; The mutant strain ACN of Pseudomonas aureofaciens ATCC 15926 produces several bromo derivatives of pyrrolnitrin . Five brominated amino- and three brominated nitrophenyl pyrrole compounds could be isolated, and their structures were established by 1H NMR, UV and mass spectroscopy . The isolated amino compounds showed no biological activity; the nitro derivatives inhibited the growth of Neurospora crassa ATCC 9276, though not as effective as pyrrolnitrin itself . 2-Carboxy-4-(2-amino-3-bromophenyl)pyrrole (X) is demonstrated to be an intermediate in the biosynthesis of brominated pyrrolnitrin; the biosynthetic pathway to bromo derivatives of pyrrolnitrin is discussed.

Eur J Biochem, 1983 Dec 1, 137(1-2), 149 - 54
Beta-agarases I and II from Pseudomonas atlantica . Substrate specificities; Morrice LM et al.; Beta-Agarase I and II were characterised by their action on agar-type polysaccharides and oligosaccharides . Beta-Agarase I, an endo-enzyme, was specific for regions containing a minimum of one unsubstituted neoagarobiose unit {3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactose}, hydrolysing at the reducing side of this moiety . Yaphe demonstrated that agar was degraded by this enzyme to neoagaro-oligosaccharides limited by the disaccharide but with a predominance of the tetramer {Yaphe, W . (1957) Can . J . Microbiol . 3, 987-993} . Beta-Agarase I slowly degraded neoagarohexaose but not the homologous tetrasaccharide . {1-3H}Neoagarohexaitol was cleaved to neoagarotetraose and {1-3H}neoagarobiitol . The highly substituted agar, porphyran was degraded to methylated, sulphated and unsubstituted neoagaro-oligosaccharides which were invariably terminated at the reducing end by unsubstituted neoagarobiose . The novel enzyme, beta-agarase II, was shown to be an endo-enzyme . Preliminary evidence indicated this enzyme was specific for sequences containing neoagarobiose and/or 6(1)-O-methyl-neoagarobiose . It degraded agar to neoagaro-oligosaccharides of which the disaccharide was limiting and predominant . Beta-Agarase II rapidly degraded isolated neogarotetraose and neoagarohexaose to the disaccharide . With {1-3H}neoagarohexaitol, exo-action was observed, the alditol being cleaved to neoagarobiose and {1-3H}neoagarotetraitol . Neoagarotetraitol was hydrolysed at 4% of the rate observed for the hexaitol . Porphyran was degraded to oligosaccharides, the neutral fraction comprising 24% of the starting carbohydrate . This fraction was almost exclusively disaccharides (22.4%) containing neoagarobiose (7.4%) and 6(1)-O-methyl-neoagarobiose (15%) . Beta-Agarase II is probably the 'beta-neoagarotetraose hydrolase' reported by Groleau and Yaphe as an exoenzyme against neoagaro-oligosaccharides {Groleau, D . and Yaphe, W . (1977) Can . J . Microbiol . 23, 672-679}.

J Bacteriol, 1983 Dec, 156(3), 1349 - 51
Transfer of pRD1 to Pseudomonas syringae and evidence for its integration into the chromosome; Vincent JR et al.; Plasmid pRD1 was conjugatively transferred from Escherichia coli to Pseudomonas syringae . Subculturing the transconjugate on a medium that selected for pRD1-determined His+ Kmr resulted in the loss of pRD1 as an extrachromosomal element as detected by agarose gel electrophoresis . DNA hybridization provided evidence for the integration of pRD1 into the P . syringae chromosome.

J Bacteriol, 1983 Dec, 156(3), 1178 - 87
Carbon monoxide-insensitive respiratory chain of Pseudomonas carboxydovorans; Cypionka H et al.; Experiments employing electron transport inhibitors, room- and low-temperature spectroscopy, and photochemical action spectra have led to a model for the respiratory chain of Pseudomonas carboxydovorans . The chain is branched at the level of b-type cytochromes or ubiquinone . One branch (heterotrophic branch) contained cytochromes b558, c, and a1; the second branch (autotrophic branch) allowed growth in the presence of CO and contained cytochromes b561 and o (b563) . Electrons from the oxidation of organic substrates were predominantly channelled into the heterotrophic branch, whereas electrons derived from the oxidation of CO or H2 could use both branches . Tetramethyl-p-phenylenediamine was oxidized via cytochromes c and a exclusively . The heterotrophic branch was sensitive to antimycin A, CO, and micromolar concentrations of cyanide . The autotrophic branch was sensitive to 2-n-heptyl-4-hydroxyquinoline-N-oxide, insensitive to CO, and inhibited only by millimolar concentrations of cyanide . The functioning of cytochrome a1 as a terminal oxidase was established by photochemical action spectra . Reoxidation experiments established the functioning of cytochrome o as an alternative CO-insensitive terminal oxidase of the autotrophic branch.

Prikl Biokhim Mikrobiol, 1983 Nov-Dec, 19(6), 744 - 50
{Properties of a bacterial enzyme preparation of alkylsulfatase}; Udilova OF et al.; A preparation of primary alkyl sulfatase was obtained from the culture of Pseudomonas species 2T/1 . It can hydrolyze alkyl sulfates, which belong to anion surface-active compounds, to sulfate ion and fatty alcohol, and as a result the harmful for biosphere property of the surface activity is gone . pH and temperature of the incubation mixture, the presence of ions of some bivalent metals and components of synthetic detergents (SD), composition of the buffer mixture and substrate concentration affect the rate of sodium dodecylsulfate (SDS) hydrolysis . The alkyl sulfatase preparation is relatively stable . The maximum rate of SDS hydrolysis was found to be at 70 degrees C . The preparation catalyzes the hydrolysis of some alkyl sulfate homologues and industrial alkyl sulfates . The temperature optimum of the preparation is 40 degrees C, the pH-optimum is 8.0-9.0.

