Pseudomonas

J Bacteriol, 1992 Feb, 174(4), 1345 - 51
DNA sequence of IS91 and identification of the transposase gene; Mendiola MV et al.; IS91 is a 1,830-bp insertion sequence that inserts specifically at the sequence CAAG or GAAC of the target and does not duplicate any sequence upon insertion (23) . By transposon mutagenesis, we have identified open reading frame 426 (ORF426; bp 454 to 1731) as the putative ORF for the transposase . It displays a cysteine-rich, potential metal-binding domain in its N-terminal region . Adjacent to ORF426, there is an ORF (ORF121) which precedes and terminally overlaps ORF426 by one amino acid . Tn1732 insertions in ORF121 do not affect the transposition frequency . IS91 has sequence similarities to IS801 from Pseudomonas syringae . Their putative transposases are 36% identical, including conservation of the cysteine-rich cluster . The information concerning IS801 insertion specificity and target duplication has been reevaluated in the light of our results.

FEBS Lett, 1992 Jan 27, 296(3), 259 - 62
Reduction of carbon monoxide to formaldehyde by the terminal oxidase of the marine bacterium Pseudomonas nautica strain 617; Arnaud S et al.; When exposed to CO, the aerobic respiratory system of the marine bacterium Pseudomonas nautica strain 617, previously reduced with dithionite, undergoes reoxidation . When dealing with the purified oxidase (dithionite reduced) exposure of the enzyme to CO induces its reoxidation (collapse of its alpha band) . Under our experimental conditions, this form of the oxidase could not be reduced again by dithionite . Addition of formaldehyde to the native oxidized enzyme resulted in full inhibition of the oxidase reduction by dithionite, presumably due to complex formation . We hypothesized a reduction of CO into formaldehyde and a locking of the active site by the reaction product . By using flash photolysis, it was possible to turn over the enzyme, accumulate the reaction product and identify it as formaldehyde . When using the membrane-bound enzyme, formaldehyde accumulated without the help of flash photolysis . This unusual reduction of CO to formaldehyde could be related to the previously reported uncommon features of the P . nautica oxidase, in particular O2 reduction into H2O2 as end product {(1989) FEBS Lett . 247, 475-479}.

J Mol Biol, 1992 Jan 20, 223(2), 415 - 26
Genetic and functional analysis of the basic replicon of pPS10, a plasmid specific for Pseudomonas isolated from Pseudomonas syringae patovar savastanoi; Nieto C et al.; The sequence of a 1823 base-pair region containing the replication functions of pPS10, a narrow host-range plasmid isolated from a strain of Pseudomonas savastanoi, is reported . The origin of replication, oriV, or pPS10 is contained in a 535 base-pair fragment of this sequence that can replicate in the presence of trans-acting function(s) of the plasmid . oriV contains four iterons of 22 base-pairs that are preceded by G+C-rich and A+T-rich regions . A dnaA box located adjacent to the repeats of the origin is dispensable but required for efficient replication of pPS10; A and T are equivalent bases at the 5' end of the box . repA, the gene of a trans-acting replication protein of 26,700 Mr has been identified by genetic and functional analysis . repA is adjacent to the origin of replication and is preceded by the consensus sequences of a typical sigma 70 promoter of Escherichia coli . The RepA protein has been identified, using the minicell system of E . coli, as a polypeptide with an apparent molecular mass of 26,000 . A minimal pPS10 replicon has been defined to a continuous 1267 base-pair region of pPS10 that includes the oriV and repA sequences.

Biochem Biophys Res Commun, 1992 Jan 15, 182(1), 14 - 9
Molecular cloning and nucleotide sequence of a pectin lyase gene from Pseudomonas marginalis N6301; Nikaidou N et al.; A pectin lyase (PNL;EC4.2.2.10) gene of Pseudomonas marginalis N6301 was cloned and expressed in Escherichia coli . We purified PNL from P . marginalis N6301 and determined N-terminal 33 amino acids sequence . From this sequence, we synthesized two oligonucleotide probes . From the analysis of Southern hybridization, 2 . 1kb EcoRI-SmaI fragment from the chromosomal DNA of P . marginalis was found to hybridize with oligonucleotide probes . Then, we cloned the fragment into pUC119 vector and transformed into E . coli DH5 alpha . A plasmid thus obtained was designated as pPNL6301 . E . coli DH5 alpha harboring pPNL6301 expressed PNL activity . The nucleotide sequence of pn1 gene in the plasmid pPNL6301 encoding PNL from P . marginalis N6301 was determined . The structural gene of pn1 consisted of 936 base pairs . An open reading frame that encodes a 34,103 dalton polypeptide composed of 312 amino acids was assigned . The molecular weight of the polypeptide predicted from the amino acid composition was close to that of PNL of P . marginalis N6301 determined . The nucleotide sequence of the 5'-flanking region of pn1 gene showed the presence of the consensus sequence of LexA binding site, Pribnow box and ribosome binding site as found in Escherichia coli . The amino acid sequence homology of PNLs and nucleotide sequence homology of pn1 gene between P . marginalis N6301 and E . carotovora Er were 60.8% and 57.2%, respectively.

FEMS Microbiol Lett, 1992 Jan 15, 69(3), 283 - 7
The detection of lipase activity in bacteria using novel chromogenic substrates; Miles RJ et al.; The propionate (Pro), decanoate (Dec) and laurate (Lau) esters of 5-(4-hydroxy-3,5-dimethoxyphenylmethylene)-2-thioxothiazoline++ +-3-acetic acid were assessed as substrates for lipase and esterase . On hydrolysis these substrates yield an intensely red coloured phenol which could be assayed at 505 nm . The Pro ester was an effective substrate for porcine esterase and was hydrolysed at a rate 20 times greater than the Lau and Dec esters . Conversely, Pseudomonas lipase had a high activity towards the Lau and Dec esters, especially in the presence of bovine serum albumin, but little activity towards the Pro ester . The Dec and Lau were used to detect lipolytic activity in Pseudomonas strains associated with milk spoilage . For this purpose, the substrates were absorbed onto filter paper disks, which were placed over bacterial colonies growing on agar plates; activity was indicated by bright red colouration of discs within 2 h . Escherichia coli colonies hydrolysed the Pro but not the Lau or Dec esters.

J Acquir Immune Defic Syndr, 1992, 5(1), 70 - 7
Activity of CD4-Pseudomonas exotoxin against cells expressing diverse forms of the HIV and SIV envelope glycoproteins; Ashorn P et al.; CD4(178)-PE40 is a genetically engineered hybrid toxin containing a portion of human CD4 linked to the translocation and ADP-ribosylation domains of Pseudomonas exotoxin A . In vitro, the molecule has been shown to selectively kill cells expressing the envelope glycoproteins of human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV), and to inhibit HIV spread . In this report we examine the activity of the hybrid toxin against cells expressing diverse forms of the HIV and SIV envelope glycoproteins, encoded by recombinant vaccinia virus vectors . The activity of CD4(178)-PE40 was found to be unaffected by mutations in the HIV-1 or HIV-2 envelope glycoprotein genes, which prevent normal proteolytic processing of the corresponding gp160 precursor molecules . Cells expressing a mutant HIV-1 envelope glycoprotein lacking most of the cytoplasmic tail of the gp41 transmembrane subunit were also sensitive to the hybrid toxin . Most interestingly, HIV-1, HIV-2, and SIVmac envelope glycoprotein molecules known to have widely differing affinities for CD4 were found to be comparably effective at mediating sensitivity to CD4(178)-PE40 . By virtue of its ability to kill infected cells, the hybrid toxin inhibited the spread of SIVmac in vitro . These results indicate that CD4(178)-PE40 is active against cells expressing HIV and SIV envelope glycoproteins with a diverse array of structural differences.

J Virol, 1992 Jan, 66(1), 190 - 6
Construction of a transducing virus from double-stranded RNA bacteriophage phi6: establishment of carrier states in host cells; Onodera S et al.; Bacteriophage phi 6 contains three double-stranded RNA (dsRNA) genomic segments . We have constructed a plasmid that contains a cDNA copy of the middle (M) segment, with a gene for kanamycin resistance (kan) inserted into the PstI site . A transcript of this cDNA was incorporated in vitro into procapsids along with natural transcripts of the S and L segments . The procapsids were coated with nucleocapsid surface protein P8 and transfected into Pseudomonas syringae pv . phaseolicola . The resulting infectious virus, phi 6 K1, was found to contain an M segment that was 1.2 kbp larger than the normal 4.1 kbp . K1 formed small, turbid plaques, and its genome was unstable . Preparations of K1 contained from about 0.1 to 10% large, clear-plaque forms of the virus which were usually missing the kan gene, and in some cases, the resulting segment M was smaller than its normal size . Cells picked from lawns of host cells infected with K1 yielded colonies that were resistant to kanamycin (Kan) . These colonies could be passaged on kanamycin-containing medium . The cells were found to contain large amounts of dsRNA corresponding to the viral genomic segments . Some strains continued to produce viable phage, while others lost this ability . One strain completely lost the small genomic segment S . Approximately 1 in 10,000 infected cells acquired the carrier state with the original phage isolate K1 . However, we isolated a viral mutant that was able to induce the carrier state in 10 to 20% of the infected cells . The ability to use drug resistance as a test for the carrier state makes this system very useful for the study of the mechanisms of induction of persistent infections.

Cancer Res, 1992 Jan 1, 52(1), 181 - 6
Characterization of the antigen (CAK1) recognized by monoclonal antibody K1 present on ovarian cancers and normal mesothelium; Chang K et al.; K1 is a monoclonal antibody that reacts with a cell surface antigen (CAK1) found in human mesothelia and nonmucinous ovarian tumors . In this article, the characteristics of the CAK1 antigen have been examined in detail . Using immunofluorescence microscopy, we have found that the CAK1 signal is removed from the cell surface by treatment with proteases or by phosphatidylinositol-phospholipase C, but not by neuraminidase and beta-galactosidase . The phosphatidylinositol-phospholipase C-released material was found to contain the CAK1 antigen which was detected by a competition radioimmunoassay . The phosphatidylinositol-phospholipase C-released CAK1 antigen was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting and found to be approximately 40 kDa protein . The CAK1-K1 antibody complex remains on the cell surface and is poorly internalized, as shown by an acid wash immunofluorescence internalization assay . An immunotoxin composed of K1 and Lys-PE40, a mutant form of Pseudomonas exotoxin lacking the cell binding domain, was not cytotoxic, supporting the conclusion that the CAK1-K1 antibody complex is not internalized . However, an immunotoxin composed of K1 and native Pseudomonas exotoxin was selectively cytotoxic to cells expressing the CAK1 antigen . This cytotoxicity is due to the fact that domain I of Pseudomonas exotoxin promotes internalization of antigens which are not internalized or bound to antibody alone . Our results suggest that CAK1 is a polypeptide that is expressed on mesothelial cells and many ovarian cancers, and that K1 may be useful as a targeting agent for the immunotherapy of human ovarian cancer.

Patol Fiziol Eksp Ter, 1992 Jan-Feb, (1), 37 - 9
{Immunotropic activity of human alpha1-glucoprotein}; Liutov AG et al.; Experiments were conducted to study the effect of alpha-acid glycoprotein on a fatal infection caused by Pseudomonas pyocyanea, growth of melanoma B-16, and transplantation of a skin graft from C57BL/6 mice to CBA mice . Injection of the agent significantly increased the anti-infection resistance in mice, suppressed growth of melanoma-16, and prolonged the survival of the skin grafts, which was evidence of marked glycoprotein immunomodulating activity.

Biotherapy, 1992, 4(4), 289 - 97
Treatment with tumour infiltrating lymphocytes and interleukin-2 in patients with metastatic melanoma: a pilot study; Baars JW et al.; Tumour infiltrating lymphocytes (TIL) were isolated and expanded from biopsy samples of 4 patients with metastatic melanoma . The patients were treated with autologous expanded TIL and continuous or bolus infusion of Interleukin 2 (IL-2) at a dose of 18 x 10(6) International Units/m2/day for 5 days starting 36-48 hours after administration of cyclophosphamide at a dose of 1 g/m2 . The number of TIL infused ranged from 10(10) to 5.56 x 10(10) cells . Two patients had stable disease (SD) lasting for 2 1/2 and 4 months respectively and they died 24 and 13 months after therapy . One patient died during therapy due to a pseudomonas septicaemia and another patient developed progressive disease (PD) . He died 3 months after the start of therapy . The side effects were substantial but most of them were reversible upon cessation of the treatment . The majority of the expanded TIL of all patients were of the CD8+ phenotype . Cutaneous metastases from two patients, removed after treatment with IL-2 and TIL, showed moderate lymphocytic infiltration also mainly of CD8+ T cells . The treatment with IL-2 and TIL is feasible, but further investigations should continue in an attempt to improve the efficacy of the therapy, to reduce toxicity and to diminish the costs and labour of the culture methods.

Bioconjug Chem, 1992 Jan-Feb, 3(1), 63 - 8
Properties of chimeric toxins with two recognition domains: interleukin 6 and transforming growth factor alpha at different locations in Pseudomonas exotoxin; Kreitman RJ et al.; Pseudomonas exotoxin (PE) is a potent cytotoxic agent that is composed of 613 amino acids arranged into three major domains . We have previously identified two positions where ligands can successfully be placed in PE to direct it to cells with specific surface receptors . One site is at the amino terminus and the other is close to but not at the C-terminus . To examine the possibility of constructing oncotoxins with two different recognition elements that will bind to two different receptors, we have placed cDNAs encoding either transforming growth factor alpha (TGF alpha) or interleukin 6 (IL6) at the 5' end of a PE gene and also inserted a cDNA encoding TGF alpha near the 3' end of the PE gene . The plasmids encoding these chimeric toxins were expressed in Escherichia coli and the chimeric proteins purified to near homogeneity . In all the new toxins, the TGF alpha near the C-terminus was inserted after amino acid 607 of PE and followed by amino acids 604-613 so that the correct PE C-terminus (REDLK) was preserved . For each chimera, the toxin portion was either PE4E, in which the cell binding domain (domain Ia) is mutated, PE40, in which domain Ia is deleted, or PE38, in which domain Ia and part of domain Ib are deleted . These derivatives of PE do not bind to the PE receptor and allow 607, 355, or 339 amino acids, respectively, between the two ligands.(ABSTRACT TRUNCATED AT 250 WORDS)

Bioconjug Chem, 1992 Jan-Feb, 3(1), 58 - 62
Rational design of a chimeric toxin: an intramolecular location for the insertion of transforming growth factor alpha within Pseudomonas exotoxin as a targeting ligand; Kreitman RJ et al.; To investigate the potential utility of Pseudomonas exotoxin (PE) in forming rationally designed chemotherapeutic agents, we inserted a cDNA encoding transforming growth factor alpha (TGF alpha) at several locations in a gene encoding a mutant full-length PE (PE4E) which does not bind to the PE receptor . After expression in Escherichia coli, we purified the chimeric toxins to near homogeneity and showed that they were specifically cytotoxic to human epidermoid, ovarian, colon, and hepatocellular carcinoma lines . Like the previously reported TGF alpha-PE40, one of the new molecules (TGF alpha-PE4E) contains the ligand at the amino terminus . Two additional chimeras (PE4E-TGF alpha and PE4E-TGF alpha-598-613) each contain TGF alpha inserted near the carboxyl terminus of PE . We show that preservation of the correct PE carboxyl-terminal amino acid sequence, REDLK, allows the toxins containing TGF alpha carboxyl inserts to retain significant cytotoxicity against target cells, since another molecule (PE4E-TGF alpha-ILK) containing a nonfunctional carboxyl-terminal sequence was over 100-fold less active . The chimeric toxins with TGF alpha had the same binding affinity for the EGF receptor whether the ligand occupied the amino or carboxyl position . Molecules with TGF alpha near the carboxyl position were consistently less active against target cells but also less toxic to mice than those with TGF alpha at the amino terminus, indicating both types of molecules might be therapeutically effective . Our results establish that a ligand can be placed near the carboxyl terminus of PE, within the portion of the toxin that translocates to the cytosol . The amino-terminal position in such molecules is then available for the placement of other targeting ligands.

Z Naturforsch {C}, 1992 Jan-Feb, 47(1-2), 26 - 32
{Biogenesis of Pseudomonas siderophores: the proof of analogous structures of a pyoverdin-desferriferribactin pair (1)}; Budzikiewicz H et al.; When grown in an iron-deficient medium Pseudomonas aptata produces both a desferri-ferribactin and a pyoverdin . The identical sequence of the peptide chain confirms the hypothesis that desferri-ferribactins are the biogenetic precursors of pyoverdins.

J Biochem (Tokyo), 1992 Jan, 111(1), 8 - 15
Primary structures of the genes, faoA and faoB, from Pseudomonas fragi B-0771 which encode the two subunits of the HDT multienzyme complex involved in fatty acid beta-oxidation; Sato S et al.; Three enzyme activities involved in fatty acid beta-oxidation, i.e., those of enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and 3-oxoacyl-CoA thiolase, are exhibited by one multienzyme complex (HDT) composed of two molecules each of two peptides in Pseudomonas fragi . Using specific antisera against the two subunits of HDT, we isolated the genes encoding the subunits of HDT and designated them "faoA" (for the alpha-subunit) and "faoB" (for the beta-subunit) . Their complete nucleotide sequences were determined and it was revealed that faoA and faoB, both with individual putative S.D . sequences at suitable positions, formed a cluster, in that order . The amino acid sequences deduced from the nucleotide sequences of the two genes indicated that the alpha-subunit, encoded by faoA, is a polypeptide of 715 amino acid residues, and that the beta-subunit, encoded by faoB, consists of 390 amino acid residues lacking the first methionine of the primary product encoded by faoB . Immunoblotting of cell lysates prepared from Escherichia coli transformants carrying plasmids which possess the faoA and/or faoB gene with antisera against the subunits of HDT showed that both the faoA and faoB genes were transcribed and translated in E . coli . The overall activities of 2-enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase were increased in the E . coli cells transformed with the plasmid possessing the faoA gene, suggesting that both the hydratase and dehydrogenase activities may be exhibited by the alpha-subunit of HDT.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biochem (Tokyo), 1992 Jan, 111(1), 16 - 9
Induction of enzymes involved in fatty acid beta-oxidation in Pseudomonas fragi B-0771 cells grown in media supplemented with fatty acid; Sato S et al.; Induction of the enzymes involved in fatty acid beta-oxidation in Pseudomonas fragi B-0771 cells grown in a medium containing straight chain saturated fatty acids was studied . The acyl-CoA dehydrogenase (ACDH) activity was induced during the exponential phase in cells grown in palmitic acid-supplemented medium, reached a maximum at the early stationary phase, and then gradually decreased thereafter . Changes in the overall activities of 2-enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase, both existing on the multienzyme complex (HDT) involved in fatty acid beta-oxidation, were similar to that in ACDH activity . Straight chain saturated fatty acids having more than 6 carbon atoms could induce both the ACDH and HDT activities, and C13-C15 fatty acids caused the greatest induction of both activities . Changes in the overall activities of 2-enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase correlated with that in the amount of the alpha-subunit of HDT during the entire culture period in the medium containing palmitic acid . Surprisingly, the stoichiometry of the alpha- and beta-subunit proteins of HDT was not maintained into the stationary phase culture, though the genes encoding the alpha- and beta-subunits are tandemly coded in bacterial genomic DNA.

Biofactors, 1992 Jan, 3(3), 173 - 84
Protein toxin inhibitors of protein synthesis; Perentesis JP et al.; Two classes of extremely toxic proteins kill eukaryotic cells by covalently modifying unique structural features of components that are essential for protein synthesis . Intoxication by these proteins results from the entry of a catalytic fragment into the cytoplasm . One class is typified by diphtheria toxin and Pseudomonas exotoxin A . The catalytic component of these toxins ADP-ribosylates and inactivates elongation factor 2 which is an essential participant in protein synthesis . This modification occurs at a unique post-translational histidine derivative, diphthamide, that is present in the ribosomal binding site of the elongation factor . The two toxins differ in their molecular organization but appear to possess identical reaction mechanisms and very similar active sites . The other class contains two types of toxins typified, respectively, by alpha-sarcin, a member of a family of fungal toxins, and ricin, a member of a group of closely related plant proteins collectively termed ribosome-inactivating proteins . The catalytic components of the two types of toxins in this second class inactivate the large ribosomal subunit through two different hydrolytic alterations of 23-28S RNA . alpha-Sarcin and its congeners act as a specific endonuclease whereas ricin and its congeners act as a specific N-glycosidase . These hydrolytic cleavages occur at a pair of adjacent nucleotides within a highly conserved sequence near the 3' terminus of 23-28S RNA . The covalent integrity of this region of RNA is essential to elongation factor-dependent ribosomal functions and is located within the ribosomal binding domain of these factors . Both of these classes of toxins are being employed as 'magic bullets' to eliminate pathological cells . By combining the catalytic component of these toxins with various cell targeting components, useful and specific anticancer and immunomodulatory agents have been created.

Carbohydr Res, 1992 Jan, 223, 255 - 61
Properties of the enzyme expressed by the Pseudomonas saccharophila maltotetraohydrolase gene (mta) in Escherichia coli; Zhou JH et al.; The maltotetraohydrolase gene (mta) from Pseudomonas saccharophila was expressed in Escherichia coli JM109 . Maltotetraohydrolase was produced mostly (approximately 90%) in the periplasmic space . The amino-terminal amino acid sequence and molecular weight of the recombinant enzyme were identical with those of the native enzyme, and there was no significant difference in the substrate specificity and modes of action . This system for maltotetraohydrolase expression is useful for studies of the structure and function of the enzyme.

Endocr Res, 1992, 18(1), 51 - 8
A bacterial binding site which binds human chorionic gonadotropin but not human luteinizing hormone; Carrell DT et al.; Exposed sites on Pseudomonas maltophilia (ATC #1637), which bind both human chorionic gonadotropin (hCG) and a native hCG-like ligand with equal high affinity, are described . This high-affinity binding site (Kd = 1.3 x 10(-10) binds hCG, but does not bind human luteinizing hormone (hLH), nor related human glycoprotein hormones, thyrotropin, and follicle stimulating hormone . A lower affinity, 2.3 x 10(-9), is also described which binds hLH and hCG equally well . This is the first description of a high-affinity binding site in nature, which distinguishes hCG from hLH.

Clin Infect Dis, 1992 Jan, 14(1), 359 - 60
Peritonitis caused by Pseudomonas putrefaciens in patients undergoing continuous ambulatory peritoneal dialysis; Dan M et al.; Three cases of peritonitis caused by Pseudomonas putrefaciens in patients undergoing continuous ambulatory peritoneal dialysis are described . In two cases asymptomatic colonization of the dialysate preceded overt infection . All patients responded successfully to standard antibiotic therapy with gentamicin or ofloxacin . This is the first report of peritonitis caused by P . putrefaciens.

Intern Med, 1992 Jan, 31(1), 50 - 4
An autopsy case of Crow-Fukase syndrome which developed 18 years after the first manifestation of plasmacytoma; Sakemi H et al.; A 57-yr-old woman developed Crow-Fukase syndrome 18 yr after resection of plasmacytoma of the rib . Irradiation applied to the relapsed plasmacytoma and systemic chemotherapy alleviated symptoms and signs, but the tumor relapsed in the unirradiated cervical lymph node and she died of Pseudomonas pneumonia during chemotherapy 3 yr after diagnosis . Biopsy of the lymph node revealed proliferation of IgG-lambda-positive atypical plasma cells while autopsy revealed plasmacytoma remnant in the pleura of the affected side 21 yr before . No amyloid was found on autopsy . Crow-Fukase syndrome can develop long after the origination of plasmacytoma.

Rev Mal Respir, 1992, 9(2), 145 - 53
{Antibiotics in aerosols}; Pascal S et al.; The treatment of bronchopulmonary infections using antibiotics administered by aerosol should enable a better local concentration of the drug to be achieved whilst reducing the side effects . The parameters of the aerosol kinetics in the airways lead to an interaction of the physico-chemical characteristics of the molecule, the material used for the aerosolisation and also the conditions on inhalation . Intrabronchial concentrations are lower than after endotracheal instillation but remain 10 to 40 times greater than after parenteral administration . Antibiotic aerosols are relatively well tolerated but should be used with care and caution in patients susceptible to bronchial hyperreactivity . Aerosol treatment is currently little used outside those patients with mucoviscidosis and dilatation of the bronchi . In mucoviscidosis aerosol antibiotics are used in the acute situation to achieve cures of infection and in the chronic situation to prevent colonisation by Pseudomonas . Antibiotic aerosols are efficacious in the treatment of acute infectious episodes but do not seem to carry any additional clinical benefit which is superior to antibiotics administered parenterally . It could nonetheless constitute a useful alternative in simplifying treatment in the home . This style of treatment remains to be further explored in a more complete fashion in bronchial dilatation.

Eur J Clin Pharmacol, 1992, 42(4), 395 - 9
FARMAGUIDA: a databank for the analysis of the Italian drug market and drug utilization in general practice; Montanaro N et al.; FARMAGUIDA, a databank of drugs marketed in Italy (2,596 pharmaceutical substances corresponding to 10,448 products), permits analysis of the nature and value of the drugs prescribed . It contains coded pharmaceutical and administrative information, an original classification, as well as indicators of the therapeutic status of each drug . The FARMAGUIDA classification was built hierarchically according to a three-level pattern: the first level (42 categories) corresponds to major pharmacological groups; the second level (157 groups) gathers drugs having similar clinical indications and/or pharmacological actions; and the third level (246 subgroups) classifies drugs according to chemical structure and/or the mechanism of action . Drugs not falling into well-established pharmacotherapeutic criteria (e.g . neurotropics or liver protectants) are classified into separate subgroups . Two larger groupings were also formulated: THER (11 headings), a utilization-oriented arrangement in which each heading also contained the corresponding placebo-like drugs, and PHARM (14 headings), a rational pharmacological arrangement, in which all placebo-like drugs were relegated into a separate set . The following quality indicators were created: DOC, which defines five degrees of documentation of clinical efficacy according to major textbooks of pharmacology and therapeutics; CLASS, which groups DOC values for a more simple evaluation of prescription data; PREP, which distinguishes monocomponent preparations from fixed-dose combinations, and also provides coded information about the rationale for the combination; HOSP, which hallmarks drugs that should be reserved for in-patients, e.g . anti-pseudomonal antibiotics . The composition of the list of reimbursable drugs, the Italian National Formulary (NF; 5782 products in 1990) was analyzed according to the FARMAGUIDA classification and indicators.(ABSTRACT TRUNCATED AT 250 WORDS)

J Basic Microbiol, 1992, 32(3), 209 - 14
Biosynthesis of pyrrolnitrin . Incorporation of 13C, 15N double-labelled D- and L-tryptophan; Zhou P et al.; Experiments on the incorporation of D- and L-{alanine-3-13C,2-15N}tryptophan into the antibiotic pyrrolnitrin in Pseudomonas aureofaciens confirmed earlier conclusions about the conversion of L-tryptophan into pyrrolnitrin . They also demonstrated that a fraction of the D isomer is incorporated without breakage of the 15N-carbon bond, consistent with the operation of a second pathway from D-tryptophan to pyrrolnitrin . Cell-free experiments confirmed the conversion of 3-(o-aminophenyl)pyrrole into aminopyrrolnitrin but failed to detect enzymatic oxidation of the latter to pyrrolnitrin.

Arch Microbiol, 1992, 158(6), 412 - 7
Maleylacetate reductase of Pseudomonas sp . strain B13: dechlorination of chloromaleylacetates, metabolites in the degradation of chloroaromatic compounds; Kaschabek SR et al.; The maleylacetate reductase of 3-chlorobenzoate-grown cells of Pseudomonas sp . strain B13 has been purified 50-fold . The enzyme converted 2-chloromaleylacetate to 3-oxoadipate with temporary occurrence of maleylacetate; 1 mol of chloride was eliminated during the conversion of 1 mol of 2-chloro- and 2,3-dichloromaleylacetate; 2 mol of NADH were consumed per mol of 2-chloro- and 2,3-dichloromaleylacetate while only 1 mol was necessary to catalyze the conversion of maleylacetate or 2-methylmaleylacetate . The maleylacetate reductase failed to use fumarylacetate as a substrate . The role of the enzyme in the chloroaromatics degradation is discussed.

Microbiol Immunol, 1992, 36(8), 899 - 904
Prevalence of antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens in military personnel in Sabah and Sarawak, Malaysia; Embi N et al.; Sera from 420 military personnel serving in Sabah and Sarawk, Malaysia, were tested for antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens by enzyme-linked immunosorbent assay procedure (ELISA) . Data showed that 54.4% of serum samples were positive for antibodies to P . pseudomallei exotoxin and 65.7% were positive for antibodies to the whole cell antigens . Samples gave much lower titers for anti-exotoxin antibodies compared to titers against crude whole cell antigens . The incidence of antibody to exotoxin was highest in the age groups ranging from 26 to 32 years, where the positive rates were higher than 40% and 30% for military personnel serving in Sarawak and Sabah, respectively.

Yi Chuan Xue Bao, 1992, 19(4), 355 - 61
{Studies of plasmids of Pseudomonas maltophilia}; Shen P et al.; Eighteen strains of P . maltophilia were screened for the occurrence of plasmid using four different methods . Five of them were harbor plasmids . The results of plasmid detection in different growth phase of P2 strain showed that the highest amount of plasmids in the strain was observed in stationary growth phase . The characteristics of plasmid of P . maltophilia P2 was investigated by methods of agarose gel electrophoresis, restriction endonucleases analysis, determination of molecular weight . The results indicated that P . maltophilia P2 contained only one type of plasmid, its molecular weight was 4.4 x 10(6) dalton and that the plasmid had single BamHI . PstI . XbaI EcoRI . HindIII sites . Thus, the plasmid of P . maltophilia P2 may be developed into a fine cloning vector.

Chin J Biotechnol, 1992, 8(1), 15 - 22
Restriction mapping and localization of GL-7-ACA acylase gene; Yang Y et al.; This paper presents the results about the restriction mapping of recombinant plasmids pMR5 and pMR6 containing GL-7-ACA acylase gene from Pseudomonas sp . 130, gene localization and its expression under the control of different promoters, tet, tac or lac/tac, in Escherichia coli . The analysis of gel electrophoresis of pMR5 cleaved with several kinds of restriction enzymes indicated that there is no sites of EcoRI, HindIII and ClaI but the presence of following sites: one HpaI, two XhoI, three EamHI and four PstI on the cloned gene fragment . The restriction maps of pMR5 and pMR6 were determined by comparative digestion of various endonucleases . The gene of GL-7-ACA acylase was localized on a 3.0kb fragment of B2-B3-HpaI from the studies on a serial subcloning . Expression of subclones pMR9, pMR10 and pMR11 in E . coli was compared . Higher yield of acylase was obtained when the gene fragment was placed downstream of the tac promoter . The expression of Pseudomonas gene in E . coli was also discussed.

J Clin Lab Anal, 1992, 6(6), 405 - 9
Streptomyces: a superior source for cholesterol oxidase used in serum cholesterol assay; Lolekha PH et al.; The present study compared three cholesterol oxidase sources (Nocardia, Streptomyces, and Pseudomonas sps.) for serum cholesterol assay . We found cholesterol oxidase isolated from Streptomyces was superior than those isolated from Nocardia and Pseudomonas sps . Performances of the reagent contained Streptomyces cholesterol oxidase was excellent and comparable to the performances obtained from reagent contained Nocardia cholesterol oxidase . Moreover, the reagent containing Streptomyces cholesterol oxidase had the lowest cost and had the longest shelf-life ($U.S . 0.17/mL, 10 weeks) compared to the reagent contained Nocardia ($U.S . 0.50/mL, 8 weeks) or Pseudomonas ($U.S . 0.20/mL, 6 weeks) cholesterol oxidase.

J Ocul Pharmacol, 1992 Spring, 8(1), 83 - 90
The effects of transferrin receptor antibody, transferrin receptor antibody bound to Pseudomonas exotoxin and transforming growth factor-alpha bound to Pseudomonas exotoxin on human tenon's capsule fibroblast proliferation; Smyth RJ et al.; Pharmacological agents which modulate the wound healing process by the inhibition of proliferation of fibroblasts may improve the success of glaucoma filtration surgery . Since cell proliferation is essential to the wound healing process, we targeted the surface receptors that are associated with proliferating cells . We present the effects of three such agents-purified mouse anti-human transferrin receptor monoclonal antibody 42/6 (anti-TfR-42/6), anti-transferrin monoclonal antibody bound to a Pseudomonas exotoxin (anti-TfR-PE40) and transforming growth factor-alpha Pseudomonas exotoxin (TGF-alpha-PE40)--on human fibroblasts from Tenon's capsule . The inhibition of human subconjunctival fibroblast proliferation by anti-TfR-42/6 (with a concentration up to 25 micrograms/ml) and by anti-TfR-PE40 and TGF-alpha-PE40 (both with a concentration range of 5000-0.00001 micrograms/ml) was determined by colorimetric (OD), and cell counting (CC) assays over a 9-day period . Neither anti-TfR-42/6 nor anti-TfR-PE40 had an antiproliferative effect on the fibroblasts . TGF-alpha-PE40 demonstrated an antiproliferative effect in a dose response manner . The mean 50% inhibitory dose (ID50) by OD was 32.91 micrograms/ml, while the ID50 by CC was 27.88 micrograms/ml . EGF was used as a negative control for TGF-alpha-PE40 toxin . The inhibitory effect of the toxin conjugate was completely blocked by the addition of 1000 micrograms/ml of EGF . These in vitro studies show that TGF-alpha-PE40 may be useful in modulating the proliferation of human ocular fibroblasts; they also give some indication of drug dosages for future in vivo testing.

DNA Seq, 1992, 2(4), 269 - 71
Nucleotide sequence of a 2 kb plasmid from Pseudomonas cepacia implicated in the degradation of phenylcarbamate herbicides; Gaubier P et al.; The complete nucleotide sequence of a very small plasmid whose presence and level in Pseudomonas cepacia have been linked to herbicide resistance is presented . The structural features of the plasmid are discussed.

J Appl Bacteriol, 1992 Jan, 72(1), 71 - 9
Efficacy of copper and silver ions with iodine in the inactivation of Pseudomonas cepacia; Pyle BH et al.; Alternatives to chlorination of water have been sought for reasons which include trihalomethane formation, possible bacterial regrowth, the high concentrations of chlorine required in certain circumstances, and the taste, odour and bodily irritation in chlorine-treated water . Electrolytically generated Cu and Ag ions at low levels, in addition to very low chlorine concentrations, have been suggested as an alternative to routine chlorination . We have examined the combination of Cu and Ag ions with low levels of iodine . Pseudomonas cepacia was grown either in rich medium or under nutrient restriction prior to disinfection . Survival of the organism and its ability to regrow after treatment as well as the effects of varying buffers, metal ion and iodine concentrations were determined . Low concentrations of metal ions (100 ppb Cu and 11 ppb Ag) and iodine (200 ppb) were more effective than either metal ions or iodine alone against Ps . cepacia grown on rich agar or in low nutrient buffer . After iodination, buffer-grown suspensions recovered to their original cell concentrations within 7 d . When Cu and Ag ions were used with or without iodine, regrowth was prevented . The results show that low concentrations of Cu and Ag in combination with iodine permit effective disinfection of bacteria after cultivation on either rich media or under nutrient restriction . These results, along with published data, suggest that the combination of these metals with halogenation may have applications in the disinfection of both recreational and potable water.

