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 b