J Infect, 1983 Nov, 7(3), 256 - 63
Nosocomial infections by chlorhexidine solution contaminated with Pseudomonas pickettii (Biovar VA-I); Kahan A et al.; Over a period of 10 days, six patients in a cardiac intensive care unit developed Pseudomonas pickettii (biovar VA-I) septicaemia after installation of a venous catheter . The organism was also recovered from all the vials of the aqueous solution of 0.05 per cent chlorhexidine ('Hibitane') prepared with contaminated bidistilled water . There were no further cases of infection when the use of this water was prohibited.

Rev Infect Dis, 1983 Nov-Dec, 5 Suppl 5, S985 - 91
Inhibition of the activity of pseudomonas toxin by methylamine; FitzGerald D et al.; Methylamine at a concentration of 20 mM protected mouse LM cell fibroblasts from the action of pseudomonas toxin . Nearly total protection was observed when cells were pretreated with amine before the addition to toxin and amine, when amine and toxin were added simultaneously, or when amine was added up to 30 min after toxin binding . Later addition of methylamine afforded partial protection of the monolayers . Using electron microscopy, we observed that toxin initially bound diffusely to the cell surface but rapidly moved to coated pits and was internalized after cells were warmed to 37 C . Methylamine blocked the clustering of toxin into coated-pit areas of the membrane but did not alter the overall level of toxin internalization . It is suggested that pseudomonas toxin enters mammalian cells by receptor-mediated endocytosis and that methylamine alters the entry process . In those instances in which partial protection was seen when methylamine was added after toxin internalization, the primary amine may be functioning by inactivation of lysosomal processing of the toxin.

J Lipid Res, 1983 Nov, 24(11), 1500 - 11
The degradation of cholesterol by Pseudomonas sp . NCIB 10590 under aerobic conditions; Owen RW et al.; The metabolic pathway of cholesterol degradation by bacteria has not been completely established . Several possible intermediates have not been identified and many pathway delineations have not involved the use of the cholesterol molecule per se and just one bacterial species . The bacterial degradation of cholesterol by Pseudomonas sp . NCIB has been studied . Major biotransformation products included cholest-5-en-3-one, cholest-4-en-3-one, 26-hydroxycholest-4-en-3-one, androsta-1, 4-dien-3-17-dione, cholest-4-en-3-one-26-oic acid, chol-4-en-3-one-24-oic acid, pregn-4-en-3-one-20-carboxylic acid, and pregna-1, 4-dien-3-one-20-carboxylic acid . Studies with selected intermediates have enabled the elucidation of a comprehensive pathway of cholesterol degradation by bacteria.

Appl Environ Microbiol, 1983 Nov, 46(5), 1182 - 6
Regulation of 2,4,5-trichlorophenoxyacetic acid and chlorophenol metabolism in Pseudomonas cepacia AC1100; Karns JS et al.; The expression of the degradative genes encoding 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 2,4,5-trichlorophenol (2,4,5-TCP), and pentachlorophenol (PCP) dechlorination in a 2,4,5-T-degrading strain of Pseudomonas cepacia was examined during growth on alternate carbon sources . The dechlorination mechanisms for all three compounds were expressed in 2,4,5-T- and 2,4,5-TCP-grown cells but were not expressed in cells grown on succinate, glucose, or lactate . The addition of 2,4,5-TCP or PCP to cells grown on succinate or lactate resulted in the expression of the 2,4,5-TCP dechlorination mechanism in resting cells after 1-h lag . This expression was prevented by the presence of chloramphenicol in the resting cell suspension . Succinate-plus-PCP-grown resting cells preincubated with 2,4,5-TCP fully induced the trichlorophenol dechlorination system and partially induced the PCP dechlorination system . Preincubation of succinate-plus-PCP-grown resting cells with PCP induced neither the 2,4,5-TCP nor the PCP dechlorinating system . Succinate-grown resting cells converted 2,4,5-T to 2,4,5-TCP even in the presence of chloramphenicol . Thus, the data indicate that the enzyme(s) which converts 2,4,5-T to 2,4,5-TCP is constitutively expressed, whereas those that convert 2,4,5-TCP to central intermediates are induced by 2,4,5-TCP but not by 2,4,5-T or PCP and are repressed in the presence of an alternate carbon source.

Appl Environ Microbiol, 1983 Nov, 46(5), 1176 - 81
Metabolism of Halophenols by 2,4,5-trichlorophenoxyacetic acid-degrading Pseudomonas cepacia; Karns JS et al.; Resting cells of 2,4,5-trichlorophenoxyacetic acid-grown Pseudomonas cepacia AC1100 were able to completely and rapidly dechlorinate several chlorine-substituted phenols, including 2,4,5-trichlorophenol, 2,3,4,6-tetrachlorophenol, and pentachlorophenol . Several other trichlorophenols were only partially dechlorinated . The evidence suggests that 2,4,5-trichlorophenol is an intermediate in the degradation of 2,4,5-trichlorophenoxyacetic acid by strain AC1100 . Moreover, although strain AC1100 was isolated by selection for growth on a chlorinated aromatic compound, brominated and fluorinated analogs were efficiently dehalogenated by strain AC1100 resting cells, whereas an iodinated analog was poorly dehalogenated.