Int J Syst Bacteriol, 1992 Jan, 42(1), 107 - 19
Transfer of several phytopathogenic Pseudomonas species to Acidovorax as Acidovorax avenae subsp . avenae subsp . nov., comb . nov., Acidovorax avenae subsp . citrulli, Acidovorax avenae subsp . cattleyae, and Acidovorax konjaci; Willems A et al.; DNA-rRNA hybridizations, DNA-DNA hybridizations, polyacrylamide gel electrophoresis of whole-cell proteins, and a numerical analysis of carbon assimilation tests were carried out to determine the relationships among the phylogenetically misnamed phytopathogenic taxa Pseudomonas avenae, Pseudomonas rubrilineans, "Pseudomonas setariae," Pseudomonas cattleyae, Pseudomonas pseudoalcaligenes subsp . citrulli, and Pseudomonas pseudoalcaligenes subsp . konjaci . These organisms are all members of the family Comamonadaceae, within which they constitute a separate rRNA branch . Only P . pseudoalcaligenes subsp . konjaci is situated on the lower part of this rRNA branch; all of the other taxa cluster very closely around the type strain of P . avenae . When they are compared phenotypically, all of the members of this rRNA branch can be differentiated from each other, and they are, as a group, most closely related to the genus Acidovorax . DNA-DNA hybridization experiments showed that these organisms constitute two genotypic groups . We propose that the generically misnamed phytopathogenic Pseudomonas species should be transferred to the genus Acidovorax as Acidovorax avenae and Acidovorax konjaci . Within Acidovorax avenae we distinguished the following three subspecies: Acidovorax avenae subsp . avenae, Acidovorax avenae subsp . cattleyae, and Acidovorax avenae subsp . citrulli . Emended descriptions of the new taxa are presented.

J Bacteriol, 1992 Jan, 174(1), 279 - 90
Purification and some properties of 2-halobenzoate 1,2-dioxygenase, a two-component enzyme system from Pseudomonas cepacia 2CBS; Fetzner S et al.; The two components of the inducible 2-halobenzoate 1,2-dioxygenase from Pseudomonas cepacia 2CBS were purified to homogeneity . Yellow component B is a monomer (Mr, 37,500) with NADH-acceptor reductase activity . Ferricyanide, 2,6-dichlorophenol indophenol, and cytochrome c acted as electron acceptors . Component B was identified as an iron-sulfur flavoprotein containing 0.8 mol of flavin adenine dinucleotide, 1.7 mol of iron, and 1.7 mol of acid-labile sulfide per mol of enzyme . The isoelectric point was estimated to be pH 4.2 . Component B was reduced by the addition of NADH . Red-brown component A (Mr, 200,000 to 220,000) is an iron-sulfur protein containing 5.8 mol of iron and 6.0 mol of acid-labile sulfide . The isoelectric point was within the range of pH 4.5 to 5.3 . Component A could be reduced by dithionite or by NADH plus catalytic amounts of component B . Component A consisted of nonidentical subunits alpha (Mr, 52,000) and beta (Mr, 20,000) . It contained approximately equimolar amounts of alpha and beta, and cross-linking studies suggested an alpha 3 beta 3 subunit structure of component A . The NADH- and Fe(2+)-dependent enzyme system was named 2-halobenzoate 1,2-dioxygenase, because it catalyzes the conversion of 2-fluoro-, 2-bromo-, 2-chloro-, and 2-iodobenzoate to catechol . 2-Halobenzoate 1,2-dioxygenase exhibited a very broad substrate specificity, but benzoate analogs with electron-withdrawing substituents at the ortho position were transformed preferentially.

Biosci Biotechnol Biochem, 1992 Jan, 56(1), 76 - 80
Cloning and nucleotide sequence of the maltopentaose-forming amylase gene from Pseudomonas sp . KO-8940; Shida O et al.; The gene coding for the maltopentaose-(G5)-forming amylase of Pseudomonas sp . KO-8940 was cloned into Escherichia coli and its nucleotides were sequenced . It was expected that a long open reading frame composed of 1,842-bp that encoded 614 amino acid residues for secretory precursor polypeptide including the typical signal sequence with an NH2-terminal was the gene . An extract of Escherichia coli carrying the cloned G5-forming amylase gene had amylolytic activity with which produced only G5 from starch, the same as that of the donor strain enzyme . In the deduced primary structure of this enzyme, the four conserved regions of many alpha-amylases were found, and the COOH-terminal portion of this enzyme showed high homology with other raw starch digesting amylases.

Adv Perit Dial, 1992, 8, 269 - 75
The impact of peritonitis on CAPD results; Viglino G et al.; The impact of peritonitis on CAPD results was evaluated in 1990 pts (mean age +/- SD:58.4 +/- 14.8 yrs, 55.9% males), treated in 30 centres participating in Italian PD Study Group, during 1980-89, with an overall observation period of 3953 years (mean +/- SD 24.1 +/- 22.3 months) . The incidence of peritonitis decreases from 1.21 (1980-84) to 0.48 (1985-89) ep/year (overall:0.68) with a significant (P < 0.001) reduction of the probability of developing the first peritonitis episode (FPE) through the same periods . The probability of developing FPE and the relative risk of peritonitis were significantly lower (P < 0.001) in pts for whom CAPD has been the first treatment (80.1%); on the contrary these parameters did not gain significant difference according to sex, age 65 years, diabetes or cardiovascular disease . As far as the organisms responsible for peritonitis are concerned a significant reduction of S . epid . and an increase of S . aureus, other Gram pos . and Pseudomonas was observed in the second 5-yr periods . Peritonitis episodes caused catheter removal in 8.2% of cases and were associated with catheter infection in 10.8% of cases . Peritonitis accounted for 24.2% of hospitalization causes and for 6.7% and 30.0% of death and of drop-out respectively . The probability of death and drop-out was significantly high (p < 0.001) in pts with a peritonitis incidence > 1 ep/year than in those with < 0.5 ep/year . The probability of drop-out due to peritonitis was not higher in diabetic or older patients.

Biometals, 1992 Summer, 5(2), 73 - 80
Plasmid mediated metal and antibiotic resistance in marine Pseudomonas; Rajini Rani DB et al.; Pseudomonas sp isolated from the Bay of Bengal (Madras coast) contained a single large plasmid (pMR1) of 146 kb . Plasmid curing was not successful with mitomycin C, sodium dodecyl sulfate, acridine orange, nalidixic acid or heat . Transfer of mercury resistance from marine Pseudomonas to Escherichia coli occurred during mixed culture incubation in liquid broth at 10(-4) to 10(-5) ml(-1) . However, transconjugants lacked the plasmid pMR1 and lost their ability to resist mercury . Transformation of pMR1 into E . coli competent cells was successful; however, the efficiency of transformation (1.49 x 10(2)Hgr transformants microsgm-1 pMR1 DNA) was low . E . coli transformants containing the plasmid pMR1 conferred inducible resistance to mercury, arsenic and cadmium compounds similar to the parental strain, but with increased expression . The mercury resistant transformants exhibited mercury volatilization activity . A correlation existed between metal and antibiotic resistance in the plasmid pMR1.

Acta Cient Venez, 1992, 43(6), 349 - 54
Characterization of polycyclic aromatic hydrocarbons degradative soil Pseudomonas; Fuenmayor SL et al.; Nine Pseudomonas strains, able to degrade polycycle aromatic hydrocarbons (PAHs), were isolated from enriched cultures with naphthalene, as carbon source, and soil samples from a land farming process applied on oil sludge, as inocula . Degradative tests showed that all the strains were capable to catabolize naphthalene (Nah) and phenanthrene (Phn) . U2 strain transferred the selected function (Nah) to P . aeruginosa T1 (Hgr Oct+), however some of the transconjugants lost the Oct character, suggesting that it is of plasmidic nature . T1 derivatives as well the wild strains U28 and U31 transferred Nah function to P . putida AC165 . All of the examined transconjugants also catabolized phenanthrene, suggesting that Nah and Phn functions in U2, U28, and U31 strains are linked and probably encoded by transferable plasmids.

Microbios, 1992, 70(283), 139 - 44
A unique type of GABA binding by Mycobacterium leprae; Prabhakaran K et al.; Neurotropism is one of the unusual properties of Mycobacterium leprae . The organism contains glutamic acid decarboxylase that generates gamma-amino-butyric acid (GABA) which is an inhibitory neurotransmitter . The binding of GABA by M . leprae in vitro was studied by using 3H-GABA as substrate . The bacteria had high-affinity binding sites for the amino acid . The uptake was a specific saturable process with a Km of 66.7 pM, pH optimum of 7.3 and a temperature optimum of 37 degrees C . The binding did not seem to be time-dependent, being complete in about 5 min . None of the known antagonists and agonists of GABA uptake by neurons, showed any significant effect on M . leprae; the receptors in the bacteria are apparently of a non-neuronal type, and different from those reported in spermatozoa and Pseudomonas.

Nephrol Dial Transplant, 1992, 7(4), 333 - 9
Endotoxin transfer through dialysis membranes: small- versus large-pore membranes; Vanholder R et al.; In this in-vivo study, dialysate and serum endotoxin was evaluated before and after haemodialysis with small-pore (PS400) and large-pore (PS600) polysulphone dialysers, and before and after haemodiafiltration with the PS600 filter . The source of the endotoxin was the presence in dialysate of Pseudomonads at a concentration of 10(3)-10(4) CFU/ml . Endotoxin was measured by a modified chromogenic limulus amoebocyte lysate (LAL) assay . In spite of dialysate endotoxin concentrations greater than 100 pg/ml, no changes in pre- versus posttreatment LAL reactivity were observed in PS400 dialysis and PS600 haemodiafiltration . In contrast, PS600 haemodialysis was related to an increase in serum LAL reactivity from 1.3 +/- 1.5 to 3.8 +/- 2.0 pg/ml (n = 15, P less than 0.01), and five patients (33.3%) showed a post-dialysis value in excess of 5 pg/ml . Our data are consistent with the absence of in-vivo endotoxin transfer during haemodialysis with small-pore dialyser membranes, and during haemodiafiltration with membranes with larger pores . An increase in LAL reactivity during haemodialysis with membranes with larger pores is, however, present, presumably due to the occurrence of backdiffusion/filtration with that specific strategy.

Acta Microbiol Hung, 1992, 39(2), 181 - 91
Production and regulation of a thermostable protease by Pseudomonas sp . B45; Chakraborty R et al.; A Pseudomonas sp . produced an extracellular thermostable protease which required induction by peptone . Growth of the organism and the production of protease was optimum at 30 degrees C . The enzyme was subjected to catabolite repression by glucose . Both chloramphenicol and rifamycin completely abolished protease production indicating de novo synthesis of the enzyme . Leucine, lysine, histidine and glycine enhanced the protease production considerably and they were the most effective when added during the active period of production . Glucose repression could not be relieved by addition of leucine.

Microbiol Immunol, 1992, 36(12), 1239 - 49
Identification of Oklahoma isolate as a strain of Pseudomonas pseudomallei; Yabuuchi E et al.; Based on the morphological, physiological, biochemical and nutritional characteristics, cellular fatty acid and lipid composition, ubiquinone-8 as the major respiratory quinone, guanine-plus-cytosine content of DNA, DNA-DNA homology value, and sequence alignment of 16S rRNA nucleotides, Oklahoma isolate was reidentified as a strain of Pseudomonas pseudomallei.

Zh Mikrobiol Epidemiol Immunobiol, 1992, (9-10), 10 - 3
{A unique marker of Pseudomonas from the first group of rRNA homology--selective sensitivity to the bacteriostatic action of barium ions}; Sivolodskii EP; On the basis of the study of the bacteriostatic action of chlorides and nitrates of barium and 24 other metals on 18 Pseudomonas species of all groups of rRNA homology and on 49 bacterial species belonging to 25 other genera, unique selective sensitivity to the bacteriostatic action of Ba2+ ions has been established in Pseudomonas of the first group of rRNA homology . The marker of barium sensitivity is in line with changes in the classification of Pseudomonas and is of interest for their further more precise systematization . The test for barium sensitivity of bacteria helps simplify the identification of Pseudomonas of the first group of rRNA homology and makes the identification more accurate.

Bioseparation, 1992, 2(6), 375 - 83
Production and purification of salicylate monooxygenase from Pseudomonas cepacia ATCC 29351; Ramsay JR et al.; Salicylate monooxygenase (EC: 1.14.13.1) has been produced and purified from Pseudomonas cepacia ATCC 29351 which has the ability to utilise salicylate as a sole carbon source . The bacterium was grown on a defined medium containing 2% (w/v) casamino acids and 0.15% (w/v) yeast extract at 25 degrees C; salicylate monooxygenase production was induced by the presence of up to 0.7% (w/v) sodium salicylate, to a level of approximately 2% of the soluble cell protein . The enzyme was purified over 50-fold, with a recovery of about 40%, by a combination of ion exchange and hydrophobic interaction chromatography . The purified enzyme had a specific activity of 14-15 U mg-1 protein and was essentially homogeneous.

Eur J Biochem, 1991 Dec 18, 202(3), 1217 - 22
Characterization of the epoxide hydrolase from an epichlorohydrin-degrading Pseudomonas sp; Jacobs MH et al.; An epoxide hydrolase was purified to homogeneity from the epichlorohydrin-utilizing bacterium Pseudomonas sp . strain AD1 . The enzyme was found to be a monomeric protein with a molecular mass of 35 kDa . With epichlorohydrin as the substrate, the enzyme followed Michaelis-Menten kinetics with a Km value of 0.3 mM and a Vmax of 34 mumol.min-1.mg protein-1 . The epoxide hydrolase catalyzed the hydrolysis of several epoxides, including epichlorohydrin, epibromohydrin, epoxyoctane and styrene epoxide . With all chiral compounds tested, both stereoisomers were converted . Amino acid sequencing of cyanogen bromide-generated peptides did not yield sequences with similarities to other known proteins.

Biochem Pharmacol, 1991 Dec 11, 42 Suppl, S93 - 8
Carbonyl reduction of metyrapone in human liver; Maser E et al.; Carbonyl reduction was investigated in cytosolic and microsomal fractions of human liver using the ketone metyrapone as a substrate . The cytosolic enzyme has a stronger preference for NADPH over NADH than the microsomal enzyme: the former shows only 14% of the NADPH-supported activity while the latter exhibits 36% activity with NADH . Barbitone and quercitrin, the classic inhibitors of carbonyl reductases, do not affect metyrapone reduction in either fraction . Dicumarol and indomethacin, the specific inhibitors of NAD(P)H: quinone-oxidoreductase and dihydrodiol dehydrogenase, respectively, only slightly decreased metyrapol formation . In contrast, 5 alpha-dihydrotestosterone, the active form of the androgen steroid testosterone, inhibited metyrapone reduction very strongly in the microsomal fractions and is postulated to be the physiological substrate of the enzyme . This resembles the situation in mouse liver {E . Maser and K . J . Netter, Biochem Pharmacol 38: 3049-3054, 1989} where microsomal metyrapone reductase was inhibited by steroids and the purified enzyme was demonstrated to mediate androsterone oxidation . Immunoblot analysis revealed antigenic cross-reaction of antibodies against the 34 kDa metyrapone reductase from mouse liver microsomes with the homologous protein in human liver microsomes pointing to structural homologies between the respective enzymes of the two species . These results--together with previous findings, which have shown that there exist functional as well as structural relationships between microsomal mouse liver metyrapone reductase and 3 alpha-hydroxysteroid dehydrogenase from Pseudomonas testosteroni {E . Maser, U . Oppermann and K . J . Netter, Eur J Pharmacol 183:1366, 1990}--suggest that metyrapone reduction in human liver microsomes might be catalysed by a microsomal hydroxysteroid dehydrogenase.

Biochemistry, 1991 Dec 10, 30(49), 11579 - 84
Q-band ENDOR spectra of the Rieske protein from Rhodobactor capsulatus ubiquinol-cytochrome c oxidoreductase show two histidines coordinated to the {2Fe-2S} cluster; Gurbiel RJ et al.; Electron nuclear double resonance (ENDOR) experiments were performed on 14N (natural abundance) and 15N-enriched iron-sulfur Rieske protein in the ubiquinol-cytochrome c2 oxidoreductase from Rhodobactor capsulatus . The experiments proved that two distinct nitrogenous ligands, histidines, are undoubtedly ligated to the Rieske {2Fe-2S} center . The calculations of hyperfine tensors give values similar but not identical to those of the Rieske-type cluster in phthalate dioxygenase of Pseudomonas cepacia and suggest a slightly different geometry of the iron-sulfur cluster in the two proteins.

FEBS Lett, 1991 Dec 2, 294(1-2), 11 - 5
A model of the copper centres of nitrous oxide reductase (Pseudomonas stutzeri) . Evidence from optical, EPR and MCD spectroscopy; Farrar JA et al.; Nitrous oxide reductase (N2OR), Pseudomonas stutzeri, catalyses the 2 electron reduction of nitrous oxide to di-nitrogen . The enzyme has 2 identical subunits (Mr approximately 70,000) of known amino acid sequence and contains approximately 4 Cu ions per subunit . By measurement of the optical absorption, electron paramagnetic resonance (EPR) and low-temperature magnetic circular dichroism (MCD) spectra of the oxidised state, a semi-reduced form and the fully reduced state of the enzyme it is shown that the enzyme contains 2 distinct copper centres of which one is assigned to an electron-transfer function, centre A, and the other to a catalytic site, centre Z . The latter is a binuclear copper centre with at least 1 cysteine ligand and cycles between oxidation levels Cu(II)/Cu(II) and Cu(II)/Cu(I) in the absence of substrate or inhibitors . The state Cu(II)/Cu(I) is enzymatically inactive . The MCD spectra provide evidence for a second form of centre Z, which may be enzymatically active, in the oxidised state of the enzyme . Centre A is structurally similar to that of CuA in bovine and bacterial cytochrome c oxidase and also contains copper ligated by cysteine . This centre may also be a binuclear copper complex.

J Clin Oncol, 1991 Dec, 9(12), 2095 - 103
Clinical evaluation of intraperitoneal Pseudomonas exotoxin immunoconjugate OVB3-PE in patients with ovarian cancer; Pai LH et al.; OVB3-PE is an immunotoxin composed of a murine monoclonal antibody reactive with human ovarian cancer and conjugated to Pseudomonas exotoxin (PE) . Twenty-three patients with refractory ovarian cancer were treated intraperitoneally (IP) with escalating doses of OVB3-PE to study toxicity, pharmacokinetics, antiimmunotoxin antibody formation, and antitumor response . Dose-limiting CNS toxicity occurred after repeated doses at 5 and 10 micrograms/kg . Other non-dose-limiting toxicities included transient elevation of liver enzymes, fever, and gastrointestinal toxicity . Pharmacokinetics of IP and serum OVB3-PE were determined in 16 patients . Peak peritoneal fluid levels exceeded the in vitro median effective dose at all doses tested . At doses of 1 to 2 micrograms/kg, the immunotoxin concentration in the peritoneal fluid remained constant for up to 8 hours and dropped to negligible levels after 12 hours . At the 5 and 10 micrograms/kg doses, levels remained high for up to 24 hours (greater than 100 ng/mL) and then gradually decreased and became undetectable (less than 4 ng/mL) after 72 hours . Serum levels of OVB3-PE were also analyzed in 16 patients . At doses of 1 micrograms/kg and 2 micrograms/kg, serum levels were not detectable (less than 5 ng/mL) . However, after doses of 5 or 10 micrograms/kg, peak serum level occurred at 24 hours after each dose and dropped to negligible levels by 72 hours . Sera from 12 patients were analyzed for anti-PE antibodies and antibodies to mouse immunoglobulin (HAMA) . All patients developed antibodies against PE within 14 days of therapy . Domain II of PE appeared to be the most immunogenic portion of the PE molecule . HAMA was detected on day 14 of therapy in nine patients, on day 21 in two, and on day 28 in one patient . No clinical antitumor responses were observed . We conclude that IP OVB3-PE at dose levels of 5 micrograms/kg (x 3) and 10 micrograms/kg (x 2) is accompanied by dose-limiting toxic encephalopathy . Neurologic toxicity is likely to be due to crossreactivity of OVB3 to normal human brain tissue, which was not appreciated during preclinical screening.

Ir Med J, 1991 Dec-1992 Jan, 84(4), 121 - 4
Cystic fibrosis in adolescents and adults; Mulherin D et al.; A cystic fibrosis (CF) clinic for adults was established in 1977 . We have reviewed the data on 164 patients who attended between 1977 and 1989 . Twenty four patients had died, 11 being over 20 years of age at the time of death . Of the 140 patients still alive, 61% were male and 53% were aged over 20 years . Only 55% were diagnosed by one year and 88% by ten years . Almost all patients had respiratory symptoms and sputum culture yielded pseudomonas species in 69% . Other respiratory problems included major haemoptysis and pneumothorax, each in 10% . We found a wide range of respiratory impairment among older patients . Among 33 patients aged over 23 years, the mean (+/-S.D.) percent predicted FEV1 and FVC were 53.3% (+/- 18%) and 71.4 (+/- 20%) respectively . Mean weight in this group was 92.5% (+/- 14) of predicted . Malabsorption occurred in most patients and meconium ileus equivalent occurred in 34% . Other complications were clinical hepatomegaly (16%), diabetes mellitus (9%) and arthropathy (20%) . Most patients were taking continuous antibiotics by mouth (89%) and by nebuliser (48%), beta-2 agonists by inhaler (57%) and oral steroids (29%) . Almost all were taking multivitamins, pancreatic replacement therapy and multiple nutritional supplements . The number of CF "bed days" grew 12 fold since 1979 and the mean stay in hospital was double the hospital mean . The economic impact was such that over 1/4 of the annual hospital antibiotic budget was expended on CF patients.

J Biochem (Tokyo), 1991 Dec, 110(6), 976 - 81
Chemical modification of Pseudomonas ochraceae 4-hydroxy-4-methyl-2-oxoglutarate aldolase by diethyl pyrocarbonate; Maruyama K; Diethyl pyrocarbonate inactivates Pseudomonas ochraceae 4-hydroxy-4-methyl-2-oxoglutarate aldolase {4-hydroxy-4-methyl-2-oxoglutarate pyruvate-lyase: EC 4.1.3.17} by a simple bimolecular reaction . The inactivation is not reversed by hydroxylamine . The pH curve of inactivation indicates the involvement of a residue with a pK of 8.8 . Several lines of evidence show that the inactivation is due to the modification of epsilon-amino groups of lysyl residues . Although histidyl residue is also modified, this is not directly correlated to the inactivation . No cysteinyl, tyrosyl, or tryptophyl residue or alpha-amino group is significantly modified . The modification of three lysyl residues per enzyme subunit results in the complete loss of aldolase activity toward various 4-hydroxy-2-oxo acid substrates, whereas oxaloacetate beta-decarboxylase activity associated with the enzyme is not inhibited by this modification . Statistical analysis suggests that only one of the three lysyl residues is essential for activity . l-4-Carboxy-4-hydroxy-2-oxoadipate, a physiological substrate for the enzyme, strongly protects the enzyme against inactivation . Pi as an activator of the enzyme shows no specific protection . The molecular weight of the enzyme, Km for substrate or Mg2+, and activation constant for Pi are virtually unaltered after modification . These results suggest that the modification occurs at or near the active site and that the essential lysyl residue is involved in interaction with the hydroxyl group but not with the oxal group of the substrate.

Appl Environ Microbiol, 1991 Dec, 57(12), 3679 - 82
Purification and characterization of the N-methylcarbamate hydrolase from Pseudomonas strain CRL-OK; Mulbry WW et al.; A unique cytosolic enzyme that hydrolyzes the carbamate linkage of the insecticide carbaryl (1-naphthyl N-methylcarbamate) was purified from extracts of Pseudomonas sp . strain CRL-OK . Substrates of the hydrolase include the N-methylcarbamate pesticides carbofuran and aldicarb but not the phenylcarbamate isopropyl m-chlorocarbanilate, the thiocarbamate S-ethyl N,N-dipropylthiocarbamate, or the dimethylcarbamate o-nitrophenyldimethylcarbamate.

Appl Environ Microbiol, 1991 Dec, 57(12), 3652 - 5
Limited bacterial mineralization of fungal degradation intermediates from synthetic lignin; Ruttimann C et al.; The ability of selected bacterial strains and consortia to mineralize degradation intermediates produced by Phanerochaete chrysosporium from 14C-labeled synthetic lignins was studied . Three different molecular weight fractions of the intermediates were subjected to the action of the bacteria, which had been grown on a lignin-related dimeric compound . Two consortia isolated from wood being decayed naturally by a Ganoderma species of white rot fungus (the palo podrido system) mineralized 10 to 11% of the fraction with a molecular weight of approximately 500 but less than 4% of the higher- and lower-molecular-weight fractions . The consortia mineralized 5 to 9% of the original lignins . The ability of two pseudomonads isolated earlier from lignin-rich environments to mineralize the original lignins or fungus degradation products was much lower.

Biotechnol Appl Biochem, 1991 Dec, 14(3), 357 - 64
Gellan gum biosynthetic enzymes in producing and nonproducing variants of Pseudomonas elodea; Martins LO et al.; A pathway for the synthesis of the repeating tetrasaccharide units in gellan gum from Pseudomonas elodea is proposed . The enzymes presumed to be involved in the synthesis of the activated precursors UDP-glucose, TDP-rhamnose, and UDP-glucuronic acid were detected and assayed in crude cell extracts of the gellan-producing (Gel+) P . elodea ATCC 31461 . The levels of UDP-glucose pyrophosphorylase and TDP-glucose pyrophosphorylase were higher in cells grown in media leading to higher gellan yields . Moreover, these enzymes exhibited lower values in cells of a Gel- variant, spontaneously obtained from the Gel+ wild type . The activation or repression of their synthesis is thought to be involved in the expression of the mucoid phenotype . Nevertheless, based on results here reported, the involvement of other enzymes, that catalyze steps downstream from the formation of the precursors cannot be excluded.

Exp Mol Pathol, 1991 Dec, 55(3), 203 - 16
Incorporation of circulating fibronectin into various tissues during sepsis: colocalization with endogenous tissue fibronectin; Jin HM et al.; We studied the plasma clearance and tissue incorporation of intravenously infused purified human plasma fibronectin into various tissues during a period of acute lung vascular injury induced by lethal postoperative bacteremia in sheep . Lung, liver, spleen, and heart tissue were examined for both endogenous sheep tissue fibronectin as well as the experimentally infused human fibronectin using dual-label immunofluorescence . Awake sheep (n = 4) received a postoperative iv infusion of 5 x 10(9) live Pseudomonas over a 60-min infusion interval . Bacterial challenge was started 2 hr after starting the iv fibronectin infusion of purified human plasma fibronectin (100 mg iv bolus; 4 hr iv at 100 mg/hr) . Human fibronectin displayed a biphasic rate of clearance from the plasma with entrance into lymph . Human fibronectin readily incorporated in all tissues studied, including the lung which was the focus of vascular injury . Analysis of tissue sections by dual-label immunofluorescence indicated that the exogenous human fibronectin colocalized with the endogenous sheep fibronectin . Thus, the plasma fibronectin concentration may influence the lung vascular barrier due to its incorporation into the tissue pool of fibronectin . Moreover, the plasma may serve as a reservoir for soluble fibronectin which can enter and colocalize with the insoluble tissue pool of fibronectin in various tissues.

J Bacteriol, 1991 Dec, 173(24), 7950 - 5
Cloning and sequencing of the gene for a Pseudomonas paucimobilis enzyme that cleaves beta-aryl ether; Masai E et al.; We isolated Pseudomonas paucimobilis SYK-6, which was able to degrade various dimeric lignin compounds (Y . Katayama, S . Nishikawa, M . Nakamura, K . Yano, M . Yamasaki, N . Morohoshi, and T . Haraguchi, Mokuzai Gakkaishi 33:77-79, 1987) . This metabolic process is a distinct characteristic of this bacterium, which is equipped with an enzymatic modification system for various dimeric lignin compounds involved in the tricarboxylic acid cycle . Cleavage of the beta-aryl ether linkage is essential in this process, because this linkage is the most abundant (approximately 50%) in lignin . Here, we report the isolation and characterization of the beta-etherase gene, which contains an open reading frame of 843 bp and which we call ligE . This gene was expressed in Escherichia coli, and the enzyme had the same kinetic properties as the P . paucimobilis SYK-6 enzyme.

J Bacteriol, 1991 Dec, 173(24), 7841 - 7
A gene cluster required for coordinated biosynthesis of lipopolysaccharide and extracellular polysaccharide also affects virulence of Pseudomonas solanacearum; Kao CC et al.; Bacterial cell surface components can be important determinants of virulence . At least three gene clusters important for extracellular polysaccharide (EPS) biosynthesis have been previously identified in the plant pathogen Pseudomonas solanacearum . We have found that one of these gene clusters, named ops, is also required for lipopolysaccharide (LPS) biosynthesis . Mutations in any complementation unit of this cluster decreased EPS production, prevented the binding of an LPS-specific phage, and altered the mobility of purified LPS in sodium dodecyl sulfate-polyacrylamide gel electrophoresis . However, restoration of LPS biosynthesis alone was not sufficient to restore virulence to the wild-type level, suggesting that EPS is important for pathogenesis.

Epidemiol Infect, 1991 Dec, 107(3), 577 - 84
Sudden unexplained death syndrome--a new manifestation in melioidosis?
Yap EH, Chan YC, Goh KT, Chao TC, Heng BH, Thong TW, Tan HC, Thong KT, Jacob E, Singh M.
The indirect haemagglutination (IHA) test using sensitized turkey erythrocytes and the indirect immunofluorescence assay (IgM-IFA) was confirmed to be sensitive in the detection of a recent or current Pseudomonas pseudomallei infection in 19 culture-confirmed Singapore melioidosis patients . All were found to have antibody titres from 4 to 32768 in the IHA test and 10 to 320 in the IgM-IFA test . When these tests were employed on sera from 16 immigrant Thai construction workers who died of sudden unexplained death syndrome (SUDS) and 73 healthy Thai fellow workers, 93.8% and 68.8% of SUDS cases had IHA titre of greater than or equal to 4 and IgM-IFA titre of greater than or equal to 10 respectively, in contrast to 39.7% and 12.3% found among healthy Thai workers . These data indicate that at the time of death, most of the SUDS patients had an active infection with P . pseudomallei, possibly resulting from reactivation of a latent infection . The aetiological role of P . pseudomallei as the major cause of SUDS is discussed.

Biochemistry, 1991 Nov 12, 30(45), 10858 - 65
Studies of the catalytic mechanism of an active-site mutant (Y14F) of delta 5-3-ketosteroid isomerase by kinetic deuterium isotope effects; Xue LA et al.; delta 5-3-Ketosteroid isomerase (EC 5.3.3.1) from Pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereoselective transfer of the 4 beta-proton to the 6 beta-position . The rate-limiting step has been shown to be the concerted enolization of the enzyme-bound substrate comprising protonation of the 3-carbonyl oxygen by Tyr-14 and abstraction of the 4 beta-proton by Asp-38 {Xue, L., Talalay, P., & Mildvan, A . S . (1990) Biochemistry 29, 7491-7500} . Primary, secondary, solvent, and combined kinetic deuterium isotope effects have been used to investigate the mechanism of the Y14F mutant, which lacks the proton donor and is 10(4.7)-fold less active catalytically than the wild-type enzyme . With {4 beta-D}androst-5-ene-3,17-dione as a substrate in H2O, a lag in product formation is observed which approaches, by a first-order process, the rate observed with protonated substrate . With the protonated substrate in D2O, a burst in product formation is detected by derivative analysis of the kinetic data which approaches the rate observed with the 4 beta-deuterated substrate in D2O . The absence of such lags or bursts with the protonated substrate in H2O or with the 4 beta-deuterated substrate in D2O, as well as the detection of buffer catalysis by phosphate at pH 6.8, indicates that one or more intermediates dissociate from the enzyme and partition to substrate 31.6 times faster than to product.(ABSTRACT TRUNCATED AT 250 WORDS)

Prikl Biokhim Mikrobiol, 1991 Nov-Dec, 27(6), 845 - 9
{Fibrinolytic activity of bacteria from Pseudomonas genus}; Imshenetskii AA et al.; A strain of the genera Pseudomonas genera was found to possess hemolytic, fibrinolytic and thrombolytic activities . The fibrinolytic activity of the lyophilized unpurified preparation was 900 conventional units/mg . After incubation in the blood plasma, the activity completely remained . The preparation (1 microgram/ml, 750 micrograms of protein) obtained by precipitation with ammonium sulfate (80% saturation) completely lysed in vitro human blood thrombi for 50 min . The strain studied can find practical applications in medical industry.

Mikrobiologiia, 1991 Nov-Dec, 60(6), 67 - 71
{Oxidation of dibenzofuran by Pseudomonas strains harboring plasmids of naphthalene degradation}; Selifonov SA et al.; Pseudomonas strains harboring plasmids pBS3, pBS4, NAH7 were shown to carry out initial transformation of dibenzofurane to 4-{2'-(3'-hydroxy)-benzofuranyl}-2-keto-3-butenic acid due to broad substrate specificity of the enzymes of naphthalene catabolism nahA, nahB, nahC and nahD . These strains did not grow on dibenzofurane because of the inability of the enzyme nahE to split pyruvate of 4-{2'-(3' hydroxy)-benzofuranyl}-2-keto-3-butenic acid, which leads to accumulation of the latter . The strains harboring plasmids pBS2 and NPL-1 are not capable of any transformation of dibenzofurane.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Nov, (11), 39 - 41
{The immunological response in volunteer donors immunized with a Pseudomonas vaccine}; Makarenko TA et al.; The trial of experimental vaccine consisting of protective protein antigens of P . aeruginosa cell wall was carried out on 114 volunteers . The vaccine proved to be faintly reactogenic and induced the formation of specific humoral immunity in 98% of the volunteers who retained a high level of anti-P . aeruginosa antibodies in their blood for up to 5 months (the term of observation after the course of immunization was over.

Res Microbiol, 1991 Nov-Dec, 142(9), 995 - 1003
Phenotypic heterogeneity of Pseudomonas syringae van Hall; Gardan L et al.; The study of phenotypic properties of 108 strains of Pseudomonas syringae pv . syringae van Hall isolated from Cherry laurel (50 strains) and various host plants (58 strains) and 53 strains of other pathovars of P . syringae and fluorescent Pseudomonas showed that the majority of the strains (91/108) were clustered in one phenon (phenon 14) containing strains commonly considered as P.s . pv . syringae . The present type strain of P.s . pv . syringae was distantly related to phenon 14 . Other pathovars of P . syringae constituted 13 discrete phenons.

J Med Assoc Thai, 1991 Nov, 74(11), 526 - 30
Lymphomatoid granulomatosis with upper airway obstruction: a case report; Prapphal N et al.; A case of lymphomatoid granulomatosis in a previously healthy 13-year-old Thai girl presenting with right sixth cranial nerve palsy and severe upper airway obstruction was reported . Cranial nerve palsy later disappeared spontaneously but the patient developed multiple pulmonary nodules and cavity leading to pulmonary insufficiency . Her course was complicated with septicemia which limited the use of corticosteroid and cytotoxic drugs . She finally expired with pseudomonas sepsis in addition to pulmonary and liver involvement . This is the first case of lymphomatoid granulomatosis in a child ever reported in Thailand . Lymphomatoid granulomatosis should be included in the differential diagnosis of upper airway obstruction with pulmonary nodules and cavity and multi-organ involvement in children.

J Am Podiatr Med Assoc, 1991 Nov, 81(11), 608 - 12
Hallux hammer toe secondary to pseudomonas osteomyelitis; Lavery LA et al.; The authors present two cases of resultant hallux hammer toe secondary to the definitive treatment of hallux sesamoidal osteomyelitis . Pseudomonas osteomyelitis developed in both cases following puncture wounds to the first metatarsophalangeal joint complex . The authors also review the literature on pseudomonas osteomyelitis secondary to puncture wounds and the development of hallux hammer toe after removal of the involved sesamoid bones.