J Clin Microbiol, 1983 Nov, 18(5), 1073 - 8
Cellular fatty acid composition of Pseudomonas marginata and closely associated bacteria; Dees SB et al.; The cellular fatty acid compositions of the lectotype strain and four clinical isolates of Pseudomonas marginata were determined by gas-liquid chromatography and compared with 11 strains of the Centers for Disease Control Pseudomonas-like group 2, which are similar to P . marginata in a number of conventional biochemical tests . Isolates of P . marginata were readily distinguished from Pseudomonas-like group 2 by the presence of a C17:0 cyclopropane acid and hydroxy acids 3-OH-C14:0, 2-OH-C16:0, 3-OH-C16:0, and 2-OH-C18:1, whereas strains of Pseudomonas-like group 2 contained C16:1 delta 9 as a major acid with small amounts of 3-OH-C12:0 and 2-OH-C14:0 acids . Our data show that cellular fatty acid composition provides useful additional information that can be combined with selected conventional tests to provide a more reliable and rapid identification of P . marginata and related bacteria.

Ann Microbiol (Paris), 1983 Nov-Dec, 134B(3), 411 - 9
{Detection of glycosidases in Pseudomonas of the fluorescent group: relation between serotype and glycosidase activities in P . aeruginosa}; Hansen W et al.; Fluorescent Pseudomonas species (P . aeruginosa, P . fluorescens, P . putida) were tested for the presence of glycosidase activities (alpha-D-glucosidase, beta-D-glucosidase, alpha-D-galactosidase, beta-D-galactosidase, beta-xylosidase, alpha-D-mannosidase, alpha-L-fucosidase, beta-L-fucosidase, beta-D-glucuronidase and N-acetyl-beta-D-glucosaminidase) . Some of the investigated glycosidases were always absent, while N-acetyl-beta-D-glucosaminidase was constantly present in all strains; 3 glycosidase activities were observed in association or separately . Serotype O11 of P . aeruginosa was found to be homogeneous with respect to some of those enzymatic activities . Search for beta-D-galactosidase, alpha-D-glucosidase and beta-D-glucosidase may be of diagnostic value in epidemiologic studies of P . aeruginosa.

Ann Surg, 1983 Nov, 198(5), 654 - 62
Lung vascular permeability after reversal of fibronectin deficiency in septic sheep . Correlation with patient studies; Saba TM et al.; Plasma fibronectin deficiency and opsonic dysfunction exist in critically ill septic surgical, trauma, and burn patients with multiple organ failure . Fibronectin deficiency can be reversed by infusion of fresh plasma cryoprecipitate . The influence of therapy with human cryoprecipitate on lung vascular permeability in septic sheep with plasma fibronectin deficiency following surgery was evaluated . Additionally, selected studies on pulmonary function in septic surgical and trauma patients after infusion of plasma cryoprecipitate were completed . In patients, ventilation-perfusion balance appeared to improve as measured by the multiple inert gas elimination technique . With the lung lymph fistula preparation in fibronectin deficient sheep, infusion of human plasma cryoprecipitate (10 units; 250 ml) delayed the onset and minimized the increase in lung vascular permeability during postoperative Pseudomonas sepsis (5 X 10(9) bacteria, I.V.; 5 X 10(10) bacteria, I.P.) . For example, in a first group of sheep, the transvascular protein clearance (TPC) at 2 hrs in septic sheep (n = 4) treated with only saline (volume control) was 20.1 +/- 3.1 ml/hr, compared to 11.23 +/- 0.83 ml/hr in the sheep (n =a 4) treated with fibronectin-rich cryoprecipitate (p less than 0.05) . In a second group of sheep, cryoprecipitate depleted of fibronectin by affinity chromatography was used as the control solution . It also did not manifest this protective effect with respect to lung vascular permeability . Thus, at 2 hrs the lymph flow (Qlym) was 30.2 ml/hr and the transvascular protein clearance (TPC) was 18.0 ml/hr in septic sheep given fibronectin-deficient cryoprecipitate . In contrast, in the fibronectin-rich cryoprecipitate treated sheep, the Qlym was 14.8 ml/hr and the TPC was 8.12 ml/hr . It is suggested that fibronectin may influence lung vascular integrity during sepsis following surgery and trauma.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1983 Nov, 255(4), 479 - 88
Micromethod for carbon substrate assimilation by Pseudomonas maltophilia; Freney J et al.; Assimilation of 60 carbon sources by 114 Pseudomonas maltophilia strains, identified by conventional methods, was studied by a standardized micromethod (API strip) . Automatic reading of growth intensity was done at 1, 2 and 4 days . Growth kinetics of reference strain NCTC 10257 were also studied and showed evidence of different growth according to the substrates . The results agreed well with nutritional patterns reported . This micro-method seems suitable for daily use by laboratories because of purity and standardization of substrates, facility of use, accurate and automatic reading, and the great number of assimilation tests possible.

Vet Microbiol, 1983 Nov, 8(6), 611 - 5
Evaluation of the API 20E and Microbact 24E systems for the identification of Pseudomonas pseudomallei; Thomas AD; One-hundred isolates of Pseudomonas pseudomallei were used to evaluate the API 20E and Microbact 24E rapid identification systems . The API 20E system identified 50% of the isolates using the revised 1979 Manual only, and 63% when referral was made to the computer centre . A higher identification rate (69 and 87%, respectively) was achieved with a longer incubation period of 96 h . The Microbact 24E system identified 84% of the isolates as P . pseudomallei using the revised 1983 Manual, and 100% when referral was made to the computer centre . The Microbact 24E system would appear to be a reliable system for the identification of P . pseudomallei.