Mol Microbiol, 1991 Nov, 5(11), 2763 - 76
New approaches in genome analysis by pulsed-field gel electrophoresis: application to the analysis of Pseudomonas species; Grothues D et al.; A general method for the evaluation of macrorestriction fragment patterns is presented and its applicability to the taxonomy of bacteria is demonstrated for 32 Pseudomonas species . Strains were differentiated at the species and subspecies level by genome size and macrorestriction fragment fingerprints of the chromosome that had been separated on pulsed-field gels . The relatedness of bacteria was ascertained from the similarity of AsnI, DraI, SpeI, SspI or XbaI fragment patterns . In general, the dendrograms calculated from the genome fingerprints corresponded with the phylogenetic classification obtained from phenotypic marker or nucleic acid hybridization analysis, but several exceptions were noted . The techniques and algorithms presented herein are generally applicable to the genome analysis of bacteria, lower eukaryotes, and DNA fragments cloned in yeast artificial chromosomes.

Pneumologie, 1991 Nov, 45(11), 910 - 2
{A 13-year-old boy with a mild form of cystic fibrosis and heterozygote gene mutation for Delta F508}; Hiort O et al.; We report a 13-year old boy with a chronic pseudomonas bronchitis who was first diagnosed as having cystic fibrosis at this age because of an elevated sweat chloride employing pilocarpine-iontophoresis . He is heterozygote for the gene mutation Delta F508 . We point out the often moderate course of illness in compound heterozygotes.

Nippon Seikeigeka Gakkai Zasshi, 1991 Nov, 65(11), 1120 - 30
{Roentgenological and pathological studies on the development of discitis in canine models}; Ohno R; Discitis was experimentally induced in 42 dogs by intradiscal injections of bacterial suspensions and sequentially studied by X-rays and histopathology up to 24 weeks . 1 . A narrowing of the serpentine intervertebral disc space was seen roentgenologically in the Pseudomonas and E . coli groups . 2 . Histologically, the acute inflammation began to subside in eight weeks, at which time new bone formation started to appear, and fusion of the adjacent vertebrae became apparent in eight weeks . The degree of the disease process was more advanced in the Staph . aureus group and less severe in the Pseudomonas group . The E . coli group lay in between . 3 . The inflammatory process seemed to be confined in the disc for a week after the injection, during which time the cartilagenous cells in the nucleus pulposus underwent atrophy and degeneration . This resulted in direct exposure of the cartilagenous plate to the infection, causing invasion of the inflammatory process into the vertebral body . 4 . The presence of the epiphyseal line, however, seemed to act as a barrier to hinder the inflammatory process invading the vertebral body.

Mikrobiol Zh, 1991 Nov-Dec, 53(6), 66 - 70
{The level of spontaneous phage production and sensitivity to melioidosis phages of museum cultures of Pseudomonas pseudomallei}; Denisov II et al.; The phage-producing activity and sensitivity of museum cultures of a melioidosis agent to melioidosis phages have been studied . Most cultures show spontaneous phage-production, though its level differs in strains . Some of the melioidosis cultures demonstrate lack of phages . The fact that these strains have different sensitivity to chloroform lysate of cultures producing phages indicates their possible lysogenic state . Electron microscopy has shown the presence of at least tow morphological types of phages in P . pseudomallei C-141 . These data confirm existence of polysogeny in the melioidosis agent.

Can J Microbiol, 1991 Nov, 37(11), 880 - 4
Environmental factors affecting the antagonism of Pseudomonas cepacia against Trichoderma viride; Upadhyay RS et al.; Antagonistic activity of the bacterium Pseudomonas cepacia against Trichoderma viride was greatly influenced by nutritional and environmental conditions . Xylose and trehalose strongly enhanced the antifungal activity of P . cepacia, whereas mannitol and glucose had little effect . The carbon sources that enhanced the antagonistic activity also inhibited sporulation of T . viride . Antagonism of P . cepacia was enhanced by ammonium nitrogen; however, with nitrite or nitrate there was only a little antagonism . The antagonism of P . cepacia was optimal at pH 5.0 . Although P . cepacia showed maximum antagonism against T . viride at 37 degrees C, the antagonism was fairly good at temperatures as low as 18 degrees C, indicating that there is a broad range of temperature for the antifungal activity of P . cepacia.

Mol Plant Microbe Interact, 1991 Nov-Dec, 4(6), 553 - 62
Gene-for-gene interactions between Pseudomonas syringae pv . phaseolicola and Phaseolus; Jenner C et al.; The gene for cultivar-specific avirulence to Phaseolus vulgaris cv . Tendergreen in races 3 and 4 of Pseudomonas syringae pv . phaseolicola was isolated and sequenced . Genomic clones from libraries of race 3 in pLAFR1 and race 4 in pLAFR3, which altered the phenotype of race 5 from virulent to avirulent in Tendergreen, were found to possess a common approximately 15-kb region of DNA that contained the determinant of avirulence . Subcloning and insertion mutagenesis with Tn1000 located an avirulence gene within a 1.4-kb BglII/HindIII DNA fragment in races 3 and 4 . Comparison of the nucleotide sequences of regions of DNA that confer avirulence confirmed that both races have an identical gene for avirulence (designated avrPph3) comprising 801 base pairs (bp) and predicted to encode a cytoplasmic protein of 28,703 Da . A sequence, TGCAACCGAAT, 91% homologous to the motif found in promoter regions of avrB and avrD from P . s . pv . glycinea was located 89-99 bp upstream of the start of the open-reading frame 1 . Hybridization experiments showed that avrPph3 was not plasmid-borne and was absent from isolates of P . s . pv . phaseolicola races 1, 2, 5, 6, 7, and 8, P . cichorii, P . s . pvs . coronafaciens, glycinea, maculicola, pisi, syringae, and tabaci . Cosegregation studies of crosses between cultivars resistant (Tendergreen) and susceptible (Canadian Wonder) to races 3 and 4 and transconjugants of race 5 confirmed that a gene-for-gene relationship controls specificity in the interaction between Tendergreen and races 3 and 4 of P . s . pv . phaseolicola.

J Bacteriol, 1991 Nov, 173(22), 7219 - 27
Cloning, sequencing, and expression of the Pseudomonas testosteroni gene encoding 3-oxosteroid delta 1-dehydrogenase; Plesiat P et al.; Pseudomonas testosteroni ATCC 17410 is able to grow on testosterone . This strain was mutagenized by Tn5, and 41 mutants defective in the utilization of testosterone were isolated . One of them, called mutant 06, expressed 3-oxosteroid delta 1- and 3-oxosteroid delta 4-5 alpha-dehydrogenases only at low levels . The DNA region around the Tn5 insertion in mutant 06 was cloned into pUC19, and the 1-kbp EcoRI-BamHI segment neighbor to the Tn5 insertion was used to probe DNA from the wild-type strain . The probe hybridized to a 7.8-kbp SalI fragment . Plasmid pTES5, which is a pUC19 derivative containing this 7.8-kbp SalI fragment, was isolated after the screening by the 1-kbp EcoRI-BamHI probe . This plasmid expressed delta 1-dehydrogenase in Escherichia coli cells . The 2.2-kbp KpnI-KpnI segment of pTES5 was subcloned into pUC18, and pTEK21 was constructed . In E . coli containing the lacIq plasmid pRG1 and pTEK21, the expression of delta 1-dehydrogenase was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) . The induced level was about 40 times higher than the induced level in P . testosteroni . Delta 1-Dehydrogenase synthesized in E . coli was localized in the inner membrane fraction . The minicell experiments showed that a 59-kDa polypeptide was synthesized from pTEK21, and this polypeptide was located in the inner membrane fraction . The complete nucleotide sequence of the 2.2-kbp KpnI-KpnI segment of pTEK21 was determined . An open reading frame which encodes a 62.4-kDa polypeptide and which is preceded by a Shine-Dalgarno-like sequence was identified . The first 44 amino acids of the putative product exhibited significant sequence similarity to the N-terminal sequences of lipoamide dehydrogenases.

Biochem Biophys Res Commun, 1991 Oct 31, 180(2), 545 - 51
A proper amino terminus of diphtheria toxin is important for cytotoxicity; Chaudhary VK et al.; A series of deletions and substitutions were made at the 5' end of the gene fusion between the first 388 codons of diphtheria toxin (DT) and a cDNA encoding human IL2 . The chimeric protein (DT388-IL2) was expressed and purified from E . coli and found to be very cytotoxic to a human T cell line, HUT 102, that expresses a large number of IL2 receptors . Deletion of the first five amino acids of DT resulted in a non-cytotoxic chimeric protein that had both ADP-ribosylation activity and IL2 receptor binding activity . Deletion of the first two amino acids of DT had little effect on cytotoxicity, while deletion of the first four amino acids or of two acidic residues at positions 3 and 4 greatly reduced cytotoxicity . Unexpectedly, a mutant containing a single leucine in place of the first two amino acids (gly, ala) was 2-3 fold more active . The amino terminus of DT may participate in the translocation of the A chain to the cytosol in a manner similar to Pseudomonas exotoxin (PE) in which a specific C-terminal sequence has been proposed to be involved in its cytotoxicity.

Proc Natl Acad Sci U S A, 1991 Oct 15, 88(20), 8915 - 9
Copper resistance in Pseudomonas syringae mediated by periplasmic and outer membrane proteins; Cha JS et al.; Copper-resistant strains of Pseudomonas syringae pathovar tomato accumulate copper and develop blue colonies on copper-containing media . Three of the protein products of the copper-resistance operon (cop) were characterized to provide an understanding of the copper-resistance mechanism and its relationship to copper accumulation . The Cop proteins, CopA (72 kDa), CopB (39 kDa), and CopC (12 kDa), were produced only under copper induction . CopA and CopC were periplasmic proteins and CopB was an outer membrane protein . Leader peptide sequences of CopA, CopB, and CopC were confirmed by amino-terminal peptide sequencing . CopA, CopB, and CopC were purified from strain PT23.2, and their copper contents were determined . One molecule of CopA bound 10.9 +/- 1.2 atoms of copper and one molecule of CopC bound 0.6 +/- 0.1 atom of copper . The Cop proteins apparently mediate sequestration of copper outside of the cytoplasm as a copper-resistance mechanism.

Biochemistry, 1991 Oct 15, 30(41), 10034 - 42
A monolayer and bulk study on the kinetic behavior of Pseudomonas glumae lipase using synthetic pseudoglycerides; Deveer AM et al.; A heat-stable lipase from Pseudomonas glumae was purified to homogeneity . Its positional and stereospecific properties were investigated and compared with those of the well-known porcine pancreatic lipase . The kinetic properties of both enzymes were determined by use of six isomeric synthetic pseudoglycerides all composed of a single hydrolyzable fatty acyl ester bond and two lipase-resistant groups: one acylamino and one ether function . Two enzyme assay techniques were applied: a detergent-free system, the monomolecular surface film technique, and the pH-stat technique using clear micellar solutions of substrate in the presence of Triton X-100 . Regarding the cleavage of primary ester bonds, P . glumae lipase possesses no stereopreference . In contrast, a large stereopreference in favor of the R-isomer is found for the hydrolysis of secondary ester bonds . Secondary ester bonds are efficiently cleaved by the lipase, which makes it of potential interest for enzymatic synthetic purposes . For the hydrolysis of this R-isomer a correlation between the experimental catalytic turnover rate and the binding constant for micelles was observed . The kinetic data of P . glumae lipase have been analyzed in terms of the scooting and hopping models for the action of lipolytic enzymes {Upreti, G.C., & Jain, M.K . (1980) J . Membr . Biol . 55, 113-121} . The results presented in this study are best explained by assuming that glumae lipase leaves the interface after a limited number of catalytic cycles.

Biochim Biophys Acta, 1991 Oct 11, 1080(1), 68 - 77
Structural and functional features of Pseudomonas cytochrome c peroxidase; Ellfolk N et al.; The secondary structure of Pseudomonas cytochrome c peroxidase (ferrocytochrome c: hydrogen-peroxide oxidoreductase, EC 1.11.1.5) has been predicted from the established amino acid sequence of the enzyme using a Chou-Fasman-type algorithm . The amount of alpha-helicity thus obtained is in agreement with previously obtained results based on circular dichroic measurements at far UV . The two heme c moieties of the enzyme have earlier been shown to have widely different characteristics, e.g., the redox potentials of the hemes differ with about 600 mV, and carry out different functions in the enzyme molecule . The structural comparisons made in this study enlighten the observed functional differences . The first heme in the polypeptide chain, heme 1, has in its environment a folding pattern generally encountered in cytochromes . In the region of the sixth ligand, however, profound differences are noted . The cytochromal methionine has been replaced by a lysine with a concomitant lowering of redox-potential thus making peroxidatic activity possible . Around heme 2, extra amino acid residues have been added to the peroxidase as compared with Rhodospirillum molischianum cytochrome c2 core structure in the 20's loop . After completion of the cytochromal fold around heme 2 an additional tail consisting of 25 residues is linked . This tail shows no stabilizing elements of secondary structure, but contains a strongly hydrophobic segment which suggests a possible membrane contact site of this extrinsic membrane protein . Heme 2 is concluded to have a cytochromal function in the molecule . To further elucidate the functional properties of the enzyme, a noncovalent two-fragment complex was produced by specific cleavage of the peroxidase by Pseudomonas elastase . The complex was studied with respect to its properties to the native enzyme . The two-fragment complex of Pseudomonas peroxidase retains the overall conformation of the native enzyme showing, however, no heme-heme interaction . Thus, a comparison of the properties of the native enzyme with those of the two-fragment complex permitted some conclusions to be drawn on the structure of the enzyme as well as the mechanism of heme-heme interaction . From the present results we conclude that the two distal heme surfaces in the peroxidase are oriented toward each other . This structural arrangement allows an inter-heme communication in the enzyme molecule and it also forms the structural basis for the enzyme mechanism . The structural comparisons also give insight into the evolution of an ancestral cytochrome c into an efficient peroxidase that has a versatile control mechanism in heme-heme interaction.

Proc Natl Acad Sci U S A, 1991 Oct 1, 88(19), 8616 - 20
B3(Fv)-PE38KDEL, a single-chain immunotoxin that causes complete regression of a human carcinoma in mice; Brinkmann U et al.; The genes encoding the heavy- and light-chain Fv regions of the monoclonal murine antibody B3, which recognizes a carbohydrate antigen on the surface of many human carcinomas, were cloned by PCR techniques and used to generate single-chain immunotoxins containing Pseudomonas exotoxin (PE) . The light and heavy chains were connected by a flexible linker to form a single-chain antigen-binding protein, B3(Fv), which was in turn fused to truncated forms of PE lacking the cell-binding domain . The single-chain Fv and two different B3(Fv) immunotoxins, B3(Fv)-PE40 and B3(Fv)-PE38KDEL, were expressed in Escherichia coli and the single-chain immunotoxins were purified to near homogeneity . Both recombinant immunotoxins were shown to be cytotoxic specifically to carcinoma cell lines that express the B3 antigen on their surface; B3(Fv)-PE38KDEL was significantly more active . Furthermore, intravenous administration of B3(Fv)-PE38KDEL caused complete regression of human epidermoid carcinomas growing subcutaneously in immunodeficient mice.

Antiviral Res, 1991 Oct, 16(3), 267 - 79
Effects of a soluble CD4 and CD4-Pseudomonas exotoxin A chimeric protein on human peripheral blood lymphocytes: lymphocyte activation and anti-HIV activity in vitro; Rubino KL et al.; Recombinant sCD4-based proteins were evaluated for their effects on antigen-stimulated proliferation of human peripheral blood mononuclear cells (PBMC) and for antiviral activity against PBMC infected with human immunodeficiency virus (HIVD34) . Two sCD4-based proteins were solubilized, refolded, and purified to homogeneity from recombinant E . coli and consisted of the 178 amino-terminal residues of CD4 fused with the translocating and catalytic domains of Pseudomonas exotoxin A (sCD4-PE40) or 183 amino-terminal residues of CD4 (sCD4-183); a third sCD4 consisting of 369 amino acids of CD4 was purified from recombinant mammalian cells for comparative purposes (sCD4-369) . Increasing molar concentrations of these sCD4s were evaluated for inhibition of PBMC proliferation induced by alloantigen (MLR), by tetanus toxoid (TTOX), or in response to crosslinking with antibody to CD3 (OKT3) . In addition, the concentrations of each protein required to inhibit replication of the HIVD34 isolate in primary PBMC was determined by quantitation of HIV p24 antigen released into supernatant fluids by infected cells . By comparing antiviral activity with anti-proliferative activity a relative estimate of the selectivity index for each recombinant sCD4 was determined . Proliferation of PBMC in response to alloantigen or OKT3 was less sensitive to inhibition than proliferation induced by TTOX, and the selectivity indices estimated for sCD4-PE40 were 170, 170 and 17, respectively . The selectivity index for sCD4-183 was greater than 350 under all assay conditions . Comparative evaluation of alloantigen-stimulated proliferation with antiviral activity of sCD4-183 versus sCD4-369 suggested that the E . coli-derived sCD4-183 may have a higher selectivity index under these conditions than its mammalian cell-derived counterpart.

J Biochem (Tokyo), 1991 Oct, 110(4), 520 - 5
Purification and characterization of a new NAD(+)-dependent enzyme, L-tartrate decarboxylase, from Pseudomonas sp . group Ve-2; Furuyoshi S et al.; A new enzyme, L-tartrate decarboxylase, was found in cells of Pseudomonas sp . group Ve-2 . The enzyme was purified to homogeneity and characterized . The enzyme requires K+, Mg2+, and NAD+ for L-tartrate decarboxylation . The dependence of the enzymatic decarboxylation on NAD+ suggests that the decarboxylation involves redox reactions of the substrate . The enzyme catalyzes NAD(+)-linked oxidative decarboxylation of D-malate as well . The enzyme is composed of four subunits with identical molecular weight (Mr 40,000) . The apparent Michaelis constants for L-tartrate and NAD+ are 1.1 mM, respectively . The cofactor requirements and the physical properties of the enzyme were similar to those of L-tartrate dehydrogenase-D-malate dehydrogenase from Rhodopseudomonas sphaeroides, and tartrate dehydrogenase from P . putida.

Biol Chem Hoppe Seyler, 1991 Oct, 372(10), 915 - 22
Purification and characterization of 2-halocarboxylic acid dehalogenase II from Pseudomonas spec . CBS 3; Morsberger FM et al.; 2-Halocarboxylic acid dehalogenase II from Pseudomonas spec . CBS 3 (EC 3.8.1.2), which had been cloned in E . coli Hb 101 was purified to electrophoretic homogeneity from crude extracts of E . coli Hb 101 clone 1164 . Ammonium sulfate fractionation and three subsequent chromatographic purification steps yielded a pure enzyme in a 230-fold enrichment . The relative molecular masses as determined by gelfiltration on Superose 12 and SDS-polyacrylamide gel electrophoresis were 64,000 Da for the holoenzyme and 29,000 Da for the subunit . The isoelectric point, determined by isoelectric focusing, was at pH 6.2 . Substrate specificity towards chlorinated and brominated substrates was limited to short chain monosubstituted 2-halocarboxylic acids . Fluorocompounds were not converted . The reaction proceeded best at a pH above 9.5 and at a reaction temperature of 40-45 degrees C.

J Gen Microbiol, 1991 Oct, 137 ( Pt 10), 2281 - 6
Purification and properties of an ethylene-forming enzyme from Pseudomonas syringae pv . phaseolicola PK2; Nagahama K et al.; A novel ethylene-forming enzyme that catalyses the formation of ethylene from 2-oxoglutarate was purified from a cell-free extract of Pseudomonas syringae pv . phaseolicola PK2 . It was purified about 2800-fold with an overall yield of 53% to a single band of protein after SDS-PAGE . The purified enzyme had a specific activity of 660 nmol ethylene min-1 (mg protein)-1 . The molecular mass of the enzyme was approximately 36 kDa by gel filtration and 42 kDa by SDS-PAGE . The isoelectric point and optimum pH were 5.9 and ca . 7.0-7.5, respectively . There was no homology between the N-terminal amino acid sequence of the ethylene-forming enzyme of Ps . syringae pv . phaseolicola PK2 and the sequence of the ethylene-forming enzyme of the fungus Penicillium digitatum IFO 9372 . However, the two enzymes have the following properties in common . The presence of 2-oxoglutarate, L-arginine, Fe2+ and oxygen is essential for the enzymic reaction . The enzymes are highly specific for 2-oxoglutarate as substrate and L-arginine as cofactor . EDTA, Tiron, DTNB {5,5'-dithio-bis(2-nitrobenzoate)} and hydrogen peroxide are all effective inhibitors.

J Antimicrob Chemother, 1991 Oct, 28(4), 561 - 8
Dosage adjustment and clinical outcomes of long-term use of high-dose tobramycin in adult cystic fibrosis patients; Li SC et al.; A two-phase study was undertaken designed to investigate the impact of computer-aided drug monitoring on tobramycin concentrations and clinical outcomes in adult patients with cystic fibrosis . In phase one, a baseline (historical control) study of drug use patterns was performed . During the second phase, patients admitted for intravenous treatment with tobramycin for acute exacerbations of pseudomonal pulmonary infections were randomly allocated to one of two schedules . Group A patients had tobramycin dosage regimens decided by clinicians based on pre-existing protocols using serum tobramycin assay data determined three times weekly . Group B patients had dosage regimens determined by a computerized pharmacokinetic predictive program using both population-based pharmacokinetic parameter estimation and fitting of serum concentration-time data using Bayesian regression . The agreed therapeutic target was a peak serum tobramycin concentration of 8-10 mg/L and a trough concentration of 1-2 mg/L . There was a major difference between the two groups comparing the number of paired trough and peak concentrations within the target concentration ranges (group A-14%; group B-34.7%, chi 2 test, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Mil Med, 1991 Oct, 156(10), 520 - 7
Risk factors for infection in fracture war wounds (1973 and 1982 wars, Israel)
Simchen E, Raz R, Stein H, Danon Y.
The development of post-surgical wound infection was compared in two groups of soldiers who sustained fractures or amputations on the battlefield during the first month of the 1982 and 1973 wars . Risk factors for the development of post-surgical wound infection were sought . In the 1982 group, numbering 184, the four variables independently associated with infection were multiple operations during the follow-up period; drains inserted in the first operation; extensive tissue loss; and blood transfusion during the first operation . For the 1973 group, numbering 130, the significant variables were multiple operations; amputations (highly correlated with extensive tissue loss); injury involving other body systems in addition to the fracture; and open drains . The high risk associated with open drains in both wars raises doubt about their usefulness . The main distinction between injuries of the two wars was the high prevalence (72.3%) of multi-system injuries in 1973 versus low prevalence (29.2%) in 1982 . Overall infection rates were similar (30.5% and 31.5%), but infections at the site of the fracture were twice as high in 1982 . Pseudomonas was the most common single species of bacteria isolated from infected wounds (26% in 1982, 33.6% in 1973) . It appeared in the wounds relatively late, 10-14 days after admission.

Jpn J Med Sci Biol, 1991 Oct-Dec, 44(5-6), 225 - 37
Substrate response in acid phosphatase activity of Pseudomonas pseudomallei and Pseudomonas cepacia, with special reference to tyrosine phosphatase; Kanai K et al.; The substrate response in acid phosphatase activity of Pseudomonas pseudomallei and Pseudomonas cepacia was examined with different phosphate esters including hexose phosphates and phosphoaminoacids in a whole cell assay system . The enzymatic activity against each substrate was evaluated in terms of percent activity to that against para-nitrophenyl phosphate set as 100 . A remarkable finding was that the phosphatase reaction was the highest with phosphotyrosine or phosphoserine as substrate showing 180% activity . This tyrosine phosphatase activity was resistant to heating at 60 C for 20 min and inhibited greatly by 0.1% ZnCl2 . Pseudomonas cepacia showed the same pattern of substrate response and the same characteristics of tyrosine phosphatase activity.

Jpn J Med Sci Biol, 1991 Oct-Dec, 44(5-6), 195 - 211
Fatty acid profile and acid phosphatase activity of fresh isolates of Pseudomonas pseudomallei; Kondo E et al.; Eighty-one fresh isolates of Pseudomonas pseudomallei from melioidosis patients were subjected to the analysis for the fatty acid composition by gas-liquid chromatography (GLC) and pH-dependent pattern of nonspecific phosphatase activity . All the test strains were identical in the GLC profile showing the three peaks of characteristic hydroxy acids (3-OH 14:0, 2-OH 16:0, 3-OH 16:0) and the two prominent peaks of cyclopropane acids (17:0 delta, 19:0 delta) . They had also basically the same pH-dependent curves of the enzymatic activity with paranitrophenyl phosphate as substrate, showing two to three peaks or shoulders only in the acidic side of the curve . These two biochemical characteristics could differentiate P . pseudomallei distinctly from P . aeruginosa, but not from P . cepacia.

Zh Mikrobiol Epidemiol Immunobiol, 1991 Oct, (10), 8 - 12
{The role of the surface antigens of Pseudomonas pseudomallei in the pathogenesis of melioidosis}; Piven' NN et al.; As established with the use of electron-immunochemical techniques, glycoprotein antigen 6 is the outer membrane component of P . pseudomallei cell wall, while glycoprotein antigen 8 is localized on the cell surface as a capsule-like formation . Antigen 6 plays no perceptible role in the realization of the pathogenic properties of the infective agent, but serves as a reliable sign in the differentiation of P . pseudomallei strains into serovars . Subcultures, defective in the synthesis of antigen 8, have sharply reduced virulence for laboratory animals . As revealed in this study, the pathogenetic action of antigen 8 is linked with its pronounced antiphagocytic function . Thus, antigen 8 is considered to be one of the key pathogenicity factors of P . pseudomallei.

FASEB J, 1991 Oct, 5(13), 2843 - 9
Cytotoxic activity of chimeric proteins composed of acidic fibroblast growth factor and Pseudomonas exotoxin on a variety of cell types; Siegall CB et al.; Chimeric proteins composed of acidic fibroblast growth factor (acidic FGF) and several forms of Pseudomonas exotoxin (PE) that cannot bind to the PE receptor have been produced in Escherichia coli by expressing chimeric genes in which DNA encoding acidic FGF is fused to various mutant forms of PE . These acidic FGF-PE fusion proteins were found to be cytotoxic to a variety of tumor cell lines including hepatocellular (PLC/PRF/5 and HEPG2), prostatic (LNCaP), colon (HT29), and breast (MCF-7) carcinomas at concentrations of 1-70 ng/ml . The cytotoxic effects of acidic FGF-PE were FGF-receptor specific as demonstrated by competition with excess acidic FGF and by showing that acidic FGF-PE bound to the FGF receptor with the same affinity as acidic FGF . Furthermore, the cell-killing activity of acidic FGF-PE was toxin-mediated, as an acidic FGF-PE mutant, which does not possess ADP-ribosylation activity, failed to kill cells . These findings demonstrate that acidic FGF-PE is a potent cytotoxic molecule that can be targeted to FGF receptor-bearing cells . Because acidic FGF is a potent angiogenic molecule, cytotoxic acidic FGF-PE chimeras may have utility as anti-angiogenic agents . These molecules could be helpful in determining the functional role of FGF receptors in cellular processes.

Appl Biochem Biotechnol, 1991 Oct, 31(1), 59 - 73
Detoxification of organophosphate pesticides using a nylon based immobilized phosphotriesterase from Pseudomonas diminuta; Caldwell SR et al.; A partially purified phophotriesterase was successfully immobilized onto nylon 6 and 66 membranes, nylon 11 powder, and nylon tubing . Up to 9000 U of enzyme activity was immobilized onto 2000 cm2 of a nylon 6 membrane where 1 U is the amount of enzyme necessary to catalyze the hydrolysis of 1.0 mumol of paraoxon/min at 25 degrees C . The nylon 66 membrane-bound phosphotriesterase was characterized kinetically where the apparent Km value for the immobilized enzyme was 0.35 mM . This is 5-6 times higher than that observed for the soluble enzyme . However, nylon immobilization limited the maximum rate of paraoxon hydrolysis to less than 10% of the value measured for the soluble enzyme . The addition of the cosolvent, methanol, resulted in an increase in the apparent Km value for paraoxon hydrolysis but concentrations up to 40% had no negative effect on the catalytic effectiveness with the soluble or immobilized phosphotriesterase . Based on the kinetic analysis, methanol appears to be a competitive inhibitor for both forms of enzyme . The nylon powder immobilized enzyme was shown to be stable for at least 20 mo . The immobilization of the phosphotriesterase onto nylon provides a practical method for the detoxification of organophosphate pesticides.

Appl Environ Microbiol, 1991 Oct, 57(10), 2928 - 34
Genetic analysis of the antifungal activity of a soilborne Pseudomonas aureofaciens strain; Vincent MN et al.; Pseudomonas aureofaciens Q2-87 produces the antibiotic 2,4-diacetophloroglucinol (Phl), which inhibits Gaeumannomyces graminis var . tritici and other fungi in vitro . Strain Q2-87 also provides biological control of take-all, a root disease of wheat caused by this fungus . To assess the role of Phl in the antifungal activity of strain Q2-87, a genetic analysis of antibiotic production was conducted . Two mutants of Q2-87 with altered antifungal activity were isolated by site-directed mutagenesis with Tn5 . One mutant, Q2-87::Tn5-1, did not inhibit G . graminis var . tritici in vitro and did not produce Phl . Two cosmids were isolated from a genomic library of the wild-type strain by probing with the mutant genomic fragment . Antifungal activity and Phl production were coordinately restored in Q2-87::Tn5-1 by complementation with either cosmid . Mobilization of one of these cosmids into two heterologous Pseudomonas strains conferred the ability to synthesize Phl and increased their activity against G . graminis var . tritici, Pythium ultimum, and Rhizoctonia solani in vitro . Subcloning and deletion analysis of these cosmids identified a 4.8-kb region which was necessary for Phl synthesis and antifungal activity.

Biochem J, 1991 Oct 1, 279 ( Pt 1), 105 - 9
Lupanine hydroxylase, a quinocytochrome c from an alkaloid-degrading Pseudomonas sp; Hopper DJ et al.; Lupanine 17-hydroxylase, the first enzyme in the pathway for bacterial degradation of the alkaloid, lupanine, was purified from a Pseudomonas sp . The enzyme acts by initial dehydrogenation of the substrate, and cytochrome c was used as electron acceptor in assays . It had an Mr of 66,000 by ultracentrifuge studies and 74,000 by gel filtration . The visible absorption spectrum was that of a cytochrome c, and a stoicheiometry of one haem group per molecule of enzyme was calculated . SDS/PAGE gave a single band of Mr 72,000 containing the haem group . The enzyme also contained pyrroloquinoline quinone (PQQ), which could be removed by isoelectric focusing . The apoenzyme was reconstituted to full activity with addition of PQQ, and a stoicheiometry of one molecule of PQQ per molecule of enzyme was calculated . Steady-state kinetics gave values of 3.6 microM for the Km for lupanine, 21.3 microM for the Km for cytochrome c and 217 s-1 for the Kcat.

Cytotechnology, 1991 Oct, 7(2), 103 - 12
Adaptation of hybridoma cells to higher ammonia concentration; Matsumura M et al.; Using two mouse-mouse hybridoma cell lines, the response to ammonia step and serial changes was investigated in batch and continuous cultures with serum-free medium . The inhibitory effect of ammonia on cell growth depended on the cultivation mode, and differed markedly between cell lines . The cell line, 4C10B6 producing IgG monoclonal antibody against Pseudomonas, showed a high adaptation ability to ammonia . The 4C10B6 cells could grow under ammonia concentration as high as 21 mmol/l NH4Cl with a viability of 80% in the continuous culture with serial increase in ammonia concentration . Whereas, in the batch culture with ammonia step change the cell growth completely ceased at 12 mmol/l NH4Cl . The other cell line, TO-405 producing IgG monoclonal antibody against hepatitis B surface antigen, could not adapt to ammonia, and the cell growth did not occur at 9 mmol/l NH4Cl even under the ammonia serial change.

J Biol Chem, 1991 Sep 25, 266(27), 18135 - 40
Purification, characterization, and molecular cloning of lactonizing lipase from Pseudomonas species; Ihara F et al.; An extracellular lipase catalyzing the synthesis of macrocyclic lactones in anhydrous organic solvents was purified to homogeneity from Pseudomonas nov . sp . 109, and characterized . The lipase showed a pI of 5.3 on isoelectric focusing and a Mr of 29,000 +/- 1,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis . With respect to substrate specificity, optimum chain length for acyl moiety varied depending on the type of reaction catalyzed: C18 in monomer lactone formation, C11 or shorter in dimer lactone formation, and C8 in ester hydrolysis . The amino-terminal 19 amino acid residues of the purified lipase were determined as Ser-Thr-Tyr-Thr-Gln-Thr-Lys-Tyr-Pro-Ile-Val-Leu-Ala-His-Gly-Met-Leu-Gly- Phe, and the gene encoding the lipase was identified by hybridization to a synthetic 20-nucleotide probe, cloned, and sequenced . Nucleotide sequence analysis predicted a 311-amino acid open reading frame, a putative ribosome-binding site, and a 26-amino acid sequence at the amino terminus of the sequence that is not found in the mature protein . This 26-amino acid sequence has many of the characteristics common to known signal peptides . The lipase gene encoded a sequence of Val-Asn-Leu-Ile-Gly-His-Ser-His-Gly-Gly which is very well conserved among lipases, and showed 38-40% overall homology to the amino acid sequences of lipases from Pseudomonas fragie and Pseudomonas cepacia, but showed little homology to those of other lipases, suggesting that some structural features are required for catalyzing macrocyclic lactone synthesis in organic solvents and are restricted to lipases of the Pseudomonas origin.

FEBS Lett, 1991 Sep 23, 290(1-2), 224 - 6
Identification of 4-chlorobenzoyl-coenzyme A as intermediate in the dehalogenation catalyzed by 4-chlorobenzoate dehalogenase from Pseudomonas sp . CBS3; Loffler F et al.; The intermediate in the reaction catalyzed by 4-chlorobenzoate dehalogenase from Pseudomonas sp . CBS3 was identified as 4-chlorobenzoyl-CoA . One component of 4-chlorobenzoate dehalogenase worked as a a 4-chlorobenzoyl-CoA ligase catalyzing the formation of 4-chlorobenzoyl-CoA from 4-chlorobenzoate, coenzyme A and ATP . This intermediate was detected spectrophotometrically and by HPLC . 4-chlorobenzoyl-CoA was the substrate for the dehalogenase component, which catalyzed the conversion to 4-hydroxybenzoate with concomitant release of coenzyme A.

Am J Med, 1991 Sep 16, 91(3B), 252S - 255S
Isoenzyme analysis of Pseudomonas cepacia as an epidemiologic tool; Carson LA et al.; Multilocus enzyme electrophoresis has successfully been used to establish basic marker systems for the epidemiologic analysis of a variety of bacterial pathogens . This study was done to determine the efficacy of this technique for characterizing Pseudomonas cepacia, using 31 known-related strains isolated during an outbreak of infections involving intrinsically contaminated povidone-iodine solution, and five outbreak-unrelated strains used in serotyping of P . cepacia . Crude cell extracts were analyzed by starch gel electrophoresis for electrophoretic variants using 13 enzyme substrates; esterase bands were detected using an additional four substrates . The 31 outbreak strains had identical isoenzyme patterns for all enzymes examined . Five electrophoretic types were obtained for the serotyping strains; electrophoretic mobilities of one of the five strains corresponded to the patterns obtained for the outbreak strains . These results suggest that enzyme electrophoretic typing may be a useful adjunct to other typing methods used in epidemiologic analyses of P . cepacia infections.