Am J Dis Child, 1983 Nov, 137(11), 1044 - 7
Ceftriaxone for the treatment of serious infections; Steele RW et al.; Ceftriaxone is an investigational cephalosporin with a half-life of five to eight hours . In an uncontrolled study, we evaluated its efficacy and safety in 30 pediatric and 12 young adult patients with serious bacterial infections . This agent was administered to children at a dosage of 50 to 75 mg/kg/day intravenously in two divided doses . Those with CNS infections received 100 mg/kg/day . In adults, the dosage was 1 g either once or twice daily . The diseases we treated included pneumonia (17), sepsis (eight), ventriculoperitoneal shunt infections (three), osteomyelitis (three), brain abscess (two), peritonitis (two), and miscellaneous (seven) . Clinical cures were achieved in all cases, although one child with cystic fibrosis and Pseudomonas pneumonia had persistent colonization in his sputum . No serious side effects were observed . Although not the agent of choice for many of these pathogens, ceftriaxone appears to represent an important alternative to therapy.

Biochim Biophys Acta, 1983 Oct 28, 748(2), 194 - 204
Coordination of the heme iron in the low-potential cytochromes c-553 from Desulfovibrio vulgaris and Desulfovibrio desulfuricans . Different chirality of the axially bound methionine in the oxidized and reduced states; Senn H et al.; The coordination geometry at the heme iron of the cytochromes c-553 from Desulfovibrio vulgaris and Desulfovibrio desulfuricans was investigated by 1H-nuclear magnetic resonance and circular dichroism spectroscopy . Individual assignments were obtained for heme c and the axial ligands . From studies of nuclear Overhauser enhancements the axial histidine imidazole ring orientation relative to the heme group was found to coincide with other c-type cytochromes . In contrast, a new structure was observed for the axial methionine in the reduced cytochromes c-553 . This includes S chirality at the iron-bound sulfur atom, but compared to cytochromes c-551 from Pseudomonads and Rhodopseudomonas gelatinosa and cytochrome c5 from Pseudomonas mendocina, which also contain S-chiral methionine, a different spatial arrangement of the gamma- and beta-methylene groups and the alpha carbon of methionine prevails . For the ferricytochromes c-553 R chirality was found for the iron-bound sulfur . This is the first observation of different methionine chirality in different oxidation states of the same c-type cytochrome.

J Biol Chem, 1983 Oct 25, 258(20), 12409 - 12
Amino acid sequence of cytochrome c-553 from Desulfovibrio vulgaris Miyazaki; Nakano K et al.; The complete amino acid sequence of cytochrome c-553 from Desulfovibrio vulgaris Miyazaki has been determined . The protein has a single polypeptide chain containing 79 amino acid residues and has the typical characteristics of small mitochondrial cytochromes . Contrary to the expectation that the amino acid sequence of cytochrome c-553 from D . vulgaris Miyazaki is closely related to that from D . vulgaris Hildenborough, the two are not alike, except for the 20 NH2-terminal residues and the 4 carboxyl-terminal residues; however, the tryptic peptides obtained from the two cytochromes are similar to each other . The sequence of cytochrome c-553 from D . vulgaris Miyazaki resembles that of Pseudomonas cytochrome c-551 . The phylogenetic situation of Desulfovibrio cytochrome c-553 in the phylogenetic tree of the cytochrome c super-family is discussed.

Eur J Biochem, 1983 Oct 3, 135(3), 553 - 8
beta-agarases I and II from Pseudomonas atlantica . Purifications and some properties; Morrice LM et al.; The agarose-degrading system of Pseudomonas atlantica has been re-examined . In addition to the previously reported extracellular endo-beta-agarase {Yaphe, W . (1966) in Proceedings 5th International Seaweed Symposium, pp . 333-335} a second, membrane-bound endo-enzyme activity, beta-agarase II has been discovered . These two enzymes act in concert to degrade agarose to neoagarobiose {3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactose} and also to degrade partially 6-O-methylated agarose to neoagarobiose and 6(1)-O-methyl-neoagarbiose . Novel assays were devised for beta-agarase II and the associated disaccharidase, neoagarobiose hydrolase . These allowed the critical purification of beta-agarase I and II . beta-Agarase I was purified 670-fold from the bacterial medium by a new method using ammonium sulphate precipitation and gel filtration on Sephadex G-100 . The enzyme was resolved from the small amount of extracellular beta-agarase II . Dodecylsulphate/polyacrylamide gel electrophoresis indicated a homogeneous protein and a molecular weight of 32000 . Activity was observed against agar over the pH range 3.0-9.0 and optimally at pH 7.0 . The enzyme could be used indefinitely at 30 degrees C but only for up to 2 h at 40 degrees C . beta-Agarase II was partially purified (5-fold) from the soluble fraction of disrupted cells by chromatography on Sephadex G-100, hydroxyapatite and DEAE-Sepharose CL-6B . This preparation was free of beta-agarase I and disaccharidase . beta-Agarase II was stimulated by NaCl, optimally in the range 0.10-0.20 mol dm-3 (2.4-fold the activity at 0.010 mol dm-3 NaCl) . Alkali earth metal (0.002 mol dm-3 CaCl2 or 0.005 mol dm-3 MgCl2) gave 1.2-fold the normal activity . Optimum activity was over pH 6.5-7.5.