Infect Immun, 1991 Sep, 59(9), 2870 - 9
Effect of site-directed mutagenic alterations on ADP-ribosyltransferase activity of the A subunit of Escherichia coli heat-labile enterotoxin; Lobet Y et al.; Previous studies of the S1 subunit of pertussis toxin, an NAD(+)-dependent ADP-ribosyltransferase, suggested that a small amino-terminal region of amino acid sequence similarity to the active fragments of both cholera toxin and Escherichia coli heat-labile enterotoxin represents a region containing critical active-site residues that might be involved in the binding of the substrate NAD+ . Other studies of two other bacterial toxins possessing ADP-ribosyltransferase activity, diphtheria toxin and Pseudomonas exotoxin A, have revealed the presence of essential glutamic acid residues vicinal to the active site . To help determine the relevance of these observations to activities of the enterotoxins, the A-subunit gene of the E . coli heat-labile enterotoxin was subjected to site-specific mutagenesis in the region encoding the amino-terminal region of similarity to the S1 subunit of pertussis toxin delineated by residues 6 through 17 and at two glutamic acid residues, 110 and 112, that are conserved in the active domains of all of the heat-labile enterotoxin variants and in cholera toxin . Mutant proteins in which arginine 7 was either deleted or replaced with lysine exhibited undetectable levels of ADP-ribosyltransferase activity . However, limited trypsinolysis of the arginine 7 mutants yielded fragmentation kinetics that were different from that yielded by the wild-type recombinant subunit or the authentic A subunit . In contrast, mutant proteins in which glutamic acid residues at either position 110 or 112 were replaced with aspartic acid responded like the wild-type subunit upon limited trypsinolysis, while exhibiting severely depressed, but detectable, ADP-ribosyltransferase activity . The latter results may indicate that either glutamic acid 110 or glutamic acid 112 of the A subunit of heat-labile enterotoxin is analogous to those active-site glutamic acids identified in several other ADP-ribosylating toxins.

Chest, 1991 Sep, 100(3), 875 - 7
Fatal pulmonary aspergillosis presenting as acute eosinophilic pneumonia in a previously healthy child; Ricker DH et al.; A previously healthy boy presented with cough and diffuse pulmonary interstitial infiltrates . Acute eosinophilic pneumonia was diagnosed by bronchoalveolar lavage in the absence of a demonstrable infectious etiologic agent . Corticosteroid therapy resulted in immediate improvement but was followed by respiratory distress and death from invasive aspergillosis and Pseudomonas cepacia sepsis.

Carbohydr Res, 1991 Sep 2, 216, 495 - 504
Structure of the glycocalyx polysaccharide of Pseudomonas fragi ATCC 4973; Parolis LA et al.; The structure of the exocellular glycocalyx polysaccharide of Pseudomonas fragi ATCC 4973, a bacterium implicated in the spoilage of meat, has been determined using hydrolysis, methylation analysis and 1D- and 2D-n.m.r . spectroscopy . The polysaccharide, which aids in the adhesion of the cells to each other and to the meat tissue, has the regular repeating unit ----4)-3-O-{(R)-1-carboxyethyl}-alpha-D-Glcp-(1----3)-beta-D-ManpNAc+ ++- (1----4)-beta-D-Glcp-(1---- . Random partial O-acetylation occurred in some preparations of the polysaccharide.

Infect Dis Clin North Am, 1991 Sep, 5(3), 467 - 84
Pneumonia in chronic obstructive lung disease; Griffith DE et al.; Despite the apparent common occurrence of pneumonia in patients with chronic obstructive pulmonary disease (COPD), there are little firm data on incidence, etiology, diagnostic procedures, and therapy in these patients . It appears that traditional respiratory pathogens such as the pneumococcus are declining in importance while "new" pathogens such as Pseudomonas sp., Moraxella catarrhalis, and Legionella sp . are becoming more important . The diagnosis of a specific etiologic agent is difficult in COPD and can be aided by obtaining specimens bronchoscopically . Directed therapy is optimal; however, empiric therapy is frequently unavoidable.

Nippon Geka Gakkai Zasshi, 1991 Sep, 92(9), 1292 - 5
{Surgical infection and its backgrounds}; Hikida S et al.; The backgrounds of surgical infection of esophageal cancer patients are studied for host factors (relation of nutrition and immune response) and parasite factors surgical parts which are easily contaminated, the differences of serum concentration with 3 methods of antibiotics administration, and the relations about the bacteria in mouth and upper esophagus in operation and bacteria which were found in the respiratory system after operation) . The results are as follows . The preoperative nutritional support was effective to rapid recovery of cytological immune functions . But in the post-operative infected cases, immunological recovery delayed and they nutritionally needed BCAA, glutamine, and essential fatty acids . For the study of parasite factors, surgical wounds (neck, chest wall, abdominal wall) were more easily contaminated than other parts except digestive tract . Continuous administration of antibiotics, 1g/hr x 4 was most excessive in three methods . The bacteria in the mouth and upper esophagus in operation and those in airway after operation, were not correlated . Furthermore, antibiotics-resistant bacteria which were Pseudomonas, MRSA were found in the airway . These results indicate that nutritional support is important for the host defense system and antibiotics should be administered considering each operative process . And about the postoperative respiratory infection, protection of hospital infection is important.

Transplantation, 1991 Sep, 52(3), 470 - 4
Treatment of corneal allograft rejection with the cytotoxin IL-2-PE40; Herbort CP et al.; IL-2-PE40 is a recombinant chimeric protein composed of IL-2, fused to a modified pseudomonas exotoxin . This molecule is extremely toxic to activated T cells expressing high-affinity IL-2R . We used this new molecule for selective immunosuppression to treat corneal allograft rejection in the rat, using Fisher and Lewis rats, a strain combination differing only in medial and minor histocompatibility antigens . The effect of IL-2-PE40 on the immunologic response was studied using both a heterotopic corneal graft model and orthotopic grafts . At the dose of 0.31 micrograms/g given intraperitoneally every 12 hr, IL-2-PE40 produced a significant reduction of both total lymph node cells and cytotoxic-T-cell (CTL) activity in draining lymph nodes (DLN) of heterotopically grafted animals . IL-2-PE40 treatment also significantly reduced the clinical rejection score and cumulative rejection rate (CRR) in orthotopic grafts and appears to be a very effective immunosuppressive agent.

Am Rev Respir Dis, 1991 Sep, 144(3 Pt 1), 580 - 5
Human neutrophil elastase and elastase/alpha 1-antiprotease complex in cystic fibrosis . Comparison with interstitial lung disease and evaluation of the effect of intravenously administered antibiotic therapy; Meyer KC et al.; In cystic fibrosis (CF), extracellular lung matrix is progressively damaged, neutrophils invade the air spaces, and activated neutrophils may release large amounts of neutrophil elastase (NE) . Although alpha 1-antiprotease (alpha 1-AP) binds and inactivates NE and is the major antielastase of the lower respiratory tract, antielastase defenses may be overwhelmed in CF, leading to progressive lung damage . To determine whether the ability of alpha 1-AP to neutralize NE is impaired in CF, we compared NE activity in bronchoalveolar lavage (BAL) fluid and human neutrophil elastase/alpha 1-antiprotease (NE/alpha 1-AP) complex in both BAL fluid and peripheral blood serum from patients with CF, normal volunteers, and patients with interstitial lung disease . We detected a considerable amount of NE activity in BAL fluid from all but one patient with CF but none in that from normal volunteers or from patients with interstitial lung disease . Although in interstitial lung disease there was a significant correlation between increased NE/alpha 1-AP complex in BAL or peripheral blood and the degree of neutrophil influx, NE/alpha 1-AP complex was disproportionately low in CF BAL compared with significantly elevated values in serum . These data suggest that in CF, alpha 1-AP-mediated defense against free NE in the lower respiratory tract is significantly impaired, and high levels of uncomplexed, enzymatically active, NE are present in CF respiratory secretions . To determine whether intravenously administered antipseudomonal antibiotic therapy for exacerbations of CF lung disease diminished the amount of free NE in respiratory secretions, we used BAL to investigate the effect of such therapy on neutrophils and NE in patients with CF colonized with pseudomonads.(ABSTRACT TRUNCATED AT 250 WORDS)

J Bacteriol, 1991 Sep, 173(17), 5406 - 13
Molecular characterization of nosA, a Pseudomonas stutzeri gene encoding an outer membrane protein required to make copper-containing N2O reductase; Lee HS et al.; A Pseudomonas stutzeri gene (nosA) encoding an outer membrane protein was cloned into the broad-host-range vector pRK290 and expressed in a mutant lacking the protein . Deletion analysis identified the approximate extent of the nosA region which was sequenced, and it was found to contain an open reading frame encoding 683 amino acids including a presumed signal sequence of 44 amino acids . The putative processed form had a molecular weight of 70,218, characteristics typical of outer membrane proteins, and considerable amino acid sequence homology with Escherichia coli BtuB . A short stretch of amino acids was homologous with the E . coli TonB-dependent outer membrane proteins, BtuB, IutA, FepA, and FhuA, suggesting a homologous function: interaction with a periplasmic protein or uptake of a specific substrate.

J Bacteriol, 1991 Sep, 173(17), 5260 - 5
Isolation, sequence, and expression in Escherichia coli of the Pseudomonas sp . strain ACP gene encoding 1-aminocyclopropane-1-carboxylate deaminase; Sheehy RE et al.; Pseudomonas sp . strain ACP is capable of growth on 1-aminocyclopropane-1-carboxylate (ACC) as a nitrogen source owing to induction of the enzyme ACC deaminase and the subsequent conversion of ACC to alpha-ketobutyrate and ammonia (M . Honma, Agric . Biol . Chem . 49:567-571, 1985) . The complete amino acid sequence of purified ACC deaminase was determined, and the sequence information was used to clone the ACC deaminase gene from a 6-kb EcoRI fragment of Pseudomonas sp . strain ACP DNA . DNA sequence analysis of an EcoRI-PstI subclone demonstrated an open reading frame (ORF) encoding a polypeptide with a deduced amino acid sequence identical to the protein sequence determined chemically and a predicted molecular mass of 36,674 Da . The ORF also contained an additional 72 bp of upstream sequence not predicted by the amino acid sequence . Escherichia coli minicells containing the 6-kb clone expressed a major polypeptide of the size expected for ACC deaminase which was reactive with ACC deaminase antiserum . Furthermore, a lacZ fusion with the ACC deaminase ORF resulted in the expression of active enzyme in E . coli . ACC is a key intermediate in the biosynthesis of ethylene in plants, and the use of the ACC deaminase gene to manipulate this pathway is discussed.

Postgrad Med, 1991 Sep 1, 90(3), 169 - 70, 173
Pseudomonas folliculitis from a health club whirlpool; Breitenbach RA; In public whirlpools, low disinfectant levels and inadequate monitoring are clearly a public health concern . Implementing guidelines published by the Centers for Disease Control seems to be the most reasonable approach to preventing dermatitis in persons who use these facilities . Physicians should be on the alert for well-demarcated rashes that may be associated with improperly maintained whirlpools.

Hua Xi Yi Ke Da Xue Xue Bao, 1991 Sep, 22(4), 372 - 5
{Preparation of monoclonal antibodies against chorionic gonadotropin receptor and study of its characteristics}; Han S et al.; We reported the production of monoclonal antibodies (McAbs) against chorionic gonadotropin hormone (CG) receptor by fusing spleen cells of BALB/c mice which had been immunized by purified bacteria (Pseudomonas maltophilia) CG receptor with mouse myeloma line SP2/0 . Four hybridoma cell lines secreting CG receptor McAbs were obtained (ED490 DG390, AB890 and GE590) . The titers of specific antibodies of both mice ascites and culture supernatant were 10(-2)-10(-6) and 1-10(-2) respectively, by solid phase ELISA . Double-Immunodiffusion test showed that the McAb GE590 was IgG1, and the McAbs ED490, DG390 and AB890 were IgG2b Immunoprecipitation indicated that 125I-HCG could bind the HCG receptor which had reacted with McAbs ED490, AB890 and DG390, suggesting that they may recognize the receptor with different antigenic determinant . Interaction of McAb GE590 with the receptor showed that the increased concentration of GE590 was in inverse proportion to the amount of 125I-HCG binding receptor, indicating that both the McAb and 125I-HCG could recognize a common site of receptor and that increased concentration of McAb GE590 may induce some change in conformation and structure of the receptor . Our study suggested that these McAbs may be used for studying structure of CG receptor.

Haematologica, 1991 Sep-Oct, 76(5), 424 - 5
Severe neutropenia in a patient with large granular lymphocytosis: prolonged successful control with cyclosporin A; Garipidou V et al.; A case of severe neutropenia associated with large granular lymphocytosis in a 40-year-old female is described . The patient, with no findings of an underlying systemic disorder, had suffered from recurrent life threatening, mainly pseudomonal, infections for about two years, despite the various regimes tried . During the last twelve months cyclosporin A treatment resulted in a significant increase in absolute neutrophil counts, concomitant with a remarkable decrease in bone marrow infiltration by GLs and almost normal counts of GLs in the peripheral blood . During this time she has remained completely free from infectious episodes . The mechanisms involved remain to be determined.

Antibiot Khimioter, 1991 Sep, 36(9), 31 - 4
{Sensitivity of Pseudomonas mallei to fluoroquinolones and their efficacy in experimental glanders}; Batmanov VP; Thirteen strains of P . mallei were found to be highly sensitive to ciprofloxacin and ofloxacin and resistant to norfloxacin . Ciprofloxacin and ofloxacin showed a high efficacy on models of malleus in guinea pigs and hamsters . The animals were infected with various strains of P . mallei . In the in vitro experiments, ciprofloxacin proved to be the most active and efficient . Norfloxacin appeared to be inefficient.

Appl Environ Microbiol, 1991 Sep, 57(9), 2540 - 3
Production and characterization of N-acyl-D-glutamate amidohydrolase from Pseudomonas sp . strain 5f-1; Sakai K et al.; N-Acyl-D-glutamate amidohydrolase from Pseudomonas sp . strain 5f-1 was inducibly produced by D isomers of N-acetylglutamate, glutamate, aspartate, and asparagine . The enzyme has been purified to homogeneity by DEAE-cellulose, (NH4)2SO4 fractionation, and chromatofocusing followed by gel filtration on a Sephadex G-100 column . The enzyme was a monomer with molecular weight of 55,000 . The enzyme activity was optimal at pH 6.5 to 7.5 and 45 degrees C . The isoelectric point and the pH stability were 8.8 and 9.0, respectively . N-Formyl, N-acetyl, N-butyryl, N-propionyl, N-chloroacetyl derivatives of D-glutamate and glycyl-D-glutamate were substrates for the enzyme . At pH 6.5 in 100 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer at 30 degrees C, a Km of 6.67 mM and a Vmax of 662 mumol/min/mg of protein for N-acetyl-D-glutamate were obtained . None of the metal ions stimulated the enzyme activity . Na+, K+, Mg2+, and Ba2+ acted as stabilizers . Hg2+, Cu2+, Zn2+, Fe3+, and EDTA were strongly inhibitory.

Analyst, 1991 Sep, 116(9), 919 - 22
Simultaneous determination of toxic metabolites by linear combination derivative spectrophotometry; Lin LM et al.; A linear combination derivative spectrophotometric method is described . The method overcomes the problem of overlapping in derivative spectrophotometry and allows the maximum use of quantitative information . In addition, the method can be used to increase the selectivity, sensitivity and accuracy of the simultaneous analysis of multicomponent mixtures . The application of the method to the simultaneous determination of bongkrekic acid and toxoflavin, the toxic metabolites produced by Pseudomonas farinofermentans, is described.

Nihon Kyobu Shikkan Gakkai Zasshi, 1991 Sep, 29(9), 1190 - 4
{Diffuse panbronchiolitis in two brothers with different clinical courses}; Makiguchi K et al.; Diffuse panbronchiolitis in two brothers is reported . The elder brother aged 46, was admitted in May 1983 due to severe dyspnea and productive cough, which had gradually worsened over several years . He had severe hypoxemia and hypercapnia . He died at age 47 of respiratory failure due to pseudomonas infection despite antibiotic therapy . The younger brother, at age 41, was admitted in March 1983 due to fever, productive cough, and abnormal shadows on chest X-ray films . He showed mild hypoxemia and his symptoms improved with antibiotic treatment . Since then he has been followed as an outpatient for over 7 years while taking 400 mg of Erythromycin per day, and he has had no exacerbation . These two cases had different clinical courses despite the facts that both had similar conditions of chronic sinusitis and appeared to be exposed to no special environmental or occupational hazards . These facts suggest that not only intrinsic factors, such as defenselessness of airways, but extrinsic factors such as viral, mycoplasmal, or bacterial infection may act together on the mechanisms of the onset and progression of diffuse panbronchiolitis.

J Gen Microbiol, 1991 Sep, 137 ( Pt 9), 2231 - 9
Molecular genetics of Pseudomonas syringae pathovar pisi: plasmid involvement in cultivar-specific incompatibility; Bavage AD et al.; A mutant (PF24) of the race 1 strain, 299A, of Pseudomonas syringae pv . pisi has been characterized in terms of its interactions with pea (Pisum sativum) cultivars . The mutant showed a changed reaction (avirulence to virulence) with a group of pea cultivars, including cvs . Belinda and Puget, previously thought to contain resistance genes R1 and R3 . Avirulence towards cv . Puget was restored by transfer of any one of five cosmid clones from a race 3 (strain 870A) gene library to a rifampicin-resistant derivative of PF24 . These observations were in agreement with a revised race-specific resistance genotype for Belinda and similar cultivars comprising a single resistance gene, R3 . An incompatible interaction was observed between strain PF24 and cvs . Vinco (postulated to harbour race-specific resistance genes R1, R2, R3 and R5) and Hurst's Greenshaft (R4 and possibly R1), indicating that the mutant retains at least one avirulence gene (A1 or A1 and A4) . Mutant PF24 showed loss of a cryptic plasmid (pAV212) compared with its progenitor, strain 299A . A subclone (pAV233) of one of the race 3 restoration clones showed strong hybridization with similar-sized digestion fragments in race 3 plasmid DNA, confirming the A3 gene to be plasmid-borne . Strong cross-hybridization was also observed with a single 3.27 kb EcoRI fragment of plasmid DNA present in strain 299A but absent from strain PF24 . This is consistent with the corresponding A3 determinant being located on pAV212 in the race 1 strain 299A . The novel avirulence gene corresponding to A3 in strain 870A is provisionally designated avrPpi3.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS Res Hum Retroviruses, 1991 Sep, 7(9), 741 - 50
Soluble CD4-PE40 is cytotoxic for a transfected mammalian cell line stably expressing the envelope protein of human immunodeficiency virus (HIV-1), and cytotoxicity is variably inhibited by the sera of HIV-1-infected patients; Pitts TW et al.; Sera were obtained from 50 individuals infected with human immunodeficiency virus type 1 or from HIV-1-uninfected individuals before or after vaccination with recombinant gp160 . These sera were evaluated for activity antagonistic to the cell-killing activity of the chimeric Pseudomonas exotoxin hybrid protein, sCD4-PE40 . For these studies, Chinese hamster ovary (CHO) cells were transfected with a chimeric plasmid encoding the tat, rev, and envelope genes of HIV-1 and a cell line was selected for stable expression of the envelope glycoproteins at the cell surface (CHO-env) . Cytotoxicity of sCD4-PE40 for CHO-env in the presence or absence of added human serum was quantitated spectrophometrically following enzymatic reduction of a tetrazolium bromide within the mitochondria of viable cells (MTT assay) . Several HIV+ sera inhibited the cytotoxic activity of sCD4-PE40; the antagonist had properties consistent with those of immunoglobulins in that it was heat stable, absorbed by protein A, and reversible by increasing the concentration of sCD4-PE40 . Of 15 HIV+ sera which strongly reacted with gp120, 11 (73%) also potently inhibited sCD4-PE40 cytotoxicity, and cytotoxicity was inhibited by sera from some HIV- individuals after, but not before, immunization with gp160 . These data suggested a role for antibody to gp120 in the antagonistic activity . However, not all sera with antibody to gp120 antagonized sCD4-PE40 cytotoxicity and high levels of antagonist activity were frequently (40%) found in HIV+ sera lacking immunoblot-detectable antibody to gp120, or antibody to either CD4 or PE40 . Grouping of the HIV+ sera according to the patients' absolute number of CD4+ cells revealed that the degree of inhibition of sCD4-PE40 cytotoxicity approached a Gaussian distribution, suggesting that persons with CD4+ cell counts between 200 and 700/mm3 may be more likely to possess significant levels of serum antagonist . This data have implications for the clinical development of sCD4-PE40 or other sCD4-based therapeutics in the management of HIV-1 infection.

Plant Mol Biol, 1991 Sep, 17(3), 409 - 13
Differential regulation in tobacco cell suspensions of genes involved in plant-bacteria interactions by pathogen-related signals; Godiard L et al.; Six cDNA clones whose corresponding mRNAs accumulate early during the hypersensitive reaction in tobacco leaves have been classified into 2 groups according to their maximum levels of accumulation in an incompatible versus a compatible interaction with Pseudomonas solanacearum . We present evidence that, at least in the first stages of the interaction, tobacco cell suspensions retain the ability to respond differentially to compatible and incompatible isolates of P . solanacearum . In addition, studies on the effect of a fungal elicitor on the accumulation of the mRNAs corresponding to the cDNA clones in cell suspensions indicate that only one group of genes responds to this treatment.

Basic Res Cardiol, 1991 Sep-Oct, 86(5), 411 - 21
Mechanisms in acute septic cardiomyopathy: evidence from isolated myocytes; Werdan K et al.; Although often not considered, the heart is one of the targets of multiple organ failure in sepsis and septic shock, with myocardial depression being a prominent component of this "acute septic cardiomyopathy" . Hypotheses concerning the etiology of this depression are increasingly elucidated on a cellular level, including dysfunction of the beta-adrenoceptor/G protein/adenylate cyclase system, calcium channel blockade by cardiodepressant factor, contractile impairment by activated leucocytes, as well as inhibition of protein synthesis by Pseudomonas exotoxin A . In the search for "mechanisms of myocardial depression in sepsis", isolated cardiomyocytes may play a role as research tools with respect to: a) discrimination between direct and indirect cardiodepressant effects; b) identifying not only the acute, but also chronic toxin- and mediator-induced cardiodepression; c) clarification of the mechanism of action of cardiodepressant bacterial toxins and sepsis mediators; d) establishment of in vitro models of leucocyte-mediated cardiodepression in sepsis.

Appl Environ Microbiol, 1991 Sep, 57(9), 2497 - 501
Effect of sodium chloride on transport of bacteria in a saturated aquifer material; Gannon J et al.; Determinations were made of the influence of NaCl concentration, cell density, and flow velocity on the transport of Pseudomonas sp . strain KL2 through columns of aquifer sand under saturated conditions . A pulse-type boundary condition was used . The experiments were conducted by using 0.3-m-long Plexiglas columns with an internal diameter of 0.05 m . When a 1-h pulse of a 0.01 M NaCl solution containing 10(8) cells per ml was added at a flow rate of 10(-4) m s-1, the bacterial density in the effluent never exceeded 2.2% of the density of cells added, and only 1.5% of the bacteria passed through the aquifer material . In contrast, when the bacteria were applied in distilled water, the relative cell density in the effluent approached 100%, and 60% of the bacteria were transported through the aquifer solids . Under these conditions, the breakthrough of Pseudomonas sp . strain KL2 was slower than chloride . When the flow rate was 2.0 x 10(-4) m s-1, the cell density in the effluent reached 7.3% of that added in 0.01 M NaCl solution, but only 3.9% of the bacteria were transported through the aquifer particles . On the other hand, the density in the effluent approached 100% of that added in deionized water, and 77% of the added bacteria were recovered . When the density of added cells was 10(9) cells per ml at a flow rate of 10(-4) m s-1, the densities in the effluent reached 70 and 100% of those added in salt solution and deionized water, respectively, and 44 and 57% of the bacteria were transported through the aquifer solids.(ABSTRACT TRUNCATED AT 250 WORDS)

Agric Biol Chem, 1991 Sep, 55(9), 2349 - 57
Cloning, nucleotide sequencing, and expression in Escherichia coli of a lipase and its activator genes from Pseudomonas sp . KWI-56; Iizumi T et al.; A lipase gene (lip) and its activator gene (act) on a 2.9 kb BglII-EcoRI fragment from Pseudomonas sp . KWI-56 were cloned in Escherichia coli using pUC19 as a vector plasmid . From the sequencing results, the open reading frames of the lip and the act were found to contain 1092 and 1032 nucleotides, respectively . The act existed downstream of the lip with the same orientation . When the lip was expressed in E . coli using the lac promoter on the pUC plasmid vector, the lipase activity of E . coli carrying both the lip and the act was 200-fold greater than that carrying only the lip . This result suggested the act was important in the expression of the lip in E . coli.

Gene, 1991 Aug 30, 105(1), 43 - 9
Molecular cloning and expression of the beta-hydroxysteroid dehydrogenase gene from Pseudomonas testosteroni; Genti-Raimondi S et al.; The structural gene (hsd) of the Pseudomonas testosteroni encoding the 17 beta-hydroxysteroid dehydrogenase has been cloned using the cosmid vector pVK102 . Escherichia coli carrying recombinant clones of hsd, isolated by immunological screening, were able to express the biologically active enzyme, as measured by the conversion of testosterone into androstenedione . Subcloning experiments, restriction and deletion analysis, and site-directed insertion mutagenesis showed that the hsd gene is located within a 1.3-kb HindIII-PstI restriction fragment . A 26.5-kDa protein encoded by a recombinant plasmid containing this Ps . testosteroni DNA restriction fragment was detected by SDS-PAGE analysis of in vitro {35S}methionine-labeled polypeptides.

J Immunol Methods, 1991 Aug 9, 141(2), 187 - 97
Isolation of a 30 kDa immunoglobulin binding protein from Pseudomonas maltophilia; Grover S et al.; We have demonstrated that Pseudomonas maltophilia (ATCC No . 13637) possesses an exposed, immunologically accessible protein which binds to the Fc region of several species of immunoglobulins . Whole bacteria suspensions were incubated for 18 h with purified 125I-labelled antibodies with and without added non-labelled immunoglobulins . The suspensions were centrifuged for 30 min and the pellet containing bacteria was assessed for radioactivity . Using this crude assay, the whole organism bound 125I-labelled rabbit and mouse immunoglobulins and the purified Fc portion of human IgG . All of these labelled preparations were competitively displaced by unlabelled rabbit and mouse immunoglobulins, and Fc of human IgG, as well as human immunoglobulin subclasses . The organism was sonicated to solubilize this immunoglobulin binding protein . Using this sonicated preparation, it was shown that unlabelled Fc of IgG, unlabelled mouse and rabbit immunoglobulins, all competitively displaced 125I-labelled human Fc of IgG in a dose-response manner . A partially purified protein was prepared by Sephacryl S-300 followed by Sephadex G-100 column chromatography . This preparation was incubated with 125I-Fc gamma and with the following purified unlabelled preparations: F(ab')2 of IgG, Fc of IgG, murine monoclonal IgA, IgG1, IgG2, IgG3, and IgG4 . All except F(ab')2 of IgG produced dose response competitive displacement . The molecular weight, as estimated by SDS-PAGE and Western blot, was 30,000 daltons . In Western blots, Fc gamma, murine monoclonal IgA, and human immunoglobulin subclasses, all showed affinity for the immobilized protein . Human F(ab')2 fragments did not show affinity for the protein . Radioiodinated pseudomonal Ig-binding protein showed affinity for human IgG coupled to Sepharose, and was displaced by unlabelled pseudomonal Ig-binding protein . Scatchard analysis of binding showed two binding affinities: two distinct types of Ig-binding proteins were obtained, a high affinity with Kd = 1.54 x 10(-10) and a lower affinity with Kd = 2.36 x 10(-8) . This immunoglobulin binding protein may be useful in immunoglobulin purification or identification.

Mol Gen Genet, 1991 Aug, 228(1-2), 294 - 9
Novel mercury resistance determinants carried by IncJ plasmids pMERPH and R391; Peters SE et al.; HgCl2 resistance (Hgr) in a strain of Pseudomonas putrefaciens isolated from the River Mersey was identified as plasmid-borne by its transfer to Escherichia coli in conjugative matings . This plasmid, pMERPH, could not be isolated and was incompatible with the chromosomally integrated IncJ Hgr plasmid R391 . pMERPH and R391 both express inducible, narrow-spectrum mercury resistance and detoxify HgCl2 by volatilization . The cloned mer determinants from pMERPH (pSP100) and R391 (pSP200) have very similar restriction maps and express identical polypeptide products . However, these features show distinct differences from those of the Tn501 family of mer determinants . pSP100 and pSP200 failed to hybridize at moderate stringency to merRTPA and merC probes from Tn501 and Tn21, respectively . We conclude that the IncJ mer determinants are only distantly related to that from Tn501 and its closely homologous relatives and that it identifies a novel sequence which is relatively rare in bacteria isolated from natural environments.

Circulation, 1991 Aug, 84(2), 778 - 87
Cytotoxic effects of a recombinant chimeric toxin on rapidly proliferating vascular smooth muscle cells; Epstein SE et al.; BACKGROUND . Restenosis after percutaneous transluminal coronary angioplasty is associated with activation of medial smooth muscle cells (SMCs); they proliferate, migrate to the subintima, and narrow the vessel lumen . Cancer cells often express more cell surface receptors than do normal cells . This has allowed tumor cells to be specifically targeted using cytotoxic agents . We have examined whether a similar concept can be applied to rapidly proliferating but nontransformed SMCs . Pseudomonas exotoxin (PE; MW, 66 kDa) is a potent toxin that kills cells by inhibiting protein synthesis; its toxicity is diminished when its cell recognition domain is deleted to produce a 40-kDa protein (PE40) . METHODS AND RESULTS . A complementary DNA encoding transforming growth factor alpha (TGF alpha) was ligated to that encoding PE40 and the chimeric toxin TGF alpha-PE40, which is cytotoxic to cancer cells displaying epidermal growth factor (EGF) receptors, was expressed in Escherichia coli . The ability of this toxin to kill proliferating SMCs was tested . When cells were seeded at low density (2,500 cells/cm2) and grown in medium supplemented with 10% fetal bovine serum, they were found to be rapidly proliferating; these cells were very sensitive to the cytotoxic effects of TGF alpha-PE40 (ID50, 4.0 +/- 0.17 ng/ml) . In contrast, cytotoxicity was 30-fold less (ID50, 125 +/- 23 ng/ml; p less than 0.0004) when cells were in a quiescent state (grown in medium supplemented with 0.5% fetal bovine serum) . CONCLUSIONS . Competition studies using excess EGF indicated that the cytotoxic effects of TGF alpha-PE40 are specifically mediated by the EGF receptor . EGF receptor binding analysis demonstrated that rapidly proliferating SMCs display 10-fold more EGF receptors than do quiescent SMCs in vitro . Thus, a chimeric toxin targeted toward the EGF receptor can selectively kill rapidly proliferating SMCs . Whether this toxin or other chimeric toxins directed against other cell surface receptors will effectively inhibit SMCs proliferating in vivo or be useful in preventing restenosis remains to be determined.

J Bacteriol, 1991 Aug, 173(15), 4836 - 41
Extracellular lipase of Pseudomonas sp . strain ATCC 21808: purification, characterization, crystallization, and preliminary X-ray diffraction data; Kordel M et al.; A procedure for the purification of a very hydrophobic lipase from Pseudomonas sp . strain ATCC 21808 was elaborated by avoiding the use of long-chain detergents in view of subsequent crystallization of the enzyme . The purification procedure included chromatography on Q-Sepharose in the presence of n-octyl-beta-D-glucopyranoside, Ca2+ precipitation of fatty acids, and Octyl-Sepharose chromatography . The enzyme was purified 260-fold to a yield of 35% and a specific activity of 3,300 U/mg . The molecular weight was determined as 35,000; a polyacrylamide gel under nondenaturing conditions revealed a band at 110,000, and the isoelectric point proved to be at 4.5 to 4.6 . The lipase crystallized with different salts and ethylene glycol polymers in the presence of n-octyl-beta-D-glucopyranoside and one alkyloligooxyethylene compound (CxEy) in the range from C5E2 to C8E4 . The crystals diffract to a resolution of about 0.25 nm . Precession photographs revealed that they belong to space group C2 with lattice constants of a = 9.27 nm, b = 4.74 nm, c = 8.65 nm, and beta = 122.3 degrees, indicating a cell content of one molecule per asymmetric unit of the crystal . In hydrolysis of triglycerides, the lipase showed substrate specificity for saturated fatty acids from C6 to C12 and unsaturated long-chain fatty acids . Monoglycerides were hydrolyzed very slowly . The N-terminal sequence is identical to that of the lipase from Pseudomonas cepacia . Treatment with diethyl-p-nitrophenylphosphate affected the activities toward triolein and p-nitrophenylacetate to the same extent and with the same velocity.

J Bacteriol, 1991 Aug, 173(15), 4587 - 94
Genetic organization and regulation of a meta cleavage pathway for catechols produced from catabolism of toluene, benzene, phenol, and cresols by Pseudomonas pickettii PKO1; Kukor JJ et al.; Plasmid pRO1957 contains a 26.5-kb BamHI restriction endonuclease-cleaved DNA fragment cloned from the chromosome of Pseudomonas pickettii PKO1 that allows P . aeruginosa PAO1c to grow on toluene, benzene, phenol, or m-cresol as the sole carbon source . The genes encoding enzymes for meta cleavage of catechol or 3-methylcatechol, derived from catabolism of these substrates, were subcloned from pRO1957 and were shown to be organized into a single operon with the promoter proximal to tbuE . Deletion and analysis of subclones demonstrated that the order of genes in the meta cleavage operon was tbuEFGKIHJ, which encoded catechol 2,3-dioxygenase, 2-hydroxymuconate semialdehyde hydrolase, 2-hydroxymuconate semialdehyde dehydrogenase, 4-hydroxy-2-oxovalerate aldolase, 4-oxalocrotonate decarboxylase, 4-oxalocrotonate isomerase, and 2-hydroxypent-2,4-dienoate hydratase, respectively . The regulatory gene for the tbuEFGKIHJ operon, designated tbuS, was subcloned into vector plasmid pRO2317 from pRO1957 as a 1.3-kb PstI fragment, designated pRO2345 . When tbuS was not present, meta pathway enzyme expression was partially derepressed, but these activity levels could not be fully induced . However, when tbuS was present in trans with tbuEFGKIHJ, meta pathway enzymes were repressed in the absence of an effector and were fully induced when an effector was present . This behavior suggests that the gene product of tbuS acts as both a repressor and an activator . Phenol and m-cresol were inducers of meta pathway enzymatic activity . Catechol, 3-methylcatechol, 4-methylcatechol, o-cresol, and p-cresol were not inducers but could be metabolized by cells previously induced by phenol or m-cresol.

Cancer Res, 1991 Aug 1, 51(15), 4001 - 7
Therapy of human cervical carcinoma with monoclonal antibody-Pseudomonas exotoxin conjugates; Roffler SR et al.; Pseudomonas exotoxin A (PE) linked to the F(ab')2 fragment of 1H10, a murine monoclonal antibody recognizing a carbohydrate epitope of a glycoconjugate expressed on the surface of human cervical carcinoma tumor cells, was evaluated for in vitro and in vivo activity . PE can kill cells by ADP-ribosylating elongation factor 2 thus inhibiting protein synthesis . Disulfide- as well as thioether-linked immunotoxins (1H10-PE) killed cervical carcinoma cells in vitro and were 20-160 times more inhibitory to target than to control cells . Cell killing was antibody mediated as demonstrated by the reduction of 1H10-PE growth inhibition to target CaSki cells by free 1H10 F(ab')2 . In addition, a control antibody immunotoxin was nontoxic to CaSki cells . Thioether-linked 1H10-PE administered either i.v . or i.p . suppressed the growth of established solid s.c . cervical carcinoma tumors xenografted in nude mice for over 30 days . Treatment with antibody alone or a control immunotoxin had no significant effect on tumor growth . Administration of immunotoxin i.p . was associated with less toxicity than administration i.v., but i.v . injections were more effective at suppressing the growth of established solid tumors.

J Biochem (Tokyo), 1991 Aug, 110(2), 169 - 72
A resonance Raman study on a reaction intermediate of Pseudomonas L-phenylalanine oxidase (deaminating and decarboxylating); Suzuki H et al.; Resonance Raman (RR) spectra of purple intermediates of L-phenylalanine oxidase (PAO) with non-labeled and isotopically labeled phenylalanines as substrates, i.e., {1-13C}, {2-13C}, {ring-U-13C6}, and {15N}phenylalanines, were measured with excitation at 632.8 nm within the broad absorption band around 540 nm . The spectra obtained resemble those of purple intermediates of D-amino acid oxidase (DAO) . The isotope effects on the 1,665 cm-1 band with {15N} or {2-13C}phenylalanine indicate that the band is due to the C = N stretching mode of an imino acid derived from phenylalanine, i.e., alpha-imino-beta-phenylpropionate . The intense band at 1,389 cm-1 is contributed to by the CO2- symmetric stretching and C-CO2- stretching modes of alpha-imino-beta-phenylpropionate . The 1,602 cm-1 band, which does not shift upon isotopic substitution of phenylalanine, corresponds to the 1,605 cm-1 band of DAO purple intermediates and was assigned to a vibrational mode associated with the C(10a) = C(4a) - C(4) = O moiety of reduced flavin . These results confirm that PAO purple intermediates consist of the reduced enzyme and an imino acid derived from a substrate, and suggest that the plane defined by C(10a) = C(4a) - C(4) = O of reduced flavin and the plane containing H2+N = C - CO2- of an imino acid are arranged in close contact to each other, generating a charge-transfer interaction.

Appl Environ Microbiol, 1991 Aug, 57(8), 2420 - 5
Distribution, clearance, and mortality of environmental pseudomonads in mice upon intranasal exposure; George SE et al.; When introduced intranasally, P . cepacia AC1100 (approximately 10(8) CFU/animal) and P . aeruginosa AC869 (approximately 10(3) CFU/animal) were readily cleared from the mouse . However, a approximately 10(7)-CFU dose of AC869 persisted for 14 days . Strain AC869 had a 50% lethal dose of 2.7 x 10(7) CFU . Slight morbidity occurred in animals treated with approximately 10(7) CFU of AC869 or approximately 10(8) CFU of AC1100.

Postgrad Med J, 1991 Aug, 67(790), 764 - 6
Melioidosis in a patient from Bangladesh; Kibbler CC et al.; A 54 year old Bangladeshi man presented with a history and chest X-ray appearances suggestive of pulmonary tuberculosis . Following deterioration 4 weeks later, he required ventilation . Although a blood culture isolate was subsequently found to be Pseudomonas pseudomallei, it was initially misidentified and dismissed as a contaminant . Further cultures demonstrated the organism, but the patient died, despite treatment with ceftazidime . The case illustrates the importance of taking a detailed travel history and having a high index of suspicion in patients from South East Asia and the Indian sub-continent, including Bangladesh, where the disease has not previously been considered endemic.

J Gen Microbiol, 1991 Aug, 137 ( Pt 8), 2025 - 32
Bacterial metabolism of 3-chloroacrylic acid; Hartmans S et al.; Two bacterial strains were isolated with 3-chloroacrylic acid (CAA) as sole source of carbon and energy . Strain CAA1, a Pseudomonas cepacia sp., was capable of growth with only the cis-isomer of CAA . Strain CAA2, a coryneform bacterium, utilized both isomers of CAA as sole source of carbon and energy . Strain CAA1 contained cis-CAA hydratase and strain CAA2 contained two hydratases, one with cis-CAA hydratase activity and one with trans-CAA hydratase activity . The product of the hydratase activities with CAA was malonate semialdehyde . In both strains malonate semialdehyde was subsequently decarboxylated by a cofactor-independent decarboxylase yielding acetaldehyde and CO2.

Antimicrob Agents Chemother, 1991 Aug, 35(8), 1635 - 40
Pseudomonas pseudomallei resistance to beta-lactam antibiotics due to alterations in the chromosomally encoded beta-lactamase; Godfrey AJ et al.; Pseudomonas pseudomallei, the causative agent of melioidosis, is generally susceptible to some of the newer extended-spectrum cephalosporins or to combinations of a beta-lactam and clavulanic acid, a beta-lactamase inhibitor . Resistance to these agents may, however, emerge during treatment . We report on alterations in the chromosomal beta-lactamase associated with the development of resistance . Three resistance patterns resulted from three different mechanisms in the strains investigated . Derepression of the chromosomal enzyme resulted in a general increase in the MICs of all of the beta-lactams tested . The second mechanism observed was an insensitivity to inhibition of the beta-lactamase by clavulanic acid . In this case, the level of susceptibility to beta-lactams as independent entities remained unchanged . The final "resistance" pattern occurred in a patient treated with ceftazidime and resulted in a beta-lactamase that was capable of hydrolyzing this antibiotic at detectable levels, but with reduced efficacy against other beta-lactams . The net result was a strain that was generally susceptible to all of the beta-lactams tested except ceftazidime . In all cases, the level of susceptibility to antibiotics other than beta-lactams remained unchanged . Such variability found within one genus over a relatively short time course suggests that treatment of infections caused by this organism should be carefully monitored to detect susceptibility alterations to the chosen therapy.

Mol Gen Genet, 1991 Aug, 228(1-2), 1 - 8
Regulation of iron assimilation: nucleotide sequence analysis of an iron-regulated promoter from a fluorescent pseudomonad; O'Sullivan DJ et al.; An iron-regulated promoter was cloned on a 2.1 kb Bg/II fragment from Pseudomonas sp . strain M114 and fused to the lacZ reporter gene . Iron-regulated lacZ expression from the resulting construct (pSP1) in strain M114 was mediated via the Fur-like repressor which also regulates siderophore production in this strain . A 390 bp StuI-PstI internal fragment contained the necessary information for iron-regulated promoter expression . This fragment was sequenced and the initiation point for transcription was determined by primer extension analysis . The region directly upstream of the transcription start point contained no significant homology to known promoter consensus sequences . However the -16 to -25 bp region contained homology to four other iron-regulated pseudomonad promoters . Deletion of bases downstream from the transcriptional start did not affect the iron-regulated expression of the promoter . The -37 and -43 bp regions exhibited some homology to the 19 bp Escherichia coli Fur-binding consensus sequence . When expressed in E . coli (via a cloned transacting factor from strain M114) lacZ expression from pSP1 was found to be regulated by iron . A region of greater than 77 bases but less than 131 upstream from the transcriptional start was found to be necessary for promoter activity, further suggesting that a transcriptional activator may be required for expression.

Biochemistry, 1991 Jul 30, 30(30), 7438 - 44
Limits of diffusion in the hydrolysis of substrates by the phosphotriesterase from Pseudomonas diminuta; Caldwell SR et al.; The catalytic mechanism for the enzymatic hydrolysis of a series of paraoxon analogues by the phosphotriesterase from Pseudomonas diminuta has been determined . The Bronsted plots relating the pKa of the leaving group to the observed kinetic parameters, Vmax and V/Km, are both nonlinear . This observation is consistent with a change in the rate-limiting step from chemical to physical events as the pKa of the leaving group is decreased . This conclusion is confirmed by the effects of solvent viscosity on Vmax and V/Km for the same series of analogues . The data were fitted to the scheme E k1A in equilibrium k2 EA k3----EP k7----E'P k9----E + products where EA is the enzyme-substrate complex, EP is the enzyme-product complex, E'P is the enzyme-product complex after a viscosity-independent unimolecular reaction, and the values for k1, k2, k7, and k9 are 4.1 X 10(7) M-1 s-1, 2550 s-1, 3370 s-1, and 5940 s-1, respectively . The magnitude of the chemical step, represented by k3, is dependent on the pKa of the leaving group phenol as predicted by the Bronsted equation (log k3 = beta pKa + C) where beta = -1.8 and the constant (C) = 17.7 . The magnitude of beta indicates that the transition state for substrate hydrolysis is very product-like.

Biochemistry, 1991 Jul 23, 30(29), 7154 - 9
Analysis of sequences in domain II of Pseudomonas exotoxin A which mediate translocation; Siegall CB et al.; Pseudomonas exotoxin (PE) contains 613 amino acids that are arranged into 3 structural domains . PE exerts its cell-killing effects in a series of steps initiated by binding to the cell surface and internalization into endocytic vesicles . The toxin is then cleaved within domain II near arginine-279, generating a C-terminal 37-kDa fragment that is translocated into the cytosol where it ADP-ribosylates elongation factor 2 and arrests protein synthesis . In this study, we have focused on the functions of PE which are encoded by domain II . We have used the chimeric toxin TGF alpha-PE40 to deliver the toxin's ADP-ribosylating activity to the cell cytosol . Deletion analysis revealed that sequences from 253 to 345 were essential for toxicity but sequences from 346 to 364 were dispensable . Additional point mutants were constructed which identified amino acids 339 and 343 as important residues while amino acids 344 and 345 could be altered without loss of cytotoxic activity . Our data support the idea that domain II functions by first allowing PE to be processed to a 37-kDa fragment and then key sequences such as those identified in this study mediate the translocation of ADP-ribosylation activity to the cytosol.

Proc R Soc Lond B Biol Sci, 1991 Jul 22, 245(1312), 23 - 30
The putative single-stranded DNA-binding protein of the filamentous bacteriophage, Ifl . Amino acid sequence of the protein and structure of the gene; Carne A et al.; The protein product corresponding to the gene located in the region of the coliphage Ifl genome shown to contain the code for the single-stranded DNA (ssDNA)-binding proteins of all filamentous phages so far studied has been isolated from infected bacterial cells and its amino acid sequence determined . The mature protein contains 95 amino acids (calculated molecular mass 10553 Da) . Its sequence corresponds to that predicted from the DNA sequence but lacks the initiating methionine residue . Although there is little direct sequence homology between the phage Ifl protein and the ssDNA-binding proteins of the other filamentous phages that have been studied, computer-based comparisons of various physical and structural parameters showed that the phage Ifl protein contains a domain that is closely related to domains in the coliphage T4 gene 32 protein and the Pseudomonas phage Pfl ssDNA-binding protein and suggest that the Ifl protein does have a ssDNA-binding function although we were unable to show this directly.

J Mol Biol, 1991 Jul 5, 220(1), 17 - 8
Crystallization and preliminary crystallographic analysis of carboxypeptidase G2 from Pseudomonas sp . strain RS-16; Lloyd LF et al.; Carboxypeptidase G2, a zinc metalloenzyme isolated from Pseudomonas sp . strain RS-16, which catalyses the hydrolytic cleavage of reduced and non-reduced folates to pteroates and L-glutamate, has been crystallized from polyethylene glycol (average Mr 4000) by vapour diffusion . The crystal symmetry is monoclinic C2, with unit cell dimensions a = 206 A, b = 82 A, c = 116 A and beta = 118 degrees . The molecular mass and volume of the unit cell suggest that there are two dimers of the enzyme in the asymmetric unit . The crystals diffract to at least 3.0 A and are suitable for X-ray structure analysis.

Nature, 1991 Jul 4, 352(6330), 70 - 3
Defective acidification of intracellular organelles in cystic fibrosis; Barasch J et al.; The phenotype of cystic fibrosis (CF) includes abnormalities in transepithelial transport of Cl- (refs 1-5), decreased sialylation and increased sulphation and fucosylation of glycoproteins, and lung colonization with Pseudomonas . It is not apparent how these abnormalities are interrelated, nor how they result from loss of function of the CF gene-encoded transmembrane regulator (CFTR) . We have previously shown that that the pH of a secretory granule is regulated by the vesicular conductance for Cl- (ref . 11) . Here we find defective acidification in CF cells of the trans-Golgi/trans-Golgi network, of prelysosomes and of endosomes as a result of diminished Cl- conductance . Sialytation of proteins and lipids is reduced and ligand traffic altered . These abnormalities can result from defective acidification because vacuolar pH regulates glycoprotein processing and ligand transport . The CF phenotype is similar to that of alkalinized cells and acidification-defective mutatants.

Ann Surg, 1991 Jul, 214(1), 24 - 30
Differential pathophysiology of bacterial translocation after thermal injury and sepsis; Jones WG 2nd et al.; Bacterial translocation (BT) occurs transiently after thermal injury and may result from an ischemic intestinal insult . To evaluate continued intestinal ischemia in the ongoing BT associated with sepsis after injury, rats were randomized to (1) 30% burn injury with Pseudomonas wound infection (BI), (2) BI + fluid resuscitation (BI + Fluid), (3) BI after allopurinol pretreatment to inhibit xanthine oxidase (BI + Allo), or (4) BI after azapropazone pretreatment to inhibit neutrophil degranulation (BI + Aza) . On postburn days (PBD) 1, 4, and 7, animals were studied for evidence of BT and intestinal lipid peroxidation . BI + Fluid, BI + Allo, and BI + Aza significantly (p less than 0.05) reduced rates of BT and ileal lipid peroxidation acutely after thermal injury (PBD 1) compared to BI . All four groups had equally high rates of BT associated with the onset of sepsis (PBDs 4 and 7), without evidence of further intestinal lipid peroxidation . These data indicate that the chronic gut barrier failure associated with sepsis after injury occurs independently of continued intestinal ischemia.

Rev Infect Dis, 1991 Jul-Aug, 13(4), 642 - 3
Cellulitis due to Pseudomonas putrefaciens: possible production of exotoxins; Chen SC et al.; Pseudomonas putrefaciens has been described as a rare cause of both lower-limb cellulitis and septicemic illness with significant morbidity . We report a case of P . putrefaciens infection in a patient with refractory lower-limb cellulitis and ulceration complicated by thrombocytopenia, hypotension, and mental obtundation in the apparent absence of bacteremia . This scenario raises the possibility of significant production of exotoxins by P . putrefaciens in vivo.

J Appl Physiol, 1991 Jul, 71(1), 317 - 21
Inhibition of pulmonary surfactant function by phospholipases; Holm BA et al.; Previous studies have shown that respiratory failure associated with disorders such as acute pancreatitis correlates well with increased levels of phospholipase A2 (PLA2) in lung lavages and that intratracheal administration of PLA2 generates an acute lung injury . In addition, bacteria such as Pseudomonas have been shown to secrete phospholipase C (PLC) . We studied the effects of these phospholipases on pulmonary surfactant activity using a pulsating bubble surfactometer . Concentrations greater than or equal to 0.1 unit/ml PLA2 destroyed surfactant biophysical activity, increasing surface tension at minimum bubble size from less than 1 to 15 mN/m . This surfactant inactivation was predominantly related to the effect of lysophosphatidylcholine on the surface film, although the fatty acids released with higher PLA2 concentrations also had a detrimental effect on surfactant function . Similarly, as little as 0.1 unit PLC increased the surface tension at minimal size of an oscillating bubble from less than 1 to 15 mN/m, an effect that could be mimicked by the addition of dipalmitin to surfactant in the absence of PLC . Moreover, lower, noninhibitory concentrations (0.01 unit/ml) of PLA2 and PLC increased the sensitivity of surfactant to other inhibitory agents, such as albumin . Thus, relatively low concentrations of PLC and PLA2 can cause severe breakdown of surfactant function and may contribute significantly to some forms of lung injury.

J Bacteriol, 1991 Jul, 173(13), 4007 - 12
In vitro deletion mapping of the viral strand replication origin of Pseudomonas bacteriophage Pf3; Luiten RG et al.; The origin of viral strand replication of the filamentous bacteriophage Pf3 has been characterized in Escherichia coli by in vitro deletion mapping techniques . The origin region was functionally identified by its ability to convey replicative properties to a recombinant plasmid in a polA host in which the replication origin of the vector plasmid is not functional . The origin of Pf3 viral strand replication is contained within a DNA sequence of 139 bp . This sequence covers almost completely one of the intergenic regions of the Pf3 genome, and it specifies both replication initiation and termination functions . Although no nucleotide sequence homology is present between the Pf3 origin of viral strand replication and that of the E . coli filamentous phages Ff (M13, f1, and fd) and IKe, their map positions and functional properties are very similar.

Arch Biochem Biophys, 1991 Jul, 288(1), 169 - 76
Affinity purification and characterization of 2,4-dichlorophenol hydroxylase from Pseudomonas cepacia; Radjendirane V et al.; 2,4-Dichlorophenol hydroxylase, a flavoprotein monooxygenase from Pseudomonas cepacia grown on 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole source of carbon, was purified to homogeneity by a single-step affinity chromatography on 2,4-DCP-Sepharose CL-4B . The enzyme was eluted from the affinity matrix with the substrate 2,4-dichlorophenol . The enzyme has a molecular weight of 275,000 consisting of four identical subunits of molecular weight 69,000 and requires exogenous addition of FAD for its complete catalytic activity . The enzyme required an external electron donor NADPH for hydroxylation of 2,4-dichlorophenol to 3,5-dichlorocatechol . NADPH was preferred over NADH . The enzyme had Km value of 14 microM for 2,4-dichlorophenol, and 100 microM for NADPH . The enzyme activity was significantly inhibited by heavy metal ions like Hg2+ and Zn2+ and showed marked inhibition with thiol reagents . Trichlorophenols inhibited the enzyme competitively . The hydroxylase activity decreased as a function of increasing concentrations of Cibacron blue and Procion red dyes . The apoenzyme prepared showed complete loss of FAD when monitored spectrophotometrically and had no enzymatic activity . The inactive apoenzyme was reconstituted with exogenous FAD which completely restored the enzyme activity.

Appl Environ Microbiol, 1991 Jul, 57(7), 1950 - 5
Multiplication of Legionella spp . in tap water containing Hartmannella vermiformis; Wadowsky RM et al.; A model was developed to study the multiplication of various Legionella spp . in tap water containing Hartmannella vermiformis . Tap water cultures prepared with the following components were suitable for the multiplication studies: Legionella spp., 10(3) CFU/ml; H . vermiformis, 10(4.4) cysts per ml; and killed Pseudomonas paucimobilis, 10(9) cells per ml . Cocultures were incubated at 37 degrees C for at least 1 week . The following legionellae multiplied in tap water cocultures in each replicate experiment: L . bozemanii (WIGA strain), L . dumoffii (NY-23 and TX-KL strains), L . micdadei (two environmental strains), and L . pneumophila (six environmental strains and one clinical isolate) . Growth yield values for these strains were 0.6 to 3.5 log CFU/ml . Legionellae which did not multiply in replicate cocultures included L . anisa (one strain), L . bozemanii (MI-15 strain), L . micdadei (a clinical isolate), L . longbeachae, (one strain), and L . pneumophila (Philadelphia 1 strain) . L . gormanii and an environmental isolate of L . pneumophila multiplied in only one of three experiments . None of the legionellae multiplied in tap water containing only killed P . paucimobilis . The mean growth yield (+/- standard deviation) of H . vermiformis in the cocultures was 1.2 +/- 0.1 log units/ml . H . vermiformis supports multiplication of only particular strains of legionellae, some of which are from diverse origins.

Appl Environ Microbiol, 1991 Jul, 57(7), 1935 - 41
Mutants of Pseudomonas cepacia G4 defective in catabolism of aromatic compounds and trichloroethylene; Shields MS et al.; Pseudomonas cepacia G4 possesses a novel pathway of toluene catabolism that is shown to be responsible for the degradation of trichloroethylene (TCE) . This pathway involves conversion of toluene via o-cresol to 3-methylcatechol . In order to determine the enzyme of toluene degradation that is responsible for TCE degradation, chemically induced mutants, blocked in the toluene ortho-monooxygenase (TOM) pathway of G4, were examined . Mutants of the phenotypic class designated TOM A- were all defective in their ability to oxidize toluene, o-cresol, m-cresol, and phenol, suggesting that a single enzyme is responsible for conversion of these compounds to their hydroxylated products (3-methylcatechol from toluene, o-cresol, and m-cresol and catechol from phenol) in the wild type . Mutants of this class did not degrade TCE . Two other mutant classes which were blocked in toluene catabolism, TOM B-, which lacked catechol-2,3-dioxygenase, and TOM C-, which lacked 2-hydroxy-6-oxoheptadienoic acid hydrolase activity, were fully capable of TCE degradation . Therefore, TCE degradation is directly associated with the monooxygenation capability responsible for toluene, cresol, and phenol hydroxylation.

Mol Gen Genet, 1991 Jul, 227(3), 401 - 10
The use of subtractive hybridization to obtain a DNA probe specific for Pseudomonas solanacearum race 3; Cook D et al.; Pseudomonas solanacearum, the causal agent of bacterial wilt, has been classified into three races based on host range and into five biovars based on physiological properties . Strains of race 3 belong exclusively to biovar 2 and primarily affect potatoes . Although this race is thought to have originated in the Andean highlands, it has unusual physiological properties that make it a potential threat to potatoes grown at the cooler latitudes worldwide . Consequently, there is need for a rapid and sensitive method for detection of race 3 strains . We have used subtractive hybridization to enrich for race 3-specific DNA sequences in total race 3 genomic DNA, and thereby obtained a 2 kb clone homologous to DNA from all 28 race 3 strains tested, but with only five of 90 non-race 3 strains . In addition, two larger regions of the genome, containing a minimum of 23 kb of DNA, are also specific for race 3 . Deletion of this DNA did not affect virulence . This race 3-specific DNA is a potentially useful diagnostic tool for the detection of race 3 strains.

Biochem Int, 1991 Jul, 24(5), 793 - 9
Isolation of two endo-beta-N-acetylglucosaminidases with different specificities from Pseudomonas sp; Takegawa K et al.; Two endo-beta-N-acetylglucosaminidases (PI and PII) have been isolated from the culture fluid of Pseudomonas sp . The substrate specificity of the PI enzyme was very similar to that of Endo-H from Streptomyces plicatus . On the contrary, the PII enzyme had a novel substrate specificity that degraded both high-mannose type and hybrid type oligosaccharides derived from ovalbumin, and the core structure of complex type oligosaccharides derived from human transferrin and porcine pancreatic lipase.

ASAIO Trans, 1991 Jul-Sep, 37(3), M256 - 7
Microbially infected thrombus in animals with total artificial hearts; Chiang BY et al.; In a retrospective study of 330 animals with total artificial hearts (TAH), 103 (31%) had microbially infected thrombi (MIT) . The incidence of MIT approximated 75% in the animals surviving more than 100 days . The most common pathogen isolated from animals with MIT was Pseudomonas . Most thrombi appeared to have originated from valve junctions and connectors . Methods to prevent MIT should be aimed at eliminating thrombus formation by improved design and materials and controlling the route of bacterial colonization . These findings suggest that bacterial interaction with the thrombus, device-related bacterial colonization, host immunomodulation, and gut barrier function after TAH implantation need further study.

J Bacteriol, 1991 Jul, 173(13), 4182 - 7
The major outer membrane protein of Acidovorax delafieldii is an anion-selective porin; Brunen M et al.; The major outer membrane protein (Omp34) of Acidovorax delafieldii (formerly Pseudomonas delafieldii) was purified to homogeneity and was characterized biochemically and functionally . The polypeptide has an apparent molecular weight (Mr) of 34,000, and it forms stable oligomers at pH 9.0 in the presence of 10% octylpolyoxyethylene or 2% lithium dodecyl sulfate below 70 degrees C . The intact protein has a characteristic secondary structure composition, as revealed by Fourier transforming infrared spectroscopy (about 60% beta sheet) . These features and the amino acid composition are typical for porins . The purified Omp34 is associated with 1 to 2 mol of lipopolysaccharide per mol of the monomer . Pore-forming activity was demonstrated with lipid bilayer experiments . Single-channel and selectivity measurements showed that the protein forms highly anion-selective channels . The unusual dependence of the single-channel conductance on salt concentration suggests that the porin complexes bear positive surface charges, accumulating negatively charged counterions at the pore mouth.

J Infect Dis, 1991 Jul, 164(1), 133 - 6
Ribotype stability of serial pulmonary isolates of Pseudomonas cepacia; LiPuma JJ et al.; Eighty-three isolates of Pseudomonas capacia were recovered from respiratory secretions from 12 chronically colonized cystic fibrosis patients and examined by ribotype analysis . In 9 patients, the ribotype of the cultured P . cepacia remained unchanged throughout the entire period of observation, indicating chronic pulmonary colonization with a single strain . In each of the remaining 3 patients, two genetically distinct strains were detected among serial P . cepacia isolates . No significant change in clinical condition was correlated with the change in identity of the colonizing strain . In control experiments, the stability of strain ribotype was demonstrated among isolated that had been subcultured 100 times in vitro and among isolates recovered from chronically colonized mice . These data demonstrate the utility of ribotype analysis and indicate that most chronically colonized cystic fibrosis patients harbor a single strain of P . cepacia for prolonged periods.

J Bacteriol, 1991 Jul, 173(13), 4124 - 32
Cloning and expression of the tabtoxin biosynthetic region from Pseudomonas syringae; Kinscherf TG et al.; Pseudomonas syringae BR2, a causal agent of bean wildfire, was subjected to Tn5 mutagenesis in an effort to isolate mutants unable to produce the beta-lactam antibiotic tabtoxin . Three of the tabtoxin-minus (Tox-) mutants generated appeared to have physically linked Tn5 insertions and retained their resistance to the active toxin form, tabtoxnine-beta-lactam (T beta L) . The wild-type DNA corresponding to the mutated region was cloned and found to restore the Tn5 mutants to toxin production . The use of cloned DNA from the region as hybridization probes revealed that the region is highly conserved among tabtoxin-producing pathovars of P . syringae and that the region deletes at a relatively high frequency (10(-3)/CFU) in BR2 . The Tox- deletion mutants also lost resistance to tabtoxinine-beta-lactam . A cosmid designated pRTBL823 restored toxin production and resistance to BR2 deletion mutants . This cosmid also converted the tabtoxin-naive P . syringae epiphyte Cit7 to toxin production and resistance, indicating that pRTBL823 contains a complete set of biosynthetic and resistance genes . Tox- derivatives of BR2 did not produce disease symptoms on bean . Clones that restored toxin production to both insertion and deletion mutants also restored the ability to cause disease . However, tabtoxin-producing Cit7 derivatives remained nonpathogenic on bean and tobacco, suggesting that tabtoxin production alone is not sufficient to cause disease.

Agric Biol Chem, 1991 Jul, 55(7), 1913 - 8
Molecular cloning and characterization of the fusaric acid-resistance gene from Pseudomonas cepacia; Utsumi R et al.; Fusaric acid-resistance genes (fus) were isolated from Pseudomonas cepacia . The nucleotides of the 5437 base pairs containing the fus genes were sequenced.

J Biol Chem, 1991 Jun 25, 266(18), 11705 - 13
In vivo 13C and 15N NMR studies of methylamine metabolism in Pseudomonas species MA; Jones JG et al.; Pseudomonas species MA was grown with methylamine as a sole source of carbon and nitrogen enabling the total flow of carbon and nitrogen into this organism to be simultaneously monitored in vivo using 13C and 15N NMR . {13C}Methylamine was rapidly and extensively incorporated into the methyl group of N-methylglutamate during high oxygenation of the cell suspension, but when the oxygenation rate was lower, a significant portion was also found in the methyl group of gamma-glutamylmethylamide . At later times the carbon label was found in intermediates of the serine assimilation pathway, with glutamate derived from the tricarboxylic acid cycle being the most abundant product . Incorporation of {15N}methylamine was only detected as N-methyl{15N}glutamate, but when protein synthesis was inhibited, the label was also detected in the amino nitrogen of glutamate . When oxygenation rates were lower, the 15N-labeled methylamine was found in the methylamide group of gamma-glutamylmethylamide in addition to being incorporated into N-methylglutamate . gamma-Glutamylmethylamide formation was linked to the overall energy state of the cell and was not affected by inhibition of the carbon assimilation pathway . Neither 5-hydroxy-N-methylpyroglutamate nor N-methyl-alpha-ketoglutaramate were detected to any significant extent . A mechanism was proposed for the role of gamma-glutamylmethylamide in the regulation of endogenous nitrogen supplies in this organism.

Gene, 1991 Jun 15, 102(1), 143 - 4
Nucleotide sequence of IS402 from Pseudomonas cepacia; Ferrante AA et al.; IS402, a transposable gene-activating element isolated on the basis of its ability to increase expression of the Tn1 bla gene in Pseudomonas cepacia, was cloned from pTGL52 into the vector, pBluescript KS+, and its nucleotide (nt) sequence was determined . This 914-bp element had terminal inverted repeats of 17 bp with a single mismatch, and upon insertion into Tn1 generated a direct target duplication of 3 bp . Comparison of its nt sequence with the GenBank and EMBL databases indicated that IS402 is unrelated to previously described bacterial IS elements.

J Bacteriol, 1991 Jun, 173(12), 3803 - 6
The cis-acting regulatory element of the mvaAB operon of Pseudomonas mevalonii; Wang YL et al.; DNA upstream of the transcription start site of the mvaAB operon of Pseudomonas mevalonii, which encodes 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (EC 1.1.1.88) and HMG-CoA lyase (EC 4.1.3.4), contains a cis-acting regulatory element which functions in the response to mevalonate . The regulatory element resides within a 36-bp region located from 48 to 84 bp upstream of the transcription start site of mvaA . This location was inferred from the beta-galactosidase activities of P . mevalonii harboring plasmid-encoded mvaA-lacZ fusions induced by mevalonate and by DNA gel retardation and competition assays . While protein from P . mevalonii grown on mevalonate produced a band shift, protein from cells grown on succinate gave no band shift, even when mevalonate was added . The operator contains three 10-bp direct repeats with the consensus sequence TGGGTACAGT, which may be important for regulation of the mvaAB operon.

J Bacteriol, 1991 Jun, 173(12), 3795 - 802
Purification and characterization of a 1,2-dihydroxynaphthalene dioxygenase from a bacterium that degrades naphthalenesulfonic acids; Kuhm AE et al.; 1,2-Dihydroxynaphthalene dioxygenase was purified to homogeneity from a bacterium that degrades naphthalenesulfonic acids (strain BN6) . The enzyme requires Fe2+ for maximal activity and consists of eight identical subunits with a molecular weight of about 33,000 . Analysis of the NH2-terminal amino acid sequence revealed a high degree of homology (22 of 29 amino acids) with the NH2-terminal amino acid sequence of 2,3-dihydroxybiphenyl dioxygenase from strain Pseudomonas paucimobilis Q1 . 1,2-Dihydroxynaphthalene dioxygenase from strain BN6 shows a wide substrate specificity and also cleaves 5-, 6-, and 7-hydroxy-1,2-dihydroxynaphthalene, 2,3- and 3,4-dihydroxybiphenyl, catechol, and 3-methyl- and 4-methylcatechol . Similar activities against the hydroxy-1,2-dihydroxynaphthalenes were also found in cell extracts from naphthalene-degrading bacteria.

Endocrinology, 1991 Jun, 128(6), 3096 - 104
Isolation of a 48.5-kDa membrane protein from Pseudomonas maltophilia which exhibits immunologic cross-reaction to the beta-subunit of human chorionic gonadotropin; Grover S et al.; In separate studies we have shown that Pseudomonas maltophilia (American Type Culture Collection 13637) possesses an immunoglobulin Fc-binding protein . We have found that this protein prevents the application of immunoassays using monoclonal antibodies to study possible production of a CG-like material by this bacteria . Employing an immunoglobulin saturation technique to block this protein as well as a zwitterion detergent membrane solubilization technique, we now report the isolation of a membrane protein from Pseudomonas maltophilia which shows immunological relationships to the beta-subunit and carboxyl tail of human pregnancy CG . This pseudomonas immunoreactive material produced dose-response curves in the following CG immunoassays: 1) a polyclonal rabbit anti-CG equilibrium assay, 2) carboxyl tail CG equilibrium assay, and 3) two CG equilibrium assays using monoclonal antibodies . The pseudomonas CG-like protein did not react in equilibrium assays for human TSH, human LH, or free alpha-subunit of CG . The pseudomonas CG-like protein was purified by affinity chromatography . The purified protein showed only 0.25% cross-reaction with pregnancy CG in the monoclonal antibody equilibrium assays . Furthermore, the purified protein showed no binding to rat testicular CG/LH receptors, but showed avid binding to the pseudomonas CG-binding protein previously described by Richert and Ryan . The pseudomonas protein showed no binding to Concanavalin-A, which avidly binds pregnancy CG . When assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, this protein had a mol wt of 48,500 daltons, which is larger than the mol wt of the unglycosylated beta-subunit of pregnancy CG . We conclude that pseudomonas contain a protein that has partial homology to the beta-subunit of pregnancy CG . This material does not bind to mammalian CG receptors, but does bind to a pseudomonas CG-binding site . The latter suggests it has a function, as yet unknown, in pseudomonas.

Cancer Res, 1991 Jun 1, 51(11), 3011 - 7
Expression of high-affinity interleukin 4 receptors on murine sarcoma cells and receptor-mediated cytotoxicity of tumor cells to chimeric protein between interleukin 4 and Pseudomonas exotoxin; Puri RK et al.; The presence of interleukin 4 receptor (IL-4R) on methylcholanthrene (MCA-106, MCA-102, and MC-38)- and viral DNA (G-2TS and 14-2TS)-induced murine sarcoma cells was demonstrated . MCA-106 tumor cells express about 500 to 1348 (median, 800) interleukin 4 (IL-4) binding sites/cell with a dissociation constant (Kd) of 115 +/- 26 pM (mean +/- SD, n = 4) . By Northern blot analysis, tumor cells exhibited a single mRNA species of 3.9 kilobases . Other murine sarcoma (MCA-102), colon adenocarcinoma (MC-38), G-2TS, and 14-2TS tumor cells express low numbers of IL-4R . By immunoperoxidase staining, 81 to 92% of the cells from fresh MCA-106 tumors were positive for IL-4 receptors, while only 7 to 10% of tumor-infiltrating cells were Thy 1.2 and less than 1% Mac-1 positive . Using a chimeric protein composed of IL-4 and Pseudomonas exotoxin (IL-4-PE40), we observed that IL-4-PE40 was cytotoxic (determined by inhibition of protein synthesis by {3H}leucine uptake) to MCA-106 tumor cells in a dose-dependent manner . A nonchimeric protein (PE40) that cannot bind to the IL-4R did not inhibit protein synthesis in tumor cells . A chimeric mutant protein (IL4-PE40 asp553) that can bind to IL-4 receptors but does not have the capability to inhibit protein synthesis was not cytotoxic to tumor cells . These studies strongly suggest that IL-4R on murine MCA-106 sarcoma cells is internalized when occupied by IL-4 PE40 . Furthermore, a neutralizing antibody (11B11) to IL-4 completely abolished the protein synthesis-inhibitory activity of IL-4-PE40 . G-2TS tumor cells which expressed low numbers of IL-4 receptors were not vulnerable to cytotoxicity by IL-4-PE40 . Taken together, these data suggest that IL-4 receptor may be a target for IL-4-toxin therapy.

Cancer Res, 1991 Jun 1, 51(11), 2808 - 12
Antitumor activity of a transforming growth factor alpha-Pseudomonas exotoxin fusion protein (TGF-alpha-PE40); Pai LH et al.; TGF-alpha-PE40 is a chimeric protein composed of transforming growth factor alpha (TGF-alpha) linked to a modified Pseudomonas toxin from which the cell recognition domain has been deleted (PE40) . TGF-alpha-PE40 has been shown to have cytotoxic effects on human cancer cell lines that express the epidermal growth factor (EGF) receptor on their surface, and when given i.p., it prolongs the survival of nude mice bearing i.p . tumors . Because several normal tissues, including liver, express EGF receptors on their surfaces, it has not been clear that this agent can be used systemically to treat EGF receptor-bearing tumors . In this study, we have delivered TGF-alpha-PE40 for 7 days by continuous infusion through a miniosmotic pump placed in the peritoneal cavity of nude immunodeficient mice . Two different human cancer cell lines that express EGF receptors on their surface were implanted s.c . One was A431, an epidermoid carcinoma; the other was DU-145, a prostate carcinoma . By using this mode of continuous i.p . delivery, we were able to achieve a constant serum level of TGF-alpha-PE40 that was nontoxic to the mice and yet delayed the growth of both tumors implanted s.c . and caused partial regression of one . We conclude that it is possible to deliver TGF-alpha-PE40 systemically and achieve a therapeutic serum level in mice without major toxicity . Although side effects may be expected, this study establishes that there is a therapeutic window for this agent in the therapy of cancers with high numbers of EGF receptors.