Antibiotiki, 1983 Oct, 28(10), 729 - 33
{R-plasmids of bacteria of the genus Pseudomonas isolated from industrial sewage}; Kozlova EV et al.; A total of 132 Pseudomonas strains isolated from untreated sewage of antibiotic plants were tested . A significant number of the strains were resistant to streptomycin (77 per cent), carbenicillin (75 per cent), kanamycin (37.5 per cent) and tetracycline (23 per cent) . Eighteen conjugative and 3 nonconjugative resistance plasmids were detected in 19 strains . The genes determining the resistance to streptomycin, kanamycin and tetracycline were most frequent . The frequency of the plasmid transfer between the strains of Ps . aeruginosa (PAO) varied within 10(-3)--10(-7) per donor cell . Six plasmids belonged to group Inc P-1 . Four plasmids belonged to group Inc P-2, 3 plasmids to groups Inc P-3 and Inc P-5 and 1 plasmid to group Inc P-7.

Arch Intern Med, 1983 Oct, 143(10), 1909 - 12
Pseudomonas stutzeri bacteremia associated with hemodialysis; Goetz A et al.; Pseudomonas stutzeri bacteremia developed in six patients undergoing hemodialysis . Fever, shaking chills, nausea, and vomiting were observed . All patients recovered, although only two received specific antibiotic therapy . The infections occurred sporadically over a period of nine months . Pseudomonas stutzeri was subsequently isolated from the dialysate that circulates within the hemodialysis machine . The ultimate source was the deionized water that is combined with the liquid concentrate to form the dialysate . Pseudomonas stutzeri could be localized to the top cannister of the dialysis machine but was also isolated throughout the machine, including the bottom reservoir and the recirculating pump . The emphasis on handwashing, strict compliance with disinfection procedures, and elimination of prolonged sitting times for the filled machine after disinfection resulted in no further cases of P stutzeri infection.

Arch Dis Child, 1983 Oct, 58(10), 826 - 8
Endotoxaemia in cystic fibrosis: response to antibiotics; Taylor G et al.; In a study of 6 children with cystic fibrosis receiving intravenous antibiotics for pseudomonas lung infection, serum endotoxin values were monitored by a modification of the limulus lysate technique . The values fell with treatment, reflecting a response that was not always apparent on clinical assessment . Endotoxin concentrations may offer a more precise way of monitoring the effects of antibiotic treatment in CF patients.

J Bacteriol, 1983 Oct, 156(1), 30 - 5
Evidence for an active role of donor cells in natural transformation of Pseudomonas stutzeri; Stewart GJ et al.; The transfer of chromosomal genes in a cell mat of Pseudomonas stutzeri was ca . 10(3) times more efficient per microgram of DNA if DNA was added as a constituent of intact donor cells rather than as a solution . Such intact cell-mediated transfer appears to depend on cell contact . It is independent of the presence of plasmids in donor strains and is DNase I sensitive, thus fitting the usual definition of transformation . It is bidirectional: cells of either strain in a transformation mixture served as the donor and recipients . The donor function in cell contact transformation was inhibited by nalidixic acid but was unaffected by rifampin and streptomycin at growth-inhibiting concentrations . Concentrations of nalidixic acid sufficient to inhibit donor function completely had no effect on the ability of nalidixic acid-resistant recipients to take up DNA from solution . These experiments suggest that certain cells donate DNA to others in the cell mat: they argue against the hypothesis that the function of donor cells is merely cell lysis.

Biochim Biophys Acta, 1983 Sep 14, 747(1-2), 16 - 25
A new spatial structure for the axial methionine observed in cytochrome c5 from Pseudomonas mendocina . Correlations with the electronic structure of heme c; Senn H et al.; Cytochrome c5 from Pseudomonas mendocina has been isolated and the coordination geometry at the heme iron was investigated by 1H nuclear magnetic resonance and circular dichroism spectroscopy . Individual assignments were obtained for heme c and the axial ligands . From studies of nuclear Overhauser enhancements the axial histidine imidazole ring orientation relative to the heme group was found to coincide with that of other c-type cytochromes . In contrast, a new structure was observed for the axial methionine . This includes S chirality at the iron-bound sulfur atom, but compared to cytochromes c-551 from Pseudomonads and Rhodopseudomonas gelatinosa, which also contain S-chiral methionine, the spatial arrangement of the gamma- and beta-methylene groups and the alpha carbon of methionine is markedly different . Analysis of the electron spin density distribution in ferricytochrome c5 in the light of this new coordination geometry provides additional support for the hypothesis that the electronic structure of heme c is primarily governed by the orientation of the sp3 lone-pair orbital of the axial sulfur atom with respect to the heme plane.

Mol Biol (Mosk), 1983 Sep-Oct, 17(5), 1108 - 11
{Dependence of the melting temperature of phage DNA on the GC pair content in a solvent with low ionic strength}; Kul'ba AM et al.; Base ratio of DNA from 21 bacteriophage of Pseudomonas was determined by chemical hydrolysis and paper chromatography . Obtained values of the GC pair content were compared with melting temperature of DNA in 0,1 X SSC . The content of GC pairs correlates with melting temperature by equation %GC = 2,53 (Tm - 53,4) . The content of GC pair for DNA from 30 bacteriophages of Pseudomonas was calculated . Some speculations concerning the distribution in DNA base ratio of bacteriophages of Pseudomonas are discussed.