Mol Gen Genet, 1991 Jun, 227(2), 205 - 12
The indoleacetic acid-lysine synthetase gene of Pseudomonas syringae subsp . savastanoi induces developmental alterations in transgenic tobacco and potato plants; Spena A et al.; The iaaL gene of Pseudomonas syringae subsp . savastanoi encodes an indoleacetic acid-lysine synthetase that conjugates lysine to indoleacetic acid . A chimaeric gene consisting of the iaaL coding region under the control of the 35S RNA promoter from cauliflower mosaic virus (35SiaaL) has been used to test if iaaL gene expression leads to morphological alterations in tobacco and potato . Transgenic tobacco plantlets bearing this construct have been shown to synthesize IAA-{14C}lysine when fed with {14C}lysine . In late stages of development, their leaves show an increased nastic curvature (epinasty) of the petiole and midvein, a finding suggestive of an abnormal auxin metabolism . The alteration is transmitted to progeny as a dominant Mendelian trait cosegregating with the kanamycin resistance marker . Transgenic potato plants harbouring the construct are also characterized by petiole epinasty . Moreover, 35SiaaL transgenic plants have an increased internode length in potato and decreased root growth in both tobacco and potato . An increased content of IAA-conjugates in leaf blade was found to correlate with the epinastic alterations caused by iaaL gene expression in tobacco leaves . These data provide evidence that IAA conjugation is able to modulate hormone action, suggesting that the widespread endogenous auxin-conjugating activities are of physiological importance.

Ir Med J, 1991 Jun, 84(2), 48 - 51
Cystic fibrosis in adolescents and adults; Mulherin D et al.; A cystic fibrosis (CF) clinic for adults was established in 1977 . We have reviewed the data on 164 patients who attended between 1977 and 1989 . Twenty four patients had died, 11 being over 20 years after time of death . Of the 140 patients still alive, 61% were male and 53% were aged over 20 years . Only 55% were diagnosed by 1 year and 88% by 10 years . Almost all patients had respiratory symptoms and sputum culture yielded Pseudomonas species in 69% . Other respiratory problems included major haemoptysis and pneumothorax, each in 10% . We found a wide range of respiratory impairment among older patients . Among 3 patients aged over 23 years the mean (+/- S.D.) percent predicted FEV1 and FVC were 53.3% (+/- 18%) and 71.4% (+/- 20%) respectively . Mean weight in this group was 92.5% (+/- 14) of predicted . Malabsorption occurred in most patients and meconium ileus equivalent occurred in most patients and meconium ileus equivalent occurred in 34% . Other complications were clinical hepatomegaly (16%), diabetes mellitus (9%) and arthropathy (20%) . Most patients were taking continuous antibiotics by mouth (89%) and by nebuliser (48%), beta-2 agonists by inhaler (57%) and oral steroids (29%) . Almost all were taking multivitamins, pancreatic replacement therapy and multiple nutritional supplements . The number of CF "bed days" grew 12 fold since 1979 and the mean stay in hospital was double the hospital mean . The economic impact was such that over 1/4 of the annual hospital antibiotic budget was expended on CF patients.

FEMS Microbiol Lett, 1991 Jun 1, 65(1), 25 - 9
Transformation of 3-chlorodibenzofuran by Pseudomonas sp . HH69; Harms H et al.; The dibenzofuran-degrading bacterial strain Pseudomonas sp . HH69 showed high oxidative activity towards 3-chlorodibenzofuran (3CDF) . During the co-metabolic turnover of 3CDF large amounts of 4-chlorosalicylate and temporarily small amounts of salicylate were excreted . Simultaneously a yellow colour appeared due to the excretion of two polar products . Conversion of 3CDF by a mutant, derived from Pseudomonas sp . HH69 and defective in 2,3-dihydroxybiphenyl-1,2-dioxygenase led to the formation of equal quantities of 4'-chloro-2,2',3-trihydroxybiphenyl (4'CTHBP) and 4-chloro-2,2',3-trihydroxybiphenyl (4CTHBP) . Crude extracts of the wild type transformed 4'CTHBP to 4-chlorosalicylate, whilst 4CTHBP was transformed to salicylate . Hence, we propose a non-selective initial attack on both aromatic rings of 3CDF and a degradative pathway for the resulting chlorotrihydroxybiphenyls.

Cancer Res, 1991 Jun 1, 51(11), 2831 - 6
Antitumor effects of interleukin 6-Pseudomonas exotoxin chimeric molecules against the human hepatocellular carcinoma, PLC/PRF/5 in mice; Siegall CB et al.; IL6-PE40 and IL6-PE664Glu are chimeric molecules composed of interleukin 6 (IL6) fused to a truncated form (PE40) or a full-length mutated form (PE664Glu) of Pseudomonas exotoxin . Both forms of IL6-Pseudomonas exotoxin are cytotoxic to IL6 receptor-bearing tumor cell types in culture . In this report, we show that both IL6-PE40 and IL6-PE664Glu have antitumor activity against the hepatocellular carcinoma PLC/PRF/5 implanted s.c . in nude mice . The PLC/PRF/5 tumor contains about 2300 IL6 receptors per cell . IL6-PE664Glu showed improved therapeutic efficacy when released continuously for 7 days by an osmotic pump planted i.p . than when administered by multiple daily i.p . injections . Both forms of IL6 toxin exhibited a schedule-dependent antitumor effect . These results demonstrate that IL6-Pseudomonas exotoxin can suppress the growth of cancer which overexpresses cell surface IL6 receptors.

J Neuroimmunol, 1991 Jun, 32(3), 209 - 17
Chimeric cytotoxin IL2-PE40 inhibits relapsing experimental allergic encephalomyelitis; Rose JW et al.; IL2-PE40 is a chimeric protein composed of human interleukin-2 (IL2) genetically fused to a modified form of Pseudomonas exotoxin lacking the cell recognition domain . IL2-PE40 is cytotoxic for IL2 receptor-bearing lymphocytes in culture and can inhibit activation of T cells in vivo . IL2-PE40 can significantly diminish antigen-stimulated proliferation of lymphocytes sensitized to myelin basic protein . Intraperitoneal administration of IL2-PE40 not only markedly inhibits the clinical manifestations of adoptively transferred relapsing experimental allergic encephalomyelitis but also dramatically reduces both inflammation and demyelination characteristic of the disease.

South Med J, 1991 Jun, 84(6), 800 - 1
Pseudomonal osteomyelitis of the medial sesamoid bone; Rimoldi RL et al.; Because osteomyelitis may complicate puncture wounds about the first metatarsophalangeal joint, we believe sesamoid roentgenograms are mandatory . These views may show subtle demineralization, which cannot be seen on standard films . Treatment must include excision of the sesamoid and culture-directed antibiotics.

Eur J Biochem, 1991 Jun 1, 198(2), 349 - 56
Purification and characterization of the oxidase from the marine bacterium Pseudomonas nautica 617; Arnaud S et al.; The aerobic respiratory system of the hydrocarbonoclastic marine bacterium Pseudomonas nautica 617 ends with a single terminal oxidase . It is a heme-containing membranous protein which has been demonstrated only to reduce molecular oxygen to hydrogen peroxide {Denis, M., Arnaud S . & Malatesta, F . (1989) FEBS Lett . 247, 475-479} . The purification of this oxidase was achieved in a single step through by DEAE-Trisacryl chromatography . SDS/PAGE showed the presence of four subunits . The pI was found to be 4.45 and a Mr of 130,000 was determined by gel filtration . The amino acid composition of the purified terminal oxidase has been determined . About 52% of the residues are hydrophobic, strengthening the membranous nature of this bacterial oxidase . Room temperature optical spectra are typical of heme b with a 560-nm band for the reduced form in the alpha range . The prosthetic group is made of two hemes b, one high-spin (S = 5/2, gl = 5.9, g parallel approximately 2.0), the other low-spin (S = 1/2, gz = 2.94, gy = 2.27) . No other metal centre was detected by EPR . The two hemes remained unresolved in optical spectra, even at low temperature, and throughout redox titration . They behaved potentiometrically like a one-electron, single redox couple, with Em = 87 +/- 10 mV at pH 7.2 and 293 K . The purified oxidase did not oxidize ferrocytochrome c, but displayed quinol oxidase activity both with the native quinone (2419 nmol O2.min-1.mg protein-1 and commercially available coenzyme (101.74 nmol O2.min-1.mg protein-1) . Exposure of the reduced enzyme to CO induced the collapse of alpha and beta bands as occurred during reoxidation . In contrast, NaCN and NaN3 fully inhibited the oxidase activity . Results are discussed with respect to other purified quinol oxidases.

Biochemistry, 1991 May 21, 30(20), 4991 - 7
Catalytic mechanism of an active-site mutant (D38N) of delta 5-3-ketosteroid isomerase . Direct spectroscopic evidence for dienol intermediates; Xue LA et al.; The delta 5-3-ketosteroid isomerase (EC 5.3.3.1) of Pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereospecific transfer of the 4 beta-proton to the 6 beta-position . The reaction involves two steps: (a) a rate-limiting concerted enolization, comprising protonation of the 3-carbonyl oxygen by the phenolic hydroxyl group of Tyr-14 and abstraction of the 4 beta-proton by the carboxylate group of Asp-38, and (b) rapid reketonization of the dienol, which may or may not be concerted . The active-site mutant D38N, which lacks the base responsible for proton transfer, is about 10(6.0)-fold less active catalytically than the wild-type enzyme . With the D38N mutant it was demonstrated spectroscopically that the enzymatic reaction involves the conversion of the substrate to both the dienol and its anion as tightly enzyme-bound intermediates, which are then converted much more slowly to the alpha,beta-unsaturated product . In contrast to the mechanism of the wild-type enzyme, the enolization reaction promoted by the D38N mutant is not stereospecific with respect to removal of the 4 beta-proton and shows primary kinetic isotope effects on enolization when either 4 alpha or 4 beta or both of these protons are replaced by deuterium . Kinetic isotope effects obtained with deuterated substrates, solvent, or combinations of the two indicate that, unlike in the wild-type enzyme, protonation of the carbonyl oxygen and removal of the C-4 proton are not concerted in the D38N mutant.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochem Biophys Res Commun, 1991 May 15, 176(3), 1106 - 11
Dehalogenation of 4-chlorobenzoate by 4-chlorobenzoate dehalogenase from pseudomonas sp . CBS3: an ATP/coenzyme A dependent reaction; Loffler F et al.; Pseudomonas sp . CBS3 was grown with 4-chlorobenzoate as sole source of carbon and energy . Freshly prepared cell-free extracts converted 4-chlorobenzoate to 4-hydroxybenzoate . After storage for 16 hours at 25 degrees C only about 50% of the initial activity was left . Treatment at 55 degrees C for 10 minutes, dialysis or desalting of the extracts by gel filtration caused a total loss of the activity of the 4-chlorobenzoate dehalogenase . The activity could be restored by the addition of ATP, coenzyme A and Mg2+ . If one of these cofactors was missing, no dehalogenating activity was detectable . The amount of 4-hydroxybenzoate formed was proportional to the amount of ATP available in the test system whereas CoA served as a real coenzyme . A novel ATP/coenzyme A dependent reaction mechanism for the dehalogenation of 4-chlorobenzoate by 4-chlorobenzoate dehalogenase from Pseudomonas sp . CBS3 is proposed.

J Biol Chem, 1991 May 15, 266(14), 8835 - 55
The substrate specificity of Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase . Analysis of myristic acid analogs containing oxygen, sulfur, double bonds, triple bonds, and/or an aromatic residue; Kishore NS et al.; We have explored the acyl-CoA substrate specificity of Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase (NMT) by synthesizing 81 fatty acid analogs and surveying their activity in a coupled in vitro assay containing Pseudomonas acyl-CoA synthetase and Escherichia coli-derived yeast NMT . Single oxygen or sulfur substitution for C-3 through C-13 is well tolerated by both enzymes . Detailed kinetic analyses suggest that the acyl-CoA and peptide-binding sites of NMT are relatively insensitive to placement of single group 6B heteroatoms . By contrast, di-oxygen-substituted analogs were very poor substrates, producing dramatic reductions in the affinity of NMTs peptide-binding site for a synthetic octapeptide substrate derived from the NH2-terminal sequence of a known N-myristoylprotein, the gag poly-protein precursor of human immunodeficiency virus 1 (HIV-1) . This observation provides an example of binding site cooperativity in NMT . Replacement of one oxygen with sulfur at either the 6, 9, or 12 position of dioxatetradecanoic acids results in a general increase in peptide catalytic efficiency (Vmax/Km) . An analysis of five fatty acids from octanoic to dodecanoic having terminal phenyl groups indicated that the best substrate was 10-phenyldecanoic acid even though Corey-Pauling-Koltun molecular models indicate that it has a length equivalent to that of tridecanoic acid . Six analogs having an equivalent length of 13 carbon atoms were subsequently prepared in which the phenyl group was systematically moved one methylene group closer to carboxyl . Movement of the phenyl just one carbon closer to carboxyl (producing 9-(p-methylphenyl) nonanoic acid) decreases peptide catalytic efficiency (Vmax/Km) severalfold compared to 10-phenyldecanoic acid . 10-(4-Tolyl)decanoic acid has the same relative positions of phenyl and carboxyl as 10-phenyldecanoic acid even though a methyl group is present on the phenyl ring . It produces peptide Km and Vmax values that are the same as 10-phenyldecanoic acid . Substitution of either oxygen or sulfur for a methylene group fails to override the effects noted when the phenyl group position is altered in the C-14 equivalent fatty acid series . Several fatty acids of differing chain lengths with cyclohexyl-, 2-furyl, and 2-thienyl groups at their omega termnius had activity profiles that paralleled those of the comparable phenyl-substituted compounds . Myristic acid analogs with triple bonds (beginning at positions 2 through 13), cis-double bonds (positions 3 through 13) and trans-double bond isomers (E5, E6, and E7) were also tested.(ABSTRACT TRUNCATED AT 400 WORDS)

J Biol Chem, 1991 May 5, 266(13), 8312 - 21
High heterogeneity of the exopolysaccharides of Pseudomonas solanacearum strain GMI 1000 and the complete structure of the major polysaccharide; Orgambide G et al.; The exopolysaccharide of Pseudomonas solanacearum, which is believed to play an important role in bacterial virulence, was considered by most authors as a homogeneous entity essentially composed of N-acetylgalactosamine . The present work demonstrates the high degree of heterogeneity of this exopolysaccharidic material, which consists of a high molecular weight acidic polysaccharide and a mainly noncarbohydrate structure as major subfractions . Rhamnose-rich polyoside and glucan fractions are also present as minor components . We report the complete structure of the acidic heteropolymer involving, in addition to N-acetylgalactosamine, equimolar ratios of two rare amino sugars, 2-N-acetyl-2-deoxy-L-galacturonic acid and 2-N-acetyl-4-N-(3-hydroxybutanoyl)-2,4,6-trideoxy-D-glucose . The structure of this acidic exopolysaccharide provides the first precise basis for the analysis of the correlation exopolysaccharide structure with pathogenicity in P . solanacearum.

Cornea, 1991 May, 10(3), 268 - 71
Topical norfloxacin therapy in Pseudomonas corneal ulceration; Vajpayee RB et al.; We treated 12 eyes with Pseudomonas ulcerative keratitis with topical 0.3% Norfloxacin drops (Ranbaxy Laboratories) . Excellent response was achieved in all eyes, resulting in complete ulcer resolution . The healing time varied from 12 to 20 days . There was a marked improvement over the pretreatment visual acuity in 11 of the 12 eyes . No systemic or ocular side effects were observed during the study period . The present study strongly suggests that topical norfloxacin is a potent and effective drug in the treatment of Pseudomonas ulcerative keratitis.

J Bacteriol, 1991 May, 173(9), 3017 - 20
Separation and partial characterization of the enzymes of the toluene-4-monooxygenase catabolic pathway in Pseudomonas mendocina KR1; Whited GM et al.; The route of toluene degradation by Pseudomonas mendocina KR1 was studied by separating or purifying from toluene-grown cells the catabolic enzymes responsible for oxidation of p-cresol through the ring cleavage step . Enzymatic transformations corresponding to each of the metabolic steps in the proposed degradative pathway were conducted with cell-free preparations . p-Cresol was metabolized by the enzyme p-cresol methylhydroxylase to p-hydroxybenzaldehyde . p-Hydroxybenzaldehyde was further oxidized by partially purified enzyme preparations to p-hydroxybenzoate and subsequently hydroxylated to form protocatechuate . Protocatechuate was then oxidized by ortho ring cleavage.

J Bacteriol, 1991 May, 173(9), 3010 - 6
Toluene-4-monooxygenase, a three-component enzyme system that catalyzes the oxidation of toluene to p-cresol in Pseudomonas mendocina KR1; Whited GM et al.; Pseudomonas mendocina KR1 grows on toluene as a sole carbon and energy source . A multicomponent oxygenase was partially purified from toluene-grown cells and separated into three protein components . The reconstituted enzyme system, in the presence of NADH and Fe2+, oxidized toluene to p-cresol as the first detectable product . Experiments with p-deutero-toluene led to the isolation of p-cresol which retained 68% of the deuterium initially present in the parent molecule . When the reconstituted enzyme system was incubated with toluene in the presence of 18O2, the oxygen in p-cresol was shown to be derived from molecular oxygen . The results demonstrate that P . mendocina KR1 initiates degradation of toluene by a multicomponent enzyme system which has been designated toluene-4-monooxygenase.

Mol Plant Microbe Interact, 1991 May-Jun, 4(3), 284 - 92
Differential expression of tomato proteinase inhibitor I and II genes during bacterial pathogen invasion and wounding; Pautot V et al.; Expression of proteinase inhibitor I and II genes was investigated during infection by Pseudomonas syringae pv . tomato, the causal agent of bacterial speck disease in tomato . Inoculation of leaves with P . s . pv . tomato of two inbred tomato lines that are resistant and susceptible to the pathogen resulted in the accumulation of proteinase inhibitor I and II mRNAs in this organ . Our data showed that in the lines used in this study, proteinase inhibitor II mRNAs accumulated in leaves to higher levels than proteinase inhibitor I mRNA in response to P . s . pv . tomato infection and wounding . Proteinase inhibitor II mRNAs accumulated more rapidly in disease-resistant than in disease-susceptible plants . Proteinase inhibitor I mRNAs were first detected in the disease-susceptible line during infection and wounding . In contrast to wounding, the systemic induction of these genes during pathogen ingression was limited . These data show that the plant proteinase inhibitors constitute one of the components of the plant defense system that are induced in response to bacterial pathogen invasion.

Vet Microbiol, 1991 May, 27(3-4), 277 - 82
Antibody to Pseudomonas pseudomallei exotoxin in sheep exposed to natural infection; Ismail G et al.; Specific antibody to Pseudomonas pseudomallei exotoxin was detected in sheep sera exposed to natural infection . An enzyme-linked immunosorbent assay (ELISA) was used . Serum antitoxin was present in 49.3% of sera obtained from a flock of sheep naturally exposed to P . pseudomallei infection . Among these sera, 17.0% gave titers of 10,000 . In contrast, serum antitoxin was present in only 6.0% of sera collected from sheep kept on a melioidosis-free farm . The ELISA reactivity of all positive sera could be completely absorbed with purified P . pseudomallei exotoxin . Similarly, preincubation of the exotoxin-coated wells with specific antiserum inhibited the ELISA reactivity of sheep sera . The results indicate that exotoxin is produced in vivo during infection by P . pseudomallei.

Br Vet J, 1991 May-Jun, 147(3), 256 - 69
Pyaemia in pigs; Chiew KT et al.; Pyaemia is by far the most important cause of condemnation in pigs slaughtered in Singapore abattoirs . Between 1983 and 1987, 1757 from a total of 4,899,731 pigs were condemned by meat inspectors for pyaemia, accounting for 0.036% of the total condemnations . The common post-mortem findings of affected pigs during the 5-year period are presented . Abscesses were most commonly seen in the liver (22%), spleen (21%), gastrohepatic lymph node (20%) and bronchial lymph node (17%) . Pseudomonas pseudomallei was the most predominant organism isolated, accounting for 39% of the pyaemic cases . The public health significance of abscesses in pigs is discussed with particular reference to melioidosis.

J Virol, 1991 May, 65(5), 2332 - 9
A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2; Watson DG et al.; RPE.40, a mutant CHO-K1 strain selected for resistance to Pseudomonas exotoxin A, is defective in the production of infectious alphaviruses, although viruses are taken in and processed normally (J . M . Moehring and T . J . Moehring, Infect . Immun . 41:998-1009, 1983) . To determine the cause of this defect, the synthesis of Sindbis virus proteins was examined . RPE.40 cells produced and glycosylated structural glycoprotein precursors PE2 and immature E1 normally . Mature E1 was formed, but PE2 was not cleaved to E2 and E3 . PE2 instead was modified to a higher-molecular-weight form (PE2') in which the high-mannose oligosaccharides were processed to the complex form without proteolytic cleavage . The data suggest that the cleavage which produces E2 occurs within the trans-Golgi or in post-Golgi elements and is closely associated with the addition of sialic acid residues to the asparagine-linked oligosaccharides . RPE.40 cells make and release noninfectious Sindbis virions that contain PE2' and no detectable E2 . These virions can be converted to an infectious form by treatment with trypsin . A defect in an intracellular endopeptidase activity in RPE.40 cells is postulated . Comparison of two Sindbis virus strains showed that the requirement for E2 in the virion to ensure infectivity is strain specific.

Mikrobiol Zh, 1991 May-Jun, 53(3), 9 - 14
{The immunochemical characteristics of the lipopolysaccharides of Pseudomonas syringae (pathovars atrofaciens and phaseolicola) and P . holci (serogroup VI)}; Iakovleva LM et al.; Lipopolysaccharides (LPS) of the representatives of strains of serogroup VI Pseudomonas syringae (P . syringae pv . atrofaciens 2399, pv . phaseolicola 120a, 7842 and P . holci 8299) possessing virulence and confinement to the host-plant are characterized by high serological activity in direct and cross reactions of the binary diffusion in agar, immunoelectrophoresis, passive hemagglutination and inhibition of passive hemagglutination . A supernatant and a sediment obtained after ultracentrifugation of LPS preparations possessed O-antigenic activity . O-specific polysaccharide (PS) is serologically less active than the LPS preparations . Problems of the intergroup and intragroup serological affinity in connection with the structure of O-specific PS . It is proved that the basic chain of O-specific polysaccharide (D-rhamnane) plays definite (but not a single) part in displaying antigenic properties of the whole LPS macromolecule.

J Cell Physiol, 1991 May, 147(2), 215 - 23
A toxin-resistant mouse L-cell mutant defective in protein transport along the secretory pathway; Laurie SM et al.; Using methods designed for isolation of mutants defective in receptor-mediated endocytosis, a novel L-cell mutant was obtained that exhibits resistance to three different protein toxins as well as alterations in secretion . This mutant, LEFIC, is resistant to modeccin, Pseudomonas exotoxin, and ricin . These toxins, which enter the cytoplasm via receptor-mediated endocytosis, are thought to penetrate into cells at the level of late endosomes or the trans Golgi network . Early endosomal acidification appears to be normal in the mutant based on its accumulation of iron from transferrin and its sensitivity to diphtheria toxin A chain-transferrin conjugate . Within the secretory pathway two delays in transport of vesicular stomatitis virus (VSV) G protein were observed in LEFIC: a 20-30 min delay in acquisition of Endo H resistance and a 1-2 hr delay in appearance of newly synthesized G protein on the cell surface . Movement of endogenous proteins along the secretory pathway was also affected in LEFIC . Fibronectin secretion was delayed by 15 min, and membrane proteins were delayed in arrival at the cell surface . The phenotype of LEFIC is consistent with a defect in a component or compartment shared by both the late endocytic and constitutive secretory pathways.

J Biol Chem, 1991 Apr 25, 266(12), 7496 - 502
Purification of cytochrome cd1 nitrite reductase from Pseudomonas stutzeri JM300 and reconstitution with native and synthetic heme d1; Weeg-Aerssens E et al.; Cytochrome cd1 nitrite reductase has been purified from Pseudomonas stutzeri strain JM 300 . This enzyme appears to be a dimer with a subunit molecular mass of 54 kDa and its isoelectric point is determined to be 5.4 . The N terminus of amino acid sequence has strong homology with that of nitrite reductase from P . aeruginosa . The apoprotein of this enzyme has been reconstituted with native and synthetic heme d1 . The nitrite reductase activity measured by NO and N2O gas evolution can be restored to 82% of the activity of the original enzyme when the protein was reconstituted with the native heme d1 and to 77% of the activity when reconstituted with the synthetic heme d1 . The absorption spectra of both reconstituted enzymes are essentially identical to that of the original nitrite reductase . These results further substantiate the novel dione structure of heme d1 as proposed . The loss of NO2- reducing activity in the absence of heme d1 and its restoration by addition of heme d1 provides further evidence that heme d1 plays a key role in the conversion of NO2- to NO and N2O.

Eur J Biochem, 1991 Apr 23, 197(2), 315 - 21
Comparison of the action of lipoprotein lipase on triacylglycerols and phospholipids when presented in mixed liposomes or in emulsion droplets; Rojas C et al.; We have compared the action of lipoprotein lipase on liposomes of egg yolk phosphatidylcholine containing less than saturating amounts of trioleoylglycerol (less than 3%) and emulsion droplets of the same lipids . The amounts of the two types of lipid particles (expressed in terms of phosphatidylcholine) needed to reach substrate saturation of the enzyme were similar, indicating similar binding of the lipase to these two lipid/water interfaces . With liposomes, as opposed to emulsion droplets, albumin was not necessary for continued hydrolysis of triacylglycerols, presumably because product fatty acids could be accommodated in the phospholipid bilayer . The maximal rate of trioleoylglycerol hydrolysis was more than 10-fold higher, and the ratio of trioleoylglycerol/phosphatidylcholine hydrolysis was more than 50-fold higher with the emulsion droplets . Qualitatively similar results were obtained with hepatic lipase, and a lipase from Pseudomonas fluorescence . The data suggest that the lipases remained at the interface for several catalytic cycles, and that a continued supply of substrate molecules to the active site favored triacylglycerol entry from the core of the lipid particle, rather than sliding in from the side through lateral diffusion in the surface layer.

Proc Natl Acad Sci U S A, 1991 Apr 15, 88(8), 3358 - 62
Anti-tumor activities of immunotoxins made of monoclonal antibody B3 and various forms of Pseudomonas exotoxin; Pai LH et al.; B3 is a monoclonal antibody that reacts with a carbohydrate epitope present on a variety of proteins located on the surface of many cancer cells and a limited number of normal tissues . We evaluated the cytotoxic activity of immunotoxins composed of monoclonal antibody B3 coupled to native Pseudomonas exotoxin (PE) or two recombinant forms of Pseudomonas exotoxin, PEArg57 or LysPE40, a form of PE with a deletion of the cell binding domain . All three conjugates were cytotoxic to human cell lines expressing the B3 antigen on their surface . The survival of each of the three immunotoxins in the circulation of mice was determined after administering the immunotoxin i.v . The half-life in blood of B3-PE and B3-PEArg57 was 20 hr, whereas the half-life of B3-LysPE40 was 4 hr . The short half-life of B3-LysPE40 may be due to the absence of domain I of PE . To determine the therapeutic effects of the three immunotoxins, they were given intraperitoneally to nude mice bearing subcutaneous A431 tumors . All three immunotoxins caused complete regression of 50-mm3 tumors with no toxic effects to the animals at therapeutic doses . Furthermore, substantial regression was also noted with much larger tumors . Our data indicate that the monoclonal antibody B3, when coupled to PE or recombinant forms of PE, may be useful for the treatment of tumors expressing B3 antigen . The therapeutic window was largest with B3-LysPE40, which can be administered in higher doses because it lacks sequences in domain I of PE that enable PE to bind to nontarget cells.

AIDS Res Hum Retroviruses, 1991 Apr, 7(4), 393 - 401
CD4-Pseudomonas exotoxin hybrid proteins: modulation of potency and therapeutic window through structural design and characterization of cell internalization; Winkler G et al.; Replacing the Pseudomonas exotoxin A (PE) cell binding domain with the human immunodeficiency virus (HIV) gp120 binding domain from CD4 yields a hybrid toxin (CD4-PE) with potential therapeutic use in treating acquired immunodeficiency syndrome (AIDS) . To find the most therapeutically potent combination of CD4 and PE four different hybrid toxins composed of one {CD4(122)} or two {CD4(181)} Ig-like CD4 domains and sequences of PE where the binding domain was partially {PE(392)} or completely {PE(364)} removed were constructed and expressed in Escherichia coli . The number of CD4 domains determined the binding affinity to gp120 and in cell viability assays the window between specific and nonspecific cytotoxicity . The length of PE determined the potency of the drug . The optimal hybrid toxin was composed of two Ig-like domains of CD4 and PE(392) . Investigation of the internalization mechanism of CD4-PE revealed that the hybrid toxin binds to target cells and is endocytosed within one hour . However, more than 6 hours are required for maximum translation inhibition . In contrast to PE which is inhibited by ammonium chloride treatment, cell toxicity of CD4-PE is not affected by ammonium chloride . Further investigations showed that the acid-induced hydrophobicity change which is required for membrane translocation is also observed with CD4-PE but at significantly higher pH than with PE.

J Trauma, 1991 Apr, 31(4), 523 - 9; discussion 529-30
The effect of granulocyte colony-stimulating factor (G-CSF) upon burn-induced defective neutrophil chemotaxis; Sartorelli KH et al.; Severe thermal injury results in impairment of granulocyte production and function . The ability to improve the functional capacity of neutrophils could contribute to a reduced morbidity and mortality from sepsis following thermal injury . Previous studies from this laboratory have shown that rhG-CSF increases the number of femoral marrow granulocyte progenitor cells and circulating neutrophils as well as the survival rate following burn wound infection . The studies reported here examine the effect of in-vivo administration of rhG-CSF on neutrophil chemotaxis following a burn injury and also following superimposed Pseudomonas burn wound sepsis in mice . Casein-elicited peritoneal neutrophils were harvested 72 hours after burn injury and 24 hours after infection . Chemotaxis was assessed using microchemotaxis chambers and 10(-5) M fMet-Phe as a chemoattractant . The number of neutrophils that migrated into the filter was used as an index of directed chemotaxis . Burn injury resulted in depressed chemotaxis compared with sham or sham/G-CSF-treated animals (p less than 0.05) . Administration of rhG-CSF to burned animals resulted in a level of neutrophil chemotaxis comparable with that in control animals . The presence of a burn wound infection caused no further impairment of chemotaxis . Administration of rhG-CSF to animals with a burn wound infection resulted in improved chemotaxis compared with sham, burned, and burned/infected animals . The beneficial effect of G-CSF following burn wound infections from this and previous studies appears to be a combination of expanded numbers of myeloid elements and preservation of their function.

Virology, 1991 Apr, 181(2), 589 - 94
In vitro packaging of the bacteriophage phi 6 ssRNA genomic precursors; Gottlieb P et al.; Bacteriophage phi 6 contains three segments of double-stranded RNA within a nucleocapsid . Plasmids containing cDNA copies of the large genomic segment direct the synthesis of viral proteins that assemble into procapsids in Escherichia coli or Pseudomonas phaseolicola . These structures are dodecahedral assemblages of proteins P1, P2, P4, and P7 . We report in this paper that these particles are capable of packaging viral single-stranded plus-sense RNA in vitro . The packaging reaction requires the presence of ATP or dATP . Synthesis of minus strands takes place within this filled procapsid in the presence of all four nucleoside triphosphates . Packaged ssRNA is found to be protected from added ribonuclease.

Arch Ophthalmol, 1991 Apr, 109(4), 503 - 5
Fulminant pseudomonal keratitis and scleritis in human immunodeficiency virus-infected patients; Nanda M et al.; Patients with human immunodeficiency virus infection are predisposed to fungal, parasitic, and viral infections . Bacterial infection can also be seen, although ocular bacterial infections have not been reported in patients with acquired immunodeficiency syndrome until recently . We present two cases of Pseudomonas corneoscleritis and one case of Pseudomonas keratitis in patients with human immunodeficiency virus infection that failed to respond to antibiotic treatment . Predisposing factors included extended-wear soft contact lens use in one patient and exposure secondary to Bell's palsy in another patient . All three patients had neutropenia that may have contributed to their poor response to treatment . Enucleation was required to treat two patients with overwhelming infection . Enucleation has been rarely required for treatment of corneoscleritis in immunocompetent patients treated at our institution . Pseudomonas keratitis in human immunodeficiency virus-infected patients represents a serious ocular infection requiring early diagnosis and aggressive treatment.

Mol Cell Biol, 1991 Apr, 11(4), 2200 - 5
Single-chain immunotoxins directed at the human transferrin receptor containing Pseudomonas exotoxin A or diphtheria toxin: anti-TFR(Fv)-PE40 and DT388-anti-TFR(Fv); Batra JK et al.; Two single-chain immunotoxins directed at the human transferrin receptor have been constructed by using polymerase chain reaction-based methods . Anti-TFR(Fv)-PE40 is encoded by a gene fusion between the DNA sequence encoding the antigen-binding portion (Fv) of a monoclonal antibody directed at the human transferrin receptor and that encoding a 40,000-molecular-weight fragment of Pseudomonas exotoxin (PE40) . The other fusion protein, DT388-anti-TFR(Fv), is encoded by a gene fusion between the DNA encoding a truncated form of diphtheria toxin and that encoding the antigen-binding portion of antibody to human transferrin receptor . These gene fusions were expressed in Escherichia coli, and fusion proteins were purified by conventional chromatography techniques to near homogeneity . In anti-TFR(Fv)-PE40, the antigen-binding portion is placed at the amino terminus of the toxin, while in DT388-anti-TFR(Fv), it is at the carboxyl end of the toxin . Both these single-chain immunotoxins kill cells bearing the human transferrin receptors . However, anti-TFR(Fv)-PE40 was usually more active than DT388-anti-TFR(Fv), and in some cases it was several-hundred-fold more active . Anti-TFR(Fv)-PE40 was also more active on cell lines than a conjugate made by chemically coupling the native antibody to PE40, and in some cases it was more than 100-fold more active.

Jpn J Med Sci Biol, 1991 Apr, 44(2), 51 - 62
Heat-stable and heat-labile components of nonspecific acid phosphatase detected in Pseudomonas pseudomallei; Kondo E et al.; In a whole cell assay system with p-nitrophenyl phosphate as substrate, strains of Pseudomonas pseudomallei showed a two-peak pattern in pH activity curve of acid phosphatase, suggesting the presence of two enzyme components different in pH optimum (4.2 and 5.2) . The component of 5.2 pH optimum was detected in the outer membrane fraction and the activity was resistant to heating at 70 C for 30 min . The other component of 4.2 pH optimum was heat-labile . No substantial difference was observed in the enzymatic activity between R and S type colonies.

Biochem Med Metab Biol, 1991 Apr, 45(2), 204 - 8
3-Hydroxy-3-methylglutaryldithio-coenzyme A: a potent inhibitor of Pseudomonas mevalonii HMG-CoA reductase; Wrensford LV et al.; 3-Hydroxy-3-methyl-1-thionoglutaryl-coenzyme A, a dithioester analog of 3-hydroxy-3-methylglutaryl-CoA, has been enzymatically synthesized using the HMG-CoA synthase catalyzed condensation of acetyl-CoA with 3-oxo-1-thionobutyryl-CoA . HMGdithio-CoA is a potent inhibitor of Pseudomonas mevalonii HMG-CoA reductase . Inhibition was mainly competitive with respect to HMG-CoA with a Kis of 0.086 +/- .01 microM and noncompetitive with respect to NADH with a Kis of 3.7 +/- 1.5 microM and a Kii of 0.65 +/- .05 microM in the presence of 110 microM (R.S)-HMG-CoA.

Gene, 1991 Apr, 100, 201 - 5
Transposon vectors for stable chromosomal integration of cloned genes in rhizosphere bacteria; Kaniga K et al.; A series of Tn5-based transposon-cloning vectors, in which many unique restriction sites lie within the transposon, have been constructed . These transposon vectors can be delivered, by conjugation, using a delivery vehicle containing a pBR322 replicon and the mobilization genes of plasmid RK2 . In Pseudomonas sp., this delivery vehicle acts as a suicide plasmid, permitting transposition to the chromosome to be detected . To facilitate cloning into the transposon vector, the delivery vehicle has been simplified so that the useful cloning sites in the transposon are not duplicated . As a model system for the transposition of cloned genes, the xylE (coding for catechol-2,3-dioxygenase) has been transposed to a variety of Pseudomonads . The transposon vectors should be useful when a stable single copy of a cloned gene is desired . They should be particularly advantageous for the genetic engineering of soil bacteria for environmental studies (agriculture or pollution control) where the stability of the engineered strains, in the absence of continuous antibiotic selection, may be important.

J Ind Microbiol, 1991 Apr, 7(3), 203 - 7
Biological activity of a transforming growth factor-alpha--Pseudomonas exotoxin fusion protein in vitro and in vivo; Heimbrook DC et al.; Transforming growth factor-alpha (TGF alpha)-pseudomonas exotoxin-40 (PE40) is a chimeric protein consisting of an N-terminal TGF alpha domain fused to a C-terminal 40-kDa segment of the pseudomonas exotoxin A protein . TGF alpha-PE40 exhibits the receptor binding activity of TGF alpha and the cell killing activity of PE40 . In the current study, we report that a modified TGF alpha-PE40 derivative significantly prolongs the survival of nude mice bearing tumors derived from cell lines which express the epidermal growth factor receptor (EGFR) . In addition, the therapeutic benefit of this protein is mediated by specific binding to the EGF receptor . These results indicate that a therapeutic window exists in vivo for the use of some growth factor--toxin fusion proteins as anticancer agents.

Proc Natl Acad Sci U S A, 1991 Mar 15, 88(6), 2336 - 40
Rapid identification of markers linked to a Pseudomonas resistance gene in tomato by using random primers and near-isogenic lines; Martin GB et al.; An approach to isolating DNA sequences that are linked to important plant genes is described . The strategy is based upon a recent modification of the polymerase chain reaction in which synthetic primers are used to amplify random sequences from genomic DNA . This technique, used in conjunction with near-isogenic lines (which differ only by the presence or absence of the target gene and a small region of surrounding DNA), leads to the rapid identification of sequences linked to the gene of interest . The feasibility of this method has been demonstrated by analyzing a pair of tomato near-isogenic lines that differ for a region on chromosome 5 that contains a gene (Pto) conferring resistance to Pseudomonas syringae pv . tomato . One hundred forty-four random primers were screened on these lines, and seven amplified products were identified that were present in one but not the other line . Of four products that were further investigated, three were confirmed by segregation analysis to be tightly linked to the Pto gene . Linked sequences identified by this method are useful for detecting the presence of the target gene in plant populations (e.g., in plant breeding) and, if very tightly linked, as starting points for a chromosome walk to isolate the gene . Since near-isogenic lines are a typical product of plant breeding and classical genetic studies, this method is applicable to a wide variety of species.

J Biol Chem, 1991 Mar 15, 266(8), 4869 - 77
The structural homology of amicyanin from Thiobacillus versutus to plant plastocyanins; Van Beeumen J et al.; The complete amino acid sequence of the blue copper protein amicyanin of Thiobacillus versutus, induced when the bacterium is grown on methylamine, has been determined as follows: QDKITVTSEKPVAAADVPADAVVVGIEKMKYLTPEVTIKAGETVYWVNGEVMPHNVA FKKGIVGEDAFRGEMMTKDQAYAITFNEAGSYDYFCTPHPFMRGKVIVE . The four copper ligand residues in this 106-residue-containing polypeptide chain are His54, Cys93, His96, and Met99 . The Thiobacillus amicyanin is 52% similar to the amicyanin of Pseudomonas AM1, the only other copper protein known with the same spacing between the second histidine ligand and the methionine ligand . T . versutus amicyanin contains no cysteine bridge and is more closely related to the plant copper protein plastocyanin than to the bacterial copper protein azurin . Alignment of the two known amicyanin sequences with the consensus sequence of the plastocyanins and comparison with the known three-dimensional structure of poplar leaves plastocyanin reveals that the bacterial proteins have the same overall structure with two beta-sheets packed face to face . The major structural differences between the amicyanins and the plastocyanins appear to be located in two of the five loops that connect the six identified beta-strands of the amicyanins . The first of these two loops, connecting strands F and G, contains a ligand histidine and must have a different conformation from the same loop in the plastocyanins because it is shorter by two amino acids . Further differences occur in the loop connecting the strands D and E . This loop contains only 17 residues in amicyanin whereas the corresponding loop of plastocyanin contains 25 residues . Despite these differences the amicyanins appear much closer related to the plastocyanins than to the azurins . The present findings demonstrate that the occurrence of blue copper proteins with clearly plastocyanin-like features is not restricted to photosynthetic redox chains.

Mol Microbiol, 1991 Mar, 5(3), 641 - 6
Ionizing groups in lipopolysaccharides of Pseudomonas cepacia in relation to antibiotic resistance; Cox AD et al.; Contrary to previous reports, lipopolysaccharides from Pseudomonas cepacia contain a 3-deoxyoct-2-ulosonic acid (probably a single residue) . The lipopolysaccharides contain only two phosphate residues, one of which apparently forms a phosphodiester bridge between 4-amino-4-deoxyarabinose and a glucosamine residue in lipid A . The second, unlocated phosphate residue occurs mainly as a monoester in some lipopolysaccharides, and mainly as a diester in others . All lipopolysaccharides lack pyrophosphate residues . The results support the view that the resistance of P . cepacia to cationic antibiotics stems from ineffective binding to the outer membrane, as a consequence of the low number of phosphate and carboxylate groups in the lipopolysaccharide, and the presence of the protonated aminodeoxypentose.

Rev Infect Dis, 1991 Mar-Apr, 13(2), 335 - 7
Corneal ulcer caused by Pseudomonas pseudomallei: report of three cases; Siripanthong S et al.; We report three cases of corneal ulcer caused by Pseudomonas pseudomallei . In all cases corneal trauma preceded the development of extensive ulcers, subconjunctival abscesses, and hypopyon . Treatment for a total of 8 weeks with topical and/or parenteral ceftazidime followed by amoxicillin-clavulanic acid produced resolution of infection in each case.

Rev Infect Dis, 1991 Mar-Apr, 13(2), 307 - 14
Transfusion reactions due to bacterial contamination of blood and blood products; Morduchowicz G et al.; Bacterial infections transmitted by blood or blood products, although rare, remain a serious threat to the recipient of a transfusion . We report on five cases of adverse reactions due to bacterial contamination of blood products, and we review 76 similar cases reported in the English-language literature . Most cases (70%) have been reported from the United States . Various sources of contamination have been suggested, including infection in the donor and invasion of the blood product during the process of collection, preparation, and storage . Frequent clinical manifestations are fever (80%), chills (53%), hypotension (37%), and nausea or vomiting (26%) . The overall mortality is 35% (28 of 81 patients) . In 38 patients (47%) the adverse reactions have appeared during transfusion; in the others the interval between completion of the transfusion and appearance of symptoms has ranged from 15 minutes to 17 days . A wide spectrum of bacteria have been implicated as causes of adverse reactions, with Pseudomonas species involved in 28% of episodes . Many such reactions are probably misdiagnosed or overlooked, the result being underestimation of the extent of the problem.

FEMS Microbiol Lett, 1991 Mar 1, 62(2-3), 163 - 9
Total degradation of various chlorobiphenyls by cocultures and in vivo constructed hybrid pseudomonads; Havel J et al.; Cocultures consisting of strains converting chlorobiphenyls to the respective benzoates or catechols and of chlorobenzoate degraders were investigated for the mineralization of chlorobiphenyls . Stable mixed cultures were obtained with 4-chlorobiphenyl, while those with 2-chloro- or 3-chlorobiphenyl were found to be unstable and released only low yields of chloride . When both sets of enzyme sequences were combined in one organism, Pseudomonas cepacia strain JH230, by conjugative transfer of genes of the biphenyl degradation sequence, the total degradation of 2-chloro-, 3-chloro-, 4-chloro-, 2,4-dichloro-, and 3,5-dichlorobiphenyl was achieved.

J Clin Microbiol, 1991 Mar, 29(3), 648 - 9
DNA fingerprinting by pulsed field gel electrophoresis and ribotyping to distinguish Pseudomonas cepacia isolates from a nosocomial outbreak; Anderson DJ et al.; We typed 40 isolates of Pseudomonas cepacia obtained from patients involved in a single outbreak using pulsed field gel electrophoresis and ribotyping . All isolates from the majority of the patients, 16 of 18 (89%), were included in a single group . These typing methods should aid in the clarification of the epidemiology of infection with P . cepacia.

J Gen Microbiol, 1991 Mar, 137 ( Pt 3), 477 - 81
Separate cloning and expression analysis of two protein components of 4-chlorobenzoate dehalogenase from Pseudomonas sp . C8S3; Elsner A et al.; The two protein components, II and III, of the 4-chlorobenzoate dehalogenase from Pseudomonas sp . CBS3 were cloned separately into Escherichia coli . Component II was obtained on plasmid pCBSII, containing a 3.0 kbp HindIII fragment, and component III on plasmid pCBSIIIb, containing a 1.3 kbp SalI/PstI fragment . The identities of the two components were confirmed by comparison with the authentic components from Pseudomonas sp . CBS3 . Both components were expressed constitutively in E . coli . Neither component alone showed dehalogenating activity . Only in the mixture of crude extracts from both clones was 4-chlorobenzoate dehalogenase detectable . The specific activities in E . coli crude extracts were 2.9 mU (mg protein)-1 for component II and 3.5 mU (mg protein)-1 for component III . Expression analysis by minicell experiments revealed a single polypeptide chain of 29 kDa for component III and of 31 kDa for component III.

Genes Dev, 1991 Mar, 5(3), 438 - 46
Inactivation of auxin in tobacco transformed with the indoleacetic acid-lysine synthetase gene of Pseudomonas savastanoi; Romano CP et al.; The iaaL gene of Pseudomonas syringae, subspecies savastanoi, encodes an indoleacetic acid (IAA)-lysine synthetase . To determine the effects of converting IAA to IAA-lysine in whole plants, the iaaL gene was fused to a constitutive plant promoter and introduced into tobacco plants . Biochemical analyses show that endogenous IAA is reduced by up to 19-fold in iaaL plants . Tobacco plants expressing the iaaL gene display reduced apical dominance, reduced rooting, and inhibition of vascular differentiation . The phenotypic effects of iaaL gene expression are reverted by crossing iaaL plants with plants that overproduce IAA . These data indicate that iaaL can act as an anti-auxin gene in vivo and confirm the role of auxin in the control of apical dominance, root growth, and vascular differentiation in whole plants.

Arch Intern Med, 1991 Mar, 151(3), 605 - 8
Melioidosis . Forgotten, but not gone!
Koponen MA, Zlock D, Palmer DL, Merlin TL.
Melioidosis, infection by the soil bacterium Pseudomonas pseudomallei, has the potential for prolonged latency with recrudescence into an acute, often fulminating, and fatal infection . Although the organism is never found in North America, infection is endemic in areas of southeast Asia, and populations of service personnel exposed during the Vietnam war and southeast Asian immigrants are at risk of severe recrudescent disease . Diagnosis, however, has been missed or delayed because of lack of familiarity with this disease . We present a case of recrudescent melioidosis that illustrates the difficulties encountered in diagnosis and treatment . This case involves a 76-year-old Vietnam veteran who presented with melioidosis of the bone 18 years after exposure to the organism and 10 years after a missed diagnosis of latent pulmonary disease . This case illustrates the protean nature of latent infection and the difficulty of selecting successful antibiotic therapy.

J Bacteriol, 1991 Mar, 173(5), 1654 - 62
Genetic and biochemical characterization of a Pseudomonas solanacearum gene cluster required for extracellular polysaccharide production and for virulence; Cook D et al.; Infection of host plants by Pseudomonas solanacerum results in wilting, which is thought to be due largely to the occlusion of xylem vessels by the P . solanacearum extracellular polysaccharide (EPS) that primarily consists of N-acetylgalactosamine (GalNAc) . By means of Tn3 mutagenesis, we identified a 6.5-kb gene cluster that contains five complementation units required for EPS production and virulence in this bacterium . There was positive correlation between the amount of EPS produced in culture and (i) in planta growth and (ii) virulence . Based on analysis of beta-glucuronidase-gene fusions, these genes are expressed both in broth cultures and in planta and may be constitutive . Both wild-type and mutant strains contained similar amounts of UDP-GalNAc, the predicted primary substrate for EPS synthesis . Thus, the EPS mutants we obtained should be useful in the analysis of steps in the assembly of the polysaccharide and how this process is related to virulence.

Infect Immun, 1991 Mar, 59(3), 776 - 80
Siderophore production by Pseudomonas pseudomallei; Yang HM et al.; Eighty-four strains of Pseudomonas pseudomallei isolated from patients with melioidosis were examined for siderophore production . All the strains were shown to produce siderophore both on chrome azurol S agar plates and in liquid medium under iron-deficient conditions . Chemical assays indicated that the siderophore belongs to the hydroxamate class . Addition of iron to the culture medium resulted in increased culture growth with markedly decreased yield of siderophore . Siderophore produced by strain U7 was purified by gel filtration chromatography, and the molecular weight was estimated to be 1,000 . When this partially purified siderophore was added to culture medium, it promoted iron uptake by P . pseudomallei in the presence of EDTA and enhanced growth of the organism in the presence of transferrin . We have given this siderophore the trivial name malleobactin.

Mol Cell Biol, 1991 Mar, 11(3), 1751 - 3
Substitution of foreign protein sequences into a chimeric toxin composed of transforming growth factor alpha and Pseudomonas exotoxin; Debinski W et al.; TGF alpha-PE40 is a chimeric toxin made by replacing domain Ia of Pseudomonas exotoxin (PE) with transforming growth factor alpha (TGF alpha) . We have now replaced a portion of domain Ib of PE with different polypeptides or an extra domain III of PE in transforming growth factor alpha-PE40 and maintained cell killing . Thus, TGF alpha-PE40 can be used to transport foreign protein sequences into the cytosol of cells.

Kansenshogaku Zasshi, 1991 Mar, 65(3), 277 - 85
{Effect of erythromycin on neutrophil function in chronic respiratory tract diseases with Pseudomonas infection}; Kadota J et al.; The "low dose and long term" erythromycin (EM) therapy has been reported as effective (or useful) in chronic respiratory tract disease with pseudomonas (P.) infection including diffuse panbronchiolitis (DPB), however the mode of action is still obscure . Therefore in this study we have examined the effect of EM on the interaction between P . aeruginosa and human polymorphonuclear leukocyte (PMN) in vitro . The efficiency of intracellular killing and the ability of superoxide production were employed to evaluate the PMN functions . For the first step, the following results were obtained; 1) Pretreatment of PMN with 20 micrograms/ml EM did not affect the killing ability of PMNs against opsonized P . aeruginosa of standard and clinical isolate from DPB patient . 2) Superoxide production from PMNs was observed by phagocytosis of P . aeruginosa in the presence of serum . This was not affected by exposure of PMNs to 20 micrograms/ml EM even with increased ratios of bacteria to cells . 3) Pretreatment of P . aeruginosa with 20 micrograms/ml EM before opsonization enhanced the killing ability of PMNs in both standard and clinical isolate . 4) Pretreatment of P . aeruginosa with 20 micrograms/ml EM resulted in no effect on superoxide production from PMNs by phagocytosis of the bacteria . These results indicate that EM may modify a certain step of the interaction between bacteria and intracellular host defence mechanisms . Therefore for the second step, we have investigated the susceptibility of EM-exposed bacteria to killing by the cell free (glucose oxidase-glucose) system, which will detect the enzymatic generation of hydrogen peroxide (H2O2) . The following results were observed; 1) EM-exposed bacteria was more susceptible to killing than control bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Lett Appl Microbiol, 1991 Mar, 12(3), 85 - 7
Conjugal transfer of recombinant plasmids into gellan gum-producing and non-producing variants of Pseudomonas elodea ATCC 31461; Fialho AM et al.; The conjugal transfer of recombinant plasmids into a gellan gum-producing, rifampicin-resistant strain of Pseudomonas elodea and into one of its non-producing variants, was studied in order to facilitate the cloning of gellan genes . The mobilization frequency of recombinant plasmids derived from pKT240, and of cosmid pJRD215 from Escherichia coli HB101 (hsdM), into the mucoid strain was below the values for the non-mucoid variant and decreased exponentially with plasmid size . Reducing the mating time on solid surfaces led to higher mobilization frequencies . Under optimal conditions, a gene bank (40-50 kb) constructed in the cosmid pJRD215 was efficiently mobilized into Gel- mutants during complementation experiments.

JAMA, 1991 Feb 27, 265(8), 981 - 6
Nosocomial Pseudomonas pickettii bacteremias traced to narcotic tampering . A case for selective drug screening of health care personnel; Maki DG et al.; Three patients in a university hospital developed nosocomial infusion-related Pseudomonas pickettii bacteremia . Investigation identified six additional patients who had received intravenous fluid contaminated by P pickettii but did not become ill . All nine patients had had surgery, and each of these patients but only nine of 19 operated-on control patients had received intravenous fentanyl citrate in the operating room; the mean dose given to the nine case patients was far greater than that given to control patients . Fentanyl in 20 (40%) of 50 predrawn 30-mL syringes was shown to be contaminated by P pickettii . Contamination was caused by theft of fentanyl from predrawn synringes and replacement by distilled water contaminated by P pickettii . Narcotic theft by health care personnel may cause patients to suffer pain needlessly and can also result in dire unanticipated consequences, such as nosocomial bacteremia . Whereas drug testing in the workplace is highly controversial, we believe that testing of health care personnel is indicated when drug abuse or theft is suspected.

Lancet, 1991 Feb 16, 337(8738), 392 - 4
Aerosol alpha 1-antitrypsin treatment for cystic fibrosis; McElvaney NG et al.; In cystic fibrosis neutrophil-dominated inflammation on the respiratory epithelial surface results in a chronic epithelial burden of the destructive enzyme, neutrophil elastase . alpha 1-antitrypsin (alpha 1AT), the main inhibitor of neutrophil elastase in lung, was given in aerosol form to 12 cystic fibrosis patients . It suppressed neutrophil elastase in the respiratory epithelial lining fluid (ELF) and restored the ELF anti-neutrophil elastase capacity when ELF alpha 1AT reached 8 mumol/l . This treatment also reversed the inhibitory effect of cystic fibrosis ELF on pseudomonas killing by neutrophils, which suggests that it may augment host defence in cystic fibrosis.

Biochem J, 1991 Feb 1, 273 ( Pt 3), 611 - 3
Solution behaviour of Chromobacter viscosum and Pseudomonas sp . lipases . No evidence of self-association; Simpkin NJ et al.; 1 . The size of two bacterial lipases was studied by SDS/PAGE, sedimentation velocity and sedimentation equilibrium to test for possible self-association behaviour . 2 . Mr values of selected lipases were obtained from SDS/PAGE and sedimentation-velocity measurements, together with an absolute determination by sedimentation equilibrium 3 . The Mr values obtained in a variety of aqueous solvents indicate that lipases do not self-associate in solution, suggesting the absence of surface hydrophobic patches.

J Bacteriol, 1991 Feb, 173(4), 1530 - 5
Complete nucleotide sequences and comparison of the structural genes of two 2-haloalkanoic acid dehalogenases from Pseudomonas sp . strain CBS3; Schneider B et al.; The nucleotide sequences of two DNA segments from Pseudomonas sp . strain CBS3 that code for two different haloalkanoic acid halidohydrolases were determined . Two open reading frames with coding capacities of 227 amino acids (corresponding to a molecular mass of 25,401 Da) and 229 amino acids (corresponding to a molecular mass of 25,683 Da) were identified as structural genes of 2-haloalkanoic acid dehalogenases I (dehCI) and II (dehCII) by comparison with the N-terminal amino acid sequences of these enzymes . Comparison of the two sequences revealed 45% homology on the DNA level and 37.5% homology on the amino acid level . No homology with other known protein or nucleotide sequences was found.

J Bacteriol, 1991 Feb, 173(3), 1073 - 9
Pseudomonas syringae pv . phaseolicola genomic clones harboring heterologous DNA sequences suppress the same phaseolotoxin-deficient mutants; Kamdar HV et al.; Cosmid cloning and mutagenesis were used to identify genes involved in the production of phaseolotoxin, the chlorosis-inducing phytotoxin of Pseudomonas syringae pv . phaseolicola, the causal agent of halo blight of bean (Phaseolus vulgaris L.) . Eight stable clones were isolated from a genomic cosmid library by en masse mating to 10 ethyl methanesulfonate (EMS)-induced Tox- mutants . In cross-matings, each suppressed all 10 mutants as well as an additional 70 EMS-induced Tox- mutants (and one UV-induced Tox- mutant) . On the basis of restriction endonuclease analysis and hybridization studies, the clones were grouped into three classes . Clones in a particular class shared common fragments, whereas clones in different classes did not . Clones from class I (but not classes II and III) also suppressed Tn5-induced Tox- mutants . Interposon mutagenesis and marker exchange of a representative clone from class III into the wild-type genome did not alter its Tox+ phenotype, indicating that this clone does not harbor structural or regulatory genes involved in phaseolotoxin production . We suggest that the genome of P . syringae pv . phaseolicola contains a "hot spot" in one of the functions involved in toxin production which is affected by EMS and UV and that heterologous clones are able to suppress the Tox- phenotype because their inserts encode products that are able to substitute for the product of the mutated gene . Alternatively, the inserts may contain sequences which titrate a repressor protein . In either case, the data suggest that suppression of EMS- and UV-induced mutants occurs when heterologous clones are present in multiple copies.

J Surg Res, 1991 Feb, 50(2), 111 - 8
Platelet-activating factor in porcine Pseudomonas acute lung injury; Byrne K et al.; We investigated the role of platelet-activating factor (PAF) in acute septic lung injury by examining the effects of the selective PAF antagonist SRI 63-675 and by measuring PAF in lung tissue in the porcine model . Four groups of pigs (15-25 kg) were studied: saline control (C, n = 5); Pseudomonas (Ps, n = 9), given 5 x 10(8) CFU/ml at 0.3 ml/20 kg/min intravenously over 1 hr; SRI (n = 3), given SRI 63-675 in a 40 mg/kg bolus; and SRI + Ps (n = 5) . Ps infusion produced a fulminant lung injury characterized by a threefold increase in pulmonary arterial pressure at 30 min and persistent pulmonary hypertension (P less than 0.05 vs C), a significant (P less than 0.05 vs C) decrease in arterial oxygen tension (PaO2) from 60 min, a significant (P less than 0.05 vs C) increase in extravascular lung water (EVLW) from 120 min, and a significant (P less than 0.05 vs C) increase in albumin flux determined scintigraphically as slope index at 150-180 min . Systemic arterial pressure and cardiac index (CI) decreased significantly (P less than 0.05) in the Ps group vs C at 60 and 180 min, respectively . Bolus injection of SRI 63-675 at the time of Ps infusion blocked the early pulmonary hypertension, attenuated the early and late fall in PaO2, ameliorated the increase in EVLW, and prevented the late (150-180 min) increase in albumin flux . SRI 63-675 had minimal effects on Ps-induced hypotension or alterations in CI.(ABSTRACT TRUNCATED AT 250 WORDS)

Semin Cell Biol, 1991 Feb, 2(1), 31 - 7
Redirecting Pseudomonas exotoxin; FitzGerald D et al.; Pseudomonas exotoxin (PE) is a three-domain bacterial toxin that kills mammalian cells by gaining entry to the cytosol and inactivating protein synthesis . The pathway of toxin entry includes binding to a surface receptor, internalization via coated pits and endosomes, proteolytic processing, reduction of disulfide bonds and finally the translocation of an enzymatically active C-terminal fragment to the cytosol . Once in the cytosol this fragment inhibits protein synthesis by ADP ribosylating elongation factor 2 . Because of its potency PE and its derivatives have been directed to kill various target cells . It is hoped this strategy will lead to the development of a novel kind of therapeutic agent for the treatment of various human diseases including cancer, AIDS and various immunological disorders.

Antimicrob Agents Chemother, 1991 Feb, 35(2), 341 - 4
Damage to the cytoplasmic membrane and cell death caused by dodine (dodecylguanidine monoacetate) in Pseudomonas syringae ATCC 12271; Cabral JP; Treatment of Pseudomonas syringae cells with low concentrations of the fungicide dodecylguanidine monoacetate (dodine) resulted in cell death and leakage of K+, UV-absorbing materials, and ribose-containing molecules . The results suggest that dodine causes gross and extensive damage to the cytoplasmic membrane, which is probably implicated in the death of cells.

Exp Cell Res, 1991 Feb, 192(2), 389 - 95
Disruption of the Golgi apparatus by brefeldin A inhibits the cytotoxicity of ricin, modeccin, and Pseudomonas toxin; Yoshida T et al.; We have studied the cytotoxicity of ricin in cells treated with brefeldin A (BFA), which dramatically disrupts the structure of the Golgi apparatus causing Golgi content and membrane to redistribute to the ER . BFA inhibits the cytotoxicity of ricin in Chinese hamster ovary, normal rat kidney, and Vero cells and abolishes the enhancement of ricin cytotoxicity by NH4Cl, nigericin, swainsonine, and tunicamycin or by a mutation in endosomal acidification . BFA protects cells from the cytotoxicities of modeccin and Pseudomonas toxin, but has no effect on the intoxication by diphtheria toxin . Pretreatment of BFA does not protect cells from ricin treatment in the absence of BFA . Our results suggest that ricin, modeccin, and Pseudomonas toxin share a common pathway of intracellular transport from endosomes to the Golgi region where they are released into the cytosol . In contrast, the lack of protection of Vero cells from diphtheria toxin by BFA indicates that diphtheria toxin is released from acidified endosomes without involving the Golgi region.

Antonie Van Leeuwenhoek, 1991 Feb, 59(2), 115 - 23
Molecular cloning and functional characterization of a recA analog from Pseudomonas stutzeri and construction of a P . stutzeri recA mutant; Vosman B et al.; A recombinant plasmid carrying the Pseudomonas stutzeri recA gene was isolated by complementation of the Escherichia coli recA13 mutation . Subcloning experiments showed that the gene was located on a 1500 bp PvuII-BglII fragment . The cloned gene complements an E . coli recA mutant for resistance to Methylmethanosulphonate (MMS) and UV irradiation . It was also capable of restoring the recombination proficiency in that mutant . The cloned fragment was used to construct a recA deletion mutant of P . stutzeri . This mutant too was shown to be sensitive towards MMS and UV irradiation . The mutant strain was found to be completely deficient in natural transformation with chromosomal DNA, due to the impairment in homologous recombination.

Gene, 1991 Feb 1, 98(1), 21 - 8
Gene-specific transposon mutagenesis of the biphenyl/polychlorinated biphenyl-degradation-controlling bph operon in soil bacteria; Furukawa K et al.; A transposon, Tn5-B21, was gene-specifically inserted into the chromosomal biphenyl/polychlorinated biphenyl-catabolic operon (bph operon) of soil bacteria . The cloned bphA, bphB and bphC genes of Pseudomonas pseudoalcaligenes KF707, coding for conversion of biphenyl into a ring meta-cleavage product (2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid), carried random insertions of Tn5-B21 . The mutagenized bphABC DNA, carried by a suicide plasmid, was introduced back into the parent strain KF707, resulting in the appearance of gene-specific transposon mutants by double crossover homologous recombination: the bphA::Tn5-B21 mutant did not attack 4-chlorobiphenyl, the bphB::Tn5-B21 mutant accumulated dihydrodiol, and the bphC::Tn5-B21 mutant produced dihydroxy compound . Gene-specific transposon mutants of the bph operon were also obtained for some other biphenyl-utilizing strains which possess bph operons nearly identical to that of KF707.

J Bacteriol, 1991 Feb, 173(3), 1215 - 22
Cloning and analysis of s-triazine catabolic genes from Pseudomonas sp . strain NRRLB-12227; Eaton RW et al.; Pseudomonas sp . strain NRRLB-12227 degrades the s-triazine melamine by a six-step pathway which allows it to use melamine and pathway intermediates as nitrogen sources . With the plasmid pLG221, mutants defective in five of the six steps of the pathway were generated . Tn5-containing-EcoRI fragments from these mutants were cloned and identified by selection for Tn5-encoded kanamycin resistance in transformants . A restriction fragment from ammelide-negative mutant RE411 was used as a probe in colony hybridization experiments to identify cloned wild-type s-triazine catabolic genes encoding ammeline aminohydrolase, ammelide aminohydrolase, and cyanuric acid amidohydrolase . These genes were cloned from total cellular DNA on several similar, but not identical, HindIII fragments, as well as on a PstI fragment and a BglII fragment . Restriction mapping and Southern hybridization analyses of these cloned DNA fragments suggested that these s-triazine catabolic genes may be located on a transposable element, the ends of which are identical 2.2-kb insertion sequences.

Hindustan Antibiot Bull, 1991 Feb-Nov, 33(1-4), 7 - 13
Effect of ten antibiotics on the recovery of seed-borne Pseudomonas syringae pv syringae of cowpea, Vigna unguiculata (L.) Walp var . IT 825-2246-4; Oluwadare TO et al.; The effect of ten different antibiotics: Amoxil, Ampicillin, Chloramphenicol, Cloxacillin, Cotrimexazole, Erythromycin, Nitrofurantoin, Penicillin, Tetracycline and Vibramycin on the recovery of seed-borne Pseudomonas syringae pv . syringae and on percentage seed germination of cowpea, Vigna unguiculata(L.) Walp var . IT 825-2246-4 at various temperature regimes: 25 degrees C, 30 degrees C, 35 degrees C and 40 degrees C were investigated using the Standard Blotter Method . Seeds were presoaked for 1 hour in each antibiotic at the concentration rate of 250mg/liter before plating on three layers of moist blotter papers in petri dishes . Seeds were incubated for 72 hours in temperature controlled incubators at the various temperature regimes . Seeds not treated with antibiotic served as control . In all the temperature regimes studied . Tetracycline was the most effective antibiotic in controlling the bacterium while Cotrimexazole was the least effective when compared with control . The highest percentage seed germination was recorded on seeds treated with Amoxil while the lowest percentage seed germination was recorded on seeds treated with Tetracycline . Generally, the temperature regimes used did not have any appreciable effect on the recovery of the bacterium and on percentage seed germination.

J Gen Virol, 1991 Feb, 72 ( Pt 2), 299 - 305
Inhibition of mouse retroviral disease by bioactive glutaminase-asparaginase; Roberts J et al.; Glutamine depletion strongly inhibits the replication of Rauscher murine leukaemia retrovirus (RLV) in vitro . Pseudomonas 7A glutaminase-asparaginase (PGA), capable of depleting glutamine and asparagine for prolonged periods, was used to determine the therapeutic effectiveness of glutamine depletion in mice infected with RLV or Friend virus . During PGA treatment of viraemic animals, serum reverse transcriptase activity fell to control levels and infected animals did not develop splenomegaly . The therapeutic results obtained with PGA compared favourably with those of azidothymidine given intraperitoneally at 30 mg/kg/day . Western blots performed on splenic tissue from control and treated animals indicated that glutamine depletion prevented readthrough of an amber codon at the gag-pol junction, stopping translation of viral mRNA at that point . Treatment of RLV-infected animals with PGA resulted in nearly a 200% increase in mean survival time even when therapy was initiated late in the course of the disease . To our knowledge, this is the first demonstration that a nutrient required for viral replication can be enzymically depleted in vivo to inhibit viral replication.

Genetika, 1991 Feb, 27(2), 217 - 21
{Regulation of the synthesis of 3-deoxy-D-arabinoheptuloso-7-phosphate-synthase in Pseudomonas bacteria}; Maksimova NP et al.; Regulation of synthesis of 3-deoxy-D-arabinoheptulose-7-phosphate-synthase (DAHP-synthase) was analysed in 12 strains of Pseudomonas belonging to 10 species . Repression of synthesis of DAHP-synthase was registered in half of strains under study, this being controlled by tyrosin and phenilalanine in P . aeruginosa PAO1 and PAT 2152, P . fluorescens BKMB 896, P . acidovorans BKMB 1251 and P . testosteroni BKMB 1241 and by only phenilalanine in P . maltophilia BKMB 30 . In the rest of cases (in P . putida AC29, P . stutzeri BKMB 148, P . mendocina BKMB 1299, P . marginata BKMB 1298, P . vesicularis BKMB 974 and P . fluorescens BKMB 35) the enzyme was synthesized constitutively.

Biotechniques, 1991 Feb, 10(2), 202 - 4, 206, 208-9
Pseudomonas exotoxin fusion proteins are potent immunogens for raising antibodies against P-glycoprotein; Bruggemann EP et al.; Antibodies to specific regions of human P-glycoprotein have been difficult to obtain . We developed a method to express in E . coli fusions between Pseudomonas exotoxin and specific regions of human P-glycoprotein . We used the polymerase chain reaction to amplify the desired regions of MDR1 cDNA and to introduce appropriate restriction sites . These fragments were cloned into the 3' end of the Pseudomonas exotoxin gene . With this system we produced large amounts of fusion proteins for immunizations, and we obtained positive rabbit antiserum against P-glycoprotein with most of these antigens . We now have a comprehensive panel of polyclonal antibodies against P-glycoprotein . This system should be generally useful to raise antibodies against other eukaryotic proteins that are difficult to prepare in large quantities.

FEBS Lett, 1991 Jan 28, 278(2), 244 - 6
Left handed alpha-helix formation by a bacterial peptide; Mortishire-Smith RJ et al.; The alpha-helix is a common element of secondary structure in proteins and peptides . In eukaryotic organisms, which exclusively incorporate L-amino acids into such molecules, stereochemical interactions make such alpha-helices, invariably right-handed . Pseudomonas tolaasii Paine is the causal organism of the economically significant brown blotch disease of the cultivated mushroom Agaricus bisporus (Lange) Imbach . P . Tolaasii proceduces an extracellular lipodepsipeptide toxin, tolaasin, which causes the brown pitted lesions on the mushroom cap . Circular dichroism studies on tolaasin in a membrane-like environment indicate the presence of a left-handed alpha-helix, probably formed by a sequence of 7 D-amino acids in the peptide . P . tolaasii represents the first reported example of an organism which has evolved the ability to biosynthesize a left-handed alpha-helix.