J Lipid Res, 1983 Sep, 24(9), 1109 - 18
Biotransformation of chenodeoxycholic acid by Pseudomonas species NCIB 10590 under anaerobic conditions; Owen RW et al.; The metabolism of chenodeoxycholic acid by Pseudomonas sp . NCIB 10590 under strict anaerobic conditions was studied . A range of unsaturated acidic and neutral metabolites were isolated and identified . The major acidic product was chola-4,6-dien-3-one-24-oic acid whilst the major neutral product was androsta-4,6-dien-3,17-dione . The major acidic products were 7 alpha-hydroxy-5 beta-cholan-3-oxo-24-oic acid, 3-oxo-4,6-pregnadien-20-carboxylic acid, 7 alpha-hydroxy-3-oxo-1,4-pregnadien-20-carboxylic acid, and 7 alpha-hydroxy-3-oxo-4-pregnen-20-carboxylic acid . The minor neutral products were androsta-1,4,6-trien-3,17-dione, 17 beta-hydroxyandrosta-4,6-dien-3,17-dione, 17 beta-hydroxyandrosta-1,4,6-trien-3-one, 7 alpha-hydroxyandrosta-1,4-dien-3,17-dione, and 7 alpha-hydroxyandrost-4-en-3,17-dione . In contrast to aerobic catabolism of chenodeoxycholic acid by Pseudomonas sp . NCIB 10590 in which 1,4-dienone steroids predominate, the major products described in this study are 4,6-dienone steroids . This is because of the induction of a 7 alpha-dehydroxylase enzyme under anaerobic conditions.

J Steroid Biochem, 1983 Sep, 19(3), 1355 - 62
The biotransformation of hyodeoxycholic acid by Pseudomonas sp . NCIB 10590 under anaerobic conditions; Owen RW et al.; The bacterial degradation of hyodeoxycholic acid under anaerobic conditions was studied . The major acidic product has been identified as 6 alpha-hydroxy-3-oxochol-4-ene-24-oic acid whilst the major neutral product has been identified as 6 alpha-hydroxyandrosta-1,4-diene-3,17-dione . The minor acidic products were 3,6-dioxochola-1,4-diene-24-oic acid, 3-oxochol-5-ene-24-oic acid, 3-oxochol-4-ene-24-oic acid, 3-oxochola-1,4-diene-24-oic acid and 6 alpha-hydroxy-3-oxochola-1,4-diene-24-oic acid and the minor neutral products were androst-4-ene-3,17-dione, androst-4-ene-3,6,17-trione, androsta-1,4-diene-3,6,17-trione, androsta-1,4-diene-3,17-dione, 17 beta-hydroxyandrosta-1,4-diene-3-one and 6 alpha-hydroxyandrost-4-ene-3,17-dione . Evidence is presented which suggests that under aerobic conditions, one pathway of hyodeoxycholic acid metabolism exists whilst under anaerobic conditions an extra biotransformation pathway becomes operative involving the induction of a 6 alpha-dehydroxylase enzyme . A biochemical pathway of hyodeoxycholic acid metabolism by bacteria under anaerobic conditions is discussed incorporating a scheme involving such an enzyme.

Proc Natl Acad Sci U S A, 1983 Sep, 80(17), 5315 - 9
Defective acidification of endosomes in Chinese hamster ovary cell mutants "cross-resistant" to toxins and viruses; Merion M et al.; Like many physiological ligands, several viruses and toxins enter mammalian cells through receptor-mediated endocytosis . Once internalized, the nucleic acids of several viruses and the toxic subunit of diphtheria toxin gain access to the cytosol of the host cell through an acidic intracellular compartment . In this report, we present evidence that one class of mutants of Chinese hamster ovary (CHO)-K1 cells, which is "cross-resistant" to Pseudomonas exotoxin A, diphtheria toxin, and several animal viruses, has a defect in acidification of the endosome . Cells were allowed to internalize fluorescein isothiocyanate-conjugated dextran before subcellular fractionation . Fluorescence measurements on subcellular fractions permitted measurement of the internal pH of the isolated endosomes and lysosomes . Our results show that (i) endosomes and lysosomes from CHO-K1 cells maintain an acidic pH, (ii) acidification of both endosomes and lysosomes is mediated by a Mg2+/ATP-dependent process, (iii) GTP can satisfy the ATP requirement for acidification of lysosomes but not of endosomes, and (iv) at least one class of mutants that is cross-resistant to toxins and animal viruses has a defect in the ATP-dependent acidification of their endosomes . These studies provide biochemical and genetic evidence that the mechanisms of acidification of endosomes and lysosomes are distinct and that a defect in acidification of endosomes is one biochemical basis for cross-resistance to toxins and viruses.

Infect Immun, 1983 Sep, 41(3), 998 - 1009
Strains of CHO-K1 cells resistant to Pseudomonas exotoxin A and cross-resistant to diphtheria toxin and viruses; Moehring JM et al.; We have investigated two phenotypically distinct types of mutants of CHO-K1 cells that are resistant to Pseudomonas exotoxin A due to a defect in the delivery of active toxin to the target site in the cell, elongation factor 2 . Both types contain normal levels of toxin-sensitive elongation factor-2 . Hybridization studies have shown that these cells fall into two distinct complementation groups . One group, designated DPVr, is resistant to Pseudomonas toxin, diphtheria toxin, and four enveloped RNA viruses . This group is also hypersensitive to ricin . The resistance of this group is apparently related to a defect in a mechanism for the acidification of endocytic vesicles . The other group, designated PVr, is resistant to Pseudomonas toxin and to three enveloped RNA viruses . The resistance of this group appears to be related to a defect in a cellular mechanism required for the maturation of Sindbis virus that is likewise required for the entry of active Pseudomonas toxin.

Plasmid, 1983 Sep, 10(2), 164 - 74
Molecular relationships between pseudomonas INC P-9 degradative plasmids TOL, NAH, and SAL; Lehrbach PR et al.; We have examined the extent to which the degradative plasmids SAL, NAH, and TOL of the Inc P-9 incompatibility group share common DNA sequences . The homology we observe using 32P-labeled SAL and NAH DNA probes can be assigned to six regions of the TOL (pWWO) restriction endonuclease cleavage map . At least three of these regions are probably related to transfer and replication functions, whereas a fourth region is related to the common metacleavage pathway . Restriction endonuclease maps of the SAL and NAH plasmids are derived and the relationships between these plasmids discussed.

J Clin Microbiol, 1983 Sep, 18(3), 738 - 40
O and H serotyping of Pseudomonas cepacia; Heidt A et al.; Procedures for the preparation, absorption, and titration of Pseudomonas cepacia O and H rabbit antisera are described . Seven O antigens (O1 to O7) for the slide agglutination test and five H antigens (H1, H3, H5, H6, and H7) for the agglutination and H immobilization tests were determined . Nearly 300 strains of P . cepacia isolated from hospitalized patients (a majority from Strasbourg hospitals) were serotyped . The use of P . cepacia serotyping as an epidemiological tool, especially in outbreak situations, was emphasized . Difficulties in obtaining monospecific H antisera are discussed.

J Biol Chem, 1983 Aug 10, 258(15), 9419 - 25
The bacterial oxidation of vitamin B6 . 4-Pyridoxic acid dehydrogenase: a membrane-bound enzyme from Pseudomonas MA-1; Yagi T et al.; A highly specific inducible membrane-bound 4-pyridoxic acid dehydrogenase has been solubilized and purified to apparent homogeneity from Pseudomonas MA-1 grown with pyridoxine as a sole source of carbon and nitrogen . The undenatured enzyme migrates as a single band on gel electrophoresis; denatured preparations show two barely resolved bands (Mr = 63,000 and 61,000) . Undenatured preparations aggregate readily, as evidenced by Mr values of 148,000, 470,000, and greater than 670,000 obtained by density gradient centrifugation or by gel filtration under various conditions . The enzyme contains FAD but no Fe or acid-labile S; an average minimum molecular weight of 131,000 was calculated from the FAD content . In the presence of 2,6-dichloroindophenol, the enzyme dehydrogenates 4-pyridoxic acid to the corresponding aldehyde; this reaction is not inhibited by CN- . At the pH optimum of 8.0, a Vm of approximately 7.0 mumol min-1 mg-1 and a Km of 9 microM were obtained . 2,6-Dichloroindophenol, phenazine methosulfate, and menadione are effective electron acceptors; ubiquinones are less active, while NAD, FAD, and O2 are inactive . However, in membrane fractions, oxygen supports 4-pyridoxic acid oxidation via a CN--sensitive electron transport chain, indicating that the dehydrogenase probably is coupled to ATP generation in such preparations.

J Bacteriol, 1983 Aug, 155(2), 505 - 11
Naphthalene dioxygenase: purification and properties of a terminal oxygenase component; Ensley BD et al.; Naphthalene dioxygenase from Pseudomonas sp . strain NCIB 9816 is a multicomponent enzyme system that oxidized naphthalene to cis-(1R, 2S)-dihydroxy-1,2-dihydronaphthalene . The terminal oxygenase component B was purified to homogeneity by a three-step procedure that utilized ion-exchange and hydrophobic interaction chromatography . The purified enzyme oxidized naphthalene only in the presence of NADH, oxygen, and partially purified preparations of components A and C . An estimated Mr of 158,000 was obtained by gel filtration . Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed the presence of two subunits with molecular weights of ca . 55,000 and 20,000, indicative of an alpha 2 beta 2 quaternary structure . Absorption spectra of the oxidized enzyme showed maxima at 566 (shoulder), 462, and 344 nm, which were replaced by absorption maxima at 520 and 380 nm when the enzyme was reduced anaerobically by stoichiometric quantities of NADH in the presence of the other two components of the naphthalene dioxygenase system . Component B bound naphthalene . Enzyme-bound naphthalene was oxidized to product upon the addition of components A and C, NADH, and O2 . These results, together with the detection of the presence of 6.0 g-atoms of iron and 4.0 g-atoms of acid-labile sulfur per mol of the purified enzyme, suggest that component B of the naphthalene dioxygenase system is an iron-sulfur protein which functions in the terminal step of naphthalene oxidation.

Can J Microbiol, 1983 Aug, 29(8), 867 - 73
Isolation and identification of N2-fixing Pseudomonas associated with wetland rice; Barraquio WL et al.; Semisolid yeast extract medium amended with glucose and tryptic soy agar were used to isolate aerobically N2-fixing (C2H2-reducing) heterotrophic bacteria from the root of wetland rice . The isolates were identified as Pseudomonas by gel immunodiffusion and fluorescent antibody techniques in combination with their morphological, cultural, and biochemical characteristics . The N2-fixing H2-utilizing Pseudomonas described in this paper is a new species.

Zh Mikrobiol Epidemiol Immunobiol, 1983 Aug, (8), 47 - 50
{The nature of changes in the antigenic spectrum of the causative agent of melioidosis during animal passage}; Peters MK et al.; The comparative analysis of the antigenic spectra of Pseudomonas pseudomallei museum and subcultured strains, carried out by the method of immunoelectrophoresis, has revealed that, along with an essential increase in the virulence of P . pseudomallei for white mice and changes in the morphology of colonies, a decrease in the amount of detected precipitinogens occurs in the process of subculturing . The immunoelectrophoregrams of the subcultured variants show the absence of antigens 5, 6 and the simultaneous increase of the production of antigen 8, one of the components of mucoid (in the pseudocapsule).