J Biol Chem, 1991 Jan 25, 266(3), 1526 - 33
Cosubstrate binding site of Pseudomonas sp . AK1 gamma-butyrobetaine hydroxylase . Interactions with structural analogs of alpha-ketoglutarate; Ng SF et al.; Forty-one aromatic and aliphatic analogs of alpha-ketoglutarate were studied kinetically for their interaction with the alpha-ketoglutarate binding site of gamma-butyrobetaine hydroxylase obtained from Pseudomonas sp . AK1 . Together, the compounds represent structural permutations probing the contribution of: 1) the C5 carboxyl group of alpha-ketoglutarate (domain I); 2) the C1-C2 keto acid moiety of alpha-ketoglutarate (domain II); 3) the distance between domains I and II; and 4) the spatial relationship of the two domains required for optimal interaction with the cosubstrate binding site . All compounds were competitive inhibitors for alpha-ketoglutarate (Km 0.018 mM) . Functionally, two subsites of the cosubstrate binding site were evident: subsite I for polar interaction with the C5 carboxyl group, and subsite II, comprising of two distinct cis-oriented coordination sites of the catalytic ferrous ion which interact with the C1-C2 keto acid moiety . The most efficient inhibitors were pyridine 2,4-dicarboxylate (Ki 0.0002 mM) and 3,4-dihydroxybenzoate (Ki 0.0006 mM) . Both compounds contain a carboxyl group and a chelating moiety corresponding to domains I and II of alpha-ketoglutarate, respectively . The fixed orientation of these groups in both analogs was used to assess intersubsite distance and spatial relationship required for optimal interaction with the cosubstrate binding site . Binding at subsite I and chelation at subsite II were indispensible for effective competitive inhibition . The distance between these two domains also helped determine whether attachment at the cosubstrate binding site would be catalytically productive . This was emphasized by the failure of either oxaloacetate or alpha-ketoadipinate to promote hydroxylation . Optimal interdomain distance, however, was not sufficient for cosubstrate utilization, as pyridine 2,4-dicarboxylate, with an interdomain distance identical to alpha-ketoglutarate in its staggered conformation, did not sustain hydroxylation . In the overall, these studies suggest that alpha-ketoglutarate utilization occurs in a ligand reaction at the active site ferrous ion of gamma-butyrobetaine hydroxylase . This is of particular interest since the delineated stereochemical mode of oxidative decarboxylation could generate the reactive oxo-iron species that was shown experimentally to promote gamma-butyrobetaine hydroxylation by an abstraction-recombination mechanism (Blanchard, J . S., and Englard, S . (1983) Biochemistry 22, 5922-5928; Englard, S., Blanchard, J . S., and Midelfort, C . F . (1985) Biochemistry 24, 1110-1116).

Int J Immunopharmacol, 1991, 13(2-3), 305 - 15
Administration of IL-2-PE40 via osmotic pumps prevents adjuvant induced arthritis in rats . Improved therapeutic index of IL-2-PE40 administered by continuous infusion; Lorberboum-Galski H et al.; IL-2-PE40 is a chimeric cytotoxin composed of interleukin 2 (IL-2) fused to a truncated form of Pseudomonas exotoxin (PE) that lacks its binding domain . IL-2-PE40 has been shown to exhibit therapeutic potency in several models in vivo when administered i.p . twice a day . Here we show that the continuous administration of IL-2-PE40 by an osmotic pump specifically prevents the development of adjuvant induced arthritis in rats with an improved therapeutic efficacy as compared to daily repeated i.p . injections . Stabilization of IL-2-PE40 at 37 degrees C for the continuous administration by pumps was achieved by adding NAD, the substrate for the enzyme portion of the chimeric toxin.

Mikrobiol Zh, 1991 Jan-Feb, 53(1), 90 - 2
{Capsule formation in the causative agent of glanders}; Popov SF et al.; Electron-microscopic and electron-cytochemical method were used to indicate the capability of Pseudomonas mallei (str . N 10230) to produce extracellular slime during the agent growth on the meat-peptone agar . In case of guinea pigs infection the agent forms a capsule that defends the pathogen from phagocytosis.

Mikrobiol Zh, 1991 Jan-Feb, 53(1), 48 - 53
{An analysis of the molecular forms of the lipopolysaccharide from Pseudomonas syringae pv . atrofaciens IMV K-1025}; Ovod VV et al.; Water extract and salt-EDTA extract of Pseudomonas syringae, pv . atrofaciens cells were fractionated by ultracentrifugation with following salting out of ultracentrifugal supernatant by ammonium sulphate at 55% saturation (pH 4.5) . The composition and distribution of LPS molecular forms were studied in the obtained fractions by means of electrophoresis in 10% polyacrylamide gel with 1% sodium dodecylsulphate when staining gels by silver nitrate and cumassi . It is shown that ultracentrifugal supernatant and a sediment as well as sulphate sediment contain S-LPS and R-LPS . SR-LPS is not differentiated . Sulphate supernatant does not contain the determinable amount of S-LPS but it is enriched by the proteins with molecular weights of 65-15 kDalton . S-LPS is localized in the gel area which corresponds to mobilities of polypeptides with molecular weights 130-45 KDalton and the number of monomeric links in O-specific chains of its molecules reaches 25-30 . R-LPS migrates under electrophoresis in gel to the mobility zone of polypeptides with molecular weights 14.5-16 kDalton.

Antonie Van Leeuwenhoek, 1991 Jan, 59(1), 19 - 25
Exchange of chromosomal markers by natural transformation between the soil isolate, Pseudomonas stutzeri JM300, and the marine isolate, Pseudomonas stutzeri strain ZoBell; Stewart GJ et al.; Both the soil isolate, Pseudomonas stutzeri JM300, and the marine isolate, Pseudomonas stutzeri strain ZoBell, have been shown previously to be naturally transformable . This study reports the detection of genetic exchange by natural transformation between these two isolates . Transformation frequency was determined by filter transformation procedures . Three independent antibiotic resistance loci were used as chromosomal markers to monitor this exchange event: resistance to rifampicin, streptomycin, and nalidixic acid . The maximum frequencies of transformation were on the order of 3.1 to 3.8 x 10(-6) transformants per recipient; frequencies over an order of magnitude greater than those for spontaneous antibiotic resistance, although they are lower than those observed for soil:soil or marine:marine strain crosses . This exchange was inhibited by DNase I . Transformation was observed between soil and marine strains, both by filter transformation using purified DNA solutions and when transforming DNA was added in the form of viable donor cells . The results from this study support the close genetic relationship between P . stutzeri JM300 and P . stutzeri strain ZoBell . These results also further validate the utility of P . stutzeri as a benchmark organism for modeling gene transfer by natural transformation in both soil and marine habitats.

Perit Dial Int, 1991, 11(1), 27 - 30
A randomised prospective comparison of oral ofloxacin and intraperitoneal vancomycin plus aztreonam in the treatment of bacterial peritonitis complicating continuous ambulatory peritoneal dialysis (CAPD); Cheng IK et al.; Forty six patients who developed 48 episodes of peritonitis while on CAPD were randomised to receive either oral ofloxacin or intraperitoneal (i.p.) vancomycin/aztreonam . Three patients were excluded from analysis: 2 were transferred to other hospitals and 1 was later found to have candida peritonitis . Of the remainder, 22 episodes were treated with oral ofloxacin and 23 with i.p . vancomycin/aztreonam . The primary cure rate in the oral ofloxacin and i.p . vancomycin/aztreonam group was 77.3% and 87.5% respectively . There were 3 primary failures and 2 relapses in the former and 1 failure and 2 relapses in the latter group . Two of the 4 primary failures were peritonitis episodes secondary to infection with pseudomonas species . The total number of days of hospital stay was 48 and 58 respectively in the two groups . Analysis of the cost of treatment revealed that i.p . vancomycin/aztreonam was 30 times more expensive than oral ofloxacin . Despite a slightly higher cure rate with i.p . vancomycin/aztreonam, oral ofloxacin is a more cost-effective primary treatment of bacterial peritonitis in patients on CAPD especially in countries with a limited health budget.

Appl Environ Microbiol, 1991 Jan, 57(1), 324 - 6
Resolution of 4-chlorobenzoate dehalogenase from Pseudomonas sp . strain CBS3 into three components; Elsner A et al.; Extracts of Pseudomonas sp . strain CBS3 grown with 4-chlorobenzoate as sole carbon source contained an enzyme that converted 4-chlorobenzoate to 4-hydroxybenzoate . This enzyme was shown to consist of three components, all necessary for the reaction . Component I, which had a molecular weight of about 3,000, was highly unstable . Components II and III were stable proteins with molecular weights of about 86,000 and 92,000.

Appl Environ Microbiol, 1991 Jan, 57(1), 163 - 7
Isolation of amoebae and Pseudomonas and Legionella spp . from eyewash stations; Paszko-Kolva C et al.; Forty eyewash units were sampled for protozoa, bacteria, and fungi . Total heterotrophic bacterial counts on nutrient agar and R2A agar (Difco Laboratories, Detroit, Mich.) ranged from 0 to 10(5) CFU/ml, with Pseudomonas spp . being the most frequently isolated . Total counts of 10(4) and 10(8) cells per ml were obtained with the acridine orange staining procedure . All samples were examined for Legionella spp . by direct fluorescent-antibody staining and by culturing on buffered charcoal-yeast extract agar containing alpha-ketoglutarate and glycine and supplemented with cycloheximide, vancomycin, and polymyxin B . DNA-DNA hybridization was used to confirm identification of the Legionella isolates . Legionellae were detected in 35 of 40 (87.5%) samples by direct fluorescent-antibody staining, with 3 samples yielding both Legionella spp . and amoebae . Amoebae identified as Hartmannella, Vahlkampfia, Acanthamoeba, and Cochliopodium spp . were detected in 19 of 40 (47:5%) samples . Sabouraud dextrose agar was used to obtain a crude estimate of viable fungal populations, pH, hardness, and ammonia, alkalinity, chlorine, copper, and iron contents were recorded for all water samples collected from eyewash stations; 33% of the samples had greater than or equal to 10 mg of CO2 per liter . It is concluded that eyewash stations not regularly flushed and/or cleaned and used to flush traumatized eye tissue may be a source of infection and can contaminate laboratory environments via aerosol transmission.

Cancer Detect Prev, 1991, 15(2), 137 - 43
Immunoconjugates of Pseudomonas exotoxin A: evaluation in mice, monkeys, and man; Morgan AC Jr et al.; Immunotoxins of PE were constructed with stable thioether linkages using two monoclonal antibodies to ovarian cancer, OVB-3 and NR-LU-10 . Antigens recognized by both antibodies have limited normal tissue distribution and are expressed on virtually all ovarian cancers . Both antibodies form highly potent conjugates (ID50 = 100 pg/ml) with high selectivity (greater than or equal to 4 logs) and can eliminate greater than or equal to 5 logs of tumor cells in vitro . The conjugates have been evaluated for efficacy in both ovarian and colon carcinoma ascites xenografts . In the ovarian model, the conjugates produce an increase in life span (ILS) of 200 to 300 with some cures against established but low tumor burden ascites . Increasing the tumor burden decreases efficacy and duration of responses . A lower ILS of 150 to 200 is achieved in the more aggressive colon model . However, the combination of immunotoxin with chemotherapy, which is ineffective on its own, demonstrated enhanced activity (ILS = 300) . Toxicity of the conjugates is hepatic and easily monitored by liver function tests (LDH) . Antitoxin responses are highly variable, but typically have a rapid onset and appear to be predicted by preexisting levels . Pilot clinical evaluation in ovarian cancer (intraperitoneal) is ongoing.

Rev Infect Dis, 1991 Jan-Feb, 13(1), 68 - 72
Adverse reactions to prolonged treatment with high doses of carbenicillin and ureidopenicillins; Lang R et al.; Charts were reviewed for 63 patients whose chronic pseudomonas osteomyelitis was treated with high doses of extended-spectrum penicillins for prolonged periods . The incidence of untoward drug reactions was significantly higher than expected . Carbenicillin evoked adverse reactions in 22.8% of patients . However, most of these reactions were mild, and a change of drug was required in only 5.7% of cases . No adverse drug reactions were observed with cumulative doses of less than 750 g . In contrast to carbenicillin, the ureidopenicillins were associated with adverse reactions in 67.7% of patients; most reactions were moderate to severe in intensity; a cumulative dose of greater than 250 g produced adverse reactions; and discontinuation or change of therapy was required in 51.6% of cases . The main adverse reactions to both carbenicillin and the ureidopenicillins included rash, drug fever, leukopenia, eosinophilia, thrombocytopenia, and hepatic damage.

Biotherapy, 1991, 3(1), 65 - 76
Immunotoxins; Press OW; Immunotoxins (ITs) are chimeric molecules constructed by covalently conjugating monoclonal antibodies (MoAbs) to plant or bacterial toxins (e.g . ricin or pseudomonas exotoxin) . The antibody moiety allows specific targeting of ITs to tumor-associated antigens, while the toxin moiety is responsible for cell killing by irreversible inactivation of protein synthesis . Since ITs must reach the cytosol to kill cells, the rates of endocytosis, the pathways of intracellular routing, and the rates of translocation to the cytoplasm are important determinants of the efficacy of an IT . Promising in vitro and in vivo IT results have been reported by many groups, and phase I clinical trials in cancer patients are currently underway.

J Bacteriol, 1991 Jan, 173(1), 301 - 7
Sequence domains required for the activity of avirulence genes avrB and avrC from Pseudomonas syringae pv . glycinea; Tamaki SJ et al.; avrB and avrC from Pseudomonas syringae pv . glycinea share significant amino acid homology but interact with different soybean resistance genes to elicit the hypersensitive defense reaction . Recombinant genes constructed between avrB and avrC revealed that the central regions were required for avirulence gene activity but the 5' and 3' termini were interchangeable . Recombinants involving the central regions did not yield any detectable avirulence gene activity, and no new avirulence phenotypes were observed from any of the chimeric genes . These results suggest that the protein products of avrB and avrC possess catalytic properties that are required for the avirulence phenotypes.

Hosp Pract (Off Ed), 1991, 26 Suppl 4, 18 - 21; discussion 48-50
Use of third-generation cephalosporins . Pseudomonas; Pizzo P; Although the frequency of infection with P . aeruginosa has declined in many centers treating neutropenic patients with cancer, infections still occur and can be accompanied by considerable morbidity and mortality . Furthermore, patterns of infection can change again, and Pseudomonas may reemerge . Thus, in high-risk, immunocompromised patients, adequate bacterial coverage for P . aeruginosa should be part of any empiric regimen . Third-generation cephalosporins are an important part of the therapeutic armamentarium for the empiric management of neutropenic patients . Assuming a low level of resistance at a given center, however, only ceftazidime and cefoperazone possess sufficient antipseudomonal activity to be used for monotherapy . If other third-generation cephalosporins are used, it is imperative that an aminoglycoside or an antipseudomonal penicillin be added . But even when combined with an aminoglycoside, ceftriaxone would not be a good choice for neutropenic patients . It is an important agent in non-neutropenic hosts or in other immunocompromised patients in whom infection with Pseudomonas is unlikely to occur.

Mol Microbiol, 1991 Jan, 5(1), 23 - 31
Computer modelling of the NAD binding site of ADP-ribosylating toxins: active-site structure and mechanism of NAD binding; Domenighini M et al.; Five ADP-ribosylating bacterial toxins, pertussis toxin, cholera toxin, diphtheria toxin, Escherichia LT toxin and Pseudomonas exotoxin A, show significant homology in selected segments of their sequence . Site-directed mutagenesis and chemical modification of residues within these regions cause loss of catalytic activity and of NAD binding . On the basis of these results and of molecular modelling based on the three-dimensional structure of exotoxin A, the geometry of an NAD binding site common to all the toxins is deduced and described in the paper . For diphtheria toxin, sequence similarity with exotoxin A is such that its preliminary structure can be computed by molecular modelling, whereas for the other toxins similarity appears to be restricted to the NAD binding site . Moreover, an analysis of molecular fitting of the NAD molecule into its binding cavity suggests a new model for the conformation of the bound NAD that better accounts for all available experimental information.

Clin Rev Allergy, 1991 Spring-Summer, 9(1-2), 47 - 74
Cystic fibrosis . Infection and immunity to Pseudomonas; Sorensen RU et al.; Chronic pulmonary infection with P . aeruginosa in CF may result from: 1 . An initial failure of clearance mechanisms (increased adherence) leading to the development of a highly compartmentalized inflammatory reaction; 2 . Inhibition of clearing mechanisms for bacteria present in the bronchial lumen; and 3 . A largely ineffective, and possibly damaging, hyperactivity of inflammatory cells in the lumen and bronchial wall . The special relationship between the CF host and P . aeruginos, always long-term, and frequently subtle in its complexity, needs further understanding in order to develop new strategies for the treatment of chronic lung infections with this organism.

J Basic Microbiol, 1991, 31(2), 101 - 6
Host-controlled modification and restriction as a criterion of evaluating the therapeutical potential of Pseudomonas phage; Gachechiladze KK et al.; The recently isolated phages phi ST3 and phi ST1 were compared as to their lysis behaviour in about 100 different P . aeruginosa strains . The growth of phi ST3 varies greatly in different host strains . We demonstrated one case of "non-classical", host-dependent modification and restriction . Here the capability to adsorb, and consequently to reproduce in a given host strain differs, depending on which modification the phage acquired in its former host . The DNA-containing phage phi ST1 displays stable lysis properties in the majority of the host strains . This makes phi ST1 a candidate for therapeutic phage preparations . One of the reasons for stable lysis properties is the apparent selection against recognition sites of restriction enzymes in its genome.

Cancer Res, 1991 Jan 1, 51(1), 174 - 80
Cytotoxic activity of a recombinant fusion protein between insulin-like growth factor I and Pseudomonas exotoxin; Prior TI et al.; A chimeric toxin in which the cell binding domain of Pseudomonas exotoxin was replaced with mature human insulin-like growth factor I (IGF-I) was produced in Escherichia coli . This protein, IGF-I-PE40, was cytotoxic to human cell lines derived from a variety of tumor types, with a breast carcinoma line (MCF-7) and two hepatoma lines (HEP3B and HEPG2) showing the highest sensitivity to the toxin . The specificity of IGF-I-PE40 cytotoxicity was confirmed through competition with excess IGF-I and through blockage of toxin binding using an antibody specific to the type I IGF receptor . A potential interaction between the toxin and soluble IGF-binding proteins was also demonstrated . IGF-I-PE40 may be useful in the selective elimination of cells bearing the type I IGF receptor.

J Bacteriol, 1991 Jan, 173(2), 575 - 86
Genetic and transcriptional organization of the hrp cluster of Pseudomonas syringae pv . phaseolicola; Rahme LG et al.; The hrp cluster of Pseudomonas syringae pv . phaseolicola encodes functions that are essential for pathogenicity on bean plants and for the elicitation of the hypersensitive response on resistant plants . The cluster was saturated with insertions of transposon Tn3-spice that served both as a mutagen and as a sensitive reporter of the expression of the target regions . The mutations covered a 17.5-kb segment in strain NPS3121, in which seven hrp::Tn5 insertions had been previously mapped, and regions outside this segment . The cluster is organized into seven distinct complementation groups (hrpL, hrpAB, hrpC, hrpD, hrpE, hrpF, and hrpSR) on the basis of the analysis of over 100 Tn3-spice insertions in plasmids and 43 similar insertions in the chromosome; it spans nearly 22 kb and is chromosomally located . The transcriptional orientation of all genes in the cluster was established by measuring the level of ice nucleation activity of complemented merodiploids carrying chromosomal hrp::inaZ fusions after inoculation in Red Kidney bean leaves . Although all seven loci were actively expressed in Red Kidney bean leaves, none of them was substantially expressed when the bacteria were grown in King B broth medium . Mutations in all loci, except those in hrpC, greatly reduced the ability of the bacteria to multiply in bean leaves . Mutations in the hrpC locus, although preventing the bacteria from eliciting a hypersensitive reaction on tobacco, allowed the bacteria to produce delayed and attenuated symptoms in Red Kidney bean leaves and to multiply to a level 10(2)- to 10(3)-fold lower than that of the wild-type strain . This is the first comprehensive report of the genetic and transcriptional organization of the hrp gene cluster in a phytopathogenic bacterium.

Plant Cell, 1991 Jan, 3(1), 61 - 72
Induction of Arabidopsis defense genes by virulent and avirulent Pseudomonas syringae strains and by a cloned avirulence gene; Dong X et al.; We developed a model system to study the signal transduction pathways leading to the activation of Arabidopsis thaliana genes involved in the defense against pathogen attack . Here we describe the identification and characterization of virulent and avirulent Pseudomonas syringae strains that elicit disease or resistance symptoms when infiltrated into Arabidopsis leaves . The virulent and avirulent strains were characterized by determining growth of the pathogen in Arabidopsis leaves and by measuring accumulation of mRNA corresponding to Arabidopsis phenylalanine ammonia-lyase (PAL), beta-1,3-glucanase (BG), and chalcone synthase (CHS) genes in infected leaves . The virulent strain, P . syringae pv maculicola ES4326, multiplied 10(5)-fold in Arabidopsis leaves and strongly elicited BG1, BG2, and BG3 mRNA accumulation but had only a modest effect on PAL mRNA accumulation . In contrast, the avirulent strain, P . syringae pv tomato MM1065, multiplied less than 10-fold in leaves and had only a minimal effect on BG1, BG2, and BG3 mRNA accumulation, but it induced PAL mRNA accumulation . No accumulation of CHS mRNA was found with either ES4326 or MM1065 . We also describe the cloning of a putative avirulence (avr) gene from the avirulent strain MM1065 that caused the virulent strain ES4326 to grow less well in leaves and to strongly elicit PAL but not BG1 and BG3 mRNA accumulation . These results suggest that the Arabidopsis PAL and BG genes may be activated by distinct signal transduction pathways and show that differences in plant gene induction by virulent and avirulent strains can be attributed to a cloned presumptive avr gene.

Plant Cell, 1991 Jan, 3(1), 49 - 59
Identification of Pseudomonas syringae pathogens of Arabidopsis and a bacterial locus determining avirulence on both Arabidopsis and soybean; Whalen MC et al.; To develop a model system for molecular genetic analysis of plant-pathogen interactions, we studied the interaction between Arabidopsis thaliana and the bacterial pathogen Pseudomonas syringae pv tomato (Pst) . Pst strains were found to be virulent or avirulent on specific Arabidopsis ecotypes, and single ecotypes were resistant to some Pst strains and susceptible to others . In many plant-pathogen interactions, disease resistance is controlled by the simultaneous presence of single plant resistance genes and single pathogen avirulence genes . Therefore, we tested whether avirulence genes in Pst controlled induction of resistance in Arabidopsis . Cosmids that determine avirulence were isolated from Pst genomic libraries, and the Pst avirulence locus avrRpt2 was defined . This allowed us to construct pathogens that differed only by the presence or absence of a single putative avirulence gene . We found that Arabidopsis ecotype Col-0 was susceptible to Pst strain DC3000 but resistant to the same strain carrying avrRpt2, suggesting that a single locus in Col-0 determines resistance . As a first step toward genetically mapping the postulated resistance locus, an ecotype susceptible to infection by DC3000 carrying avrRpt2 was identified . The avrRpt2 locus from Pst was also moved into virulent strains of the soybean pathogen P . syringae pv glycinea to test whether this locus could determine avirulence on soybean . The resulting strains induced a resistant response in a cultivar-specific manner, suggesting that similar resistance mechanisms may function in Arabidopsis and soybean.

J Basic Microbiol, 1991, 31(6), 403 - 11
In vivo utilization of N-(phosphonomethyl)-anilines and related substances by Pseudomonas spec . GS; Albrecht B et al.; Utilization of various phosphonates as source for phosphorus by the glyphosate degrading strain Pseudomonas spec . GS was investigated . Metabolites of phosphonate degradation were characterized indicating the cleavage of the C-P-bound as primary step of breakdown . The phosphonate N-(phosphonomethyl)-4'-nitroazobenzene-4-amine (azophon) was characterized as a suitable substrate for detection of C-P-bond splitting activity in vivo . Pseudomonas cells permeabilized by toluene treatment were also capable of phosphonate degradation whereas no in vitro activity of a putative C-P-bond cleaving enzyme was detectable after cell disruption.

Klin Wochenschr, 1991, 69 Suppl 26, 168 - 77
{Prevention with pseudomonas immune globulin in burn injury patients with inhalation trauma: does it have an effect on lung function and outcome?}; Stuttmann R et al.; In an evaluation of the effect of prophylactic application of Pseudomonas immunoglobulin on the immunoglobulin serum concentration, infection rate, lung function and mortality in major burn-trauma patients, a clinical, prospective, controlled and randomized trial along with an extensive literature review was carried out in the intensive care unit (ICU) of a major burn-trauma center at a major municipal hospital in the Federal Republic of Germany . A total of 60 patients suffering from major burn trauma were studied . Some of them exhibited inhalation injury as a secondary trauma . Inclusion criteria comprised an age of 15-60 years, burns covering 30%-70% of the body surface area, and second and third degree skin burns . In a randomized fashion, the consecutively admitted patients were assigned to either the study or the control group, each comprising 30 subjects . Study-group patients (PIG-GRP) received 250 mg/kg Pseudomonas immunoglobulin (Psomaglobin; Tropon-Cutter, Cologne, FRG) intravenously on days 3, 5, 7, 10 and 13 following the trauma, whereas controls (CON-GRP) received no prophylaxis . The immunoglobulin concentration was measured in serum on days 1, 3, 5, 7, 10, 13, 16, 19 and 28 . From day 3 to day 13, significant higher values were found in study-group patients; however, this held true only for that subgroup of subjects in each group who displayed additional inhalation injury (PIG-SUBGRP = 23; CON-SUBGRP = 16) . Immunoglobulin serum levels showed an earlier return to normal in the PIG-SUBGRP (day 7) than in the CON-SUBGRP (day 13) . Blood cultures were taken on suspicion of septicaemia . In the above-described subgroups, the number of positive blood cultures was significantly reduced in the study patients (PIG-SUBGRP, 27 bacteremic subjects among a total of 70; CON-SUBGRP, 22 among a total of 36; P = 0.0045) . In all, 21 subjects in the PIG-SUBGRP and 13 patients in the CON-SUBGRP were mechanically ventilated according to an adaptive scheme . The target value of pulmonary function was the O2 quotient (P(ALV)O2-P(ART)O2/P(ALV)O2), which was significantly closer to the normal value in the PIG-SUBGRP . Mortality was lower in the PIG-SUBGRP (34.8%, 8 patients) than in the CON-SUBGRP (50%, 8 subjects) . In conclusion, prophylaxis with Pseudomonas immunoglobulin does not appear to be beneficial to burn trauma patients in general; however, it was shown to be effective in burn-trauma patients exhibiting inhalation injury.

Chin J Biotechnol, 1991, 7(2), 93 - 104
Cloning of GL-7-ACA acylase gene from Pseudomonas sp . 130 and its expression in Escherichia coli; Yang YL et al.; Using BamHI digested and dephosphorylated pBR322 as vector, a GL-7-ACA acylase gene from Pseudomonas sp . 130 chromosomal DNA was cloned and expressed in Escherichia coli C600 . Seven positive clones were detected from 3205 recombinant plasmids with 32P-labeled oligonucleotide probes in situ hybridization . Out of them, three clones which produced active GL-7-ACA acylase were identified by radio-immunological assay, chemical test and chromatographic analysis of reaction mixture . The plasmid DNAs of recombinant pMR5, pMR6 and pMR7 were extracted . Analysis of gel electrophoresis indicated that pMR5 and pMR7 contained the same 6.8kb fragment and the size of the insert in pNR6 was 5.7 kb . The effects of various E . coli hosts on the expression of cloned GL-7-ACA acylase gene is also presented.

Biomed Biochim Acta, 1991, 50(4-6), 731 - 41
Biological and clinical relevance of the urokinase-type plasminogen activator (uPA) in breast cancer; Schmitt M et al.; Tumor cell invasion and metastasis is a multifactorial process, which at each step may require the action of proteolytic enzymes such as collagenases, cathepsins, plasmin, or plasminogen activators . An enzymatically inactive proenzyme form of the urokinase-type plasminogen activator (pro-uPA) is secreted by tumor cells which may be converted to an enzymatically active two-chain uPA-molecule (HMW-uPA) by plasmin-like enzymes . Action of proteases on pro-uPA may generate the enzymatically active or inactive high-molecular-weight form of uPA (HMW-uPA) . Some proteases (plasmin, cathepsin B and L, kallikrein, trypsin or thermolysin) activate pro-uPA by cleaving the peptide bond Lys158 and IIe159 . Other proteases (elastase, thrombin) cleave pro-uPA at different positions to yield enzymatically inactive HMW-uPA . HMW-uPA may be split into the enzymatically active LMW-uPA and the enzymatically inactive ATF (amino terminal fragment) . ATF may be cleaved between peptide sequence 20 and 40 within the receptor binding domain of uPA (GFD) . Such impaired ATF does not bind to uPA-receptors . Action of the bacterial endoproteinase Asp-N from Pseudomonas fragi mutant on pro-uPA or HMW-uPA, however, generates intact ATF which efficiently competes for binding of HMW-uPA or pro-uPA to receptors on tumor cells . High uPA-antigen content (pro-uPA, HMW-uPA, or LMW-uPA) in breast cancer tissue (not in plasma) indicates an elevated risk for the patient of recurrences and shorter overall survival . Thus pro-uPA/uPA-antigen content in breast cancer tissue serves as an independent prognostic parameter for the outcome of the disease . Cathepsin D is also an independent prognostic factor for recurrences and overall survival . High content of cathepsin D in breast cancer tumors is, however, not correlated with elevated levels of pro-uPA/uPA indicating that synthesis and release of cathepsin D and pro-uPA/uPA are independent events.

Optom Clin, 1991, 1(3), 123 - 33
Management of corneal abrasions in an extended-wear patient population; Catania LJ; Contact-lens-related corneal abrasions are a management problem for optometrists because of the risk of ulcerative keratitis, particularly from Pseudomonas . Risks and causes of abrasions should be identified for extended-wear patients, and appropriate steps should be taken to minimize the opportunity for injury . When an extended-wear patient presents with an abrasion, the clinician should take a careful history, conduct a thorough examination, and provide the necessary treatment: discontinuation of lens wear, irrigation of the cornea, use of cycloplegia and dilation of the pupil, utilization of the aminoglycoside tobramycin to treat the abrasion, no patching, and appropriate use of cold compresses and oral nonsteroidal anti-inflammatory drugs . Follow-up evaluations should be provided at 24 and 48 hours and at 3 to 5 days after the injury.

Pediatr Pulmonol Suppl, 1991, 7, 42 - 5
Early respiratory course in infants with cystic fibrosis: relevance to newborn screening; Accurso FJ et al.; Respiratory morbidity and mortality during infancy are important problems in the care of CF patients whether they are diagnosed conventionally or through newborn screening . Although the mechanisms of lung disease in CF remain to be elucidated, two potential pathophysiologic mechanisms--viral infection and undernutrition--can be associated with respiratory morbidity in infancy . Colonization of some infants with Pseudomonas and the presence of early mucus casts and cytokines in bronchoalveolar lavage suggest that pathophysiologic processes that are important in later life may begin in infancy . The early respiratory abnormalities, morbidity and mortality seen in CF indicate the need for future investigations of the respiratory course and interventional trials in infancy.

Biol Met, 1991, 4(4), 211 - 6
Effects of iron(III) analogs on growth and pseudobactin synthesis in a chromiumtolerant Pseudomonas isolate; Fekete FA et al.; The growth and siderophore production of a fluorescent Pseudomonas species isolated from soil contaminated with chromium was found to be influenced by the presence of trivalent cations . Overproduction of pseudobactin occurred when the isolate was grown in media containing 1 mM Cr(III) under iron-limited conditions but not when Fe(III) was added at 10 microM . Pseudobactin synthesis was derepressed in iron-limited cultures containing 1 mM Sc(III) or Y(III), examples of group III-B elements . We found that Al(III), Ga(III) or In(III), representative metals from group III-A, repressed synthesis of pseudobactin under iron-deficient conditions . Analogs of Fe(III) were found to inhibit growth of the Pseudomonas isolate in iron-limited media and the trivalent metals listed in order of decreasing toxicity were as follows: Ga greater than In greater than Sc greater than Cr greater than Y greater than Al . The inhibition of growth by 1 mM In(III), Sc(III) and Ga(III) was greater during iron-limited growth than in media containing 10 microM Fe(III) . These data show that, although the metal analogs of Fe(III) have similar chemical and physical characteristics, the physiological response of the fluorescent pseudomonad when grown in the presence of these metals varied markedly.

Adv Exp Med Biol, 1991, 302, 191 - 8
Effect of solute on the nucleation and propagation of ice; Charoenrein S et al.; Using the emulsion technique, we have studied nucleation of ice in aqueous solutions containing silver iodide or Pseudomonas syringae . Using a Differential Scanning Calorimeter (DSC), we determined characteristic temperatures of nucleation, and also rates of nucleation at selected temperatures . The freezing point depression induced by added solute is linearly related to the lowering of both homogeneous and heterogeneous nucleation temperature . Nucleation kinetics depend on a fifth power function of the temperature . Solute is found to affect the parameters of this relationship in different ways, dependent upon the nature of the catalytic site for ice nucleation . We have also studied the effect of composition on the linear propagation velocity (LPV) of ice in undercooled solutions contained in a U-tube . We have determined velocities in a range of concentrations of sugar solution at the same undercooling, and also as a function of undercooling . The role of added polymer has also been investigated . It is affected by the sugar concentration.

Lab Delo, 1991, (10), 61 - 3
{Characteristics of the biochemical properties of pseudomonads isolated from environmental objects}; Stavertii LV; Biochemical characteristics of 117 Pseudomonas cultures isolated from water bodies and washings off the hospital environment objects were under study, including the strains from patients' wound surfaces . Analysis of the strains' characteristics permitted referring 83.8 +/- 3.7% of them to typical ones as regards their biochemistry . 45.9 +/- 7.5% of these were P . putida, 21.4 +/- 4.8% were P . aeruginosa, 10.2 +/- 1.01% P . fluorescens strains, and the shares of P . cepacia and P . stutzeri strains were 11.2% each . 16.2% of the strains were found atypical because of lysine decarboxylase and growth at 42 degrees C; they were conditionally referred to P . putida . One third of these cultures were isolated in hospital . The detected differences may be helpful in identification of Pseudomonas.

Microbiol Immunol, 1991, 35(6), 461 - 74
Mechanism of protective effect of recombinant human granulocyte colony-stimulating factor (rG-CSF) on Pseudomonas infection; Matsumoto M et al.; Decrease in resistance to systemic Pseudomonas infection in cyclophosphamide (CPA)-induced neutropenic mice was prevented by injections of recombinant human granulocyte colony-stimulating factor (rG-CSF) . In order to explore mechanism of the prevention of CPA-induced decrease in the anti-infectious resistance by rG-CSF, CPA-treated and then rG-CSF-injected mice were inoculated i.p . with P . aeruginosa, and growth of the infecting bacteria and infiltration of leukocytes in the peritoneal cavity were determined . In the mice who had received 4 daily s.c . injections of rG-CSF from the day after CPA-injection, a large number of neutrophils were mobilized into the peritoneal cavity in response to the bacterial inoculation and growth of the infecting Pseudomonas in the cavity was markedly inhibited, whereas in CPA-induced neutropenic mice few neutrophils were mobilized and the infecting bacteria proliferated vigorously in the peritoneal cavity . These results suggest that administration of rG-CSF prevents CPA-induced neutropenia and neutrophils circulating at normal level in the number are normally mobilized into the peritoneal cavity in response to Pseudomonas inoculation, and that the mobilized neutrophils inhibit proliferation of the infecting Pseudomonas.

Infect Immun, 1991 Jan, 59(1), 407 - 14
Analysis of Pseudomonas exotoxin activation and conformational changes by using monoclonal antibodies as probes; Ogata M et al.; Pseudomonas exotoxin (PE) is a protein toxin composed of three structural domains . In its native form, the toxin is a 66,000-Mr proenzyme that must be activated to express full ADP-ribosylating activity . To study the process of activation and accompanying conformational changes, we have isolated 10 monoclonal antibodies to a 40,000-Mr fragment of the toxin (PE40) that exhibits full enzyme activity but lacks the toxin's cell-binding domain and contains amino acids 253 to 613 (comprising domains II, Ib, and III) . By using mutant PE molecules in which all of domain I and portions of domains II, Ib, and III were deleted, the locations of the epitopes for each of the antibodies were determined . Eight of these monoclonal antibodies were further characterized . Of these eight, all reacted with soluble PE40 and an interleukin-2-PE40 conjugate, but only two reacted strongly with native soluble PE . However, all eight reacted with PE after it had been immobilized on nitrocellulose or after it had been activated to express full ADP-ribosylating activity . Antibodies were also assessed for their ability to neutralize the cytotoxic activity of either PE or interleukin-2-PE40 . These antibodies should be useful as probes for monitoring the activation and processing of PE that occur during endocytosis and in determining the location of epitopes that are important for toxin activity.

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