J Steroid Biochem, 1983 Aug, 19(2), 1127 - 33
The effect of carbonyl cyanide m-chlorophenylhydrazone on steroid transport in membrane vesicles of Pseudomonas testosteroni; Culos D et al.; The uncoupler carbonyl cyanide chlorophenylhydrazone (CCCP) was an effective inhibitor of steroid transport in membrane vesicles of Pseudomonas testosteroni between 10 microM and 1 microM CCCP . At these concentrations the inhibition of steroid transport was not due to an inhibition of the 3 beta and 17 beta-hydroxysteroid dehydrogenase enzyme . CCCP also affected testosterone-dependent oxygen consumption at concentrations up to 100 microM and inhibited respiration at 0.5 and 1 microM . The effect of CCCP on testosterone-dependent oxygen consumption indicated that CCCP was acting as an uncoupler . The concurrent inhibition of testosterone transport and stimulation of testosterone-dependent oxygen consumption at 10-100 microM CCCP supported the conclusion that transport and metabolism were tightly coupled processes . When membrane vesicles were pre-incubated with CCCP for 15 min, CCCP did inhibit transport and the 3 beta and 17 beta-hydroxysteroid dehydrogenase activity . However, both transport and enzyme inhibition could be prevented by the addition of NAD+ to the incubation mixture . This indicated that CCCP exhibits the properties of a sulfhydryl reagent under pre-incubated conditions.

J Gen Microbiol, 1983 Aug, 129 (Pt 8), 2545 - 56
Specification of the conjugative pili and surface mating systems of Pseudomonas plasmids; Bradley DE; Conjugative pili were identified for representative Pseudomonas plasmids of incompatibility groups P-2, P-3, P-5, P-7, P-8, P-10, P-11, and P-13, pili for groups P-1 and P-9 having already been described in detail . FP5 pili (unclassified) were also found . In most cases pili could be characterized by electron microscopy as rigid or flexible . The majority of Pseudomonas plasmids transferred significantly better on a surface than in a liquid . Examples of all incompatibility groups were tested.

Mol Cell Biol, 1983 Jul, 3(7), 1283 - 94
Binding and uptake of diphtheria toxin by toxin-resistant Chinese hamster ovary and mouse cells; Didsbury JR et al.; We investigated two phenotypically distinct types of diphtheria toxin-resistant mutants of Chinese hamster cells and compared their resistance with that of naturally resistant mouse cells . All are resistant due to a defect in the process of internalization and delivery of toxin to its target in the cytosol, elongation factor 2 . By cell hybridization studies, analysis of cross-resistance, and determination of specific binding sites for 125I-labeled diphtheria toxin, we showed that these cell strains fall into two distinct complementation groups . The Dipr group encompasses Chinese hamster strains that are resistant only to diphtheria toxin, as well as mouse LM cells . These strains possess a normal complement of high-affinity binding sites for diphtheria toxin, but these receptors are unable to deliver active toxin fragment A to the cytosol . Cells of the DPVr group have a broader spectrum of resistance, including Pseudomonas exotoxin A and several enveloped viruses as well as diphtheria toxin . In these studies, which investigate the resistance of these cells to diphtheria toxin, we demonstrate that they possess a reduced number of specific binding sites for this toxin and behave, phenotypically, like cells treated with the proton ionophore monensin . Their resistance is related to a defect in a mechanism required for release of active toxin from the endocytic vesicle.

Eur J Biochem, 1983 Jul 1, 133(3), 673 - 84
Porphyran primary structure . An investigation using beta-agarase I from Pseudomonas atlantica and 13C-NMR spectroscopy; Morrice LM et al.; Porphyran, a highly substituted agarose from Porphyra umbilicalis was degraded by highly purified beta-agarase I from Pseudomonas atlantica . This enzyme cleaved at the reducing side of units of beta-neoagarobiose (3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-beta-D-galactopyranose) . The oligosaccharides were divided into fractions of low and high molecular weight by dialysis . The permeate (23% of total starting carbohydrate) was separated by ion-exchange into neutral and anionic fractions . Gel filtration of the neutral fraction (19%) resolved two major oligosaccharides . These were shown by 13C-NMR spectroscopy to be 6(3)-O-methyl-neoagarotetraose and 6(3),6(5)-di-O-methyl-neoagarohexaose . Gel filtration of the anionic oligosaccharides (3.3%) revealed two novel monosulphated tetrasaccharides, 6-O-sulphato-alpha-L-galacto-pyranosyl-(1 leads to 3)-beta-D-galactopyranosyl-(1 leads to 4)-3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactopyranose and its 6(3)-O-methylated derivative . The 13C-NMR data from the sulphated tetrasaccharides provided a novel reference which was used to characterise higher, partially sulphated fragments in the dialysis permeate . The fraction retained on dialysis (77%) had an average degree of polymerisation of 40 and was homologous with the high-molecular-weight anionic permeate . From 13C-NMR spectroscopy porphyran was found to comprise 49% sulphated disaccharide units and these were calculated to occur in stretches averaging 2.0-2.5 contiguous units.

Clin Orthop, 1983 Jul-Aug, (177), 172 - 5
Salvage of bilateral knee arthroplasties with Pseudomonas infection . A case report; Hershman E et al.; The treatment of infection after knee arthroplasty presents difficult medical and technical problems . Adequate control of the infection by appropriate antibiotic treatment and arthrodesis is no longer the only alternative to surgical management of the infected implant . Successful reimplantation can be achieved, although the quality of arthroplasty is often inferior to that of noninfected per primam arthroplasty.

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